Proceedings of ICBC 2019

Proceedings
International Conference
on
Bio-Commerce 2019
Volume 1 Number 1
14-15 March, 2019
Department of Zoology and Research Centre

Aditanar College of Arts & Science
Virapandianpatnam, Tiruchendur – 628 216.
Tamil Nadu, India.
Website: www.aditanarcollege.in Phone: 04639 242232

Dr. P. SUBRAMANIAM, M.Sc., Ph.D.
SECRETARY
FOREWORD
Earth has a variety of flora and fauna that can be called in a single word as Nature.
When the human became part of the evolutionary process he developed strategies to live
sophisticatedly. Then man started to disturb the natural resources which led to exploitation.
However, the advance of sixth sense has created a variety of technology, which together with
Science produced a variety of cultivable living organisms for its sustainable use. This competes
with the growing world population.
In the present scenario, a spectacular and overwhelming utilization of Bio-resources

has reached an inevitable state. As such, in order to maintain a sustainable Bio-resource
management and to generate revenue for the country and to encourage entrepreneurship,
many unexplainable efforts have been taken in relation to the utilization of Bio-resources.
Every day, Bio-resources satisfy the food requirements of the entire world. After
globalization, the term culture and serve has been modified into culture and sale. This is known
as Bio-commercialization. Many countries are dependent on this term Bio-commerce for their
annual budget. This clearly shows the importance of this word in the present world scenario.
So this program intends to empower the knowledge related to commercially important Bio-
resources and their technologies to improve the economic status of the nation at a global level.
The theme of this conference “Bio-Commerce 2019” is an urgent need for developing
countries like us. I highly appreciate the entire team of Zoology Department of Aditanar
College of Arts and Science for organizing this International program. I hope this would
definitely benefit the young minds of India.
EDITORIAL BOARD
Editor in Chief
DR. G. LAKSHMANAN
Associate Professor & Head
Department of Zoology & Research Centre
Aditanar College of Arts & Science, Tiruchendur
Editors
Dr. C. P. BALAKRISHNAN
Assistant Professor & Head
Department of Botany
Dr. T. MOHANRAJ
Assistant Professor
Department of PG Zoology
Dr. S. LINGATHURAI
Assistant Professor
Department of Zoology & Research Centre
Editorial Board Members
Dr. C. SUNDARAVADIVEL Dr. D. VASUMATHI

Associate Professor Assistant Professor
Department of Zoology & Research Department of Zoology & Research
Centre Centre
Aditanar College of Arts & Science, Aditanar College of Arts & Science,
Tiruchendur Tiruchendur
Dr. P. AROCKIA MARY FERNADEZ MISS. P. JENIFER

Assistant Professor Research Scholar
Department of Zoology & Research Department of Botany
Centre Aditanar College of Arts & Science,
Tiruchendur
©Department of Zoology and Research Centre 2019, Aditanar College of Arts
& Science, Virapandianpatnam, Tiruchendur – 628 216. Tamil Nadu, India.
This publication is in copyright. Subject to statuary exception and to the

provisions of relevant collective licensing agreements, no reproduction of any
part take place without the written permission of Department of Zoology and
Research Centre, Aditanar College of Arts & Science.
First published 2019
Printed in India
Proceedings of International Conference on Bio-Commerce - 2019
Published by Dr.G.Lakshmanan for ‘Proceedings of International

Conference on Bio-Commerce 2019’
Typeset & Printed by Robert Printers, Thoothukudi – 628 002. Ph:

99944 22806. E-mail: robertxeroxtuty@gmail.com
Proceedings of International Conference on Bio-Commerce 2019
Vol.1 Number 1, 14-15 March 2019 – ISBN 978-93-5351-470-0
Page
CONTENTS
Number
USING THE WORLD’S DEEPEST CORAL NURSERY TO EXAMINE THE

INFLUENCE OF GENOTYPE ON SURVIVAL 01
Daniel G. Merselis, ThineshThangadurai, Anthony bellantuono, Katherine E.
Dougan, Trevor Graff, William Todd, James W. Fourqurean and Mauricio
Rodriguez-Lanetty
EVALUATION OF ANTI-INFLAMMATORY AND ANTIBACTERIAL 02

ACTIVITY OF LENTINUS CLADOPUS LEV - IN VITRO
Rajendran Bhuvaneswari, Mani Vinotha, Muthukumar Abinaya, Varadharaj
Nithya
AN EVALUATION OF PEST MANAGEMENT STRATEGIES IN THE FACE 03
OF POTENTIAL EFFECTS OF CLIMATE CHANGE ON AGRICULTURAL
SECTOR
Muzafar Riyaz and S. Ignacimuthu
REVIEW ON IMPACT OF CLIMATE CHANGE IN PHOSPHATE 04
NUTRIENT MANAGEMENT
T.P Ajeesh Krishna and V. Duraipandiyan
REVIEW ON ROLE OF PHT1 FAMILY TRANSPORTERS IN 05
ACQUISITION AND REDISTRIBUTION OF PHOSPHORUS IN PLANTS
G. Victor Roch and V.Duraipandiyan
DIRECT PLANT REGENERATION FROM IN VITRO-DERIVED SHOOT 06
APEX OF FINGER MILLET AND ANALYSIS OF GENETIC FIDELITY
USING MOLECULAR MARKERS
T. Maharajan and V. Duraipandiyan
DISTRIBUTION OF PHYTOPLANKTON DIVERSITY IN AUTHOORANGAL 07
CHANNEL AT THOOTHUKUDI DISTRICT
F. Esther Isabella Eucharista and A.Shanthi
IN VITRO ANTIMICROBIAL ACTIVITY OF RHIZOPHORA MUCRONATA 08
LEAF EXTRACTS AGAINST CLINICAL PATHOGENS
J.Rajaselvam, I.Vasudhevan and T.Subi
SELECTIVE SYNTHESIS OF BENZYL PHENYL SULFOXIDE FROM 09
BENZYL PHENYL SULFIDE USING [Fe(Phen)3]3+
Pathakaraimuthu Balakumar, Subramaniam Balakumar, Perumal Subramaniam
and Sweetlin Rajula Rubavathi. D
SURVEY OF BY- CATCH OF CRAB FISHERY FROM THOOTHUKUDI 10
COAST
K. Jesima and S. Jesily
A STUDY OF MANAPADU WATER BODIES WITH REFERENCE TO 11
HYDROBIOLOGICAL AND FISHERIES ASPECTS
Devi Priya .S and L. JeyaPraba
PURIFICATION AND CHARACTERIZATION OF LECTIN FROM 12
SYZYGIUM CUMINI
P. Rama Devi , K. Nandha Devi, Arokiya Mary Fernandez and C. Babu
HIV PREVALENCE 13
C. Sundaravadivel and A. Alagu Rajasekar
A MOSQUITOCIDAL ACTIVITY OF MUSIZIN ISOLATED FROM 14

RHAMNUS WIGHTII WIGHT AND ARN
William Raja Tharsius Raja, Ganesan Pathalam, Duraipandiyan Veeramuthu
ICHTHYOFAUNAL DIVERSITY OF OKHA REEF, GULF OF KACHCHH, 15
WEST COAST OF INDIA
S. Subburaman, Nilesh joshi, D. Nagarajan, J.Nagarajan and Ramakotti
EFFECT OF YEAST ADDED DIET ON CAROTENOID CHANGES IN 16
FRESH WATER ORNAMENTAL FISH GOLD FISH (CARASSIUS
AURATUS)
R. Prema and Beena Somanat
EFFECTS OF VITAMIN C ON CARBOHYDRATE PROFILE OF SILK 17
WORM, BOMBYX MORI LXCSR2 AND PURE MYSORE
Sundara Raj J, W. Kerenhap Evangelin, S Akhil Kumar
ANTIMITOTIC ACTIVITY OF FUSINUS NICOBARICUS EXTRACT ON 18
ALLIUM CEPA ROOT TIP
P. Subavathy, R. D. Thilaga and B. Jeba Nesam
PLANT PHYTO-CHEMICALS USED AS A BIO-PESTICIDES FOR 19
INSECT PEST CONTROL
Lingathurai S and Crosslin Anto Singh
CULTURE OF OYSTER MUSHROOM (PLEUROTUS OSTREATUS) ON 20
DIFFERENT KIND OF AGRO WASTE AND EXOTIC PLANT WASTE
Lakshmanan G, Kamala Devi N.G. and Kabilan S
BIOFABRICATION OF SILVER NANOPARTICLES USING LANTANA 21
CAMARA AND ITS ANTIBACTERIAL ACTIVITY
Lakshmanan G, Kamala Devi N, Muthu Krishnan G. and G. Anantha Sundar Raj
FOURIER TRANSFORM INFRARED SPECTROSCOPY ANALYSIS OF 22
METHANOLIC LEAF EXTRACT OF GREWIA ORBICULATA ROTTL.
T. Hemalatha and A. Saravana Ganthi
EVALUATION OF ANTIBACTERIAL ACTIVITY OF PLECTRANTHUS 28
WIGHTII BENTH.
Subramonian K Saravana Ganthi A Padma Sorna Subramanian Mand Subha S.L
QUANTITATIVE PHYTOCHEMICAL ANALYSIS OFLEPIDAGATHIS 34
PUNGENS NEES.
Subha S and Saravana Ganthi A
ANTICANCER ACTIVITY OF POUZOLZIA WIGHTII BENN. STEM 43
ETHANOLIC EXTRACTS AGAINST HELA CELL LINE
M. Johnson, M. Syed Ali Fathima and A. Mohamed Ansar
PHARMACEUTICAL EVALUATION OF AVICENNIA MARINA (FORSSK) 49
VIERH.
P. Arockia Mary Fernandez and D. Vasumathi
ANALYSIS OF HEAVY METAL CONCENTRATION OF TUTICORIN 58
MANGROVE ECOSYSTEM
S.R.T.Sherly Cross and T.Mohanraj
ANTIBACTERIAL POTENTIAL AND PHYTOCHEMICAL ANALYSIS OF 66
BROWN SEAWEED STOECHOSPERMUM MARGINATUM
R.S. Raubbin and A. Pushparaj
INVESTIGATION ON ETHNO PHARMACOLOGICAL AND 75

PHYTOCHEMICAL PROPERTIES OF MEDICINAL PLANTS EUPHORBIA
HIRTA LINN. AND VITEX NEGUNDO LINN.
J. Mugil S. Abirami, M. Kannan and S. Venkatesan
DIVERSITY OF MARINE FINFISH RESOURCES OF TIRUCHENDUR 93
COAST
T.Jebarani Rajathy and T.Mohanraj
EFFECT OF PROBIOTICS ON CICHLID FISH, MAYLANDIA 101

GRESHAKEIREPRODUCTION UNDER CAPTIVE CONDITIONS
Ambika. P and A. Pushparaj
STUDIES ON THE EFFECT OF ORGANIC FERTILIZERS ON GROWTH 113
AND ECONOMIC TRAITS OF SILKWORM, BOMBYX MORI L.
C. Dyana Selinand MA. Ramani Bai
EFFECT OF VITAMINS AND MINERALS ON SILK GLAND RATIO AND 122
ECONOMIC TRAITS OF BOMBYX MORI L.
M. Thilsath Fatima Quraiza and M. Ramani Bai
STUDIES ON AQUATIC PHYTO DIVERSITY IN VAGAIKULAM POND, 132
AMBASAMUDRAM TALUK, TIRUNELVELI DISTRICT, TAMIL NADU
Paul David Selson S Darwin Paul Edison S and Amish Abragam D
STUDIES ON PHYTOPLANKTON DIVERSITY IN KEELAIDAYANKULAM 138
POND, AMBASAMUDRAM TALUK, TIRUNELVELI DISTRICT
Darwin Paul Edison S, Paul David Selson S and Amish Abragam D
TO COMPARATIVE ANALYSIS AND IMPACT OF VERMICOMPOST ON 148
VIGNA RADIATA L.(MUNG BEAN) VIGNA MUNGO (BLACK GRAM) AND
VIGNA UNGUICULATA( CHICK PEA).
P. Mathiarasi and Dr. D. Amish Abragam
GROWTH, ECONOMIC FEATURES AND FECUNDITY OF SILKWORM 165
BOMBYXMORI UNDER THE INFLUENCE OF VITAMIN B- COMPLEX
P. Kumutha and R. Kanagapriya
ANALYSIS AND DETECTION OF BREAST CANCER MAMMOGRAM 174
USING MULTI-LAYER NEURAL NETWORK
G. Sam Singh, R. Selva Priya, K. Sabana Asmin
ISOLATION AND CHARACTERIZATION OF BIOACTIVE COMPOUNDS 185
FROM FUSINUS NICOBARICUS (ROEDING, 1798)
P.Subavathy and R.D.Thilaga
GREEN SYNTHESIS OF SILVER NANOPARTICLES USING 193
ATROCARPUS HETEROPHYLLUS LEAVES AND THEIR
SUPPLEMENTARY EFFECT ON SILKWORM BIOLOGY
P.Kumutha and M. VidyaKalaivani
ANTIMICROBIAL ACTIVITY & SELECTIVITY STUDIES OF POLY (O- 204
TOLUIDINE) ZR (IV) TUNGSTOIODO PHOSPHATE NANOCOMPOSITE
ION-EXCHANGER
M.Vijayakumari
INTERACTION OF MONOTERPENES OF ESSENTIAL OIL WITH Β- 213
CYCLODEXTRIN: ANTIMICROBIAL AND ANTIOXIDANT APPLICATION
A. Antony Muthu Prabhu
GREEN APPROACH TO CORROSION INHIBITION OF MILD STEEL 219
USING EMBLICA OFFICINALIS (AMLA SEED) IN ACID MEDIUM
R. Rajkumar and C. Kavitha
SEAWEED EXTRACT OF RED ALGAE AS ORGANIC FERTILIZER AND 227
THEIR IMPACT ON GROWTH PARAMETERS, BIOMASS
PRODUCTIVITY AND ANTIOXIDANT OF AMARANTHUS TRISTIS L.
T. Kumareswari, S. Maria Victorial Rani,
SOCIO-ECONOMIC AND DEMOGRAPHIC CORRELATES AND RATE
OF CONSANGUINITY IN CERTAIN COMMUNITIES OF KANYAKUMARI
236
DISTRICT
K. Hemarana, Jayashree KV and Elango T
ECOLOGY OF TUTICORIN AND VEMBAR GROUP OF ISLANDS, GULF 255
OF MANNAR
M.Sheeba and T.Mohanraj
271
OBSERVATION ON CAPTURED MARINE FISHES OF MANAPADU
COAST
A. Vanmathi and T. Mohanraj
MORPHO-ANATOMICAL STUDIES OF RED ALGAE GRACILARIA 278
FERGUSONII J. AG
P. Jenifer, C.P Balakrishnan, and S Chidambaram Pillai
PHARMACOGNOSTIC STANDARDIZATION AND UV SPECTRUM 286
ANALYSIS OF MARINE RED ALGAE GRACILARIA SPECIES OF
MANAPAD COAST, TAMIL NADU
P Jenifer, C.P Balakrishnan, and S Chidambaram Pillai
STUDIES OF PHYSICOCHEMICAL AND BIOCHEMICAL COMPOSITION 294
OF MARINE RED ALGAE GRACILARIA FERGUSONII J. AG
P Jenifer, C.P Balakrishnan, and S Chidambaram Pillai
A COMPARISON OF THE EFFECTS OF COBALOXIMES ON 302
ANTIBACTERIAL AND ANTIFUNGAL PROPERTIES
M. Amuthaselvi, C. Balachander, A. Dayalan, S. Lingathurai and S. Ignacimuthu
ANTIFEEDANT AND TOXIC PROPERTIES OF CALOTROPIS PROCERA 312
(AITON) W.T. AITON EXTRACT ON HELICOVERPA ARMIGERA HUBNER
Bakavathiappan GA, Baskaran S and Lingathurai S
BIOMONITORING OF AIR POLLUTION AT MILLERPURAM, JUNCTION, 320
PALAYAMKOTTAI ROAD, THOOTHUKUDI BY ANALYSING SELECTED
PLANTS
M. Isakkiyammal
AMINO ACID CONTENT OF EARTHWORM EISENIA FETIDA 328
CULTURED ON AQUATIC WEED WATER HYACINTH
T. Sakthika
STUDIES ON THE GROWTH PERFORMANCE OF AZOLLA 338
PINNATAR.BR. IN DIFFERENT WATER MEDIUM
Kamala Devi N, Lakshmanan G and Mohammed Ariff Sehriff
SYNTHESIS OF SILVER NANO PARTICLES FROM EXPIRED 345
PARACETAMOL TABLETS
Kamala Devi N, Lakshmanan G and Krishna sagar V.M
SYNTHESIS OF LEAD NANO PARTICLES FROM CRAB SHELLS 354
A.Kavitha, N. Kamala Devi, B. Megala and G.Lakshmanan
SURVEY OF MARINE MOLLUSCS FROM TIRUCHENDUR LOCATIONS 363
OF THOOTHUKUDI COST, TAMIL NADU, INDIA
Kamala Devi N, Lakshmanan G and Muthu lingam S
COMMERCIALLY IMPORTANT MARINE MOLLUSCS FOR HUMAN 373
CONSUMPTION IN TUTICORIN, TAMIL NADU, INDIA
Kamala Devi N, Lakshmanan G and Petchimuthu ananth B
BIO-EFFICACY OF EUPHORBIAHIRTA L (EUPHORBIACEAE) AGAINST 384

COTTON BOLLWORM, HELICOVERPA ARMIGERA HUB.
(LEPIDOPTERA: NOCTUIDAE) IN THE LABORATORY AND
GREENHOUSE
Saranya T and Lingathurai S
SYNTHESIS OF SILVER NANO PARTICLES FROM CRAB SHELLS 391
A.Kavitha, N. Kamala Devi, B. Megala and G.Lakshmanan
DIVERSITY OF ZOOPLANKTON IN THAYAMANGALAM POND, 398
SIVAGANGAI DISTRICT, TAMIL NADU, INDIA.
Uma maheswari K, Chandran G, Rajeshkannan S, Lingathurai S and Nagarajan S
TOXICITY AND SYNERGISTIC STUDY ON AZADIRACHTAINDICA A. 403
JUSS. AND MELIAAZEDARACH L. ON CALLOSOBRUCHUS
MACULATUS FABRICIUS
D Sarasa, P C Sathya Narayanan, M Pushpalatha R Jalajagandhi, M Vidhyaand S
Lingathurai
BIOLOGICAL ACTIVITIES OF EXTRACTS FROM AZADIRACHTA INDICA 420
A. JUSSAND MELIA AZEDARACH LINN. ONSPODOPTERA LITURA FAB.
(LEPIDOPTERA: NOCTUIDAE)
D Sarasa, P C Sathya Narayanan, M Pushpalatha R Jalajagandhi, M Vidhya and S
Lingathurai
BIODEGRADATION OF SYNTHETIC PYRETHROID- CYFLUTHRIN BY 434
ENTEROBACTER ASBURIAE FROM SOIL
S. Lingathurai, P. Sivadurgadevi and P. Hemavathi
DEVELOPMENT OF VALUE ADDED PRODUCT FROM TUNA (MASI) 444
Mohamed Ramlath Sabura, S., Delighta Mano Joyce, M.I., HasnaNaziya, I.
TOXIC, FEEDING DETERRENCE, NUTRITIONAL AND 449
DEVELOPMENTAL PHYSIOLOGY OF PIPER BETLE L.(PIPERACEAE
TREATED SPODOPTERA LITURA FAB.(LEPIDOPTERA: NOCTUIDAE).
Lingathurai
BIO-EFFICACY AND DETOXIFYING ENZYME ACTIVITIES OF 469
EUPHORBIAHIRTA L. (EUPHORBIACEAE) TREATED ARMYWORM,
SPODOPTERA LITURA FAB. (LEPIDOPTERA: NOCTUIDAE)
Saranya T and Lingathurai S
PRODUCTION AND MARKETING OF PALM LEAF ARTICLES – A STUDY 475
WITH SPECIAL REFERENCE TO UDANKUDI
S. Siril Arun and M. Ruban Jesu Adaikalam

USING THE WORLD’S DEEPEST CORAL NURSERY TO
EXAMINE THE INFLUENCE OF GENOTYPE ON SURVIVAL
Daniel G. Merselis1, Thinesh Thangadurai 1, Anthony bellantuono 1, Katherine

E. Dougan 1, Trevor Graff2, William Todd 2 , James W. Fourqurean 1 and
Mauricio Rodriguez-Lanetty1
1 Department of Biological Sciences, Florida International University, Miami, FL,

USA
2 NASA Johnson Space Center, Houston, TX, USA.
ABSTRACT
Worldwide coral transplantation efforts are being undertaken to restore reefs

lost to climate change-induced bleaching and disease. Successful reef restoration
efforts to date are limited to shallow depths above 10 meters with fast growing corals,
despite the loss of many coral species across a wide range of depths. In this study, we
constructed Orbicellafaveolata and Orbicellaannularis restoration nurseries along a
depth gradient to test coral survival and growth capacity at different depths, to
facilitate restoration activities at greater depths in the future. We assessed 10 genets of
each species at 20 m and 30 m (n=15), using a total of 600 fragments. We identified
two genets which we found to be highly successful across depths. We did not find
significant differences in coral survival between depths. Out of 300 fragments at each
depth, 46% (138 fragments) survived at 30 m, while 52% (158 fragments) survived at
20m. This preliminary study identifies favorable genotypes for future transplantation
and emphasizes the importance of genotype selection during transplantation.
International Conference on Bio-Commerce 2019 Page 1

66EVALUATION OF ANTI-INFLAMMATORY AND
ANTIBACTERIAL ACTIVITY OF LENTINUS CLADOPUS LEV -
IN VITRO
Rajendran Bhuvaneswaria, Mani Vinothaa, Muthukumar Abinayab,
Varadharaj Nithyaa‫٭‬
a
Research Scholar, Pharmacognosy Lab, Department of Animal Health and
Management, Alagappa University
b
Research Scholar, Biomaterials and Biotechnology in Animal Health Lab,
Department of Animal Health and Management, Alagappa University, Karaikudi-
630003, Tamil Nadu, India.
a
‫٭‬AssistantProfessor, Pharmacognosy Lab, Department of Animal Health and
Management, Alagappa University.
*Corresponding author:dr.nithya.gopinath@gmail.com
ABSTRACT
Despite the huge diversity of antibacterial compounds, bacterial resistance to first
choice antibiotics has been drastically increasing. The present study investigated that
the edible wood putrefying mushroom Lentinus cladopus Lev mushroom was
collected from the roots of trees and identified based on morphological characteristics
through mycochemical screening. The preliminary photochemical constituents and
health promoting biomedical properties of methanolic and ethanolic extract of
Lentinus cladopus Lev mushroom was evaluated. Further, the antibacterial activity of
L.cladopus extracts showed greater inhibition against E.coli, Staphylococcus aureus,
Pseudomonas aeruginosa, Bacillus subtilis at 75μg/ml. In addition, in vitro anti-
inflammatory activity was assessed when compared with standard Diclofenac (100
μg/ml) using albumin denaturation method. From the results, the methanolic extract of
L.cladopus (66.47 % at 1000 μg/ml) showedmore significant of anti-inflammatory
activity when compared with ethanolic extract of L.cladopus. Based on the findings,
Lentinus cladopus Lev edible mushroom can be considered as a medicinal drug for
the treatment of inflammation associated diseases and pathogens and it may be
recommended for further analysis.
KEYWORDS: Lentinus cladopus Lev, antibacterial,anti-inflammatory.

AN EVALUATION OF PEST MANAGEMENT STRATEGIES IN
THE FACE OF POTENTIAL EFFECTS OF CLIMATE CHANGE
ON AGRICULTURAL SECTOR
Muzafar Riyaz1 and S. Ignacimuthu1*

1
Entomology Research Institute, Loyola College Chennai, Tamil Nadu -600 034.
*Corresponding Author:eriloyola@hotmail.com
ABSTRACT
The agrarian ecosystems are very subject to the atmospheric conditions, increments in
temperature and carbon dioxide (CO2) canenhance some product yields in a few
areas.Be that as it may, to understand these advantages, supplement levels, soil
dampness, water accessibility, and different conditions should likewise be met.
Changes in the recurrence and seriousness of pest outbreaks, dry seasons and surges
could posture challenges for agriculturists and farmers and food security. The long-
time analysis and investigations of exceptional insect communities give us vital data
about the feedback of the insects distant to the current century. Becoming acquainted
with the potential feedback of insect communities to climate change makes us
conceivable to evaluate the adjustment of pest management options and in addition to
detail our future administration protocol. Climatic change, including global warming
and extended variance, require improved examinations that can be used to assess the
peril of the current and as of recently made pest managements and techniques, and to
describe the impact of these systems on condition, productivity and profit. Each
strategy must be evaluated whether and how it is fitting in the cultivating framework
where they are to be practiced. The influence of unstable conditions on farming are
known by the time we faced some major decrease in productivity of crops, yet
substantial divergence stays in our insight into how agricultural frameworks will be
influenced by both short-and long-haul changes in climate, and what suggestions
these progressions will have for agrarian subsistence.
KEYWORDS: Pests, Climate, Global warming, Agriculture.

REVIEW ON IMPACT OF CLIMATE CHANGE IN PHOSPHATE
NUTRIENT MANAGEMENT
T.P Ajeesh Krishna1 and V. Duraipandiyan1*

1
Division of Plant Biotechnology, Entomology Research Institute, Loyola College,
Chennai-600034, India.
*Corresponding author:avdpandiyan@yahoo.co.in
ABSTRACT
Each macro- and micro-nutrient plays a unique and irreplaceable role in the optimal
growth, development and yield of plants. Next to nitrogen, phosphorus (P) is a vital
macro nutrient affecting overall plant growth and as a component of cell membranes,
P is also involved in a multitude of functions including energy transfer,
photosynthesis and many metabolisms. Climate change adversely affects soil P
availability and plant-soil microbial activities in inorganic phosphate (Pi) uptake and
remobilization. Climate change also alters the frequency and intensity of precipitation
leading to soil P leaching while both high precipitation and temperature facilitate
rapid immobilization, mineralization and weathering affecting soil P forms and
availability. The one-way transport of P from agricultural fields to various water
bodies deteriorates water quality and causes eutrophication and other ecological
damages. The indispensable role of P calls for a high resolution study on the impact of
climate changes and various plant processes of P management. Given the global P
crisis in terrestrial ecosystems, this paper aims at presenting a better understanding of
how various effects of climate change such as elevated temperature, CO 2,
precipitation, drought and agricultural P runoff affect P nutrient management and
suggests creative ways to mitigate them to ensure better plant growth, yield and food
security.
KEYWORDS: Phosphorus, Climate changes, Elevated temperature, CO 2,

Precipitation, Drought, Agricultural P runoff.

REVIEW ON ROLE OF PHT1 FAMILY TRANSPORTERS IN
ACQUISITION AND REDISTRIBUTION OF PHOSPHORUS IN
PLANTS
G. Victor Roch and V.Duraipandiyan*

*Corresponding author:avdpandiyan@yahoo.co.in
ABSTRACT
Phosphorus (P) is one of the most important nutrients for plant growth and yield. Low
availability of inorganic phosphate (Pi) in soil seriously curbs crop production, while
excessive Pi-fertilization causes economic and ecological problems. The rapid
depletion of global rock phosphate reserves calls for a high resolution research on
efficient plant Pi-management. To cope with low-Pi stress, plants have evolved
morphological, physiological, molecular and biochemical adaptations. Apart from
arbuscular mycorrhizae fungi mediated Pi-acquisition, Pi-uptake, export, utilization
and remobilization depend on transport processes mediated by membrane bound
Phosphate Transporters(PHTs), which are grouped into five families viz. PHT1,
PHT2, PHT3, PHT4 and PHT5. Among these, PHT1 family is the primary transporter
involved in the acquisition of Pi from soil and redistribution within plants. In this
review, we assess the details of PHT1 to PHT5 identified till date and correlate their
function in Pi-transport. We enlist all the PHT1s identified and characterized in
various plants including their phylogenetic relationship, expression patterns, and
induction by AMF, localization and affinity.We also discuss the present
understanding of regulation of PHT1s at transcriptional, post-transcriptional and post-
translational levels. Further exploitation of PHT1 and other family of transporters
will help overcome the problems associated with low Pi soils and assist in improving
crop yields in order to ensure global food security.
KEYWORDS: Phosphorus, Rock phosphate, Arbuscular mycorrhizae fungi,
Phosphate transporters, Global food security.

DIRECT PLANT REGENERATION FROM IN VITRO-DERIVED
SHOOT APEX OF FINGER MILLET AND ANALYSIS OF
GENETIC FIDELITY USING MOLECULAR MARKERS
T. Maharajan1 and V. Duraipandiyan1#

1
#
Corresponding author:avdpandiyan@yahoo.co.in
ABSTRACT
Finger millet (Eleusine coracana (L.) Gaertn.) belongs to the family Poaceae, and is
widely recognized as an annual herbaceous cereal crop and consumed by poor people
in Africa and Asia. It contains rich amounts of protein, minerals and nutrients as
compared to other major cereals like wheat, rice and sorghum. An efficient protocol
for the plant regeneration of finger millet was developed by the usage of shoot apex
explants through direct plant regeneration. The explants were inoculated with three
different cytokinins such as benzyl amino purine, thidiazuron (TDZ) and kinetin at
various concentrations for the optimization of multiple shoot induction conditions.
The maximum response of shoot induction was developed in Murashige and Skoog’s
(MS) medium supplemented with 4.5 µM TDZ with an average of 17.3cm shoots
after six weeks of incubation. The effect of Indole-3-butyric acid along with MS
medium was studied for root development, where an individual shoot and root length
was 8.2cm after two weeks on incubation. The present study was, inter-simple
sequence repeats primers were used to analyze clonal fidelity of in vitro regenerated
plantlets of finger millet genotype. This work also will be helpful and supportive to
the genetic improvement of these millets as successful development of transgenic
plants varieties modified for resistance to both biotic and abiotic stress in the near
future.
KEYWORDS: Finger millet, Shoot apex, In-vitro, Regeneration, ISSR.

DISTRIBUTION OF PHYTOPLANKTON DIVERSITY IN
AUTHOORANGAL CHANNEL AT THOOTHUKUDI DISTRICT
F. Esther Isabella Eucharista*and A.Shanthi
*Department of PG Zoology, Aditanar College of Arts & Science, Tiruchendur.
ABSTRACT
Fresh water is finite resource, essential for agriculture, industry and even
human existence, without fresh water of adequate quantity and quality, sustainable
development will not be possible. Water is mainly used for drinking, bathing,
fisheries and other domestic purpose. During recent years there has been increasingly
greater concern for inland fresh water resources, which are affected in different ways
by all kinds of human activities. The maximum values of physic-chemical parameters
of water at Authoorangal channel investigated during January to March 2018 were
such as turbidity 4.4±0. 307 NTU during February, temperature 23.5±0.223 0c during
March, pH 7.8±0. 183during February, dissolved oxygen (DO) 8.8±0. 326 mg/l
during January and total dissolved solids (TDS) 11.5±0.353 mg/l during March at
ranges respectively. A total number of about 130 species of phytoplankton belonging
to cyanophyceae (48 species), bacillariophyceae (33 species), chlorophyceae (32
species), euglenophyceae (7 species), dinophyceae (4 species), chrysophyceae (1
species), pyrrophyceae (1 species), ulvophyceae (1 species), pyrmnesiophyceae (1
species), noctiluciphyceae (1 species) and zygnemophyceae (1 species) were
identified. Among these cyanophyceae was the most dominant group. The result of
the present investigations revealed that there was a fluctuation in the physic-chemical
parameters of water. This will be due to climatic change, sedimentation of organic
nutrients, dumping of municipal and domestic wastes and also agricultural run-off
from the nearby agricultural field.
KEYWORDS: Physico-chemical parameters, Phytoplankton.

IN VITRO ANTIMICROBIAL ACTIVITY OF RHIZOPHORA
MUCRONATA LEAF EXTRACTS AGAINST CLINICAL
PATHOGENS
J. Rajaselvam, I.Vasudhevan and T.Subi

PG & Research Department of Zoology, Vivekananda College, Agasteeswaram,
Affiliated to Manonmaniam Sundaranar University, Tirunelveli.
ABSTRACT
The present study was aimed to evaluate the aqueous, chloroform, ethanol, butanol,
acetone and petroleum ether extracts of Rhizophora mucronata leaf on some human
pathogenic bacteria. The inhibitory effect of the Rhizophora mucronata leaf extract
was tested against Klebsiella pneumoniae, Salmonella typhi,Proteus mirabilis,
Enterobacter, Shigella flexneri, Aspergillus niger and Candida tropicalis using disc
diffusion method. Among the various microorganisms tested the maximum zone of
inhibition was observed on chloroform leaf extract against Aspergillus niger (15 mm)
and the minimum zone of inhibition was observed on chloroform and ethanol leaf
extract against Enterobacter (7mm). Thus the results revealed that Rhizophora
mucronata has potential antimicrobial agent used for the development of potent drugs.
KEYWORDS:Antimicrobial activity, Rhizophora mucronata, Disc diffusion method.

SELECTIVE SYNTHESIS OF BENZYL PHENYL SULFOXIDE
FROM BENZYL PHENYL SULFIDE USING [Fe(Phen) 3]3+
Pathakaraimuthu Balakumar1, Subramaniam Balakumar2, Perumal

Subramaniam3 and Sweetlin Rajula Rubavathi. D4
1
Dr. Sivanthi Aditanar College of Engineering, Tiruchendur 628 215, India
2
PSN College of Engineering and Technology, Tirunelveli 627 152, India
3
Aditanar College of Arts & Science, Tiruchendur 628 216, India
4
Loyola College, Nungambakkam, Chennai 600034, India
ABSTRACT
The present proposal work is on the selective synthesize of benzyl phenyl
sulfoxidefrom benzyl phenyl sulfidewithout formation of sulfone using
[Fe(Phen)3]3+as oxidant. Usually oxidation of sulphides produces mixture of sulfoxide
and sulfone. But in this method only sulfoxide is identified as the product without
further oxidation to sulfone. Benzyl phenyl sulfide (BPS) has been taken as a reactant
and [Fe(Phen)3]3+ as oxidant. The reaction was carried out at a p H of 4-5in aqueous
methanol medium. The yield of the product sulfoxide formed was studied by varying
the concentration of BPS, [Fe(Phen) 3]3+, Temperature, pH and solvent composition.
The optimum conditions for the maximum yield of sulfoxide. It has been observed
that one mole of benzyl phenyl sulfide consumed two moles of [Fe(phen) 3]3+. The
maximum yield of sulfoxide achieved in the present study was approximately 92%
whilesulfone formation being only 2 % confirming selective oxidation of sulfoxide.
The purification of the product performed using silica gel column chromatography
with EtOAc/n-hexane (1/10). The productsulfoxidewas characterized by IR and 1H-
NMR and GC-MS studies.

SURVEY OF BY- CATCH OF CRAB FISHERY FROM
THOOTHUKUDI COAST
K. Jesima* and S. Jesily

PG and Research Department of Zoology, St.Mary’s
College(Autonomous),Thoothukudi
Affiliated to Manonmaniam Sundaranar University, Abishekapatti, Tirunelveli -
627012,
Tamilnadu, India.
*
Corresponding Author: kjesifernando@gmail.com
ABSTRACT
Trawling though one of the most efficient methods of fish capture is also
found to be the most important human caused physical disturbance on the
world’s continental shelves and hence the physical destruction of ecosystems.
The main objective of the present work is to know the species composition of by-
catch from crab fishery landed out Therespuram landing centre, Thoothukudi
coast. Crab fishery in Thoothukudi coast is constituted by five commercially
important species namely Scylla serrate, Portunus pelagicus, Portunus
sanguinolentus, Charybdis natator and Charybdis feriatus. The edible species of
organisms recorded from the discard during the study period were gastropod mollusk
Turbinella pyrum and the finfishes Lutjanus eherenbergii and Nibea Maculata. Out of
the 45 species collected 11 species were of commercially important molluscs. In
the present study about 15 species of molluscs were identified from the by-catch.
In the present study 43 species of organisms were identified with unidentified corals
and sponges. In the present study, the organisms recorded from by-catch are 10
species of gastropod molluscs, 5 species of bivalve molluscs , 3 species of star
fishes, 6 species of inedible crabs, 5 species of finfishes, corals, ascidians, sea
cucumber, sea urchins and sea horse.
KEYWORDS: By-catch, bottom set gill net, crab net, star fish, finfishes,
molluscs.

A STUDY OF MANAPADU WATER BODIES WITH
REFERENCE TO HYDROBIOLOGICAL AND FISHERIES
ASPECTS
Devi Priya .S* and L. JeyaPraba

Zoology Department & Research Centre, Sarah Tucker College (Autonomous),
Tirunelveli, Tamil Nadu.
ABSTRACT
Water is one of the man’s most important natural resources. Most of living organisms
in this biosphere cannot survive for long periods without water. Fishery is an
environment depends on industry and it is based on rich blessings from sea. The
present study was under taken to study the fish diversity in Manapadu coast from
March to May 2017 along with the physico-chemical parameters Mayer’s
method(pH,salinity,chloride,EC and TDS) of water different site- River surface ,River
bottom, Seasurface, Seabottom, Estuary surface and Estuary bottom. Fish collection
was done with gill nets. The result of present investigation reveals the occurrence of
34 species belonging to 19 families .species diversity calculation was done using
SPSS , and most found in all months but low in March because of annual fish
banning and The fish diversity was dominant during summer season May. This kind
of fish diversity data and physico chemical parameter analysis is very essential to
create public awareness about the non-biodegradable materials like plastic covers,
bottles, metal glass pieces, garbage etc.,that are thrown in to the sea and to educate the
local public regarding the significance of the sea and its organism which has to be
protected from pollution. Thereby protecting one of the most important natural
resources.
KEY WORDS: Physico-chemical parameters, pH,salinity,Manapadu fish diversity.

PURIFICATION AND CHARACTERIZATION OF LECTIN
FROM SYZYGIUM CUMINI
P. Rama Devi * ˡ K. Nandha Devi1, Arokiya Mary Fernandez1 and C. Babu ²

ˡ Aditanar College of Arts and Science, Tiruchendur
² Pioneer Kumaraswamy College, Nagercoil
Corresponding Author: p.remadev@yahoo.co.in
ABSTRACT
Lections are carbohydrate binding protein, which are highly variable in their amino
acid sequence that are abundantly found in nature. There is a continuous interest in
lectin due to their biological properties than can be exploited for medical and
therapeutic purpose. In the present study novel lectin was purified and characterized
from Syzygiumcumini. The lectin was purified by gel filtration chromatography on
Sephadex G75 (1.5cm x 100 cm) performed in TBS buffer pH 7.6 fraction (2 ml each)
were monitored at A280. Protein concentration in the purified lectin was 10mg-ml-1
measure by Broadford method. The purified lectin gave a single symmetric protein
peak on gel filtration chromatography showing a molecular weight of 110KDa and
when subjected to native PAGE, showed a single protein band. The
haemagglutination activity was significantly higher when assayed against human O +
red blood cells as compared to other blood group tested. Then the thermal stability,
the lectin (1mg|ml concentration) was incubated at different temperature of 300C to
900C for the period of 50 mins. Heamagglutination activity was assessed which
revealed that the lectin was stable at pH7.6 and 500C.
KEYWORDS: Lectin, Heamagglutination, Chromatography, SDS

HIV PREVALENCE
C. Sundaravadivel and A. Alagu Rajasekar
Department of Zoology and Research Centre, Aditanar College of Arts and Science,
Tiruchendur, Tamil Nadu, India
ABSTRACT
This study investigates the prevalence and correlates of sexual activity and HIV-risk
behavior among adults with a mental disorder. Demographic, psychiatric, sexual
behavior, and substance-use data were available for 1,558 outpatients. During the past
year, 69% were sexually active and 23% engaged in risky behavior. Risk markers
included multiple sexual partners (19%), a sexually transmitted disease (4%), sex
trading (3%), injection drug use (1%), and needle sharing (<1%). Being sexually
active and being at risk for HIV infection were associated with alcohol and drug use,
psychiatric diagnoses other than schizophrenia, and younger age. Married patients
were more likely to be sexually active but less likely to engage in risk behavior.
Screening for HIV risk in psychiatric settings can identify patients who may benefit
from risk reduction programs.

A MOSQUITOCIDAL ACTIVITY OF MUSIZIN ISOLATED
FROM RHAMNUS WIGHTII WIGHT AND ARN
William Raja Tharsius Raja1, Ganesan Pathalam1, Duraipandiyan Veeramuthu1*
1
Division of Ethnopharmacology, Entomology Research Institute, Loyola College,
*
Corresponding Author: avdpandiyan@yahoo.co.in
ABSTRACT
The present study is the efficacy of Rhamnus wightii Wight.,extracts were tested
againstCulexquinquefasciatus(Say)., at various concentrations (0.5, 1.0, 1.5 and 2.0
ppm).The different extracts were screened on larvae and egg of C.
quinquefasciatus.The active ethyl acetate extractwas packed incolumn
chromatography followed by eluting the fractions. Overall 250 fractions were
collected and combinedtogether into 10 major fractions. The fractions were combined
based on the TLC profiles. Among these, fraction 2 [Hexane:Chloroform (1:1)]
showed a promising larvicidal activity. Further, the active fraction 2 purified and
subjected to spectroscopic analysis.The compound was identified as Musizin. Musizin
caused 70% larvicidal activity with LC50 and LC90 values of 1.62 and 4.51 ppm
respectively at 24 h.The larvae exhibited restless movement and seizure followed by
death; in control, the larvae presented normal movement. Musizin showed 83%
ovicidal activity against Cx. quinquefasciatus eggs after 120 h post-treatment at 2
ppm. Musizin treated eggs were contracted and the majority of the eggs did not hatch.
Musizin was tested for toxicity against non-target organism Poeciliareticulata, and no
toxicity was found against this organism. Histopathology study was conducted for
musizin treated larvae and the result revealed serious harm to the larval mid-gut cells;
in control, there was no damage in the mid-gut cells of larvae. The present study
revealed that the isolated compound Musizin could be utilized as a natural pesticide to
control vector mosquitoes.
KEYWORDS: Rhamnus wightii, Musizin, Cx. Quinquefasciatus, Larvicidal,
Ovicidal, Poeciliareticulate, Histopathology.

ICHTHYOFAUNAL DIVERSITY OF OKHA REEF, GULF OF
KACHCHH, WEST COAST OF INDIA
S. Subburaman¹*, Nileshjoshi², D. Nagarajan¹, J.Nagarajan¹ and Ramakotti¹

¹Kamaraj College, 482, Tiruchendur Road, Thoothukudi – 628003, Tamilnadu, India
²Gujarat State Biotechnological Mission, UdyogBhavan, Gandhinagar – 382011,
Gujarat
*
Corresponding Author: subburaman.mb@gmail.com
ABSTRACT
The coral reefs are one of the major marine ecosystems, because of their high
productivity. The ichthyo faunas are the major indicators of the healthy reefs.Gulf of
kachchh is one of the marine parks in India.Okha reef is located tip of the Gulf of
Kachchh, faced on Arabian Sea, west coast of India. Three villages lay on Okhareef;
they are Okha, Arambada, and Mithapur. The marine fishes collected from fishermen,
those were exclusively depending on Okha reef. The study was carried out 2015.
During this study, 39 marine fish species were identified, which belongs to 36 genus
and 36 families. However, twoclasses were observed;the class Actinopterygii (94.1%)
was the dominant class than Elasmobranchii (5.9%).Likewise, the14 orders were
observed. The order Perciformes was the highly dominant group (62.75%) than others
like Clupeiformes (7.84%), Gonorhynchiform (3.92 %), Beloniformes (3.92 %),
Siluriformes(3.92 %) and rest of them having 1.96 % equally.
KEY WORDS: Icthyofauna, Coral reef, Mithapur reef, Gulf of Kachchh.

EFFECT OFYEAST ADDEDDIET ON CAROTENOID CHANGES
IN FRESH WATER ORNAMENTAL FISH GOLD FISH
(CARASSIUSAURATUS)
R. Prema1 and BeenaSomanat2

1
Research Scholar Dept. of Zoology, Muslim Arts College, Thiruvithancode.
2
Assistant Professor Govt. Rani Anna College, Tirunelveli.
ABSTRACT
Probiotics are a cultured product or live microbial fed supplement, which
beneficially affects the host by improving its intestinal balance and health of
the host. In the present study the effect of probiotics (yeast) induced carotenoid
changes in skin, and muscle tissues of gold fish(Carassiusauratus) were
investigated. Normally pigmentation in the skin is responsible for the
colouration in the fish. Ornamental fishes are acceptable toconsumers if they
have striking and vibrant colours. Colouration which is one of the most
important factors deciding the market value of the ornamental fish. Carotenoid
especially astaxanthin has effects on many of these creatures main body
function like prevention from essential unsaturated fatty acids, prevention
from effects of ultraviolet light, immunological reactions, pigmentation. At the
end of the experimental period, biochemical studies, carotenoid content and
TLC were analyzed. It is concluded from this study that probiotic diet had
positively improved growth performance and colouration in gold fish
(Carassiusauratus). This study suggests that yeast can be included as an ingredient of
gold fish diet upto 2%. The 2% for total carotenoid content of the probiotics
supplement gold fish skin tissue was ranged from 4.2344±0.039 mg/g.(10 days)
to10.6527±0.0621mg/g(40days) wet weight and in gold fish it was ranged
2.3824±0.083(10 days) to 8.56±0.43 (40days) wet weight,respectively. Based on this
study, it was concluded that short term feeding had no adverse effect on gold fish
but long term effect needs future research further work should focus on
digestibility co-efficient from of different nutrient classes in Saccharomyces sps .
KEY WORDS: Probiotics, Supplemented diets, Carotenoid, Gold fish.

EFFECTS OF VITAMIN C ON CARBOHYDRATE PROFILE OF
SILK WORM, BOMBYX MORI LXCSR2 AND PURE MYSORE
1
Sundara Raj J, 1W. Kerenhap Evangelin, 2S Akhil Kumar
1
Department of Zoology, St. John’s College, Palayamkottai, Tamil Nadu, India
2
Departement of Animal Science, M.S University, Tirunelveli, Tamil Nadu, India
ABSTRACT
The present study has been demonstrated severe perturbations in carbohydrate profile
of the silk worm Bombyx mori, in different tissues such as Silk gland, Intestine, Fat
body and Haemolymph when fed on mulberry leaves fortified with Vitamin C in
different proportions. The experimental worms were fed thrice a day. The fortification
of leaves was done by leaf spraying method. Dietary supplementation of 1000 ppm
Vitamin C increased the Haemolymph Carbohydrate into a greater extent in the Silk
worm race Pure Mysore. Since the result achieved were considerable and could be
recommended to improve the sericulture parameters.
KEY WORDS: Supplementation, Silk worm, Haemolymph, Bombyx mori

ANTIMITOTIC ACTIVITY OF FUSINUS NICOBARICUS
EXTRACT ON ALLIUM CEPA ROOT TIP
P. Subavathy, R. D. Thilaga and B. Jeba Nesam*

PG and Research Department of Zoology, St.Mary’s College (Autonomous),
Thoothukudi
Affiliated to Manonmaniam Sundaranar University, Abishekapatti, Tirunelveli –
627012, Tamilnadu, India
*Corresponding Author: jebamofrin@gmail.com
ABSTRACT
Molluscs are a rich source for discovering novel compounds for the possible
development of new types of antibiotics for pharmaceutical use. Cancer is a class of
disease characterized by out-of-controlled cell growth. The studies on antimitotic
property with the help of onion root tips also pointed out the evidence of the presence
of antitumour agents in molluscs. Methylene chloride extract of Fusinus nicobaricus
showed maximum inhibition with 36.99% followed by hexane extract with 20.78%,
methanol extract with 17.62% and benzene extract with 10.31%. It is evident that all
extracts reduced the mitotic index significantly. The reduction in number of dividing
cells in the root meristem showed the antimitotic effects of the substances that found
in gastropod Fusinus nicobaricus extracts. Fusinus nicobaricus contains antimitotic
constituents that can stop the mitosis in anywhere of the cell cycle. They also affect
the cytoskeleton or tubulin polymerization or degradation. A new generation of
antimitotic drugs is being developed to understand how cancer cells respond to them.
So, the present study has been carried out with a view to investigate antimitotic
activity of Fusinus nicobaricus.
KEYWORDS: Fusinus nicobaricus, Antimitotic activity, Allium cepa, Methylene

chloride, Hexane

PLANT PHYTO-CHEMICALS USED AS A BIO-PESTICIDES
FOR INSECT PEST CONTROL
Lingathurai S* and Crosslin Anto Singh
*Corresponding Author: lings02@gmail.com
ABSTRACT
Plant phytochemicals have long been touted as excellent alternatives to synthetic

insecticides for pest management because botanicals reportedly posed little threat to
the environment and human communities. Pyrethrum and neem are well established
commercially. Pesticides based on plant essential oils have recently entered the
marketplace. A number of plant substances have been considered for use as insect
antifeedants, repellents, attractant, ovicidal, oviposition deterrent, and growth
regulators with insecticidal properties. The resistance developed among the target
insect population due to repeated application of chemicals in higher doses, their rising
cost and lack of eco-friendliness have contributed to the public opinion against the use
of chemical pesticides. Hence, there is a need to adopt eco-friendly agro practices for
preserving environment and protecting human health by reducing the use of toxic
chemical pesticides and replacing them by botanical pesticides to the extent possible.
The information pertaining to botanical pesticides is compiled with particular
emphasis on rotenone, pyrethrum and essential oils their mode of action is elucidated
with suitable case studies and their role in IPM is also highlighted.
KEY WORDS: Botanicals, Antifeedant, Oviposition deterrent, Ovicidal, Growth

regulation, Integrated Pest Control

CULTURE OF OYSTER MUSHROOM (PLEUROTUS
OSTREATUS) ON DIFFERENT KIND OF AGRO WASTE AND
EXOTIC PLANT WASTE
Lakshmanan G*, kamala Devi N.G. and Kabilan S

*Corresponding Author: menilax@gmail.com
ABSTRACT
Mushroom culture is one of the simplest and excellent applications of
biotechnology. Mushrooms are fungal origin and it now has universal focus and
importance because of its nutritional value and therapeutic values which can be used
for the remedy of various diseases and disorders. Mushroom cultivation uses
agriculture waste products such as paddy straw, wheat straw sugar cane magesy,
coconut pith etc., this present research is aimed to production of oyster mushroom
Pleurotus ostreatus on agro waste paddy straw. The information will help the farmers
for decision making for diversifying their cultivation as well generating extra income.
KEY WORDS: Agricultural waste, paddy straw, oyster mushroom, production

BIOFABRICATION OF SILVER NANOPARTICLES USING
LANTANA CAMARA AND ITS ANTIBACTERIAL ACTIVITY
Lakshmanan G*, Kamala Devi N, Muthu Krishnan G. and G.

Anantha Sundar Raj
Department of Zoology and Research Centre, Aditanar College of Arts
and Science, Tiruchendur, Tamil Nadu, India
ABSTRACT
In the present study, the biosynthesis of silver nanoparticles using
Lantana camara, has been successfully demonstrated. We found that the
L. camara can be a good source of synthesis of silver nanoparticles.
Large scale synthesis of silver nanoparticles can be done by eco-friendly
method as mentioned above. In this method there is no need to use high
pressure, energy, temperature and toxic chemicals as in case of chemical
and physical method. Color change occurs due to surface Plasmon
resonance during the reaction with the ingredients present in the plant
leaves extract results in the formation of silver nanoparticles which is
confirmed by UV-vis. These nanoparticles have a great application in the
field of pharmacological and electronical industries and many more. The
antibacterial activity of biologically synthesized silver nanoparticles was
evaluated against common pathogens showing effective bactericidal
activity. Hence the complimentary properties of silver nanoparticles
properties of silver nanoparticles obtained from L. camara can unfasten
newer pathways in drug development.
KEY WORDS: silver nanoparticles, Lantana camara, antibacterial

PHYSICO-CHEMICAL CHARACTERIZATION OF AGGLUTININ
FROM THE LATEX OF PEDILANTHUS TITHYMALOIDES
Sheeja . H.M 1.,Vinoliya Josephine Mary, J2., and Mary MettildaBai, S3

1
Research Scholar, 2,3Assistant professor
1,2,3
Department of Zoology, Holy Cross College (Autonomous), Nagercoil.
Tamil Nadu, India.
2
Corresponding author: vinoliya75@gmail.com
ABSTRACT
Lectins are proteins or glycoproteins isolated from viruses, bacteria, fungi, plants,
invertebrates and vertebrates with the ability to interact with carbohydrate structures and
to agglutinate cells or glycoconjugates. The latex of Pedilanthus tithymaloides was
examined for agglutinating activity. HA activity was performed with different
mammalian erythrocytes. Among different erythrocytes tested rabbit erythrocytes gave
high hemagglutination activity. The HA activity of the latex was stable between 5-8.5
pH and the temperature 0- 40◦C. Preliminary characterization of the hemagglutination of
latex showed that the activity was dependent on Ca2+ and Mg2+. Addition of EDTA and
trisodium citrate decreased the HA activity, suggesting the agglutinin to be calcium
dependent. Disappearance of agglutinability following cross adsorption revealed the
presence of a single agglutinin. The agglutinin was weakly inhibited by sugars such as
raffinose and lactose. Among the glycoprotein tested the agglutinin was potentially
inhibited by PSM.
KEY WORDS: Agglutinin, latex, Pedilanthus tithymaloides

EFFECTS OF PROBIOTICS ON HEAVY METAL
DETOXIFICATION IN ORNAMENTAL FISH (CYPRINUS CARPIO)
R.C. Ramya*, A. Palavesam

Department of Animal Science, Manonmaniam Sundaranar University, Tirunelveli-
627012, Tamilnadu, India.
*Corresponding Author: ramyarajagopalakrishnan94@gmail.com
ABSTRACT
The hazardous effect of heavy metal, cadmium and its detoxification in selected organ
of the freshwater fish common carp (Cyprinus carpio L.) was investigated. For toxicity
test, Cyprinus carpio of similar age group weighing 8.50±1.20g were procured from the
commercial hatchery, Palayamkottai, Tirunelveli. Toxicity test was performed under
laboratory condition to determine 24hr LC50 and for that fishes were exposed to various
concentrations of Cadmium metal (2.5ppm, 3ppm, 3.5ppm, 4ppm) @ ten fishes for each
concentration. A control devoid of cadmium was also maintained in the same volume of
water. LC50 value for 24 hrs was determined as 3.5ppm. In order to findout the effect of
cadmium 10% of 24 hr LC50 (0.35 ppm) was incorporated along with feed. A control
diet was also prepared without cadmium, and was given to control group. For both
control and experiment, 5 fishes were reared in 20 l glass tank. The experiment was
lasted for 21 days. And during experimentation, fishes were feed with control and
experimental diet @ 10% body weight one in a day. Also 50% water exchange was
made. The feeding experiment was carried out for 21 days, and after experimentation
During this refeeding experiment, probiotic feed was given at the rate of 0.25mg/100g.
After refeeding, gut histology was studies for 7, 14 and 21 days of experiment and it
indicated the recovery of damaged gut tissues. The present investigation evidenced that
the probiotic bacteria can cure the toxicity caused by heavy metal Cadmium and it was
found to be concentration dependent.
Keywords: Heavy metal, Cadmium, Bacillus, Histology.

FOURIER TRANSFORM INFRARED SPECTROSCOPY ANALYSIS OF
METHANOLIC LEAF EXTRACT OF GREWIA ORBICULATA ROTTL.
T. Hemalatha* and A. Saravana Ganthi

Department of Botany, Rani Anna Govt. College for Women, Tirunelveli, Tamil
Nadu, India
*Research Scholar, Manonmaniam Sundaranar University, Tiurnelveli, Tamil Nadu,
India
Corresponding Author:saran_gan@rediffmail.com
ABSTRACT
Medicinal plants have critical role in curing diversity of human diseases. Grewia orbiculata
Rottl. is a medicinal plant used to treat many diseases in folk medicine. The present study
aimed to investigate and characterize the chemical composition of the crude extracts from the
leaves extract of Grewia orbiculata. The air dried plant materials extracted with methanol and
subjected to FTIR analysis. The FTIR analysis of methanol leaf extract of G. orbiculata
confirmed the presence of amide, alcohols, phenols, alkanes, carboxylic acids,
aldehydes, alkenes, primary amines, aromatics, esters, ethers, alkyl halides and aliphatic
amines compounds, which showed major peaks. The FTIR method was performed on a
spectrophotometer system, which was used to detect the characteristic peak values and their
functional groups. The results of the present study generated the FTIR spectrum profile for
the medicinally important plants of Grewia orbiculata can be used in the industry
KEY WORDS:Grewia orbiculata; FTIR Spectroscopy; Functional groups.
INTRODUCTION
Identification of secondary metabolic fingerprint by chromatography and

spectroscopy tools provides useful information about qualitative, quantitative and the
pattern of the composition of there biomolecules. Recently, Fourier Transforms
Infrared Spectroscopy (FTIR) reveals to phytochemical profiles containing
overlapping signals from a wide array of the compounds. Grewia orbiculata samples
extracted with the help of the solvent methanol continuously was subjected to FTIR
analysis. This study was carried out to identify the reactive functional groups present
in the leaves extract of G. orbiculata. Grewia orbiculata Rottl. (Ney-c-citti in tamil)
is a small deciduous trees. The root is used to treat stomach-ache. Leaves of are used
to treat ulcers (Pullaiah et al., 2016). Bark extract of these plants can be used to treat
diarrhea, smallpox, urinary troubles, and irritation in the bladder, gonorrhea and
syphilis. Various parts of this tree are used in the treatment of eye ache, stomach ache
and spleen troubles. The powder of its bark is used to treat wounds (Umberto
Quattrocchi, 2014).
MATERIALS AND METHODS
Preparation of extract
The medicinal plant G. orbiculata was collected from Kalakad, Tirunelveli

District. The specimen was identified referring to the Flora of Presidency of Madras
(Gamble, 1915–1936) and Flora of TamilNadu Carnatic (Mathew, 1983). The
specimens were authentified by Botanical Survey of India, Coimbatore. Healthy leaf,
stem and root samples were collected and shade dried and coarsely powdered. The
500 g of the powdered plant material was defatted with petroleum ether (60-80°C)
using a soxhlet extractor. Then it is successively extracted with methyl alcohol for 72
h. The extract obtained was filtered and concentrated.
FTIR analysis
The FT-IR studies have been followed by the method described by (Jagmohan,
2005). The lyophilized resin or powered samples were mixed with dry potassium
bromide pellet (KBr) and subjected to a pressure of about 5x106 Pa in an evacuated
die to produce a clear transparent disc of diameter 13 mm and thickness 1mm. IR
spectra region 4000- 400 cm-1 were recorded at room temperature on a perkin-Elmer
fourier transform spectrometer equipped an air cooled DTGs (deuterated triglycine
sulfate) detector. For each spectrum, 100 scans were CO- added at a spectral
resolution of 4cm-1. The frequencies for all sharp bands were accurate to 0.01 cm-1.
RESULTS AND DISCUSSION
In the present study, the biochemical content of Grewia orbiculata was

investigated using FT-IR spectroscopy by monitoring different functional groups.
Figure 1 shows the representative FT-IR spectra obtained from the Grewia orbiculata
in the 4000–400 cm-1 region. The frequency ranges from 3340.48 cm-1 peaks are
represents the O-H stretching vibration, the presence of carbohydrate and amino
acids. The frequencies are 2886.27 cm-1 peaks were represents the C-H stretching
mainly: lipids (Table.1). The frequency ranges from 1626.84cm-1 peaks are represents
the Amide: C=O stretching mainly: proteins and the frequency range from 1453.26
cm-1 peaks are represents the ring C=C stretch vibration mainly: aromatic compounds.
Then the frequency ranges from 1318.25 and 1369.37 cm-1 peaks are represents the C-
F stretching mainly: sulfur compounds. The frequency ranges from 1157.21 and
1236.29 cm-1 peaks are represents the C-O stretching mainly: alcohols. The frequency
ranges from 780.15cm-1 peaks are represents the C-H bending mainly: starch. The
frequency ranges from 517.85 cm-1 peaks are represents the C–Br stretching mainly:
aliphatic bromo compounds (Table- 1).
Table 1: FT-IR frequency range and functional groups present in the leaf extract
of G. orbiculata
S. Area Intensity
N Wavelength Functional Type of vibration
o cm-1 group
.
alkyl C–Br stretching 0.045
1. 517.85 Strong
halides
Alkyl C-H bending 0.155
2. 780.15 Strong
halides
=C-O-C sym. & 0.031
3. 1032.81 Ethers asym. stretch Strong broad
Alcohol C-O stretch 0.05

4. 1157.21 Strong
Alcohol C-O stretch 0.04

5. 1236.29 Strong
Alkyl C-F stretch 1.835
6. 1318.25 Very strong
halides
Alkyl C-F stretch 1.282
7. 1369.37 Strong
halides
Aromatic ring C=C stretch 0.221
8. 1453.26 Medium strong
Compounds
Amides N-H bend 0.091
9. 1512.09 Medium strong
10 1626.84 Amides N-H bend 0.234 Medium strong

Esters C=O stretch 0.097
11 1738.71 Strong
- - 0.084
12 2313.46 -
Alkanes 1.056
13 2886.27 C-H stretch Strong
and Alkyls
14 3340.48 Alcohols O-H stretch 0.275 Strong
The various functional groups observed in the different extracts probably

indicate the presence of carbohydrates, carotenoid, amino acids, amides, starch,
phosphates, lipids, and cellulose. Spectral differences are the objective reflection of
componential differences. By using FT-IR spectrum, we can confirm the functional
constituent’s presence in the given parts and extract, identify the medicinal
materials from the adulterate and even evaluate the qualities of medicinal materials
The results of the present study coincided with the previous observations observed
by various plant biologist and taxonomist Many researchers applied the FTIR
spectrum as a tool for distinguishing closely associated plants and other organisms
(Lu et al., 2004; Ellis et al 2002; Kim et al., 2004; Lamprell et al., 2006; Rebuffo et
al., 2006). The results of the present study developed novel phytochemical
marker to identify the medicinally important plant. Further advanced spectroscopic
studies are required for the structural elucidation and identification of active
principles present in the leaves of G. orbiculata.
FT-IR spectroscopy is capable of providing strong insight into the structural

and functional alterations induced by various factors due to its high sensitivity
(Gorgulu et al., 2007). FT-IR technique was used for assessment the type of organic
and inorganic complexes in plants. FTIR analysis results proved the presence of
alcohols, phenols, alkanes, carboxylic acids, aromatics, ketones, and alkyl halides
(Bobby et al., 2012). The more intense bands occurring at O–H/N– H, C– H, C–O,
and C–CI/C–CS stretching/bending vibrations respectively indicate the presence of
amino acids, alkenes, nitrates, ethers, organic halogen compounds and carbohydrates
in plants (Manoj and Ragothaman, 1999).
Fig 1: FT-IR Spectrum of leaf extract of G. orbiculata
100
716.51
781.12
516.89
894.91
%T
616.21
1233.39
2884.35
1317.29
2312.49
3778.29
1512.09
1453.26
1748.35
1367.44
97.5
1604.66
1055.95
1106.10
92.5
3454.27
00 3500 3000 2500 2000 1750 1500 1250 1000 750 500
Sample-P2 1/
Leaf cm
CONCLUSION
The result in the present study showed that FTIR spectroscopy is a valuable
technique to fingerprint and to analyze the different biomolecules from Grewia
orbiculata extract which contains 14 peaks. Based on the peak values more functional
groups were obtained from the leaf extract of G. orbiculata.
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Bobby, MA., Wesely EG, Johnson M. spectroscopy and machine

FT-IR studies on the leaves of learning. Applied and
Albizia lebbeck Benth. Int J Pharm Environmental Microbiology 2002;
Pharm Sci. 2012; 4(3):293-6. 68:2822-2828.
Ellis DI, Broadhurst D, Kell DB, Gamble JS. 1915-1936. Tiliaceae In:
Rowland JJ, Goodacre R. Rapid Flora of the Presidency of Madras
and quantitative detection of the vol:1, London: West, Newman and
microbial spoilage of meat by Adlard.
Fourier transform infrared
Gorgulu ST, Dogan M, Severcan F. cadmium on the Red blood cells of
The characterization and Boleophthamus duosumieri (Cuv.)
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Mathew KM. 1983. The Flora of
spectroscopy. Appl Spectrosc.
Tamilnadu Carnatic. Vol I. The
2007; 61:300-308.
Rapinat Herbarium, Trichirapalli.
Kim SW, Ban SH, Chung H, Cho
Pullaiah T, Krishnamurthy KV, Bir
S, Chung HJ, Choi PS, Yoo OJ,
Bahadur. 2016. Ethnobotany of
Liu JR. Taxonomic
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discrimination of flowering
West Coast of Peninsular India.
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Fourier transform infrared Rebuffo CA, Schmitt J, Wenning M,
spectroscopy data. Plant Cell von Stetten F, Scherer S. Reliable
Reports 2004; 23:246-250. and rapid identification of Listeria
monocytogenes and Listeria
Lamprell H, Mazerolles G, Kodjo
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A, Chamba JF, Noel Y, Beuvier
network-based Fourier transform
E. Discrimination of
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Staphylococcus aureus strains from
Environmental Microbiology 2006;
different species of Staphylococcus
72:994-1000.
using Fourier transform infrared
(FTIR) spectroscopy. International Sahoo S, Chakraborti CK, Mishra SC,
Journal of Food Microbiology Nanda UN, Naik S. FTIR and XRD
2006; 108:125-129. Investigations of some
Fluoroquinolones.Int J Pharm
Lu H-F, Cheng C-G, Tang X, Hu Z-H.
Pharm Sci 2011; 3(3): 165-170.
Spectrum of Hypericum and
Triadenum with reference to their Umberto Quattrocchi, FLS. 2014. CRC
identification. Acta Botanica Sinica World Dictionary of medicinal and
2004; 46:401-406. poisonous plants. In: CRC World
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Manoj, K. & Ragothaman, G. (1999).
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Effect of mercury, copper and
Newyork.
EVALUATION OF ANTIBACTERIAL ACTIVITY OF PLECTRANTHUS
WIGHTII BENTH.
Subramonian K1 Saravana Ganthi A2Padma Sorna Subramanian M 3 and Subha

S.L4
1
Department of Botany, the MDT Hindu College, Tirunelveli, Tamil Nadu, India
2
Department of Botany, Rani Anna Govt. College for women, Tirunelveli, Tamil
Nadu, India
3
Siddha Medicinal Garden, CCRAS, Mettur Dam, Salem, Tamil Nadu, India
4
Sri LakshmiSiddha Clinic, Tirunelveli, Tamil Nadu, India
Corresponding Author:ksmaniyan26@gmail.com
ABSTRACT
Medicinal plants are traditionally used for the treatment of human infections. The
present study was undertaken to evaluate the antibacterial potential of leaf, stem and
root extracts of Plectranthus wightii Benth. against human bacterial pathogens. The
activity of plant extract and selected antibiotic was evaluated against five bacterial
pathogens including Acinetobacter baumannii (MTCC 2031), Chromobacterium
violaceum (MTCC 2431), Enterococcus faecalis (ATCC 29212), Proteus vulgaris
(MTCC 5412), Staphylococcus aureus (ATCC 29213) using disc diffusion method.
The methanolic and petroleum ether extracts showed maximum antimicrobial activity
when compared to benzene, chloroform and water extracts. The chloroform extract of
the stem powder shows maximum inhibition zone against Enterococcus faecalis (6.9
mm).Petroleum ether extracts showsmaximum activity againstProteus vulgaris and
Staphylococcus aureus. The results from the study suggest that the show Plectranthus
wightii hasantibacterial activity against different bacterial species. They could be used
as alternatives to common antimicrobial agents for treatment of bacterial infections.
KEYWORDS: Antibacterial activity, inhibition zones, Plectranthus wightii, disc
diffusion method
INTRODUCTION
Lamiaceae or Labiatae, also called as the mint family, is a family of flowering
plants. The family has a cosmopolitan distribution (Yuan et al., 2010). The
Lamiaceaecontains about 6,900 to 7,200 species (Venkateshappa and Sreenath, 2013).
They are mainly herbs and shrubs, very fragrant and rich in medicinal properties of
great worth in natural medicine and pharmacopoeia. Medicinal constituents include
the strong aromatic essential oil, tannins, saponins and organic acids. The plant has
sedative, diuretic, tonic, antispasmodic and antiseptic properties. Still, several potent
plants of Lamiaceae particularly from the rural areas are unexplored which deserve
attention and research. Plectranthus wightii Benth. is such a plant which have not
been explored extensively by the scientific world so far. Plectranthus wightiiis an
aromatic herb employed in folk medicine for treating hepatic insufficiency and
dyspepsia. Kurichia tribes residing at Vythiri taluk, Kerala, mixed the leaf paste with
turmeric powder and salt is applied externally on wounds (Devi Prasad and Shyma,
2013). Its fresh leaves have a distinct odour and are taken as infusion or aqueous
extract for healing purposes (Lorenzi and Matos, 2002). The present study was
undertaken to evaluate the antibacterial potential of leaf, stem and root extracts of
Plectranthus wightii against human bacterial pathogens.
Mature and healthy parts from the Plectranthus wightii were collected from
different specific locations in Kalakad, Tirunelveli District. The specimens were
identified referring to the Flora of Presidency of Madras (Gamble, 1915-1936) and
Flora of Tamil Nadu Carnatic (Mathew, 1983-1988). Voucher specimens of the
collections are deposited at the Herbarium of Survey of Medicinal Plant Unit, CCRAS
(Siddha), Govt. Siddha Medical College Campus, Palayamkottai, Tirunelveli, Tamil
Nadu.
Preparation of plant extracts
60 g of root, leaf and stem samples were air dried separately, ground and
extracted successively with of 400 ml of petroleum ether (60-80o C), benzene,
chloroform, methanol and water. This sequence of solvents allows for leaching of all
compounds based on their polarity. The individual fractions were collected and
concentrated to obtain crude extracts. For the purpose of experimental use each
extract sample was dissolved in respective solvent so as to get 1/10 solution.
Microbial strains
The antimicrobial activity was tested against five different microbial strains
namely Acinetobacter baumannii. (MTCC 2031), Chromobacterium
violaceum(MTCC 2431), Enterococcus faecalis (ATCC 29212), Proteus vulgaris
(MTCC 5412), Staphylococcus aureus (ATCC 29213)These bacteria were obtained
from the Microbial Type Culture Collection and Gene Bank, Chandigarh, India. The
bacterial strains were grown in Muller Hinton (MH) agar plates at 37 0 C and
maintained on nutrient agar slants, while fungi were grown at 30 0C and maintained in
Saboraud glucose agar slants. Each organism was maintained in a separate culture
media and was recovered for testing by sub-culturing on a fresh media. Inoculums of
each bacterial strain were transferred in 10 ml of Muller Hinton agar broth and
incubated overnight at 37°C.
Antibacterial assay
Antimicrobial activity was assayed by filter paper disc diffusion method. Each
sterile disc was incorporated individually with 200 - 500 µl of extract. Antibacterial
assay was conducted by the method described by Lennette (1985) with some
modification. Levomycin (10 µg / ml) was used as positive control and the solvents
which were used to dissolve the crude extracts served as negative control. The plates
were incubated at room temperature for 24 hrs. After the incubation period the
inhibition zone around the discs were measured and recorded. Three replicates for
each concentration were maintained.
Leaf, stem and root extracts of Plectranthus wightii shows inhibitory activity
against all microorganisms tested. Their inhibition zones ranged from 6.9 – 1.0 mm.
Proteus vulgaris and Staphylococcus aureus are the most susceptible strains to the
petroleum ether extract when tested by the disc diffusion method. The petroleum ether
extracts of root shows maximum activity against all the selected bacteria (4.5 mm)
except Enterococcus faecalis (3.4 mm). The chloroform extract of the stem produces
larger zone against Enterococcus faecalis (6.9 mm) and smaller zone (1.0 mm)
against Acinetobacter baumannii. The methanol extract shows pronounced
antibacterial activity by inhibiting the growth of all isolates used. Leaf extracts
exhibit good antibacterial activity and inhibited growth of Proteus vulgaris. The
aqueous extract is effective against all the bacterial cultures.
The chosen plant used in this study is used in folk medicine across villages in
Tirunelveli district to cure urinary tract infections and wound infections. As
demonstrated in Table 1 the efficacy of antimicrobial activities of crude extract of

tested plant was determined and concluding that the crude extracts of Plectranthus
wightii possess bactericidal effect on five different selected microbial strains.
Levomycin 10 µg/ml is used as positive control and the respective solvents
which are used to dissolve the crude extracts served as negative control. The positive
control Levomycin shows inhibitory zone against all selected bacterial strains. The
positive control exhibits pronounced activity against Chromobacterium violaceum (15
mm) and positive control produces comparatively larger zones than the tested plant
samples. The solvents used in the present study have not shown any inhibition zone
against all the chosen bacteria.
Table 1: Antibacterial activity of leaf, stem and root extracts of Plectranthus

wightii Benth.
S. Name of the Part Diameter of inhibition zone (mm)
No bacteria Pet. Benzene Chloroform Methanol Water
ether
1. Acinetobacter Leaf 1.8 ± 1.7 ± 1.4 ± 0.46 3.6 ± 1.6 ±
baumannii. 0.65 0.64 0.85 0.65
Stem 3.8 ± 1.4 ± 1.0 ± 0.49 3.2 ± 1.3 ±
0.52 0.69 0.56 0.56
Root 4.5 ± 1.5 ± 1.5 ± 0.54 2.4 ± 2.2 ±
0.54 0.58 0.34 0.68
2. Chromobacterium Leaf 3.4 ± 2.3 ± 1.9 ± 0.42 4.2 ± 2.4 ±
violaceum 0.36 0.45 0.25 0.58
Stem 3.4 ± 1.9 ± 1.5 ± 0.57 4.5 ± 2.1 ±
0.32 0.54 0.35 0.65
Root 4.5 ± 1.4 ± 1.6 ± 0.62 2.2 ± 1.9 ±
0.38 0.25 0.65 0.25
3. Enterococcus Leaf 3.9 ± 1.7 ± 2.3 ± 0.57 3.6 ± 3.9 ±
faecalis 0.65 0.36 0.54 0.25
Stem 3.9 ± 1.9 ± 6.9 ± 0.36 4.4 ± 1.9 ±
0.54 0.85 0.46 0.57
Root 3.4 ± 1.7 ± 2.3 ± 0.65 2.4 ± 2.3 ±
0.35 0.63 0.65 0.39
4. Proteus vulgaris Leaf 4.5 ± 2.3 ± 2.3 ± 0.42 2.4 ± 2.4 ±
0.36 0.59 0.59 0.57
Stem 5.3 ± 1.8 ± 2.8 ± 0.56 2.7 ± 1.9 ±
0.65 0.36 0.62 0.63
Root 4.5 ± 1.8 ± 1.8 ± 0.69 3.2 ± 1.6 ±
0.62 0.57 0.26 0.65
5. Staphylococcus Leaf 3.7 ± 2.1 ± 1.4 ± 0.58 3.6 ± 2.3 ±
aureus 0.36 0.85 0.35 0.32
Stem 4.5 ± 1.6 ± 1.7 ± 0.65 2.3 ± 2.3 ±
0.52 0.25 0.24 0.64

Root 4.5 ± 1.7 ± 1.9 ± 0.45 2.7 ± 1.7 ±
0.64 0.36 0.54 0.36
Values are means of three independent analyses of the extract ± standard deviation (n
= 3).
The secondary metabolites present in the plant could be responsible for some
of the observed antimicrobial activity. Normally, plants produce metabolites under
stress conditions, which are being used by the plant as a defense against diseases,
microbial attacks and herbivores. These compounds confer the plants with
antimicrobial activity. The secondary metabolites like tannin, flavonoids and steroids
showed antibacterial activities (Tona et al., 1999; Rhoda, 2001). The antimicrobial
properties of volatile aromatic oils have been recognized since development and
promotion of herbal drugs. Chami et al. (2005) reported that clove essential oil is
shown to possess a broad spectrum of antibacterial activity.
REFERENCES
Chami F Chami N Bennis S Bouchikhi Lennette EH (1985). Antibiotic
T and Remmal A (2005). Oregano susceptibility testing by a standard
and Clove essential oil induce single disc method. In: Manual of
surface alteration of Clinical Microbiology. Macmilan
Saccharomyces cerevisiae. Publishers, Washington, D.C. 978.
Phytotherapy Research. 19 (5): 405 Lorenzi H and Matos FJA (2002).
- 408. Plantas medicinais no Brasil -
Devi Prasad AG and ShymaTB (2013). nativas e exoticas. Nova Odessa:
Medicinal plants used by the tribes Plantarum.
of Vythiri taluk, Wayanad district Mathew K M (1983 -1988). The
(Kerala state) for the treatment of Flora of Tamilnadu Carnatic.I.
human and domestic animal The Rapinat Herbarium. St. Joseph
ailments. J. Medicinal Plants College, Tiruchirapalli, India.
Research.7(20): 1439 - 1451. Rhoda M Kariba (2001). Antibacterial
Gamble JS (1915 - 1936). Lamiaceae activity of Ajuge remota.
In: Flora of the Presidency of Fitoterapia 72: 177.
Madras, Adlard and Son, London. Tona Lutete K Kumbu D Ntondole and
Manga KC (1999).Antimicrobial

activity of tannins.Fitoterapia 2: Yuan YWD Mabberly DA Steane
279. and Olmstead RG (2010). Further
Venkateshappa SM and Sreenath KP disintegration and redefinition of
(2013).Potential medicinal plants Clerodendrum (Lamiaceae):
of Lamiaceae American Implications for the understanding
International J. Research in of the evolution of an intriguing
Formal, Applied & Natural breeding strategy. Taxon.59: 125 -
Sciences.3(1): 82 - 87. 133.

QUANTITATIVE PHYTOCHEMICAL ANALYSIS
OFLEPIDAGATHIS PUNGENS NEES.
Subha S and Saravana Ganthi A

Department of Botany, Rani Anna Govt. College for women, Tirunelveli, Tamil Nadu
Corresponding Author:saran_gan@rediffmail.com
ABSTRACT
Lepidagathis pungens Nees. is a spiny herb of Acanthaceae found in dry lands of
South Tamilnadu and endemic to peninsular India. The present study aimed to carry
out a preliminary phytochemical analysis of the taxa extracted in different solvents to
find out their organic constituents and to carry out a systematic quantitative
phytochemical study above the taxa. Steroids, Phenolic compounds, Saponin, Tannin
and Amino acids are predominantly present in aerial and underground part samples.
The quantitative mineral analysis showed rich in iron, calcium and magnesium
content. Quantitative phytochemical study indicated that the presence of more
flavonoids, phenoilic compounds and tannin. This study has provided some
biochemical basis for the ethanomedical use of extracts from Lepidagathis pungens in
treatment and prevention of infections. As a rich source of phytochemical
Lepidagathis pungenscan be a potential source of useful drugs.
INTRODUCTION
Chemical analysis and biochemical assays are very important aspects in
pharmacognostic evaluation of medicinal plants (Choudhury et al., 2009; Harborne,
1973). The medicinal plants are useful for healing as well as for curing of human
diseases because of the presence of phytochemical constituents (Nostro et al., 2000).
Phytochemicals are naturally occurring in the medicinal plants, leaves, vegetables and
roots that have defense mechanism and protect from various diseases. Phytochemicals
are primary and secondary compounds. Chlorophyll, proteins and common sugars are
included in primary constituents and secondary compounds have terpenoid, alkaloids
and phenolic compounds (Krishnaiah et al., 2007). Terpenoids exhibit various
important pharmacological activities i.e., anti-inflammatory, anticancer, anti-malarial,

inhibition of cholesterol synthesis, anti-viral and anti-bacterial activities (Mahato and
Sen, 1997). Terpenoids are very important in attracting useful mites and consume the
herbivorous insects (Kappers et al., 2005). Alkaloids are used as anaesthetic agents
and are found in medicinal plants (Hérouart et al., 1988). The large numbers of
publications on the chemistry of Acanthaceous taxa are on record (Adam et al., 1992;
Bratoeff and Amador, 1994; Wu Tian et al., 1995; Asano et al., 1996; Sarma and
Narayana, 1998; Singh et al., 2000). The present study aimed to analysis the
phytocinstutients of Lepidagathis pungens aerial and underground part powders.
The identified plant of Lepidagathis pungens collected from Sivanthipatti hills
near Palayamkottai, Tamil Nadu, South India. It was confirmed with voucher
specimen deposited at the Survey of Medicinal Plants Unit, Govt. Siddha Medical
College, Palayamkottai. The taxonomic features of the plant confirmed with the Flora
of Presidency of Madras (Gamble, 1915 – 1921) and The Flora Tamil Nadu Carnatic
(Mathew 1983 – 1988). The air-dried and powdered plant materials were taken in
different amber coloured bottles, extracted (by Soxlet method) in ethanol and then the
solvent were filtered off. The extract thus obtained from the plant was then subjected
to qualitative tests. Total ash was determined by employing standard methods of
analysis as described in Pharmacopoeia of India (Anonymous, 1996). The percentage
of major elements like carbon, nitrogen, phosphorus, potassium, sodium, calcium,
magnesium and sulphur was determined by the method of AOAC (1984). The trace
elements like zinc, copper, iron, manganese, boron and molybdenum were determined
by the method of Williams and Twine (1960). The minerals (N, P, K, Na and Ca)
were estimated using Flame Photometer (Spectronics Flame Photometer, India).
Alkaloids were determined by using the method of Harbone (1973). Flavonoids were
determined by the method of Boham and Kocipal-Abyazan (1974). Biochemical
estimation for Phenol (Farkes and Kiraly 1962) and Tannin (Aparna Buzarbarua,
2000) were carried out. Glycoside and serpentines were carried out on the powdered
samples using the standard procedures as given in Anonymous, AOAC (1984).
Preliminary photochemical screening
The petroleum ether, benzene, chloroform, ethyl alcohol and distilled water
extracts of Lepidagathis pungens shows that the plant contains different types of

chemical constituents and the results are present in Table 1. Steroids, Phenolic
compounds, Saponin, Tannin and Amino acids are predominantly present in aerial
and underground part samples. Reducing sugar is only present in chloroform extract.
Alkaloids are present in benzene extract of underground part. Flavonoids, Catechins
and Anthroquinone are totally absent in all the extracts. Triterpenoids are present in
petroleum ether extract of aerial part and of ethyl alcohol extract of underground part.
Sugar is present in aerial part of distilled water extract and underground part of
chloroform extract. Amino acids are present in aerial part of benzene extract and ethyl
alcohol extract and underground part of benzene extract.
Quantitative estimation of minerals
Table 2 shows the results of quantitative estimation of organic carbon,
nitrogen, phosphorus, potassium, sodium, calcium, magnesium, sulphur, zinc, copper,
iron, manganese, boron, and molybdenum in the dry powder of the aerial and
underground part of the selected taxa. The analysis yielded significant results. Except
total nitrogen and total phosphorus, all other parameters are higher in root powder
extract. The total of iron content is more in the aerial and underground part of L.
pungens (42.06, 81.26 ppm). Total boron and total molybdenum are completely
absent in aerial parts powder extract.
Calcium is essential for healthy bones, teeth and blood (Charles, 1992). The
health of the muscles and nerves depends on calcium. It helps to regulate the activity
of skeletal muscle, heart and many other tissues. Deficiency of calcium causes
rickets, osteomalacia and scurvy. The recommended daily dietary allowance of Ca for
children is between 500 and 1000 mg and 800 mg for adults. In the present study it is
observed that calcium is rich in underground parts power (4.12%), followed by aerial
parts power (2.41%).
Magnesium is the second most abundant element inside human cells and the
fourth most abundant positively charged ion in the human body (Dean, 2007; Fox et
al., 2001). DNA stability is dependent in part on magnesium. Magnesium not only
stabilizes DNA structures, it also functions as a cofactor in the repair of DNA damage
by environmental mutagens (Hartwig, 2001). The richest magnesium content is
observed in underground parts powder extract of L. pungens (4.26%).
Sulphur is a mineral that is present in every cell of the body. It plays a key
role in liver metabolism and the function of the joint cartilage and keratin of the skin

and hair. It is also critical for metabolism and anti-oxidant defence systems that
protect the aging patterns of the brain. Some of the healthiest cultures in the world
have the highest levels of sulphur in their diet (David Jockers, 2011). Current study
shows that sulphur is 0.59% in the underground dry powder extract.
Quantitative estimation of phytochemicals
Phytochemical reports on family Acanthaceae was showed the presence of are
glycosides, flavonoids, benzonoids, phenolic compounds, naphthoquinone and
triterpenoids (Awan and Aslam, 2014). Chavan et al. (2010) reported that the
medicinal value of plants lies in some chemical substances like alkaloids, flavonoids,
tannins and phenolic compounds which serve as defence against many
microorganisms, insects and herbivores. Kumari et al. (2012) also reported that the
natural phenolic, alkaloids, tannins, glycosides and flavonoids compounds function as
antioxidants.
Table 1. Preliminary phytochemical analysis of Leoidagathis pungens Phenolic compounds
Reducing sugars
Anthroquinones
Triterpenoids
Amino acids
Extract Plant part
Flavonoids
Catechins
Alkaloids
Saponins
Steroids
Tannins
Sugars
Petroleum Aerial part + + - - - - - - + + - -

ether Underground
(40 - 60o) + - - - - + - - + - - -
part
Aerial part + - - - - + - - + + - +
Benzene Underground - - - + - - - + - - +
part +
Aerial part + - + + - - - - + - - -
Chloroform Underground - + + - - - - + - - -
part +
Aerial part + - - + - + - - + - - +
Ethyl
alcohol Underground + - + - + - - - - - -
part -
Distilled Aerial part - - - + - - - - + + - -
water Underground
- - - - - - - - + + - -
part
+ present, - absent
Table 2. Comparative quantitative minerals analysis of selected plant

S. No Estimation Aerial part Underground part
1. Organic Carbon (%) 2.23 3.58
2. Total Nitrogen (%) 1.52 1.21
3. Total Phosphorus (%) 0.26 0.25
4. Total Potassium (%) 2.89 3.25
5. Total Sodium (%) 0.03 0.12
6. Total Calcium (%) 2.41 4.12
7. Total Magnesium (%) 0.62 4.26
8. Total Sulphur (%) 0.14 0.59
9. Total Zinc (ppm) 0.52 7.64
10. Total Copper (ppm) 0.06 0.72
11. Total Iron (ppm) 42.06 81.26
12. Total Manganese (ppm) 2.36 5.06
13. Total Boron (ppm) Nil 0.55
14. Total Molybdenum (ppm) Nil 0.02
Table 3. Comparative quantitative phytochemical analysis of selected plant
S. No Estimation Aerial part Underground part
1. Total Alkaloids (mg kg-1) 1.69 1.87
2. Total Flavonoids (mg kg-1) 2.36 3.12
3. Tannin (mg kg-1) 0.51 0.42
4. Lignin (mg kg-1) 0.24 0.26
5. Glycosides(mg kg-1) 0.06 0.07
6. Serpentines(mg kg-1) 0.19 0.14
7. Carbohydrates (mg/g/dry wt.) 1.32 1.54
8. Amino acids (mg/g/dry wt.) 0.26 0.65
9. Fats (mg/kg-1.) 0.03 0.02
10. Protein (mg/g/dry wt.) 0.19 0.14
11. Phenols (mg/g/dry wt.) 0.28 021
12. Terpenoids (mg kg -1) 0.04 0.01
13. Saponins (mg kg-1) 0.02 0.04
Phenol and phenolic compounds such as flavonoids have been shown to

possess significant antioxidant activity (Van, 1996). Flavonoids exhibit a broad range
of biological activity such as antimicrobial, anticancer, antiallergic as well as
antitumour properties. On the other hand, flavonoids are potent water-soluble
antioxidants and free radical scavengers, which prevent oxidative cell damage, have
strong anticancer activity (Pietta, 2000). Flavonoids in intestinal tract lower the risk
of heart disease, and as antioxidant, they provide anti-inflammatory activity (Okwu,
2004; Essiett et al., 2010).
The present study carried out on the Lepidagathis pungensrevealed the
presence of medicinal active constituents. Table 3 shows the quantitative estimation

of phytochemicals in selected plant.The highest amount of total flavonoids is reported
in underground part powder of L. pungens (3.12 mg kg-1).. Total alkaloid content is
more in underground part powder extract (1.87 mg kg -1) compared to other powder
extract selected for present study. The glycoside content is more in underground part
powder samples compared to aerial part powder samples. Similar to present
observations, from the aerial parts of Acanthus ilicifolicus, which is a member of
Acanthaceae family, different types of glycosides have been isolated (Kanchanapoom
et al., 2001).
The phenolic content in various parts of plant was studied by spectroscopic
method. In L. pungens, the phenolic content of the aerial part dry powder extract is
found to be 0.28 (mg kg -1) and in underground part powder is 0.21 (mg kg-1). Phenols
are stimulating, antiseptic, anti-infectious and detoxifying activities (Kenner and
Requena, 1996).
The tannin content of the plant was found to be more in aerial parts powder
extracts. Tannins have important roles such as stable and potent antioxidants (Trease
and Evans, 1999). Alkaloids are known to be effective for antihypertensive (Zee-
Cheng, 1997). The present observation presence of alkaloids supports the use of
selected in traditional medicine for treating hypertension.
The result of the present study offers supportive evidence that the selected
plant possess some active chemical principles which are traditionally used in
treatment of boils, purgative, anthelmintic, stimulant etc. It has authenticated the
usefulness of the chosen plant for medicinal purposes. These species could also be
seen as potential sources of useful drugs due to their rich contents of phytochemicals.
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ANTICANCER ACTIVITY OF POUZOLZIA WIGHTII BENN. STEM
ETHANOLIC EXTRACTS AGAINST HELA CELL LINE
M. Johnson, M. Syed Ali Fathima* and A. Mohamed Ansar**

Centre for Plant Biotechnology, Department of Botany,St. Xavier’s College
(Autonomous), Palayamkottai -627 002.
*Department of Botany, Sadakathullah Appa College (Autonomous), Tirunelveli –
627 011.
**Department of Pharmaceutics, S.A. Raja Pharmacy College, Vadakkangulam.
ABSTRACT
Medicinal plants are component and bundle of human society to struggle diseases,
from dawn of civilization. Plants are the largest source of herbal medicines in the
world. About 60-80 % of human beings still depend on plant based medicines as
traditional human health care system. Pouzolzia wightii belongs to family:
Utricaceae, a native of Southern Western Ghats. In India it is distributed in Tamilnadu
and Kerala. Different parts of this plant have the various uses like anti inflammatory,
wound healing, ulcers, boils. The present study reports that ethanolic extracts of
Pouzolzia wightii stem have been tested for anticancer activity. In-vitro anticancer
studies were performed against human cancer cell line (HeLa) and MTT assay was
used to analyze the cell growth inhibition. The results showed that the ethanolic
extracts of stem of Pouzolzia wightii possessed a moderate amount of anticancer
activity with an IC50 value 187.1µg/ml.
Keywords: HeLa, MTT assay, IC50
INTRODUCTION
Now a day’s utility of medicinal plants has been increasing day by day in the
present world. Compounds which are naturally derived from the herbs are safer and
easily biodegradable than the synthetic compounds. So that the problem of drug
resistance observed in synthetic drugs is also reduced (Chanchal and
Balasubramaniam, 2011). Higher plants, a source of many plan derivatives, play an
important role in health care of human beings. Therefore medicinal herbs have
received a significant interest in anti-cancer therapy. Medicinal herbs do not have any

of these drawbacks. Medicinal plants provide natural products have valuable sources
for anticancer drug discovery (Schwartsmann et al., 2002). Plants, vegetables and
herbs used in traditional and folk medicine as one of the important source of cancer
chemoprevention drug discovery and development (Abdullaev et al., 2000). Many
anticancer drugs derived from natural plant sources (Tan et al., 2006). Dietary
substances from natural products have played a vital role in creating new
chemopreventive agents (Surh, 2008). Cragg and Newman states that over 50 % of
the new drugs contain anticancer properties were isolated from natural sources.
Evaluation of phytochemical screening of herbal products are becoming popular as
sources of possible anticancer compounds (Parag et al., 2010). Cancer is a generic
term for a group of diseases that can affect any part of the body. Cancer is caused by a
number of genetic alterations. More number of natural products contain anticancer
activity on various animal models. Sixty percentageof drugs currently used for the
cancer treatment have been isolated from the natural products (Gordaliza, 2007). All
around the world medicinal plants constitute a common alternative for cancer
treatment in many countries around the world (Gerson-Cwillich et al., 2006; Tascilar,
2006). According to World Health Organization 80% of the population in developing
countries is dependent on the traditional medicinal plants as a source of drug used to
treat infection since 3000 BC( Philip, 2011; Tripathi and Tripathi, 2003; Demiray et
al., 2009). In accordance In accordance with this background, the current study was
undertaken to investigate the anti-cancer activities of P.wightii stem ethanolic extracts
against human cervical cancer cell line (HeLa).
METHODOLOGY
Anticancer activity
The human cervical cancer cell line (HeLa) was obtained from National
Centre for Cell Science (NCCS), Pune and grown in Eagles Minimum Essential
Medium containing 10% fetal bovine serum (FBS). The cells were maintained at 37
0
C, 5% CO2, 95% air and 100% relative humidity. Maintenance of cultures was
passaged weekly and the culture medium was changed twice a week.
Cell treatment procedure
The monolayer cells were detached with trypsin-ethylene diamine tetraacetic
acid (EDTA) to make single cell suspensions and viable cells were counted using a
hemocytometer and diluted with medium containing 5% FBS to give final density of

1x105 cells/ml. One hundred microlitres per well of cell suspensions were seeded into
96-well plates at plate density of 10,000 cells/well and incubated to allow for cell
attachment at 7C, 5%CO2, 95% air and 100% relative humidity. After 24 h the cells
were treated with various concentrations of theP. wightii various extracts. They were
initially dissolved in Dimethyl Sulfoxide (DMSO) and an aliquot of the sample
solutions were diluted to twice the desired final maximum test concentration with
serum free medium. Additional four serial dilutions were made to provide a total of
five sample concentrations. Aliquots of 100 µl of these different sample dilutions
were added to the appropriate wells already containing 100 µl of medium, resulting in
the required final sample concentrations. Following sample addition, the plates were
incubated for an additional 48 h at 37 C, 5% CO2, 95% air and 100% relative
humidity. The medium containing without samples were served as control and
triplicates were maintained for all concentrations.
MTT assay
3-[4,5-dimethylthiazol-2-yl]2,5-diphenyltetrazolium bromide (MTT) is a
yellow water soluble tetrazolium salt. A mitochondrial enzyme in living cells,
succinate-dehydrogenase, cleaves the tetrazolium ring, converting the MTT to an
insoluble purple formazan. Therefore,the amount of formazan produced is directly
proportional to the number of viable cells.
After 48 h of incubation, 15µl of MTT (5 mg/ml) in phosphate buffered saline
(PBS) was added to each well and incubated at 37 C for 4h. The medium with MTT
was then flicked off and the formed formazan crystals were solubilized in 100 µl of
DMSO and then the absorbance was measured at 570 nm using micro plate reader.
The percentage of cell growth was calculated with respect to controlas follows
% Cell Growth = [A] Test / [A] control x 100
The percentage of cell inhibition was determined using the following
formula: % Cell Inhibition = 100 - Abs (sample)/Abs (control) x 100.
Nonlinear regression graph was plotted between % of Cell inhibition, Log
concentration and IC50 was determined using GraphPad Prism software.

RESULTS
In vitro anticancer activity
Ethanolic extracts of Pouzolzia wightii stem
The different concentrations of P. wightii ethanolic extracts of stem were
subjected to MTT assay. The results showed moderate activity against HeLa cell line.
The cytotoxic effects of P. wightii stem against the growth of human cervical cancer
cell line were demonstrated in Fig.1. The ethanolic extracts of P. wightii stem
showed a significant activity against HeLa with an IC50 value 187. 1 µg/ml.
60
R2 = 0.9887
% Cell Inhibition
40
IC50= 187.1 µg/ml

20
0
0.0 0.5 1.0 1.5 2.0 2.5
Log10 Concentration (g/ml)
Fig.1 In vitro anticancer activity of P. wightii- Stem ethanolic extracts

DISCUSSION
Olowa and Nufieza (2013) reported that ethanolic extracts of Lantana camara
and Euphorbia hirta leaves showed potent cytotoxic activity with varied
concentrations. In the present study also, ethanolic extracts of P. wightii stem showed
LC50 value of 23.81. Dantu et al. (2012) studied the in-vitro anticancer potential of
hydroalcoholic extract of Tabernamontana divaricata flowers against HeLa cell line.
Hydroalcoholic extract of T. divaricata flowers possess moderate anticancer activity
with an IC50 value greater than 100 µg/ml. In the present study also ethanolic extracts
of P. wightii stem showed moderate anticancer activity with IC50 value 187.1 µg/ml.
Several anti-cancer studies were carried out by using various experimental
cancer model such as Dalton’s Lymphoma (DL), human breast tumor (MCF-7) and
human cervical cancer cell (HeLa) (Rosangkima and Prasad, 2004; Patel and Patel,
2011;Damle et al., 2013; Rosangkima and Jagetia, 2015). In the present study,

anticancer activity of stem ethanolic extracts were determined using MTT assay
against HeLa cell lines.
SUMMARY AND CONCLUSION
The present study suggests that the qualitative and quantitative analysis
revealed the occurrence of more number of important phytochemicals in the ethanolic
extracts might play a key role in the anticancer activity of the ethanolic extracts of
Pouzolzia wightii stem. In conclusion that further research works in ethanolic extracts
of Pouzolzia wightii stem may bringout the specific bioactive compounds responsible
for the antitumor activity. Further research on the isolation of active principles from
Pouzolzia wightii may leads to find an alternative medicine with anticancer property.
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PHARMACEUTICAL EVALUATION OF AVICENNIA MARINA
(FORSSK) VIERH.
P. Arockia Mary Fernandez and D. Vasumathi
ABSTRACT
Avicennia marina is the marine mangrove found abundantly along the coastal regions
of Thoothukudi District. It is enriched with numerous bioactive compounds having
wide range of medicinal properties. In this study, A.marina was extracted with five
different solvents viz. hexane, water, isopropanol, acetone and methanol. Different
solvent extracts of A. marina was subjected to biochemical assays viz. Total
antioxidant activity, Protein denaturation inhibition activity, Nitric oxide scavenging
activity and Metal chelating ability. Methanolic extract of A.marina shown highest
activity in all the biochemical assays. Hence, the methanolic extract can be used a
source of drug for various diseases.
KEY WORDS:Avicennia marina, Solvent extracts, Antioxidant, Protein denaturation
inhibitory, Nitric oxide scavenging and Metal chelating study.
INTRODUCTION
Plants are the very important chemical factories of nature. Plants produce a
diverse range of bioactive molecules, making them rich sources of different types of
medicine. Most of the drugs today are obtained from natural sources or semi synthetic
derivatives of natural products and used in the traditional systems of medicine. Thus it
is a logical approach in drug discovery to screen traditional natural products.
Approximately 20 % of the plants found in the world have been submitted to
pharmaceutical or biological test and a sustainable number of new antibiotics
introduced on the market are obtained from natural or semi synthetic resources
(Panda, 2000).
Phytochemicals are the natural bioactive compounds found in plants. They are
divided into two groups which are primary metabolites and secondary metabolites
according to their functions in plant metabolism. Primary metabolites comprise

common sugars, amino acids, proteins and chlorophyll while secondary metabolites
consist of alkaloids, flavanoids, tannins, saponins, terpenoids and phenolic
compounds (Edeoga et al., 2005; Prasad et al., 2012). Most of the novel bioactive
principles of plants constituted by secondary metabolites like alkaloids, terpenoids,
flavanoids, phenolic compounds, organic acids and lipids (Harborne,1998).
The present study aimed to utilize the mangrove plant which is one of the
important sources of mangrove. They are widespread in tropical and subtropical
regions, growing in the saline intertidal zones of sheltered coast lines (Chelliah, 2001;
Marium Tariq et al., 2007). Mangrove species are woody, seed bearing and highly
specialized plants (Duke et al., 1998) found coast lines of estuaries and lagoons
(Kamaruzzaman, 2011). Because of their unique adaptations mangroves thrive well in
the environment where other plants cannot grow (Shanmugapriya et al., 2012).
Mangroves are salt tolerant plants. The specific regions where plants occur are termed
as mangrove ecosystem (Kathiresan et al., 2001; Chelliah, 2001). There are seven
species of true mangroves have been identified are also recorded. Mangrove plants
also have been also used as traditional medicine in India (Dhayanithi et al., 2012;
Bharathi et al., 2011).
However during the last decade screening of mangroves for bioactive
compounds (Kokpal et al., 1990), has received high interest as a potential bioresource
for novel bioproduct leads (Ishibasi et al., 1993; Miki et al., 1994). Among all the true
mangroves of the world Avicennia marina (Forsk.) Vierh is a valuable mangrove
because of its medicinal values and abundant distribution.
Avicennia marina is commonly called white mangrove belongs to the family
Avicenniaceae. It is a small medium sized tree (3 – 11 meter) with many branches.
Extensive underground root system with Pencil root (Pneumatophores or breaking
roots) up to 90 mm long. The plant has received some attention in determining its
important chemical constituents. Phenolic compounds are secondary plant metabolites
and are involved in a wide range of specialized physiological function (Bharathi et al.,
2011; Dhayanithi et al., 2012; Borojeni et al., 2013). Previous reports suggest that this
species was useful in stabilizing banks of estuaries in salty water and that it has tannin
rich bark (Bandaranayake, 2002). In this study pharmaceutical properties of
Avicennia marina was evaluated using certain in vitro assays viz. Total antioxidant

activity, Nitric oxide scavenging activity, Protein denaturation inhibition activity and
chelating ability of metal ions.
Collection and Identification of plant Avicennia marina
The fresh leaf of mangrove plant, Avicennia marina was collected from the
estuarine region of Roche Park of Thoothukudi, South East Coast of Tamil Nadu,
India by hand picking method. The leaves were washed thoroughly thrice with tap
water and once with sterile distilled water to remove salt and sand. Then they were
shade dried for two weeks and were partially powdered using domestic blender and
stored in air tight container for further use.
Extraction
Extraction was carried out by using cold percoalation method. An amount (100
gm) of partially crushed and powdered leaves was taken separately into 1000 ml of
conical flask with Acetone, Hexane, Methanol, Isopropanol and sterilized water
individually. This set up was kept on a rotary shaker at 120 rpm for 24 hrs. After
shaking, it was filtered through eight layers of muslin cloth, 100 ml of extract was
centrifuged at 5000 x g for 15 min. Extraction solvent was evaporated and dried over
Sodium Sulphate in dessicator under vacuum.
Total antioxidant activity of A. marina by Phosphomolybdenum assay

Total antioxidant activity was estimated by phosphomolybdenum assay (Prieto
et al, 1999). A. marina extracts of different concentration ranging from 200g/ml to
1000 g/ml were taken in individual test tubes and made up to 1 ml using distilled
water and 2 ml of Molybdate reagent solution (0.6 M sulfuric acid, 28 mM sodium
phosphate and 4 mM ammonium molybdate) was added. The tubes were incubated at
95C for 90 minutes. After incubation, the tubes were cooled to room temperature for
20-30 min and the absorbance of the reaction mixture was measured at 695nm.
Experiments were done in triplicates. Ascorbic acid was used as the positive reference
standard.
Protein denaturation inhibition activity of A. marina extract
The reaction mixture (0.5ml) consisted of 0.45 ml bovine serum albumin (5%
aqueous solution) and 0.1 ml of A.marina extract at different concentration (200-1000

µg/ml) was taken in test tubes and incubated at 37ºC for 30 min (Tanford, 1968).
After cooling the samples, 2.5 ml phosphate buffer saline (pH 6.3) was added to each
tube. Turbidity was measured spectrophotometrically at 660nm. 0.5 ml distilled water
was used as blank. The percentage inhibition of protein denaturation was calculated
by the following formula,
Percent protein denaturation inhibition = Abs Control - Abs sample / Abs Control X 100
Nitric oxide scavenging activity of A. marina
Nitric oxide scavenging activity can be estimated by the use of Griess reaction
(Garrat, 1964). The compound sodium nitroprusside decomposes in aqueous solution
at physiological pH (7.2) producing nitric oxide . Under aerobic conditions, nitric
oxide reacts with oxygen to produce stable products (nitrate and nitrite). The
quantities of nitrate and nitrite can be determined using Griess reagent (1%
sulfanilamide, 2% H3P04 and 0.1% N-(1-naphthyl) ethylenediamine dihydrochloride).
Scavengers of nitric oxide compete with oxygen leading to reduced production of
nitrite ions. For the experiment, sodium nitroprusside (10mM) in phosphate buffered
saline was mixed with different concentrations (200-1000 μg/ml) of A.marina extract
and incubated at 30ºC for 2 hours. After the incubation period, 0.5 ml of Griess
reagent was added. The absorbance of the chromophore that formed during
diazotization of the nitrite with sulfanilamide and subsequent coupling with Naphthyl
ethylenediamine dihydrochloride was immediately read at 540nm. Inhibition of nitrite
formation by the plant extracts and the standard antioxidant ascorbic acid were
calculated relative to the control. All experiments were performed in triplicates and
the results were expressed as mean ± Standard Deviation.
Chelating ability of A.marinaextract on ferrous ions

The ferrous ion chelating potential of the extracts was evaluated by Dinis et al.
method. The reaction mixture contained 1.0 ml of various concentrations of the
extracts (2-10 mg/ml) and 0.05 ml of 2 mM FeCl3. The reaction was initiated by the
addition of 0.2 ml of 5 mM ferrozine. The reaction mixture was shaken vigorously
and left standing at room temperature for 10 min and the absorbance of the reaction
mixture was measured at 562 nm against a reagent blank. A lower absorbance of the
reaction mixture indicated a higher ferrous ion chelating ability. The control contained

all the reagents except sample. Gallic acid and ascorbic acid was used as standard for
comparison. The following formula was used to calculate percent inhibition.
Percent Inhibition = [(Control- Test)/control] × 100
The total antioxidant activity of A. marina extract
The total antioxidant activity is the representation of the compounds ability to
neutralize or scavenge free electrons in biological system. Activity of different
extracts of A.marina was shown in Figure 1. Methanolic extract of A. marina shown
highest activity followed by acetone, isopropanol, aqueous and hexane. Ascorbic acid
was used as control. The results indicated that methanolic extract of A.marina has
contained highest concentration of bioactive compounds with anticancer activity.
Nitric oxide scavenging activity
Nitric oxide is the important free radical cause severe damage to DNA and
proteins. The drug molecules of nitric oxide scavenging potential with less side
effects will be the better drug than chemically derived counterparts. The nitric oxide
scavenging activity of different extracts of A.marina was shown in Figure 2.
Methanolic extract of A. marina shown highest activity followed by acetone,
isopropanol, aqueous and hexane. Ascorbic acid was used as control.
Protein denaturation inhibition activity
Proteins are the essential biomolecules in human body. The destruction of
proteins results in progression of diseases such as alzheimers and Parkinson. The
drugs having protein denaturation inhibition activity be used as drugs for protein
related diseases. Protein denaturation inhibition activity of different extracts of A.
marina was shown in Figure 3. Methanolic extract of A. marina shown highest
activity followed by acetone, isopropanol, aqueous and hexane. Diclofenac was used
as control.
Metal chelating ability of A. marina
Fe (III) reduction is often used as an indicator of electron donating activity,
which is an important mechanism of phenolic antioxidant action. The reducing ability
of a compound generally depends on the presence of reductones (antioxidants), which
exert the antioxidant activity by breaking the free radical chain by donating a
hydrogen atom. Metal chelating activity of different extracts of A. marina was shown

in Figure 4. Methanolic extract of A.marina shown highest activity followed by
acetone, isopropanol, aqueous and hexane. Ascorbic acid was used as control.
Figure1. Total Antioxidant activity of A. marina extract
Figure 2. Nitric oxide scavenging activity of A.marina
Figure 3. Protein denaturation inhibition activity
Figure 4. Metal chelation activity

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biological activities of extracts.

ANALYSIS OF HEAVY METAL CONCENTRATION OF
TUTICORIN MANGROVE ECOSYSTEM
S.R.T.Sherly Cross1 and T.Mohanraj2
1
Dept of Zoology, St.Mary’s College for Women, Tuticorin – 628 001
2
Dept of Zoology, Aditanar College Arts & Science, Tiruchendur – 628 216
ABSTRACT
Mangroves are considered as the most critical habitats in the world. They form one of
the biologically important and productive ecosystems. In recent times heavy metals
are known to pose a potential threat to this sensitive biota. However, very little is
known with regard to the levels of heavy metals found in mangrove ecosystem in
India. To understand this, we analyzed the heavy metal accumulation in water,
sediment and tissue (Marcia opima) samples collected from surrounding root zone
in the Karapad bay and Korampallam creek regions of Tuticorin from March 2010 to
February 2011. Consequently the heavy metals such as Fe, Pb, Zn, Cu and Cd were
recorded during the present study. The study revealed the maximum accumulation of
Fe in the sediment and tissue samples that ranged from 1497. 2 to 201 .56 μg.g -1.
The accumulation of heavy metals was recorded in the order of: Fe>Zn>Cu>Cd>Pb.
One way ANOVA indicated statistically insignificant difference (p>0.01) in
the variation of Fe, Zn and Cu among samples.
KEY WORDS: Mangroves, ecosystem, heavy metal, sediment, accumulation
INTRODUCTION
Mangroves are salt-tolerant vegetation of tropical and subtropical intertidal
regions of the world. The mangrove ecosystem is said to be highly productive but
extremely sensitive and fragile. Besides mangroves, the ecosystem also harbours other
floral and faunal species. Asia covers the largest area of mangroves in the world, of
which India contributes nearly 3% of the global mangrove habitat (FAO, 2003). Over
the past few decades, heavy metal pollution by anthropogenic and industrial
activities is a serious concern throughout the world. The intense development

and industrialization have posed an ecological threat to the nearby mangrove
forests which have revealed elevated levels of heavy metals that decline even
the sediment quality (Sarika and Chandramohankumar, 2008; Lewis et al.,
2011). Rapid pace of industrialization and urbanization has led to introduction
of these heavy metals into the coastal ecosystem in the form of pesticides,
fertilizers, organochlorine compounds etc. The heavy metals form the building
blocks of these harmful chemical compounds and enter the organism’s body
from ambient water and sediment.
The most commonly explored heavy metal contaminants include lead,
zinc, mercury, magnesium, nickel, chromium, cadmium, and manganese
(Basamba et al., 2010). Among these, metals like lead, cadmium, chromium
and zinc are highly toxic which could be significantly associated with
bioaccumulation since they cannot be biologically degraded and instead get
concentrated within sediments (Agoramoorthy et al., 2008). Initially, heavy
metals that are introduced into the mangrove habitat are readily trapped by the
sediments (Harbison, 1981; Silva et al., 1990; Clark et al., 1997). These metals are
initially adsorbed to fine grained sediments, and are later bound more permanently as
metal sulphides beneath the sediment surface. These metal sulphides are largely
immobilised, while there are possibilities that they may be released during physical or
biological disturbance of the sediment (Clark et al., 1997). Hence, mangrove
sediments help to prevent the heavy metals from entering into the adjacent marine
ecosystems. Similarly, various faunal species that are found in the mangrove
environment also take up these heavy metals.
Numerous studies that have been carried out with special emphasis to
mangrove species and their sediments have proved them to be reliable bio-
indicators of heavy metal pollution and contamination (Defew, 2005).
Analysis of various heavy metals in the environment and the related
management strategy has recently become a matter of concern for several
researchers. Hence, the current investigation was carried out with the endeavour to
analyze heavy metal accumulation in water, sediment and tissue (Marcia opima)
samples of Karappad bay and Korampallam creek areas of Tuticorin.

Collection of sample
For the analysis of trace metals in water, sediments and tissue sample, two
mangrove ecosystems namely Karappad bay and Korampallam creek regions of
Tuticorin were selected. Monthly samples of water, sediment and tissue were
collected, for a period of twelve months from March 2010 to February 2011.
Sediment sample
The sediment samples were collected using vertical corer, and were
shade-dried to a constant weight. These sediments were ground and sieved
through mesh (0.5mm) before digestion (Guzman and Jimenez, 1992).
Tissue sample (Marcia opima)

The clams were collected manually and the soft tissues were
removed and dried at 60 C. The dried tissue were ground to a fine powder
using mortar and pestle and was stored in desiccators for further analysis.
For digestion process 20 ml of the concentrated HNO 3 and

perchloric acid were added to 5g of dried samples and the mixture was left for
24 h. The same mixture was digested on a hot plate at 120°C. Thereafter 10
ml of (10%) nitric acid was added and the constituents were transferred to 20
ml polytop vials and were allowed to stand for 24 h for residue to settle down
(Walting and Walting, 1982). The supernatant liquid was filtered through a
0.45 µm Millipore membrane filter. This filtered sample in the vial was then
analyzed in Inductive coupled plasma system (ICP) (Optical Emission
Spectrophotometer by using the instrument Optima 2100 DV and quantified
against a known standards) to estimate the concentration of heavy metals.

Heavy metals, including both essential and non-essential elements seem to
have significance in the field of ecotoxicology (Ebrahimpour and Mushrifah, 2010)
mainly due to their toxicity, level of persistence, bioaccumulation and bio-
magnification in the food chain (Yousafzai et al., 2010). Hence monitoring of heavy
metal contamination using biological samples helps to assess the quality of the aquatic

systems. Fe, Pb, Zn, Cu and Cd were the heavy metals recorded during the present
study. Among these Fe was found prevalent in the sediment and tissue (Marcia
opima) samples at both the study areas and its concentration ranged from 1497.32 to
201 .56 μg.g-1 and 64.27 to 171.24 μg.g-1 respectively. As compared to the
previous studies of Ganesan and Kannan (1995) these results are relatively
higher, revealing enhanced industrial pollution in and around Tuticorin. Out of
the three samples analyzed, the sediment sample recorded the highest Cu
(14.21 μg.g-1), Zn (26.24 μg.g-1) and Fe (201 .56 μg.g-1) concentrations. The
study also revealed that the concentration of Pb in water was higher than the
sediment and tissue samples and its mean value varied from 5.84 to 10.68
μg.g-1. This is in total contradiction to the observation made by Palanichamy
and Rajendran (2000) that recorded high concentration of Pb in the sediment
than in waters off Tuticorin. It has been envisaged that the levels of lead
within mangrove sediments in the Gulf region and others could be increasing
in the upcoming years due to impacts of oil spills and higher rate of fuel
exploitation (Shiradah, 1999; Basamba et al., 2010).
The trace metal concentration particularly Fe, Cu, Pb and Zn was
higher during the northeast monsoon (October to December) in all the
samples. A similar observation was made by Ganesan and Kannan (1995) that
affirmed higher concentration of heavy metals during monsoon seasons which
may be chiefly attributed to the inputs of land based discharges. The
concentration of Cu and Fe were higher in the sediment samples, which is
equivalent to the study made by Baskaran etal.,(2002) that also recorded high
concentration of Fe, Cu and Zn in the sediment at fly ash dumping dyke at
Tuticorin Thermal Power Plant. Chandrasekar (2001) has also observed
elevated levels of heavy metals in the sediments of Tuticorin waters. The
lowest concentration of Fe was recorded during the month of September
(1497. 2 μg.gl-1). The water sample comprised of only Cu, Cd, Pb and Zn
during this study. Cadmium was found be the lowest in concentration in water
during September (0.58 μg.gl-1). The tissues samples of Marcia opima have
accumulated Cu, Cd, Fe, Pb and Zn metals, out of which Fe was estimated in
higher concentration during the month of November (162.49 μg.gl -1). Whereas
Zn showed the lowest concentration in the tissue during May (27.29 μg.gl-1).

The One way ANOVA indicated statistically insignificant difference (p>0.01)
in the variation of heavy metals among samples.
Table1: One way ANOVA analysis for tissue sample heavy metals
Source of
SS df MS F P-value F crit
variation
Between Groups 4970.212 11 451.8375

0.305708 0.982574 1.924308
Within Groups 106416.2 72 1478.003
Total 111386.5 83
Table2: One way ANOVA analysis for water sample heavy metals
Source of
SS df MS F P-value F crit
variation
Between Groups 16.41767 11 1.492515

0.102401 0.999881 1.924308
Within Groups 1049.414 72 14.5752
Total 1065.832 83
Table3: One way ANOVA analysis for sediment sample heavy metals
Source of variation SS df MS F P-value F crit
Between Groups 93017.74 11 8456.158

0.023655 1 1.924308
Within Groups 25738101 72 357473.6
Total 25831118 83

Fig. 1: Heavy metal concentration in water sample
10
9
8
7
6
5 Fe
4
3 Pb
2
1 Zn
0
Cu
October
November
December
March' 2010
February' 2011
June
September
May
July
January
April
August
Cd
Fig. 2: Heavy metal concentration in sediment sample
2000
1500
1000
Fe
500 Pb
0 Zn
Cu
Cd
Fig. 3: Heavy metal concentration in the tissue sample (Marcia opima)
180
160
140
120
100
Fe
80
60 Pb
40 Zn
20
Cu
0
Cd
November
December
September
October
May
January
March' 2010
February' 2011
April
August
June
July

Conclusion
The findings of this investigation revealed that the heavy metals namely Fe,
Pb, Zn and Cu were found in higher concentrations in the study areas. Globally, there
has been an undeniable conformity that the reported levels of heavy metals within
mangrove sediments are increasing each year as a result of pollution and activities
caused by developmental expansion and urbanization. Therefore we need to safeguard
the mangrove forests which are a potential pollutant traps that has been very effective
in protecting the adjacent environments. Hence, periodical monitoring of heavy
metals in the aquatic ecosystems need to be ensured through safe disposal of domestic
wastes and industrial effluents.
References
1. Agoramoorthy G, Chen FA, Hsu tributaries. Int. J. Environ. Sci.
MJ (2008) Threat of heavy metal Tech., 7: 435-446.
pollution in halophytic and 4. Baskaran, M., Ramadhas, V.
mangrove plants of Tamil Nadu, and R. Santhanam, 2002. Metal
India. Environ Pollut 155: 320-326. pollution in Tuticorin coastal
2. Basamba TA, Kakudidi E, waters due to fly ash of thermal
Mutumba G, Oryem Origa H, power plant. Proc. National
Sekabira K (2010) Assessment of Seminar on Marine and Coastal
heavy metal pollution in the urban Ecosystems: Coral and
stream sediments and its Mangrove- Problems and
tributaries. Int J Environ Sci and Management Strategies. SDMRI
Tech 7: 435-446. Res. Publ., 2: 190 - 193.
3. Basamba, T.A., Kakudidi, E., 5. Chandrasekar, N., 2001. Trace
Mutumba, G., OryemOriga, H. elements in the suspended
and K. Sekabira, sediments of salt marsh area of
2010.Assessment of heavy Karapad creek, Tuticorin. Ind.
metal pollution in the urban J. Environ. Ecoplan.,5(1), pp 81
stream sediments and its - 86.

6. Clark, M.W., D. McConchie, P. 11. Lewis, M., Pryor, R. and L.
Saenger and M. Pillsworth, Wilking, 2011.Fate and Effects
1997. Hydrological controls on of Anthropogenic Chemicals in
Copper, Cadmium, Lead and Mangrove Ecosystems: A
Zinc concentrations in an review. Environ. Pollut.,159:
anthropogenically polluted 2328-2346.
mangrove ecosystem, Wynnum, 12. Palanichamy, S. and A.
Brisbane. J. Coastal Res., Rajendran, 2000. Heavy metal
13:1150-1158. concentrations in sea water and
7. Ebrahimpour, M. and Mushrifah, sediments of Gulf of Mannar
I. 2010. Seasonal variation of and Palk Bay, Southeast Coast
cadmium, copper, and lead of India. Indian J. Mar. Sci., 29:
concentrations in fish from a 116 - 119.
freshwater lake.Biol. Trace Elem. 13. Silva, C.A.R., Lacerda, L.D.
Res., 138: 190-201 and C.E. Rezende, 1990.
8. Ganesan, M. and L. Kannan, “Metals Reservoir in a Red
1995. Iron and manganese Mangrove Forest,” Biotropica,
concentrations in sea water, 22(4): 339-345.
sediment and marine algae of 14. Walting, R.J. and Walting, H.R.
Tuticorin coast, southeast coast 1982. Metal surveys in South
of India. Indian J. Mar. Sci., 24: Africans estuaries.VI Sundays
236 - 237. River. Water S.A., 8(4): 192.
9. Guzman, H.M. and C.E. 15. Yousafzai, A.M., Chivers, D.P., Kh
Jimenez, 1992. Contamination an, A.R., Ahmad, I. and Siraj,
of coral reefs by heavy metals M. 2010. Comparison of heavy
along the Carubben coast of metals burden in two freshwater
Central America. Mar. Poll. fishes Wallago attu and Labeo
Bull., 24: 554-563. dyocheilus with regard to their
10. Harbison, P., 1981. The case for feeding habits in natural
the protection of mangrove ecosystem. Pak. J. Zool., 42 (5):
swamps: geochemical 537-544.
considerations. Search 12:273-
276.

ANTIBACTERIAL POTENTIAL AND PHYTOCHEMICAL ANALYSIS OF
BROWN SEAWEED STOECHOSPERMUM MARGINATUM
R.S. Raubbin* and A. Pushparaj

Department of Zoology, TDMNS College, T.Kallikulam-627113.Tamil Nadu, India.
*Corresponding Author:rsraubbin@gmail.com
ABSTRACT
The present study was carried out to investigate the phytochemical constituents and
antibacterial potential of the Stoechospermum marginatum. The antibacterial activity
carried out against six bacterial pathogens (Gram +ve: Bacillus
subtilis,Staphylococcus aureus,Lactobacillus acidophilus;Gram -ve: Pseudomonas
aeruginosa, Escherichia coli and Proteus mirabilis). The phytochemical constituents
of aqueous extract were studied according to the standard method. The antibacterial
activity selected brown seaweed was extracted by solvents viz., acetone, ethanol,
methanol, chloroform and ethyl acetate extracts tested against pathogens by using
agar well diffusion method. The aqueous extract of Stoechospermum marginatum
showed the presence of tanins, saponins, flavonoids, Cardiac glycosides, alkaloids and
carbohydrate. The antibacterial activity results revealed that the highest activity of
21mm recorded against E. coli (100μg/ml) and lowest activity 7mm was observed
against S. aureusby using ethyl acetate as a solvent. However, ethanol extract was
found to be more effective than other solvents tested. The screening results confirmed
that phytochemical constituent of Stoechospermum marginatum has suitable
substance for antibacterial activity.
INTRODUCTION
Seaweeds are the group of plants that live either in marine or brackish water
environment. That’s represents half of the global biodiversity, its holding colossal
resource for new compounds. Seaweeds have plenty of pharmaceutical and
commercially important phytochemical constituents such as caratenoids, sterol,
glycosides, diatery fibers, tanins, saponins, terpenoids, flavonoids, glycosides,
alkaloids, carbohydrate, vitamin, minerals and amino acids. The nutrient compositions
of seaweeds are varying based on species, maturity, habits and environmental

condition. Seaweeds are consider as a source of bioactive compounds as they have the
potential of produce a great variety of secondary metabolites and its abundantly
growing in the coastal region between low tide to sub tidal region, as well as
classified under three divisions, of Chlorophyceae, Phaeophyceae and Rhodophyceae
(Sreenivasa Rao et al., 2009 and Anantharaman, 2002). Marine environment is unique
with respect to its biological and chemical diversities and represents a source of novel
antimicrobial compounds. In recent years, there are number of reports relevant to
macro algae derived compounds that have a broad range of biological activities such
as antibacterial, antifungal, antiviral, antitumor, antioxidant, antifouling,
antineoplastic, cytotoxic, anti-inflammatory and antimitotic activities (Okai, 1997,
Pushparaj et al., 2014, Kalamba and Kanica, 2003, Bouhlal et al., 2010, Devi et al.,
2011 and Priyadharshini et al., 2011).
Micro organisms are inevitably found ways of resisting the antibiotics. Thus,
aggressive action is needed to find out new drugs to control the diseases, which is
caused by organisms. The therapies of infected diseases, then the uses of
antimicrobial drugs have certain limitations because of changing patterns of microbial
resistance in pathogens and side effects they produced (Al- Haj et al., 2009). So the
researchers were preyed on to develop new resistance from terrestrial plants and
marine resource. The natural products are consumed by us in the form of food.
Moreover, new resistance will extend by using natural products, which are the good
way for improving health and immunity. There are number of reports regarding the
medicinal uses of seaweeds (Dawczynski et al., 2007 and Pushparaj et al., 2014).
Solvents are playing an important role on extracting bioactive compounds from the
different species of seaweeds. Mainly methanol, ethanol, acetone, chloroform,
benzene, ethyl acetate, petroleum ether, hexane etc., are used for the extraction
process. Likewise, using these organic solvents for antimicrobial activity always
provides a higher efficacy in extracting compounds besides heating the solvent can
enhances the mass transfer (Manivanan et al., 2011). Screening of organic extracts
from seaweeds and other marine organisms is a common approach to identify
compounds of biomedical importance. Hence, the aim of the present study was to
analyse the phytochemical constituents and evaluate the antibacterial activity of
selected brown seaweed Stoechospermum marginatum against of six pathogenic
bacteria.

Sample collection
The selected seaweed Stoechospermum marginatum (S. marginatum), belongs
to the family of Phaeophyceae was collected from Tuticorin coastal waters, Gulf of
Mannar, Tamil Nadu, South India. It was thoroughly washed with running water to
remove epiphytes, animal casting and sand particles and again washed using distilled
water and dried under shade. The shade dried sample was cut into small pieces and
powdered in a lab mixer grinder. The powdered sample was then stored in freezer for
further study.
Preparation of seaweed extract
Approximately 5 g of the powdered materials were extracted successively with
250 mL of ., acetone, ethanol, methanol, chloroform and ethyl acetate by using
Soxhlet apparatus for 8 h at room temperature not exceeding the boiling point of the
solvents. The extracts were filtered by using Whatman No. 1 filter paper and then
concentrated in vacuum at 40 °C by using hot air oven. The residues obtained were
stored in a freezer at -20 °C until further tests. The seaweed extracts were further
subjected for antimicrobial activity by agar well diffusion method.
Phytochemical analysis
The dried, powdered sample of Stoechospermum marginatum was studied the
presence of different phytochemicals like tannin, saponin, terpenoids, flavonoids,
aminoacids, cardiac glycosides, alkaloids, carbohydrates, coumarins, sterol,
glycosidesetc., according to standard procedures (Lala, 1993, Horborne, 1973 and
Brinda et al., 1981)
Test organisms
Gram-positive bacteria such as Bacillus subtilis (B. subtilis),Staphylococcus
aureus (S. aures),Lactobacillus acidophilus (L. acidophilus) andGram negative
bacteria Pseudomonas aeruginosa (P. aeruginosa), Escherichia coli (E. coli) and
Proteus mirabilis (P. mirabilis) were obtained from the Research Department of
Microbiology, VHNSN College, Virudhunagar, Tamil Nadu, India.
Antibacterial activity
The antibacterial activity was examined by agar well diffusion method. The
solvents like acetone, ethanol, methanol, chloroform and ethyl acetate were used to
collect the seaweed extract and were tested against the selected pathogens at four dose

levels (40µg/ml, 60µg /ml, 80µg /ml and 100µg/ml). Overnight grown bacterial
culture was transferred to sterile Petri plate with Mueller Hinton agar medium (Hi
Media Laboratories Limited, Mumbai, India) and was spread with sterile spreader to
create a lawn. About 5 wells of 6mm diameter were made in each plate with the help
of a sterile cork borer. Among the five, four wells were placed with the different
concentration of the extracts using sterile pipettes and remaining one well was
mentioned as control had with solvent alone followed by incubated at 37°C for 24 h.
The antibacterial activity was recorded by measuring the diameter zone of inhibition.
Triplicates were maintained for each and every test.
RESULTS
Phytochemical analysis
The phytochemical screening of Stoechospermum marginatum is tabulated in
Table 1. The aqueous extract showed the presence of 7 positive results out of 12 such
as tanin, saponin, flavonoids, Cardiac glycosides, salkowski, alkaloids and
carbohydrate.
Table 1: Phtochemical analysis of Stoechospermum marginatum aqueous extract
Sl. No. Phytochemical H2O

1 Tannin +
2 Saponin +
3 Terpenoids -
4 Flavonoids +
5 Aminoacids -
6 Cardiac glycosides +
7 Salkowski +
8 Alkaloids +
9 Carbohydrates +
10 Coumarins -
11 Sterol -
12 Glycosides -
Antibacterial activity
Different solvents like, acetone, ethanol, methanol, chloroform and ethyl
acetate extracts of Stoechospermum marginatum were tested for their antibacterial
activity against six strains of Gram-positive and Gram-negative bacteria by agar well
diffusion method. The results are tabulated in Table 2. The highest activity of 21mm
in 100µg/ml recorded against E. coli and lowest activity of 7mm against S.

aureuswhen used ethyl acetate as a solvent. Ethanol and methanol extracts showed
notable activity against all the tested pathogens, at the same time very limited results
in acetone as a solvent. The maximum activity of 14mm in 80µg/ml observed against
E.coli, ethyl acetate as a solvent and minimum activity 7mm were seen against S.
aureus, acetone and methanol were used as a solvent. In this concentration ethanol
shows activity against all the six pathogens. In 60µg/ml dose level maximum zone of
10mm and minimum zone 7mm recorded against B. subtilis, ethanol and methanol as
solvents, respectively. At the last one 40µg/ml concentration of antibacterial study
ethanol showed activity against all the tested pathogen except E.coli and other
solvents there were no activities obtained. The highest activity 9mm seen against B.
subtilis and minimum activity 7mm against L. acidophilus were recorded. There were
no one of the zone was recorded in chloroform as a solvent.
Table 2: Antibacterial activity of Stoechospermum marginatum in five different

solvents
Organisms Extracts in different concentration

40(μg/ml) 60(μg/ml) 80(μg/ml) 100(μg/ml)
Acetone Bacillus subtilis - - - -

extract Staphylococcus aureus - - 7 8
Lactobacillus acidophilus - - - -
Pseudomonas aeruginosa - - - -
Escherichia coli - - 8 11
Proteus mirabilis - - - -
Ethanol Bacillus subtilis 9 10 11 13
extract Staphylococcus aureus 8 11 13 14
Lactobacillus acidophilus 7 9 10 15
Pseudomonas aeruginosa 8 9 12 13
Proteus mirabilis 8 9 10 14
Methanol Bacillus subtilis - 7 9 12
extract Staphylococcus aureus - - 7 10
Lactobacillus acidophilus - 8 9 9
Pseudomonas aeruginosa - - - 8
Escherichia coli - 8 9 11
Chloroform Bacillus subtilis - - - -
extract Staphylococcus aureus - - - -
Lactobacillus acidophilus - - - -
Pseudomonas aeruginosa - - - -
Escherichia coli - - - -

Ethyl Bacillus subtilis - - - 8
acetate Staphylococcus aureus - - - 7
extract Lactobacillus acidophilus - - - -
Pseudomonas aeruginosa - - - 8
Proteus mirabilis - - - 10
DISCUSSION
Seaweeds are a marine group simple plant like organisms called as macro
algae. Seaweed extracts are used in many products, including foods, medicine, plant
growth hormone, shampoos and cosmetic items. They are remove carbon dioxide
from seawater by photosynthesis activity as well many animals rely for food and
shelter. They contain different vitamins, minerals, protein, trace element, iodine and
bioactive substances. Seaweeds are the only source for agar-agar, algin and
carrageenan, which are used in various industries viz., confectionary, textile,
pharmaceuticals, dairy, and paper industries (Megha et al., 2013). Antibacterial
activity of seaweeds against gram positive and gram negative bacterial has been
established by many scientists. Although a variety of solvents have been used for
screening seaweeds for antimicrobial activity, it is still uncertain what type of solvents
are the suitable for extraction of seaweeds. A few workers tried using different
solvents for screening the antibacterial activity of seaweeds (Kolanjinathan et al.,
2009, Subba Rangaiaha et al., 2010 and Manivannan et al., 2011). The result of
antibacterial activity the highest and lowest activity reported in ethyl acetate solvent.
All the tested pathogens were resistant against chloroform extracts, similarly parekh
reported that the seaweeds extracted in ethyl alcohol, acetone, and diethyl ether
showed higher activity when compared with chloroform extracts (Parekh, 1978).
Acetone extracts showed very limited activity against the pathogens tested
particularly there were no inhibition recorded against B. subtilis, L. acidophilus, P.
aeruginosaand P. mirabilis. The ethyl acetate extract was not sensitive against L.
acidophilus, as suggested by Schwarz and Noble, these bacterial strains may have
some kind of resistance mechanisms e.g. enzymatic inactivation, modification of
target sites and decrease intracellular drug accumulation or the concentration of the
compound used may not be sufficient (Schwarz and Noble, 1990). The activity of the
algae against the gram positive and gram negative bacteria showed that the presence

of broad range of antibiotic compound or probably the content of pharmacological
active contents like alkaloids, glycosides, tannins and saponins etc (Omulokoli et al.,
1997 and Milgate and Robert 2005).
The phytochemical screening of aqueous extract of Stoechospermum
marginatum revealed the presence of tannin, sapnin, flavonoides, cardiac glycosides,
salkowski, alkaloids and carbohydrates. The presence of flavonoids and tannin were
help to prevent from disease through free radical scavenging activity, besides
flavonoids enhance the activity of antitumor, anti-inflammatory, anti thrombic,
antiallergic, and antioxidants (Jiang et al., 2008). Flavonoids also known as natures
tender drug which have number of pharmaceutical and biological activities. Alkaloids,
salkowski, saponin and related active metabolites are having great medicinal value
and extensively used in pharmaceutical industry. Recently, a number of researchers
were reported about the phytochemistry of seaweeds across the world (Solomon et al.,
2012).The presence of secondary metabolites which include tanin, saponin,
flavonoids, Cardiac glycosides, salkowski, alkaloids and carbohydrate in the crude
extracts of S. marginatum suggested that the seaweed can be used as antimicrobial,
anti-parasitic, antifeedent, anti-inflammatory, antioxidant, antiallergenic,
antithrombic, anticarcenogenic and anti-ulcer agents. Further research also needed for
the separation and purification of the bioactive compounds, it will provide complete
data on the nutritive and antimicrobial potential of this seaweed.
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Screening of antibacterial activity

INVESTIGATION ON ETHNOPHARMACOLOGICAL AND
PHYTOCHEMICAL PROPERTIES OF MEDICINAL PLANTS EUPHORBIA
HIRTA LINN. AND VITEX NEGUNDO LINN.
J. Mugil2 S. Abirami 2, M. Kannan3 and S. Venkatesan4
1
Department of Microbiology, V. H. N. Senthikumara Nadar College (Autonomous),
Virudhunagar – 626001. Tamil Nadu, India.
2
Department of Microbiology, KAMARAJ College, Thoothukudi. Tamil Nadu, India.
3
Department of Zoology, V. H. N. Senthikumara Nadar College (Autonomous),
Virudhunagar – 626001. Tamil Nadu, India.
4
Department of Environmental Sciences, Periyar University, Salem. Tamil Nadu,
India.
ABSTRACT
Plants were used for medicinal purposes since time immemorial. Natural products
produced by plants have been isolated as biologically active pharmacophores. Plants
have been screened for their anti - infective properties as the probability of finding
diverse chemistries have been implicated to serve as leads for new anti - infective
drugs and using their standardized extracts as over the counter products and in the
health care products.In the present study, antimicrobial screening of two plants viz.,
Euphorbia hirta, and Vitex negundo revealed the wide spectrum of antimicrobial
properties of Euphorbia hirta leaves. Euphorbia hirta is native to India and Australia
and has been used as a traditional medicine. Antimicrobial activity of methanol and
ethyl acetate of Euphorbia hirta leaves was evaluated using Agar well diffusion
assay. Among the extracts tested methanolic extract was the most active. It exhibited
prominent antimicrobial activity against Staphylococcus aureus, Salmonella
typhimurium, Klebsiella pneumoniae, and Bacillus subtilis. Extracts evaluated were
validated against standard antibiotics (chloramphenicol and tetracycline) to know the
efficacy of the extracts.

KEY WORDS:Euphorbia hirta, Antimicrobial activity, methanol and ethyl acetate,
Agar well diffusion assay, Chloramphenicol and tetracycline.
INTRODUCTION
Medicinal plants are a source of great economic value in the Indian
subcontinent. Nature has bestowed on us a very rich botanical wealth and a large
number of diverse types of plants grow in different parts of the country. India is rich
in all the 3 levels of biodiversity, namely species diversity, genetic diversity and
habitat diversity. In India, thousands of species are known to have medicinal value
and the use of different parts of several medicinal plants to cure specific ailments has
been in vogue since ancient times.
Herbal medicine is still the main stay of about 75 - 80% of the whole
population, mainly in developing countries, for primary health care because of better
cultural acceptability, better compatibility with the human body and fewer side
effects. However, the last few years have seen a major increase in their use in the
developed world. Infectious diseases are the leading cause of death world - wide.
Antibiotic resistance has become a global concern (Westh, 2004). Now - a - days
multiple drug resistance has developed due to the indiscriminate use of commercial
antimicrobial drugs commonly used in the treatment of infectious disease (Davis,
1994). In addition to this problem, antibiotics are sometimes associated with adverse
effects on the host including hypersensitivity, immune - suppression and allergic
reaction (Ahmad et al., 1998).
This situation forced scientists to search for new antimicrobial substances.
Given the alarming incidence of antibiotic resistance in bacteria of medical
importance (Monroe et al., 2000), there is a constant need for new and effective
therapeutic agents (Bhavnani et al., 2000). Therefore, there is a need to develop
alternative antimicrobial drugs for the treatment of infectious diseases from medicinal
plants (Clark, 1996; Cordell, 2000). Several screening studies have been carried out in
different parts of the world. There are several reports on the antimicrobial activity of
different herbal extracts in different regions of the world (Nair et al., 2005).
Due to the side effects and the resistance to the pathogenic microorganisms
build against antibiotics, recently much attention has been paid to extracts and
biologically active compounds isolated from plant species used in herbal medicine

(Essawi et al., 2000). Plant based antimicrobials represent a vast untapped source of
medicines and further exploration of plant antimicrobials needs to occur.
Antimicrobials of plant origin have enormous therapeutic potential. They are effective
in the treatment of infectious diseases while simultaneously mitigating many of the
side effects that are often associated with synthetic antimicrobial (Iwu et al., 1999).
Vitex negundo
It is a wellknown medicinal plant for the treatment of various types of

disorders in the ayurvedic and folklore system of medicine in India. In this chapter the
plant leaves were extracted for ethanopharmacological studies. Vitex negundo Linn.
(Verbenaceae) commonly known as Nirkundi or Nallanocci. It is an aromatic large
shrub or small tree about 3 m in height with quadrangular branche sand almost found
throughout India, ascending to 1500 m in the outer Himalaya, fairly common in waste
lands, on road side, the banks or streams or in moist places near deciduous forests
(Shri Sawhney, 1976). It is an erect (2 - 5 m in height), slender tree with quadrangular
branchlets. The leaves have five leaflets in a palmately arrangement, which are
lanceolate, 4 - 10 cm long, hairy beneath and pointed at both ends. The bluish purple
flowers are numerous. The fruit is succulent, black and rounded when ripe having
about 4 mm in diameter.
METHODOLOGY
Collection of medicinal plants
The experimental medicinal plant Euphorbia hirta and Vitex negundo were
collected from our college campus V. H. N. S. N College, Virudhunagar and was
identified by Dr. K. Rajarathinam, Department of Botany, V. H. N. S. N College.
Then the plant parts were cleaned washed thoroughly 2 - 3 times with running tap
water and once with sterile distilled water, air dried for about one week then ground
into fine powder and stored in airtight bottles.
Extraction of selected medicinal plants
The crude plant extracts were obtained from Euphorbia hirta and Vitex
negundo by using soxhlet apparatus. Two types of solvents namely methanol and
ethyl acetate were used for extraction. About 50 g of shade - dried powder was filled
in the thimble and extracted successively with 250 ml of methanol at 60ºC and ethyl
acetate at 77ºC in Soxhlet extractor for 24 hr. The crude solvent extractswere dried at

45ºC in rotary evaporator to produce semisolid mass and stored in airtight containers
in refrigerator below 10ºC until further use. The yield percentage of each extract was
calculated as follows.
Final weight of dried extract
Percentage of yield = 100
Initial weight of powder
Percentage yield of the powdered plant Euphorbia hirta and Vitex negundo
crude extracts obtained using various solvents were shown in (Table 1). The data
revealed that among the solvent used, methanol showed more yield compared to ethyl
acetate and the yield was very less in ethyl acetate for all two plants. Out of the 50 g
of powdered Euphorbia hirta plant material, the percentage yield obtained was 12.4%
in methanol, 9% in ethyl acetate and the Vitex negundo plant powdered percentage
yield was 8.4% in methanol and 7.8% in ethyl acetate.
Phytochemical analysis of plant extract by General method
Phytochemical screening was carried out on the powdered plant material for
the presence of bioactive components such as alkaloids, flavonoids, cardiac
glycosides, phenols, and saponins (Trease and Evans, 1996) was determined as
follows,
Phytochemical analysis of plant extract by TLC method
The presence of bioactive secondary metabolites phytocompounds from the

leaves of Euphorbia hirta and Vitex negundo were qualitatively analysed by Thin
Layer Chromatography (TLC). In TLC the solid phase of silicagel was kept in hot air
oven in 1000C for 20 minutes. Silica powder was mixed with petroleum ether and
makes slurry. 20 x 20 cm TLC glass plates covered with the slurry and allowed to air
dried. After drying the plates were kept in hot air oven at 72 0C for 1 hour. After
developing the plates, the condensed filtrate was spotted using capillary tube. The
different spots were separated using different solvent mixture based on the
phytocompounds act as mobile phase. The R fvalue for each plate is then worked out
using the formula:
distance travelled by component
R f=
distance travelled by solvent
Separation of alkaloids
The powdered leaves of the selected plants were wetted with a half diluted
ammonium hydroxide and lixiviated with ethyl acetate for 24 hours at room

temperature. The organic phase was separated from the acidified filtrate and then
basified with ammonium hydroxide (pH 11-12). It is extracted with chloroform (3X),
and condensed by evaporation. The evaporated product was used for chromatography.
The alkaloid spots were separated using the solvent mixture chloroform and methanol
(15:1). The colour and hRf value of the separated alkaloids were recorded both under
ultra-violet (254 nm) and visible light after spraying with Dragendroff’s reagent.
Separation of flavonoids
One gram of the powdered leaves of the selected medicinal plants was
extracted with 10 ml methanol on water bath (600C / 5min). The filterate was allowed
to condense by evaporation. To filter the extracts a mixture of water and ethyl acetate
(10:1 ml) was added and mixed. The ethyl acetate phase retained was used for
chromatography. The flavonoid spots were separated using chloroform and methanol
(19:1) solvent mixture. The colour and hR f value of these spots were recorded under
Ultraviolet (UV 254 nm) light.
Separation of glycosides
The powdered leaves of the selected plants were extracts with 70% ethyl
alcohol on rotary shaker (180 thaws / min) for 10 hours 70% lead acetate was added
to the filtrate and centrifuged at 5000 rpm / 10 min. The supernatant was further
centrifuged by adding 6.3% sodium bi carbonate at 10,000 rpm / 10 min. The
remaining supernatant was dried, redissolved in chloroform and used for
chromatography. The glycosides were separated using ethyl acetate - methanol -
water (80:10:10) solvent mixture. The colour and hR f values of the spots were
recorded by observing under Ultraviolet (UV 254 nm) light.
Separation of phenols
The powdered leaves of the selected plants were lixiviated in methanol on
rotatory shaker (180 thaws / min) for 24 hours. The condensed filterate was used for
chromatography. The phenols were separated by using chloroform and methanol
(27:0.3) solvent mixture. The colour and the hR f values of the phenols were recorded
under visible light after spraying the plates with Folin - Ciocalteus reagents which is
heated at 80ºC for 10 minutes.

Separation of saponins
Two grams of powdered leaves of the selected plants were extracted with 10
ml 70% ethyl alcohol by refluxing for 10 min. The filtrate was condensed, enriched
with saturated n-butanol, and mixed thoroughly. The butanol was retained, condensed
and used for chromatography. The saponins were separated by using chloroform,
glacial acetic acid, methanol and water (64:34:12:8) solvent mixture. The colour and
hRf values of these spots were recorded by exposing chromatogram to the iodine
vapours.
Phytochemical analysis of plant extract by HPLC
High Performance Liquid Chromatography (HPLC) was analysed for the
selected plant extracts. It was performed on Shimadzu Spintrom HPLC - 530
available in science instrumentation centre, ANJAC College, Sivakasi. The results
were recorded.
Sample preparation for antioxident

The plant sample was ground to fine powder with a special grinder for herbal
medicine. A precisely weighed amount 0.5 g of the powder was extracted with 10 ml
of 80% methanol at 35°C for 24 hr in a shaking bath according to the literature (Cai et
al., 2004). The samples were then cooled down to the room temperature and
centrifuged at 4000 rpm for 10 min. The supernatant was recovered for the evaluation
of antioxidant capacity.
DPPH free-radical scavenging activity
DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging activity was
measured by the method of (Szabo et al., 2004). The reaction mixture contained 50 µl
of plant extract in methanol, yielding 100 µg / ml in each reaction was mixed with 1
ml of 0.1 mm DPPH in methanol and 450 µl of 50 mm Tris HCL buffer (pH 7.4).
Then 50 µl of methanol was used as the experimental control. The tubes were
incubated for 30 mins at room temperature. After 30 mins incubation the reduction in
the number of DPPH free radicals was measured at 517 nm. The decrease in optical
density of DPPH on addition of test samples in relation to the control was used to
calculate the antioxidant activity as percentage of inhibition of DPPH radical,
Absorbancy of control- Absorbancy of sample
DPPHscavenging activity % = X 100
Absorbancy of control

Where Abs control is the absorbance of DPPH + methanol; Abs sample is the
absorbance of DPPH radical + sample (i.e. extract).
Fentons hydroxyl radical scavenging activity
The scavenging activity for hydroxyl radicals was measured with Fenton
reaction. This method was recommended by Yu et al., (2004). The reaction mixture
contained 100 µl plant extracts (0.6 mM) 100 µl of deoxyribose (3 mM) in phosphate
buffer (20 mM, pH 7.4), 500 µl ferric chloride (0.1mM), 500 µl EDTA (0.1 mM), 500
µl of ascorbic acid (0.1 mM) and 500 µl of H2O2 (1 mM) and 800 µl of phosphate
buffer so that the final volume is 3 ml. After incubation for 1 hr at 37°C add 1.0 ml of
TCA (2.8%) and 1.0 ml of (thiobarbituric acid) TBA (1%) place the reaction mixture
in water bath for 20 minutes at 100°C, the absorbance of the mixture was measured at
532 nm. Ascorbate was used as standard, control test extract were replaced by
methanol.
Absorbancy of control- Absorbancy of sample

Hydroxyl scavenging activity % = X 100
Absorbancy of control
DNA nicking assay
The DNA nicking assay was performed using supercoiled pUC18 plasmid
DNA (Lee et al., 2003) using the methanol plant extracts of Vitex negundo (10 μl) in
different concentrations (100 ~1000 mg / ml) and DNA (0.5 μg) were incubated for
10 min at room temperature followed by the addition of 10 μl Fenton’s reagent ( 0 μl
of 0 μM H2O2, 500 μl of 50 μM ascorbic acid, and 800 μl of 80 μM FeCl3). The
reaction mixture was incubated for 30 min at 37°C and analysed on 1 % agarose gel
(prepared by dissolving 0.5 g of agarose in 50 ml of 1× TAE buffer) followed by
using ethidium bromide. Ellagic acid was used as a positive control (Dhan prakesh, et
al., 2007).
Evaluation of antimicrobial activity
The agar diffusion method as described by Esimone et al., (1998) was adopted
for the study. 15 ml of molten nutrient agar was seeded with 1 ml of standardized
7
broth cultures of the bacteria (1.0 x 10 cfu/ml) by introducing the broth cultures into

sterile petridishes, incorporating the molten agar, rotating slowly to ensure uniform
distribution of the microorganisms and then allowed to solidify on a flat surface. A
sterile cork borer (5 mm) was used to make wells in each plate for extracts. These
plates were labelled and 100 μl of each plant extracts was added aseptically into the
well. The plates were allowed to stand for one hour for prediffusion of the extract to
occur (Esimone et al., 1998). Plates were incubated at 37ºC for 24 hours. At the end
of incubation the plates were collected and zones of inhibition that developed were
measured. The average of the zones of inhibition was calculated. Standard antibiotics
of chloramphenicol ( 0 μg / disc) and tetracycline ( 0 μg / disc) were used as positive
control.
Percentage yield of the powdered plant Euphorbia hirta and Vitex negundo
crude extracts obtained using various solvents were shown in (Table 1). The data
revealed that among the solvent used, methanol showed more yield compared to ethyl
acetate and the yield was very less in ethyl acetate for all two plants. Out of the 50 g
of powdered Euphorbia hirta plant material, the percentage yield obtained was 12.4%
in methanol, 9% in ethyl acetate and the Vitex negundo plant powdered percentage
yield was 8.4% in methanol and 7.8% in ethyl acetate.
Table 1 Yield percentage of different solvent crude extracts of Euphorbia hirta

and Vitex negundo plants obtained by soxhlet extraction method.
Volume Raw
plants Extracted
Extraction of Percentage
Test plants Temp. plants
solvent solvent powder yield (%)
powder (g)
(ml) (g)
Euphorbia Methanol 65ºC 50 6.2 12.4

hirta Ethyl acetate 77ºC 50 4.5 9
250
Vitex Methanol 65ºC 50 4.2 8.4
negundo Ethyl acetate 77ºC 50 3.9 7.8

Phytochemical constituents such as alkaloids, flavonoids, glycosides, phenols,
Saponins and several other aromatic compounds are secondary metabolites of plants
that serve a defense mechanism against prediction by many microorganisms, insects
and other herbivores (Bonjar et al., 2004). The present study carried out on the plant
samples revealed the presence of medicinally active constituents. The phytochemical
constituents of the selected plants investigated are summarized in (Table 2). Analysis
of plant extracts revealed the presence of flavonoids, glycosides, phenols, saponins,
steroids and tannins in most of the selected plants which could be responsible for the
observed antimicrobial property. Alkaloids were absent in the selected plant extracts.
These bioactive compounds are known to act by different mechanism and exert
antimicrobial action. Tannins bind to proline rich proteins and interfere with the
protein synthesis (Shimada, 2006). Flavonoids are hydroxylated phenolic substance
known to be synthesized by plants in response to microbial infection and it should not
be surprising that they have been found in vitro to be effective antimicrobial
substances against a wide array of microorganisms. Their activity is probably due to
their ability to complex with extracellular and soluble proteins and to complex with
bacterial cell walls (Marjorie, 1999). Antimicrobial property of saponin is due to its
ability to cause leakage of proteins and certain enzymes from the cell (Zablotowicz et
al., 1996). Steroids have been reported to have antibacterial properties, the correlation
between membrane lipids and sensitivity for steroid compound indicates the
mechanism in which steroids specifically associate with membrane lipid and exerts its
action by causing leakages from liposomes (Raquel, 2007).
The plant extracts of Euphorbia hirta and Vitex negundo leaves were tested for
the presence of alkaloids, flavonoids, glycosides, phenols and saponins using (TLC)
Thin Layer Chromatography technique (Plate 1). The alkaloids were separated as
spots at the Rf value of 0.16 for Euphorbia hirta leaves, Rf value 0.46 for flavonoids,
0.06 for glycosides, 0.58 for phenols and 0.51 for saponins. Similarly the alkaloids
were separated from Vitex negundo and the Rf value is 0.11, flavonoids Rf value was
0.70, 0.15 for glycosides, 0.38 for phenols and 0.42 for saponins and the results were
tabulated in (Table 3).
The qualitative HPLC plant extract profile were detected at a wavelength of

254 nm and its sharpness of the peaks and proper baseline, Rt min, percent area and

heights were recorded (Fig. 1). The plant Euphorbia hirta showed a single major peak
at 2.843 retention time in methanol extract and 2.397 in ethyl acetate extact. In Vitex
negundo the single major peak was seen at 2.067 retention time in methanol extract
and three novel peaks at 2.070, 2.207 and 2.393 retention time in ethyl acetate extract.
The HPLC studies confirmed the bioactive compounds presenting in the sample.
Table 2 Preliminary phytochemical analysis of Euphorbia hirta and Vitex

negundo
Alkaloids
Flavonoids
Glycosides
Saponins
Steroids
Tannins
Wagner’
Mayor’s
d orff’s
Dragen
Plant species
s test
test
test
Euphorbia
+ + - + - - + +
hirta
Vitex negundo + + + - + - + -
(+) indicates presence of active constituents; (-) indicates absence of active
constituents
Table 3 TLC profile of Euphorbia hirta and Vitex negundo

Name of the secondary Rf value of Rf valueof
metabolic compounds Euphorbia hirta Vitex negundo
Alkaloids 0.16 0.11
Flavonoids 0.46 0.70
Glycosides 0.66 0.13
Phenols 0.58 0.38
Saponins 0.51 0.42

Plate 1 The plates showed the TLC profile of the phytocompounds present in
Euphorbia hirta and Vitex negundo
Figure 1 HPLC analysis of Euphorbia hirta Figure 2 HPLC analysis of Euphorbia hirta
methanol extract ethyl acetate extract

Figure 3 HPLC analysis of Vitex negundo Figure 4 HPLC analysis of Vitex negundo
methanol extract ethyl acetate extract
The presence of flavonoids, phenols and tannins in all the plants is likely to be
responsible for the free radical scavenging effects observed. Flavonoids and tannins are
phenolic compounds and plant phenolics are a major group of compounds that act as primary
antioxidants or free radical scavengers.
The DPPH test provides information on the reactivity of the test compounds with a
stable free radical. DPPH gives a strong absorption band at 517 nm in visible region. When
the odd electron becomes paired off in the presence of a free radical scavenger, the absorption
reduces and the DPPH solution is decolourised as the colour changes from deep violet to light
yellow. The degree of reduction in absorbance measurement is indicative of the radical
scavenging (antioxidant) power of the extract. The crude extract of Vitex negundo appeared
to be as potent antioxidant capacity with a maximum inhibition of 84.97% at 350 µl, at the
same concentration Euphorbia hirta showed 64.24% of inhibition. While comparing the free
radical scavenging activity of Vitex negundo, it showed better antioxidant capacities than
Euphorbia hirta (Figure 5).
Hydroxyl radical is very reactive and can be generated in biological cells through
Fenton’s reaction. The hydroxyl radical is the most reactive of the reactive oxygen species,
and it induces severe damage in adjacent biomolecules (Gutteridge, 1984). The hydroxyl
radical can cause oxidative damage to DNA, lipids and proteins (Spencer et al., 1994). The
scavenging activities of different plant extract were shown in (Table 6). Figure 6 showed that
methanolic extract of Vitex negundo exhibited highest of 96.12% and least by Euphorbia
hirta leaves. Hence, the Vitex negundo plant extract can be considered as a good scavenger of
hydroxyl radicals and the Euphorbia hirta exhibit moderate scavenging activity.
The DNA protective potential of Vitex negundo was studied in pUC18 DNA
protection assay. The effect of the fractions was compared with standard antioxidant
compound gallic acid. The concentration of fractions used was 100 – 1000 mg / ml. Both the

plant extract protected the DNA from hydroxyl radicals generated by Fenton’s reaction. DNA
nicking assay was showed a significant reduction in the formation of nicked DNA. Extracts
of Vitex negundo effectively prevented DNA nicking.
The present work also revealed that the extract from the leaves of these medicinal
plants possesses potent antioxidant activity presumably because of its phytochemical
constituents. These facts justify the medicinal use of the plant for the treatment of various
diseases.
Table 7 Analysis of antioxidant activity of plant extract by DPPH method
Plant extract of various concentration

Test
Plant name
control 150 µl 200 µl 250 µl 300 µl
Euphorbia 0.86 0.81 0.71 0.69
hirta (55.44%) (58.03%) (63.21%) (64.24%)
1.93
0.54 0.43 0.37 0.29
Vitex negundo
(72.02%) (77.72%) (80.82%) (84.97%)
Table 8 Analysis of antioxidant activity of plant extract by Fenton’s method
Test Plant extract of various concentration

Plant name
control
150 µl 200 µl 250 µl 300 µl
Euphorbia 0.492 0.439 0.374 0.323
hirta (60.32%) (64.59%) (69.83%) (73.95%)
1.24
0.204 0.161 0.081 0.048
Vitex negundo
(83.54%) (87.01%) (93.46%) (96.12%)
Based on their morphological and biochemical characters the clinical pathogens were
identified as Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae,
Salmonella typhimurium, Bacillus subtilis, and Staphylococcus aureus (Table 7).Plants such
as Euphorbia hirta and Vitex negundo are well known herbs used in ayurvedic traditional
medicine for their effectiveness against wide range of diseases.

Fig. 5 DPPH radical scavenging activity of Fig. 6 Hydroxyl radical scavenging activity
Euphorbia hirta and Vitex negundo of Euphorbia hirta and Vitex negundo
100 100
% of inhibition
90 90
% of inhibition
80 80
70 E.hirta 70 E.hirta
60 60
50 50
150µl
200µl
250µl
300µl
V.negun V.negund
do o
Concentration of plant extract Concentration of plant extract
Table 6 Antibacterial activity of methanol and ethyl acetate extracts of screened

medicinal plants
Diameter of the Zone (mm)

Euphorbia hirta Vitex negundo
Test organisms Ethyl
Methanol Methanol Ethyl acetate
acetate
extract extract extract
extract
Escherichia coli 10 12 19 13
Pseudomonas
15 17 18 10
aeruginosa
Klebsiella pneumoniae 20 15 16 10
Salmonella typhi 25 15 12 11
Staphylococcus aureus 35 26 19 10
Bacillus subtilis 21 22 18 10
Table 8 Inhibitory effect of standard antibiotics
Antibiotics zone of inhibition (mm)

Bacteria Chloramphenicol Tetracycline
0 μg / disc 0 μg / disc
Staphylococcus aureus 10 9
Klebsiella pneumoniae 11 10
Bacillus subtilis 9 8
Pseudomonas aeruginosa 10 9
Escherichia coli 9 10
Salmonella typhi 8 9

The methanolic extract of Euphorbia hirta exhibited pronounced activity against
B. subtilis (21 mm), K. pneumoniae (20 mm) and Salmonella typhi (25 mm), highly activity
against the gram-positive organism Staphylococcus aureus (35 mm), low activity against
Pseudomonas aeruginosa (15 mm) and E. coli (10 mm). Comparing the methanolic extract,
the ethyl acetate Euphorbia hirta showed low antimicrobial activity (Figure 8). The ethyl
acetate extract of Euphorbia hirta active highly against Bacillus subtilis (22 mm) and low
activity in Escherichia coli.
Vitex negundo showed lower responsible in both the solvent when compared with
Euphorbia hirta (Figure 9).The methanol extract of Vitex negundo show somewhat similar
activity against E. coli (19 mm), P. aeruginosa (18 mm), S. aureus (19 mm), B. subtilis (22
mm) and low activity in Salmonella typhi. Euphorbia hirta ethyl acetate showed lower
antimicrobial activity likewise the Vitex negundo ethyl acetate extract also showed lower
zone inhibition ranges from 10 - 13 mm. The standard antibiotic chloramphenicol and
tetracycline was found to have pronounced zone of inhibition effect between 8 – 10 mm at
the concentration of 30 µg / disc (Table 6.3).
Fig 8. Antimicrobial activity of methanol Fig 9. Antimicrobial activity of methanol
and ethyl acetate extracts of Euphorbia hirta and ethyl acetate extracts of Vitex negundo
430 2105
Zone of inhibition
Zone of inhibition
210 50
0
(mm)
(mm)
Microorganisms
Microorganisms V.negundo Methanol

E.hirta Methanol E.hirta Ethyl acetate V.negundo Ethyl acetate
The observed antibacterial properties corroborate its use in traditional medicine.
Traditionally, extracts of the plant are used in sore and wound healing, as ear drop for boils in
the ear and treatment of boils. They are also used in the control of diarrhoea and dysentery
(Igoli et al., 2005). The large zones of inhibition of Euphorbia hirta methanol extract
exhibited against S. aureus which is also corroborates with the findings of Braude (1982)
which is used in the treatment of sores, bores and open wounds. S.aureus have been
implicated in cases of boils, sores and wounds, also the moderate growth inhibition against E.
coli justifies its use in the control of diarrhoea and dysentery. E. coli is the common cause of
traveller’s diarrhoea and other diarrheagenic infections in humans.

It is very necessary to introduce new and biologically safe and active drugs
eco‐friendly in nature and effective as antimicrobial agents. Usually medicinal plants contain
several phytochemical compounds, which are very much necessary to control the growth of
the microorganisms. From this study, it can be concluded that crude extract of methanol and
ethyl acetate of leaf and flower exhibited potential bactericidal properties. Present
investigations together with previous studies provide support to the antibacterial properties of
Euphorbia hirta and Vitex negundo.
From our investigation of screening different plant species, the results obtained
confirm the therapeutic potency of some plants used in traditional medicine. In addition,
these results form a good basis for selection of candidate plant species for further
phytochemical and pharmacological investigation. The overall findings of this study form a
good basis to select this plant for futher phytochemical and pharmacological investigation
and suggest that the methanol extract contain certain constituents with antibacterial that can
be used for therapy of infectious diseases and cancer. Our future studies to isolate these active
phytochemicals and determine their activities against microorganisms and different cancer
cell lines, are in PROGRESS.
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169-176.

DIVERSITY OFMARINE FINFISH RESOURCES OF TIRUCHENDUR
COAST
T.Jebarani Rajathy1 and T.Mohanraj2

1
Research Scholar, CAS in Marine Biology, Annamalai University, Parangipettai.
2
Dept of Zoology, Aditanar College of Arts & Science, Tiruchendur.
ABSTRACT
The present study was carried out to assess the diversity of marine fin fishes of
Tiruchendur coast from January 2017 to April 2017. 80 Species of fishes belonging to 32
families were evidenced during the study period. The families: Lutjanidae, Serranidae,
Lethrinidae and Carangidae comprised the maximum number of species. Hook & line is the
predominantly used gear type in the study area. The dominant and most frequently observed
fishes include Carangids, Snappers, Emperors, Groupers and Barracudas. The One-way
analysis of variance (ANOVA) computed from the collected data revealed (p>0.05) that there
is significant difference between the groups.
KEY WORDS:diversity, dominant, gear, analysis of variance, species
INTRODUCTION
Fishing in India is a very important economic activity and a flourishing sector with
varied resources and potentials; it is a home to more than 10 percent of the global fish
diversity. Fish consumption is on an increase in many countries particularly India, where
fishery has emerged as a major trade with an annual turnover of more than ₨.220 billion,
accounting for 1.4% of the total GDP (Ayyapan and Biradar, 2000). Accordingly, India
ranked seventh in the Marine capture fish producing countries and has witnessed a growth
rate of 15.1% during the years 2003-2012 (FAO, 2014). And during the financial Year 2014-
2015, India has been the world’s second largest fish producing country after China, with
current estimated total output of 9.58 million tonnes (David, 2016). The marine fishery
resources from the Indian seas are harvested using more than 35 different types of craft gear

combinations. Major gears used in the marine fisheries sector are trawl nets, gill nets, bag
nets, hooks & lines and seines.
The state of Tamil Nadu has a long and glorious tradition of maritime activities.The
chief fishery resources of the state includes the pelagic sardines, seerfish, tunas, mackerel,
sharks, carangids, baaracudas, wolf herring; the demersal perches such as groupers, goat
fishes, snappers, croakers, sharks, rays, skates, coral fishes, threadfin breams, silverbellies;
and the shell fishes like chanks, squids, cuttle fish, shrimps, crabs and lobsters. Shore seines,
boat seines, trawl nets and hook & line, are the principal gears operated.Of the total marine
fish production (3.97 lakh tonnes), the share of demersal variety is about 2.26 lakh tonnes and
pelagic variety accounts for 1.71 lakh tonnes (Ilavarasan and Veerachamy, 2013).Moreover,
Thoothukudi district accounts for 11.08% of total marine fish catch of the state (Anon,
2002).The Gulf of Mannar (GoM) is a very important biodiversity region in the east coast.It
known to be richly gifted in terms of finfish diversity i.e. about 538 sp. has been documented
in the GoM region (Venkataraman and Wafar, 2005); both finfish and shell fish species are
rich in this area.The main aim of this investigation is to make a comprehensive study on the
diversity of the landed fin fishes of Tiruchendur coast.
A checklist of the landed fin fishes at Tiruchendur coast was made from January 2017
to April 2017. The survey was performed on weekly basis for a period of four months. The
specimens were photographed at the landing centre and were identified using the FAO
manual, Book of Smith’s fishes and Fishbase.org. The different fishing crafts and gears
utilized in the study area were also recorded. One way ANOVA was computed using the data
collected from the study area.
RESULTS
During the study period 80 species of fishes belonging to 32 families were recorded
namely: Serranidae, Pomacentridae, Monacanthidae, Balistidae, Caesionidae, Carangidae,
Chirocentridae, Scatophagidae, Cynoglossidaae, Chaetodontidae, Lethrinidae, Nemipteridae,
Lutjanidae, Mullidae, Haemulidae, Plotosidae, Priacanthidae, Rachycentridae, Ariidae,
Scombridae, Siganidae, Sphyraenidae, Terapontidae, Synodontidae, Labridae, Scaridae,
Clupeidae, Erangulidae, Tetradontidae, Acanthuridae, Holocentridae and Leiognathidae
(Table1). Both pelagic and demersal fishes were recorded during the study.Lutjanidae
comprised the diverse taxa with 10 different species, followed by Serranidae, Carangidae and

Lethrinidae. A high abundance of Lethrinus species was observed during the study, which
was mainly due totheir availability throughout the year in this coastal region. They have a
high economic value in the market. Apart from the local market the fishes such as:
Lethrinuslentjan, Lethrins nebulosus, Lethrinus elongatus, Lethrinus ornatus, Epinephelus
malabaricus, Cephalopholis spp., Carangids, Scarus spp., etc. are being exported.
Of the several types of traditional fishing methods, the hooks and line is one of the
most dominant and economically viable fishing techniques to exploit large pelagic, column
and demersal fishes. In the study area also Hook & line has been extensively used that yields
the maximum number of species (Fig.1). Apart from the hook & line, gill net and sardine gill
nets are also employedfor fishing. The One-way analysis of variance (ANOVA) computed
from the collected data for a significance level of 5%, shows P>0.05 i.e. the p-value is 0.064
which is greater than 0.05, hence the null hypothesis is accepted. This means that there is
statistically significant difference between the families and the species recorded during the
study (Table 2).
DISCUSSION
The present study focuses on the capture data of fin fishes of Tiruchendur coast for a
period of 4 months i.e., from January 2017 to April 2017. About 80 species of fishes
belonging to 32 families were identified; of which pelagic fishes were higher in number in
comparison to the demersal. The wider distribution of the pelagic species is indirectly
influenced by the wind through mixing of the surface waters (Cury and Roy, 1989).
Thecoastal and pelagic ecosystems are comparatively productive zones depending upon the
availability of nutrients, sunlight and stability for phytoplankton production (Cury and Roy,
1989; Bakun and Weeks, 2004). Wind forcing causes the upwelling in the coastal zone and
thereby enhances the productivity of the coastal and pelagic fishery. Hence pelagic fishes
were encountered in huge numbers in the study area.
Out of the 32 families of fin fishes, the fishes belonging to the family Lethrinidae and
Lutjanidae were captured in massive quantity, due to their extensive occurrence in this
coastal region. They hold a high economic value in the local market as well as exported
overseas. Murugan et al., (2014), have studied the occurrence of the snappers from the Gulf
of Mannar region. The species Lutjanus argentimaculatus, L. malabaricus, L. rivulatus and L.
stellatus fetched good income when compared to other species. Their study also reported that

the most frequently caught snappers were Lutjanus fulviflamma and L. fulvus, in coral reef
fish trap and gillnet.
There are about 57,000 hook & line units in India. It is considered as the second
dominant gear for marine fishing, next to drift/gill net, with the maximum number in Tamil
Nadu (39%) followed by Orissa (27%) and Andhra Pradesh (19%) (Anon, 1981). Similarly,
in the present study area, Hook & line is the predominantly used gear for the capture of small
as well as larger fishes. Hooks of various sizes are used for specific target fishes. The catch
composition for hook & line was dominated by predatory fishes, specially the indicator
species like Groupers, snappers, parrot fish, seer fish, carangids and emperor fishes. The hook
Nos. 8, 7 and 5 are utilized for capturing larger fishes including the Carangids, Parrot fishes,
groupers, seer fishes that are mainly intended for export. The other smaller fishes are caught
using hook of sizes 9, 10, 12, 13 & 14. The hook Nos.5 and 6 have been reported as ideal
hook sizes for capturing perches in Veraval waters (Kartha et al., 1973).
One impetus for fisheries development in many developing countries including India
is its potential for export and foreign exchange earnings. The export of marine products has
steadily grown over the years. The price of the fishes in the study area is driven by the
Stakeholders (Purchasing agents/processing unit) and the price rate varies, since the selling of
fishes is done by auction mode.
Table 1: Shows list of fish species recorded during the study period(January-April 2017)
Sl.
No. Family Common name Scientific name
1 Ariidae Arius sp.
2 Acanthuridae - Acanthurus sp.
3 Balistidae Orangelined triggerfish Balistapus undulates
Starry triggerfish Abalistes stellaris
- Balistes sp.
Masked triggerfish Sufflamen fraenatus
Red-toothed triggerfish Odonus niger
4 Caesionidae Gold band fusilier Caesio caerulaurea
- Caesio sp.
Wide band fusilier Pterocaesio lativittata
- Pterocaesio sp
5 Carangidae Horseye jack Caranx latus
Bigeye trevally caranx sexfasciatus
- Caranx sp.
Shrimp scad Alepes djedaba

- Decapterus sp.
Needle scaled queenfish Scombroides tol
- Scombroides sp.
Yellowtail amberjack Seriola lalandi
6 Chirocentridae Whitefin wolf herring Chirocentrus nudus
7 Cynoglossidae Bengal tonguesole Cynoglossus cynoglossus
Malabar tonguesole Cynoglossus macrostomus
8 Chaetodontidae Pennant coralfish Heniochus acuminatus
9 Clupeidae Indian oil sardine Sardinella longiceps
10 Erangulidae Indian anchovy Stolephorus indicus
11 Holocentridae Crowned squirrelfish Sargocentron diadema
12 Haemulidae Painted sweetlips Plectorhincus picus
- Plectorhincus sp1
- Plectorhincus sp2
13 Leiognathidae Stripped ponyfish Leiognathus fasciatus
- Leiognathus sp.
14 Lethrinidae Longface emperor Lethrinus elongates
Thumbprint emperor Lethrinus harak
Pink ear emperor Lethrinus lentjan
Pangled emperor Lethrinus nebulosus
Ornate emperor Lethrinus ornatus
- Lethrinus sp.
15 Lutjanidae Twin spot snapper Lutjanus bohar
Black spot snapper Lutjanus fulviflamma
Black tail snapper Lutjanus fulvus
- Lutjanus sp.
Humpback red snapper Lutjanus gibbus
Blue stripe snapper Lutjanus kasmira
Bigeye snapper Lutjanus lutjanus
Five-lined snapper Lutjanus quinquilineatus
Blubber lip snapper Lutjanus rivulatus
- Lutjanus sp.
16 Labridae Peacock wrasse Xyrichtys pavo
17 Monacanthidae Unicorn leatherjacket Aluterus monoceros
Scribbled leatherjacket Aluterus scriptus
18 Mullidae Indian goatfish Parupeneus indicus
Goldband goatfish Upeneus moluccensis
19 Nemipteridae Delagoa threadfin bream Nemipterus bipunctatus
Oblique-barred monocle bream Scolopsis xenochrous
White-cheek monocle bream Scolopsis vosmeri
20 Plotosidae Stinging eel catfsh Plotosus nkunga
21 Priacanthidae Moontail bullseye Priacanthus hamrur
22 Pomacentridae Blackspot sergeant Abudefduf sordidus

23 Rachycentridae Cobia/Black kingfish Rachycentron canadum
24 Serranidae Blue lined hind Cephalopholis Formosa
Tomato hind Cephalopholis sonnerati
Honey comb grouper Epinephelus merra
Brown hind reefcod Epinephelus undulosus
Malabar grouper/greasy grouper Epinephelus malabaricus
Foursaddle grouper Epinephelus spilotoceps
- Epinephelus sp.
Mackerel tuna Euthynnus affinis
Yellow edged lyretail Variola louti
25 Scatophagidae Spotted scat Scatophagus argus
26 Scombridae Indian mackerel Rastrelliger kanagurta
27 Siganidae White spotted Rabbitfish Siganus canaliculatus
28 Sphyraenidae Pickhandle barracuda Sphyraena jello
Obtuse barracuda Sphyraena obtusata
Yellowtail barracuda Sphyraena flavicauda
29 Synodontidae Bluntnose lizardfish Trachinocephalus myops
30 Scaridae Daisy parrotfish Scarus sordidus
Blue-barred parrotfish Scarus ghobban
Tricolor parrotfish Scarus tricolor
31 Tetradontidae Silver-cheeked toadfish Lagocephalus sceleratus
32 Terapontidae Tiger perch Terapon jarbua
Figure1: Shows Gear wise landing of fishes at Tiruchendur coast
25
20
No. of Families
15
10
0
Hook & line Gill net Sardine gill net
Gears used
Table2: One way ANOVA calculated between families and species reported from the study area.

Source of SS df MS F P-value F crit
Variation
Between 3784.5 1 3784.5 5.115341 0.064397 5.987378
Groups
Within Groups 4439 6 739.8333
Total 8223.5 7
CONCLUSION
Globally there is a surge of interest in designating areas of the seas as marine

reserves and protected areas to maintain and conserve marine species and habits threatened
by human activities. There is a growing consensus that living marine resources require more
stringent protection. Crises facing many marine ecosystems are increasing and attracting
more public attention. Hence, there is widespread concern among policy makers, scientists
and the public at large about the current status and uncertain future of marine ecosystems.
The present investigation is therefore an initial step towards preserving and protecting the
marine ecosystem. Further studies are requiring to be carried out in the study area to know
the better understanding of the current status and occurrence of the fin fish resources.
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EFFECT OF PROBIOTICS ON CICHLID FISH, MAYLANDIA

GRESHAKEIREPRODUCTION UNDER CAPTIVE CONDITIONS

Ambika.* P and A. Pushparaj**
*Assistant Professor of Biological Science, Keins College of Education for Women, Vallioor,
TN.
** Assistant Professor of Zoology, T.D.M.N.S. College, T.Kallikulam – 627 113, TN.
ABSTRACT
The present investigation was designed to evaluate the efficacy of probiotic B. subtilis on
reproductive performance of freshwater ornamental ice blue cichlid fish, Maylandia
greshakei under captive conditions. The probiotic feed R1 fed fish exhibited the maximum
average fecundity per female (20.23±10.29), followed by R2 (19.9±9.46), R3 (18±9.39) and
R4 (17.81±9.5). A significant (P<0.05) decline in the average fecundity per female
(15.23±7.39) was observed in fish fed the control feed (experimental group R0). The fish fed
with the probiotic feeds R1 shows the higher fry survival (19.19±9.39), weight (0.74±0.07),
length (41.31±1.1), fry weight (0.0024±0.0004) and fry length (6.36±0.54) followed by R2
survival (19.9±9.46), weight (0.72±0.10), length (39.68±1.15), fry weight (0.0023±0.0003)
and fry length (6.29±0.58), R3 survival (18±9.39), weight (0.71±0.14), length (39.63±1.88),
fry weight (0.0022±0.002) and fry length (6.08±0.52) and R4 survival (17.81±9.5), weight
(0.68±0.11), length (39.57±1.22), fry weight (0.0024±0.0003) and fry length (6.16±0.68). All
the parameters exhibited significant differences (P<0.05) within the probiotic diet fed fish.
Likewise, the higher total fecundity was observed of fish fed the probiotic feeds R1 has the
higher range (20.23) followed by R2 (19.9), R3 (18) and R4 (17.81) were also significantly
higher (P<0.05) than fish fed the control diet R0 (15.23). All the probioticfed fish exhibited
significantly lower. The probiotic feed R1 fed fish exhibited the maximum average hatching
and survival rate per female (97.75 and 97.41), followed by R2 (93.25 and 91.80), R3 (89.25
and 88.78) and R4 (85.33 and 84.22) throughout the experimental period. A significant
(P<0.05) decline in the average hatching and survival rate per female (83.25 and 82.25) was
observed in fish fed the control feed (R0).
KEY WORDS: Probiotics, Cichlids, Spawning, Fecundity, survival rate
INTRODUCTION

In aquaculture, nutrition like lipid, protein, fatty acids, vitamin E, vitamin C and
carotenoids influencing various reproduction process such as fertilization, larval development
and fecundity (Izquierdo et al., 2001). Nutrition plays a significant role on growth and
reproductive potential of aquarium fish and various live feeds have been used for fish rearing.
Various nutrient supplements like hormones, nutrient mixture, antibiotics,
chemotherapeutants and herbal products are used for brood stock of ornamental fish. But in
general, care should be taken to use of these agents such as increased risk of suppression of
the beneficial microbial activity in the intestinal tract of the breeders. Hence, the breeders
easily become prone to disease by the opportunistic pathogens. However, the indiscriminate
use of antibiotics and chemotherapeutants for improved health and nutrition has been
criticized because their use has created a lot of problems with drug resistance bacteria,
toxicity and accumulation both in fish and environment (Ghosh et al., 2008).
A probiotic is defined as a live microbial adjunct which has a beneficial effect on the
host by altering the host associated or ambient microbial community, by ensuring improved
quality of its favorable environment. Based on this definition, probiotics may include
microbial community that prevents pathogens from multiplying in the gastro intestinal tract,
superficial structures and in the culture environment of the cultured species.
Now-a-days, probiotics have been increasingly used in the biological control to
prevent diseases in aquaculture. Currently, commercial products are available in liquid or
powder forms and various technologies have been developed for improvement. Dosta et al.
(2012) to isolated and identified the 16Sr DNA, bacteria with probiotic capabilities from the
digestive tract of Pterophyllum scalare and evaluate their ability to adhere to the intestinal
epithelium using immunohistochemical techniques and bacteriological analysis. Martinez
Cruz et al., (2012) listed the in applications of probiotics in aquaculture. In this context the
present investigation was designed to evaluate the efficacy of probiotic B. subtilis on
reproductive performance of freshwater ornamental ice blue cichlid fish, Maylandia
greshakei.
Experimental Design
The experimental animal M. greshakei (Ice blue cichlid) is an omnivorous freshwater
ornamental fish was chosen for the present study. About 4 months old juveniles of ice blue
cichlid (M. greshakei) were purchased from a commercial fish farm (Saravanan fish farm) in
Sawyerpuram at Thoothukudi District. The collected fishes were transported to laboratory in

a polythene bag with oxygenated water. M. greshakei having a length 3 - 4 cm
approximately were randomly selected for the study. The chosen fishes were transferred to a
cement water tank and acclimatized to the laboratory conditions for a period of 15 days. After
that the experimental animal were transferred to the plastic troughs and water was changed
once in two days. The experimental diets were feed thrice a day (9am, 1pm and 5pm). Water
quality parameters like pH, dissolved oxygen, temperature, total alkalinity and ammonia were
measured periodically following standard methods (APHA, 1998).
Probiotics
A commercial probiotic Bacillus subtilishas been selected for the present
investigation. The probiotic B. subtilis is a Gram-positive, catalase-positive bacterium, found
in soil and the gastrointestinal tract of ruminants and humans.
Probiotic Feed Preparation
The selected probiotic B. subtilis was supplemented to the basal diet of 100gm at the
levels of 0.2, 0.4, 0.6 and 0.8g. The basal diet was prepared by using the following
ingredients like wheat flour, soybean, groundnut oil cake, fish meal and corn flour. The
experimental diets were prepared by thoroughly mixing the dry ingredients with water, steam
sterilized and after cooling 0.2, 0.4, 0.6 and 0.8 gm of probiotic B. subtilis were added to the
basal diet and soft dough was prepared. This was then passed through a mincer and the
obtained pellets were dried properly. To avoid fungal infection, these pellets were packed in
an air tight polythene bags. The proximate compositions like moisture, protein, lipid and ash
of all probiotic feeds and control feed were determined using standard procedures of AOAC
(1990).
Experimental Condition
The feeding trial was conducted in laboratory in circular plastic tubs for 60 days.
Before feeding the experimental feed, the initial weight of all the fishes were noted. Fishes
were divided into four groups and the groups were fed with different concentrations of 0.2
(R1), 0.4 (R2), 0.6 (R3) and 0.8 gm (R4) of probiotic supplemented feeds during the
experimental period (Table 3.1). The experimental feeds were given to the fishes twice a day
and the control group was also maintained. The experimental fish were fed with feed at 5% of
their body weight and also fed with two split doses throughout the experimental period. To
prevent the water quality, excess feed should be avoided and also the unutilized feed and
fecal matter were collected before every morning and water was changed once in every two
days.

Parameters Analyzed
Reproductive parameters such as Relative Fecundity (RF), Gonado Somatic Index
(GSI), Fry Survival, dead fry, deformed fry, weight and length of fry and weight and length
of adult by using the following formula. All data obtained from experiments were analyzed
by one-way analysis of variance (ANOVA) to compare the significance between control and
experimental diets. Significance was considered at 5% (P<0.05) level.
Total fry production throughout experimental period
Relative Fecundity
Mean weight of female in Gram
Gonado Somatic Index Ovary weight

X 100
Body weight
Total live fry after time
Fry Survival X100
Total fry production
Total Number of fry
Hatching Rate X 100
Experimenta l period
Total Number of fishes introduced
Survival Rate X 100
Number of fish survival
Statistical Analysis
Data are presented as means ± standard deviation (SD). The effect of dietary organic
acids on reproductive parameters was analyzed by one-way analysis of variance (ANOVA).
Multiple comparisons among means were made with Duncan’s new multiple range tests and
significance was set at P<0.05. All the statistical analysis was performed using the software
SPSS (Version 16).
RESULTS
Feed
The proximate composition of experimental probiotic diets and control feed are
presented in Table 1. The viability of the probiotic strain in probiotic incorporated feeds with
days of storage is given in Table 2. In the control feed, no B. subtiliswas found throughout the
storage period.
The fish were fed by Prepared Granulated Feed (PGF) revealed the best growth.
However, there were no significant differences (P>0.05) among the LEW and PGF.
Accordingly, the best weight gain (3.80 ± 0.43 g) was obtained in this treatment. The
proportion of growth in DG was lower than other groups, but there were not any significant
differences between DG and DT.
Assessment of reproductive performance

The table 3 shows that the results of reproductive performance of different
experimental groups of M. greshakei. The average weight and length of spawning females
were highest in fish of experimental group R1 and differed significantly (P<0.05) from the
spawning females of the other experimental groups. The average fecundity and fry survival in
M. greshakei increased with an increase in the concentration of probiotic supplementation in
feed. The probiotic feed R1-fed fish exhibited the maximum average fecundity per female
(20.23 ±10.29), followed by R2 (19.9 ±9.46), R3 (18 ±9.39) and R4 (17.81 ±9.5). A
significant (P<0.05) decline in the average fecundity per female (15.23 ± 7.39) was observed
in fish fed the control feed (experimental group R0). The fish fed with the probiotic feeds
(experimental groups R1, R2, R3 and R4) recorded significantly higher (P<0.05) survival,
weight and length of fry. The length of fry also exhibited significant differences (P<0.05)
within the probiotic feed-fed fish. Likewise, the GSI of fish fed the probiotic feeds R1, R2,
R3 and R4 were also significantly higher (P<0.05) than fish fed the control feed. All the
probiotic-fed fish exhibited significantly lower (Table 4).
The table 5 and 6 shows that the average hatching and survival rate of the
experimental fish. The probiotic feed R1-fed fish exhibited the maximum average hatching
and survival rate per female (97.75 and 97.41), followed by R2 (93.25 and 91.80), R3 (89.25
and 88.78) and R4 (85.33 and 84.22) throughout the experimental period. A significant
(P<0.05) decline in the average hatching and survival rate per female (83.25 and 82.25) was
observed in fish fed the control feed (experimental group R0).
Table 1: Proximate composition of control and different experimental diets
Proximate
R0 R1 R2 R3 R4
composition
Moisture 34.20±4.35 34.74±4.11 34.50±4.16 34.13±4.72 35.28±4.61
Protein 4.26±0.09 5.28±0.12 5.44±0.16 5.20±0.18 4.91±0.11
Lipid 10.82±0.19 10.45±0.18 10.52±0.30 10.83±0.27 10.54±0.31
Ash 8.90±0.18 8.75±0.17 9.06±0.11 9.07±0.12 8.41±0.21
Values are represented as mean ±.
Table 2: Log counts g _1 of probiotic strain in probiotic incorporated feeds during

storage
Feed 0th day of storage 30th day of storage 60th day of storage
R1 8.62 ± 0.18 8.59 ± 0.31 7.46 ± 0.16
R2 7.60 ± 0.11 7.62 ± 0.06 6.81 ± 0.04
R3 6.63 ± 0.23 6.60 ± 0.13 6.29 ± 0.19
R4 5.66 ± 0.09 5.64 ± 0.2 5.02 ± 0.25

Values are mean± SD for three samples of each feed.
Table 3: Reproductive performance of different experimental groups
Experime Experimental Groups

nt groups
R0 R1 R2 R3 R4
Fecundity 15.23±7.39c 20.23±10.29a 19.9±9.46ab 18±9.39ab 17.81±9.5b
Fry 12.91±5.15c 19.19±9.39a 18.66±8.94ab 16.77±9.62b 16.45±9.24b
survival
Dead fry 2.32±2.69b 1.04±1.49a 1.24±1.72a 1.23±1.76a 1.36±1.81a
Deformed 0.32±0.79b 0.1±0.31a 0.15±0.36ab 0.24±0.53ab 0.21±0.54ab
fry
Weight 0.71±0.07ab 0.74±0.07a 0.72±0.10ab 0.71±0.14c 0.68±0.11b
(g)
Length 39.17±1.05a 41.31±1.1a 39.68±1.15a 39.63±1.88b 39.57±1.22a
(mm)
Fry 0.0019±0.0002b 0.0024±0.0004a 0.0023±0.0003a 0.0022±0.002a 0.0024±0.0003a
weight (g)
Fry length 5.82±0.76d 6.36±0.54a 6.29±0.58ab 6.08±0.52c 6.16±0.68bc
(mm)
GSI (%) 8.13±0.44b 9.25±0.56a 9.4±0.5a 9.1±0.39a 8.84±0.36ab
Average per female. Mean±SD values with different letters in each row are significantly
(P<0.05) different.
Table 4: Spawning details of M. greshakei during the study period
Study Total Number of eggs/female

Pair
Period fecundity Max Min Average
R0 60 8 17 09 15.23
R1 60 12 22 18 20.23
R2 60 10 20 17 19.9
R3 60 9 18 14 18
R4 60 9 18 12 17.81
Table 5: Average hatching rate of M. greshakei during the study period

Temp No. of Hatching Rate
Pair Average
(oC) Spawn 1 2 3 4 5 6 7 8 9 10 11 12
R0 29±2 8 84 83 80 81 82 87 80 82 - - - - 83.25
R1 29±2 12 96 97 98 100 97 96 98 98 96 97 97 96 97.75
R2 29±2 10 90 91 93 95 94 93 94 91 93 94 - - 93.25
R3 29±2 9 87 89 88 89 91 90 91 88 87 - - - 89.25
R4 29±2 9 84 86 85 88 87 85 83 84 86 - - - 85.33
Table 6: Average Survival rate of M. greshakei during the study period
Temp No. of Survival Rate

Pair Average
(oC) Spawn 1 2 3 4 5 6 7 8 9 10 11 12
R0 29±2 8 82 83 82 83 81 80 82 85 - - - - 82.25
R1 29±2 12 96 97 98 97 99 98 98 98 99 96 96 98 97.41
R2 29±2 10 90 91 92 93 94 94 92 93 90 89 - - 91.80
R3 29±2 9 89 88 87 89 89 89 85 90 90 - - - 88.78
R4 29±2 9 85 84 83 85 87 86 84 80 84 - - - 84.22
DISCUSSION
Feed
The proximate compositions like the crude protein, fat content, moisture, ash content
etc in all the test feeds were on par with the recommended levels (crude protein 30-45% and
crude lipid 4-8%) proposed for feeding ornamental fish (Adhikari, 2008). The incorporation
of probiotics (B. subtilis) into the feed base in all probability resulted in an increase in the
protein content of all the probiotic feeds (R1, R2, R3 and R4) than that of the control feed.
The counts of B. subtilisdecreased on an average by 1 log unit in all the probiotic
incorporated feeds after 90 days of storage period. Gildberg et al., (1998) also observed a
reduction in counts of probionts from109 to107 g-1 after storage for about a month at 4oC.
Similarly, Robertson et al. (2000) also reported only a gradual reduction of the probiont
Carnobacteriumsp in feed from 5.70 x107 - 1.20 x106 over a 6-month storage period. The
high viability of B. subtilisin feed is due to its ability to form endospores. Junge et al., (2000)
observed 100% viability of B. subtilisspores at 5% moisture content after 6 weeks of storage
at 54oC.

Assessment of reproductive performance
The reproductive performance of the experimental fish in terms of high fecundity,
high GSI, high fry survival, reduction in fry mortality and deformity and higher average
weight and length of fry were increased after the incorporation of probiotics in the feed. The
dietary administration of an indigenous spore-forming Bacillus probiont resulted in an
elective probiont colonization and proliferation in the host digestive tract (Rengpipat et al.,
2000). Probiotic bacteria established in the gut enhance brood stock and larval nutrition by
synthesizing essential nutrients (proteins and essential fatty acids) and enzymes (amylase,
protease and lipase) (Irianto and Austin 2002). Probiotic bacteria in the fish intestine
enhances host enzyme secretion by the superior maturation of fish intestinal secretory cells
(Tovar et al., 2002), which increases the digestive efficacy of the complex proteins and lipids
included in the diet, thus increasing the rate at which they can be assimilated by the host
animal. This finding is similar to that obtained by De Schrijver and Ollevier (2000), who
investigated protein digestion in juvenile Scophthalmus maximusand showed that
supplementation of the diet with a potential probiont, Vibrio proteolyticus, resulted in
increased digestion and absorption of protein, particularly in the distal portion of the
gastrointestinal tract. The nutrients like proteins and fatty acids are very essential constituents
of the yolk and their presence in diet consequently supports good oocyte development and
maturation and a higher rate of vitellogenesis (Dahlgren, 1980) and the essential fatty acids
also supply energy to sustain the spawning activities. Probiotic bacteria also produce B group
vitamins (Goldin and Gorbach 1992) and supply of the same and certain unknown stimulants
(Coves et al., 1990) could have played a key role in the elevated reproductive performance of
the probiotic feed-fed fish. However, except for X. helleri, no direct correlation was
demonstrated between fecundity and size of female fish. Tamaru et al., (2001) reported that
age of brood stock in X. helleriaffects fry production and that optimal fry production was
obtained from brood stock of 6 - 12 months in age or 6 cm – 7 cm total length. Similarly in
this study, more fry production was observed in the higher weight group M. greshakei.
Dharmaraj and Dhevendran (2010) evaluated the efficacy of Streptomyces as a
probiotic feed for the growth of ornamental fish, Xiphophorus helleri. Hernandez et al.
(2010) analyzed the effects of the commercial probiotic, Lactobacillus casei on the growth
and protein content of skin mucus and stress resistance of juveniles of the Porthole live bearer
Poecilia gracilis (Poecilidae). A study was conducted to examine the effects of the probiotic
Lactobacillus rhamnosus, as a feed additive on zebrafish (Danio rerio) fecundity (Gioacchini

et al., 2010). Effect of diet (fatty acid and protein) content during spawning season on
fertility, eggs and larvae quality of common porgy was studied by Abrenouch et al., (2010).
Effect of probiotic immunogen on reproductive performance in female platy
(Xiphophorus maculatus) was carried by Abasali and Mohamad (2011). Mansa and Allah
(2011) studied the effect of dietary vitamin C on reproductive performance of a fresh water
ornamental species the platy (Xiphophorus maculatus). Nekoubin et al., (2012) studied the
effect of symbiotic (Biomin imbo) on fecundity and reproduction factors of zebra fish (Danio
rerio).
The oviparous breeding strategy was exhibited by the experimental fish, which means
egg laying, egg fertilization, followed by hatching of eggs, release of free-swimming fry into
the water is external (Siciliano, 1972). The average numbers of dead and deformed fry were
found to vary significantly (P<0.05) in all the experimental diets with the higher values being
recorded in control feed-fed fish and the lower values in probiotic feed fed fish. The average
weight and length of released fry were also significantly (P<0.05) more in the probiotic feed-
fed fish, when compared to the control diet fed fishes (Irianto and Austin, 2002). Studies have
shown the importance of balancing the composition of dietary unsaturated fatty acids
(arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid) in fish to ensure
enhanced larval quality (Porter and Bromage, 2003). Likewise in the present study also
essential fatty acids plays a key role to supply the energy for spawning activities. In the
present study, the reduced numbers of dead and deformed fry in the probiotic diet fed fish
was observed because of the synthesis of B group vitamins, particularly thiamine (vitamin
B1) and vitamin B12 by the probiont B. subtilis. Several earlier studies had demonstrated the
beneficial effect of thiamin (vitamin B1) in reducing the mortality of progeny in the Atlantic
salmon (Robertson et al., 2000) and early mortality syndrome in the feral lake trout (Brown
et al., 1998) and in the Pacific salmon (Hornung et al., 1998).
Care should be taken when the probiotic levels administered to fish to avoid
overdosing and under dosing with resultant lower efficacy and unnecessary costs. From this
study, we can conclude that a probiotic concentration of 10 6 - 108 cells g-1 was sufficient for
enhanced reproductive performances. However, the use of a higher concentration of probiotic
cells did not always yield significantly better results.
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STUDIES ON THE EFFECT OF ORGANIC FERTILIZERS ON
GROWTH AND ECONOMIC TRAITS OF SILKWORM, BOMBYX
MORI L.
C. Dyana Selin1and MA. Ramani Bai2

1
Asst.Professor, Department of Zoology, Muslim Arts College, Thiruvithancode,
Kanyakumari District, Tamil Nadu.
2
Associate professor, Department of Zoology, Muslim Arts College, Thiruvithancode,
ABSTRACT
The domesticated silkworm Bombyx mori is a monophagous insect that can be raised on fresh
mulberry leaves. Mulberry is a sole food plant for silkworm. Good quality leaf production in
mulberry is highly dependent on supply of various input. Application of inorganic fertilizers
though increased the yield substantially but cannot sustain the soil fertility status. Recently a
great attention was drawn towards the application of bio organic farming to avoid the heavy
use of agrochemical that resulted in numerous environmental troubles. A field experimental
study was conducted during 2014-2015. The experimental results related the effect of organic
fertilizers specially VM+PM had a significant effect on growth
KEYWORDS: Poultry Manure, Vermicompost, Bombyx mori, Organic Fertilizer
INTRODUCTION
Mulberry is the sole food plant of silkworm, Bombyx mori L. plays vital role in the
growth and development of silkworm and in turn the silk production. Leaf quality and
quantity not only influence the silkworm growth and development, but also the cocoon
production, quantity and quality of raw silk. Nearly 70 per cent of silk protein produced by
silkworm is derived directly from proteins of mulberry leaves.
Yield and quality of mulberry leaf are influenced by agronomic practices such as
spacing, irrigation, fertilizer schedules and pruning practices apart from mulberry variety and
environmental conditions viz., season, temperature, humidity, duration of sunshine hours, soil
type etc. Introduction of high yielding mulberry varieties and high intensity cropping system

initiated to meet the increased demand for leaf with high nutritive value for feeding the
silkworm. The nutritional status of mulberry leaves can be improved by enriching them with
extra nutrients in order to increase larval growth and improve cocoon characteristics
(Sengupta et al. 1992). According to Miyashita (1986), mulberry leaf contributes to an extent
of 38.20 per cent for successful cocoon crop production.
Unilateral usage of heavy doses of chemical fertilizers upset the availability of
different plant nutrients and resulted in widespread deficiencies in mulberry field including
micronutrients (Krishna and Bongale, 2001). The present study was undertaken the individual
and combined effect of vermicompost manure, poultry manure as their influence on growth
and economic traits of silkworm, B.mori.
MATERIAL AND METHODS
The MR2 mulberry variety was cultivated on separate pots and supplied with
vermicompost manure, poultry manure of individual and combined mixture. After one month,
the organic fertilizers were applied in the following forms; Control, VM (Vermicomposed
Manure), PM (Poultry Manure), VM+PM (Vermicomposed Manure 25g + Poultry Manure
25g).
The organic fertilizers were supplied 15 days intervals. After of six months, leaves
were collected from the control and experimental mulberry plants for leaves analyses. The
amount of leaf protein, carbohydrate and lipid were estimated by the following methods of
Lowry et al., 1951; Sciefter et al., 1950 and Folch et al., 1957 respectively.
The first instar larvae were divided into eight groups. Each group consists of six
replicates of 30 worms in each. The Control, VM, PM, VM+PM, treated leaves was supplied
to the B.mori larvae throughout the larval period. The silkworm larvae were reared following
the shelf rearing method of Krishnaswami et al., 1978 in the rearing house.
After the second moult, the third instar larvae were divided into six group. Each group
consisted of six replicates of 30 worms in each. The first group was given normal feedings 4-
5 times a day and treated as the control. The remaining group was considered as the
experimental batch. They were fed with Poultry, vermicompost, poultry + vermicompost
enriched mulberry leaves. The growth rate (%) and economic traits such as cocoon weight
(mg), pupal weight (mg), shell weight (mg), shell ratio (%), filament length (m), denier,
fibroin (%) and sericin (%) were analyzed by the method of Sonwalker, 1993. Data were
statistically analyzed by Zar, 1984.
RESULTS

The biochemical constituents of M.indica leaves supplied with organic manure is
presented in Table 1. In control, the biochemical content of mulberry leaves was protein
(26.30±0.59 mg/g,) carbohydrate (17.35±0.45 mg/g) and lipid (2.60±0.06 mg/g). Maximum
41.45±0.55 mg/g and minimum 26.80±0.69 mg/g amount of protein were observed in the
mulberry leaves, when mulberry plant was supplied with VM+PM respectively. The
carbohydrate 27.31±0.51 mg/g and lipid 4.65±0.07 mg/g content were high, when the soil
supplied with VM+PM respectively.
Organic manure supplemented to the silkworm larvae for the extra synthesis of silk.
Table 2 shows the growth rate (%) of B.mori fed with organic manure enriched mulberry
leaves. The maximum growth rate (%) (440.77±28.05) percentage was observed when the
larvae fed with mulberry leaves enriched with VM+PM fertilizer, when compared to control
322.77±18.30 %.
Table 1: Biochemical constituents in mulberry leaves enriched with organic manure
Treatments Protein (mg/g) Carbohydrate (mg/g) Lipid (mg/g)
Control 26.30±0.59 17.35±0.45 2.60±0.06
VM 31.46±0.42 18.25±0.50 3.24±0.07
(19.61) (5.18)* (24.61)
PM 30.26±0.60 20.16±0.55 3.02±0.06
(15.05) (16.19) (16.15)
VM+PM 41.45±0.55 27.31±0.51 4.65±0.07
(57.60) (57.40) (78.84)
Percent deviation over control values in parentheses*not significant
All other deviations significant at P < 0.05 (t-test)
Table 2: Growth rate (%) of B.mori larvae fed with mulberry leaves enriched with
organic manure
Treatments Instars Growth rate (%)
IV V
Control 382.70±23.60 1615.00±43.20 322.77±18.30
VM 406.40±27.40 2020.00±50.60 397.53±19.40
(6.19)* (25.07) (23.16)
PM 386.40±27.05 1920.00±38.60 397.40±24.85
(0.96)* (18.88) (23.12)
VM+PM 420.30±22.60 2270.00±40.10 440.47±28.05
(9.82) (40.55) (36.46)
N = 30;Percent deviation over control values in parentheses*not significant
All other deviations significant at P < 0.05 (t-test)

The economic traits of B.mori larvae fed with organic manure enriched mulberry
leaves are presented in Table 3. The maximum cocoon weight, pupal weight, shell weight,
shell ratio, filament length and fibroin percentage and denier was 1680±41.68 mg,
1405±30.41 mg, 275±21.79 mg, 16.36±1.05 per cent, 902.25±22.80 m, 81.75±1.13 per cent
and 3.01±0.08, respectively, when larvae fed with VM+PM.
Table 3: Economic traits of B.mori larvae fed with organic manure enriched mulberry
leaves
Economic Traits Treatments

Control VM PM VM+PM
Cocoon Weight 1380±32.59 1625±27.38 1540±39.37 1680±41.68
(mg) (17.75) (11.59) (21.73)
Pupal Weight 1170±23.71 1361±26.36 1305±46.01 1405±30.41
(mg) (16.32) (11.53) (20.08)
Shell Weight 210±13.69 262±11.51 234±19.17 275±21.79
(mg) (24.76) (11.42) (30.95)
Shell Ratio (%) 15.21±0.75 16.12±0.67 15.19±1.12 16.36±1.05
(5.98)* (-0.13)* (7.56)*
Filament length 745.10±35.17 885.30±26.45 878.10±31.34 902.25±22.80
(m) (18.81) (17.84) (21.09)
Fibroin (%) 71.20±1.185 78.45±1.25 75.70±1.40 81.75±1.13
(10.18) (6.32)* (14.81)
Sericin (%) 28.80±1.185 21.55±1.25 24.30±1.40 18.25±1.13
(-25.17) (-15.62) (-36.63)
Denier 2.32±0.05 2.62±0.04 2.80±0.07 3.01±0.08
(12.93) (20.68) (29.74)
N=30; Percent deviation over control values in parentheses*not significant All other
deviations significant at P < 0.05 (t.-test)
DISCUSSION
In the present study, the biochemical components such as, proteins, carbohydrate and
lipid contents were increased, when larvae fed with organic fertilizers. The protein content
varied significantly under different supplementation of organic manure. The higest protein
content in silkgland (67.68 mg/g) was recorded, when mulberry leaves administered with
VM+PM. This work was agreed with Mondal et al. (2007) suggested that the silkworm
B.mori produce massive amount of silk protein during the final stage of larval development
and this protein are stored in the middle silkgland. Easwaran and Mariselvi (2016) reported
that the integration of vermicompost with inorganic fertilizers tended to increase the yield of
crops vize potato, rape seed, mulberry and marigold over other traditional composts. In the

present study, growth rate (%) were increased, when larvae fed on mulberry leaves treated
with VM+PM (vermicomposed and poultry manure) significantly gained more larval weight,
as compared to those fed on control mulberry plant. Earlier study indicated the quality and
quantity of mulberry can be increased by adopting the physiological manipulations through
the foliar sprays vermin wash and cow dung wash (Kurtz 1950). The different concentrations
of (100, 150 and 200 ppm) vermin wash and cow dung wash improve the biochemical
parameters and yield of mulberry.
Woods (1999) observed that fifth instar larvae reared on essential sugar, amino acids,
proteins and vitamins for its normal growth, survival and also for the silk growth in this work
the maximum amount of protein 56.28 mg/ml in the haemolymph of B.mori was observed,
when the soil was applied with vermicomposed + poultry fertilizers. Fatbody and muscles
protein 42.95 mg/g and 55.63 mg/g were increased with Poultry manure and VM+PM
fertilizers respectively. Okoli and Nweke (2015) observed that the effect of poultry manure
and mineral fertilizer on the growth performance and quality of cucumber fruits.
Jadhav et al. (2000) reported that the application of organic fertilizers, and in their
combination revealed that the carbohydrate, and crude protein percentages of mulberry leaves
increased which in turn significantly increased in larval body weight, silk gland weight,
cocoon yield and silk ratio. Maximum cocoon weight, pupal weight, shell weight, shell ratio
per cent, filament length, fibroin and denier were 21.73 mg/g, 20.08 mg/g, 30.95 mg/g,
7.56%, 21.09 m, 14.81 % and 29.74 respectively, when B.mori larvae fed with VM+PM
(vermicompost + poultry manure) treated mulberry leaves. The present finding are in
agreement with those of Vishwanath et al. (1997), Jadhave et al. (2000) who reported that the
nutritional sources effects not only the growth and development of the silkworm larvae but
also its final silk produce.
Vermicompost is not only a rich organic manure but also a very good quality soil
additive. It is aerobically degraded organic manure that has undergone chemical
disintegration by the enzymic activity in the gut of earthworms and also by the enzymes of
the microbial populations (Kale, 1992). It is also rich in growth stimulators with plant
hormone-like properties of Auxin and Gibberllin. Plants grow well in vermicompost
supplemented soil as soil nutrients are made inoculants more readily accessible to them
(Kale, 2006). It was also suggested by Kerenhap et al. (2007), who studied the influence of
different organic manure on the growth of mulberry and silkworm. The present analysis
showed that combination VM+PM (vermicomposed manure 25g + poultry manure 25g)

resulted maximum in growth, biochemical and economic parameters of larvae and treated
mulberry plants.
CONCLUSION
Mulberry plants were treated with organic manures vermicompost manure, poultry
manure, vermicompost manure + poultry manure by soil application method. In addition to
organic fertilizers to mulberry plant were used to enhance the growth and economic
characteristics of silkworm, B.mori. The protein, carbohydrate and lipid content in mulberry
leaves were increased, when vermicompost + poultry manure applied. The maximum
moisture content was observed in mulberry plant treated with vermicompost + poultry
manure. The biochemical content of organic fertilizers treated mulberry plants showed high,
when compared to the control mulberry treated leaves.
The sustainable production of mulberry leaf and cocoon crop is entirely dependent on
the maintenance of the soil fertility of mulberry garden. B.mori larvae fed with mulberry
leaves, supplied with different individual and combined organic fertilizers. It is very much
clear that, there is appreciable improvement in the cocoon shell weight, shell ratio percentage,
filament length and denier due to soil application of organic fertilizers in the present study.
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EFFECT OF VITAMINS AND MINERALS ON SILK GLAND RATIO
AND ECONOMIC TRAITS OF BOMBYX MORI L.
M. Thilsath Fatima Quraiza1 and M. Ramani Bai2

1
Asst.Professor, Department of Zoology, Muslim Arts College, Thiruvithancode,
2
Associate professor, Department of Zoology, Muslim Arts College, Thiruvithancode,
ABSTRACT
The silkworm nutritive requirements are very different and the most of it is supplied by
feeding on mulberry leaves. Although the mulberry leaves is complete diet for silkworm it is
possible that some deficiencies occur for different reasons. The supplementation of the leaves
results higher yield because the production of good quality and quantity of silk depends on
larval nutrition and healthiness of the larva, which are partially influenced by the nutritive
value of mulberry leaves. In order to investigate the effects of supplementary nutrients on
silkworm, B.mori, an experiment was conducted with vitamins and minerals treatments.
Vitamins such as ascorbic acid and folic acid (1, 2 and 3%) treated mulberry leaves fed
through first to fifth instar, one of the four normal feeding per day was substituted with
treated leaves. This same pattern was conducted in the mineral treatments such as MgSO4 and
Kcl2 (2, 4 and 6 %). The supplementation of the leaves was done by spraying the treatments
on them. These treatments resulted in a significant increase in silk gland ratio (%) and
economical parameters such as cocoon weight, pupal weight, shell weight, shell ratio and
filament length when compared with normal control. Maximum silk gland ratio (21.15±1.27
%) was observed with 1 per cent folic acid which had the most enhancements in the fifth day
of 5th instar larvae. These vitamin treatments are resulted the economic parameters such as
cocoon weight, pupal weight, shell weight, shell ratio and filament length was high, when the
larvae fed with 2 per cent folic acid. In these mineral treatments, the same economic
parameters were significantly increased, whenB.mori larvae fed with 4 per cent KCl2. Such
studies provide substantial evidence for practical application vitamins and minerals for
qualitative and quantitative improvements in silk production.
KEY WORDS: Bombyx mori, ascorbic acid, folic acid, MgSO4 and KCl2.

INTRODUCTION
A variety of nutrients, minerals, vitamins, hormones and other exogenous modulators
were successfully applied in sericulture with a view to stimulate growth, metabolism and silk
production in B. mori (Laskar and Datta, 2000). Vitamins are one of the organic compounds
used by organisms and limited amount of them is essential for natural performance.
Generally, the vitamins present in the mulberry leaves satisfy minimum needs of silkworm,
but the amount of these vitamins in mulberry leaves depends on different climate, seasons,
mulberry varieties and the use of fertilizers in the field (Etebari, 2002). All insects require a
variety of minerals and trace elements as micronutrients. Mineral nutrition has been neglected
compared with other nutrients and the quantitative requirements for insects are largely
unknown. However, caterpillars are known to require appreciable amount of potassium and
magnesium (Lock and Nichol, 1992).
Various researches have been carried out on the diet supplementation of mulberry
leaves fed to silkworms. This supplementation includes vitamins such as ascorbic acid,
thiamine, niacin, folic acid and multivitamins (Etebari et al., 2004). Its nutrients are very easy
to digest protein, carbohydrates, 50 different minerals and trace minerals, beta-carotene,
chlorophyll, fatty acid, and many other nutrients. Supplementation of ascorbic acid to
silkworm larvae has increased the fecundity, cocoon yield and filament length (Sarker et al.,
1995). Ascorbic acid (1.5 per cent) enriched mulberry leaves resulted in higher filament
length, weight and denier values of B.mori (Babu et al., 1992). Dietary supplementation of
folic acid to silkworm larvae resulted significant increase in economic traits and post cocoon
parameters (Nirwani and Kaliwal, 1996).
The supplementation of leaves results in higher yield because the production of good
quality and quantity of silk depends on larval nutrition and healthiness of the larvae, which
are partially influenced by the nutritive value of mulberry leaves (Ito, 1978). The nutritional
status of the mulberry leaves can be improved by enriching them with vitamins and other
nutrients. Fortification of mulberry leaves with complementary compounds was found to
increase the larval growth and post cocoon characteristics (Etebari and Fazilati, 2003).
Ascorbic acid has many important functions in the animal body. It is a powerful antioxidant,
protecting against oxidative damage to DNA, membrane lipids and proteins. The absence of
ascorbic acid in the diet of first and second instar larvae postponed growth and development
of silkworm (Etebari et al., 2004).The increase in cocoon shell weight might be due to the

protein conversion efficiency of the silk gland which might be resulted from the increased a
variability of the folic acid as reported by Hamano (1989).
Another point of view larval and shell weight enhanced due to mineral salts
(Chakraborthy and Medda, 1977). Narasimhamurthy and Govindappa (1988) reported a
significant increase in cocoon weight due to cobalt supplementation. Supplementation with
potassium dichromate increase not only in the larval weight and size of the silk gland, but
also in the pupal weight, cocoon weight, shell weight as well as in the filament length, as
against a decrease of the larval duration (Bhoopathy and Gunasegar, 1998).
The present study has been aimed to find out effective nutritional supplementary
compounds (vitamins: ascorbic acid, folic acid and minerals: MgSO 4 and KCl2) which mostly
enhance the silk gland ratio and economic parameters in 1 and 2 per cent vitamins and 4 per
cent minerals treated B.mori treated groups.
This investigation was carried out on mulberry silkworm, B.mori. Disease Free
Layings (DFLs) of B.mori (PMXCSR2) were obtained from the State Government Sericulture
Centre at Konam, Nagercoil Town and were incubated at 27 0C in ant proof racks at 70-80%
humidity. The emerging caterpillars were transferred to clean bamboo baskets (25cm
diameter and 5cm deep) with a scaffolding of paraffin paper (Krishnaswamy, 1978). The
young caterpillars were fed with ad libitum mulberry leaves (V1). The caterpillars were
maintained in oven dried trays. Properly disinfected mulberry leaves were supplied. The
caterpillars were carefully observed and monitored for their general good health.
Experimental Design
The first instar larvae were selected randomly and grouped into 13 batches for the
experimental and control, each group consisting of 5 replicates with 30 silkworms. Require
concentration of vitamins and minerals were prepared in distilled water shown below.
Vitamins, such as, ascorbic and folic acid are selected as supplements, purchased from U.K
Medical Shop, Nagercoil in the form of tablets. Samples are dissolved in distilled water to
obtain 1, 2 and 3 per cent solutions, respectively. The different concentrations of vitamin
solutions were uniformly sprayed on fresh mulberry leaves and were dried by air- condition,
through first to fifth instar one of the four normal feeding per day was substituted with the
vitamins treated leaves. Control larvae were fed with untreated leaves and the another
treatment have two minerals, such as, Magnesium Sulphate (MgSO 4) and Potassium Chloride
(KCl2) are selected as supplements, purchased from Global Scientific Suppliers, Nagercoil.

Samples are dissolved in distilled water to obtain 2, 4 and 6 per cent solutions, respectively.
The same vitamin treatment patterns are carried out in the mineral treatment.
Silk gland ratio was calculated and tabulated using the following formula,
Silk gland weight (mg)
Silk gland ratio = X 100
Matured larval weight (mg)
The cocoons were harvested on the fourth day after spinning and the cocoon characters were
recorded in experimental and control groups. Assessment of various cocoon parameters was
made as follows (Sonwalker, 1993). All the data were analyzed statistically by ANOVA and
t-test. (Zar, 1984).
RESULTS
Table 1 show silk gland ratio in B.mori larvae fed with vitamins. Maximum silk gland
ratio (21.15±1.27 %) was observed with 1 per cent folic acid when compared to control
(14.36±0.83 %). Table 2 shows silk gland ratio in B.mori larvae fed with minerals. Maximum
silk gland ratio (16.08±1.12 %) was observed with 4 per cent KCl2, when compared to control
(14.36±0.83 %).
The economic parameters of B.mori fed with ascorbic acid are presented in Table 3.
Maximum cocoon weight, shell weight, shell ratio, filament length were 1288.00±79.94 mg,
240.00±12.81mg, 18.63±1.51% and 711.72±52.37 m respectively, when B.mori larvae fed
with 1 per cent ascorbic acid and maximum pupal weight (1060.00±69.24 mg) were observed
in larvae fed with 2 per cent ascorbic acid and these parameters decreased, when the larvae
fed with 3 per cent ascorbic acid (22.6, 21.12, 33.00, 8.15 and -0.63 per cent respectively).
Table 4 shows the economic parameters of B.mori fed with folic acid. These economic
parameters were high, when the larvae fed with 2 per cent folic acid (1330.00±75.11 mg,
1060.00±66.53 mg, 270.00±15.65 mg, 18.79±0.93 % and 857.14±69.11 m respectively).
Table 1: Silk gland ratio (%) of B.mori larvae fed with vitamins
Conc. (%) Ascorbic acid Folic acid
Matured larval Silk gland Silk gland Matured Silk gland Silk gland
weight (mg) weight (mg) ratio (%) weight (mg) weight (mg) ratio (%)
Control 880.00± 126.40± 14.36± 880.00± 126.40± 14.36±
57.14 9.22 0.83 57.14 9.22 0.83
1 1559.00± 253.10± 16.23± 1183.10± 250.31± 21.15±
91.60 14.11 0.74 86.67 19.35 1.27
(74.69) (100.09) (13.01) (33.34) (97.88) (47.25)
2 1240.34± 187.22± 15.09± 1403.62± 290.41± 20.69±
65.27 16.03 1.02 72.05 21.07 1.14
(39.63) (48.04) (5.08)* (57.59) (129.56) (44.05)

3 1110.96± 160.43± 14.44± 1112.94± 227.04± 20.40±
80.54 8.45 0.90 81.68 15.68 0.47
(25.40) (26.88) (0.55)* (25.62) (79.50) (42.03)
N=30; Per cent deviation over control values in parentheses * not significant All other
deviations significant at P≤ 0.05 (t-test)
Table 2: Silk gland ratio (%) of B.mori larvae fed with minerals
MgSO4 KCl2
Concentration Matured Silk gland Matured Silk gland
Silk gland Silk gland
(%) larval weight weight weight
ratio (%) ratio (%)
weight (mg) (mg) (mg) (mg)
880.00± 126.40± 14.36± 880.00± 126.40± 14.36±
Control
57.14 9.22 0.83 57.14 9.22 0.83
1432.24± 200.51± 14.00± 1443.04± 220.73± 15.29±
2 78.01 12.17 9.78 64.56 10.77 1.07
(60.74) (58.59) (-2.50)* (61.93) (74.52) (6.47)*
1843.07± 278.14± 15.09± 1741.25± 280.09± 16.08±
4 56.01 13.53 1.01 84.99 12.39 1.12
(105.93) (119.87) (5.08)* (94.73) (121.41) (11.97)
1170.37± 186.57± 15.94± 1066.51± 160.14± 15.01±
6 108.51 9.27 0.87 44.27 10.29 1.05
(31.94) (47.53) (10.99) (20.51) (26.65) (4.52)*
N=30; Per cent deviation over control values in parentheses * not significant
All other deviations significant at P≤ 0.05 (t-test)
Table 3: Economic traits of B.mori fed with ascorbic acid

Concentration (%)
Parameters Control 1 2 3
Cocoon weight 1288.00±79.94 1280.00±80.19 1196.00±62.77
970.00±71.27
(mg) (31.8) (31.0) (22.6)
Pupal weight 1048.00±72.59 1060.00±69.24 996.00±56.33
820.00±66.31
(mg) (27.36) (28.68) (21.12)
Shell weight 240.00±12.81 220.00±13.04 200.00±16.87
150.00±14.81
(mg) (59.40) (46.20) (33.00)
Shell ratio 18.63±1.51 17.18±1.92 16.72±1.57
15.46±1.34
(%) (20.51) (11.11) (8.15)*
Filament length 711.72±52.37 690.25±35.74 460.31±22.54 (-
463.31±33.54
(m) (52.16) (47.65) 0.63)*
N=30; Per cent deviation over control values in parentheses* not significant

Table 4: Economic traits of B.mori fed with folic acid
Concentrations (%)
Cocoon
1050.00±82.14 1330.00±75.11 1030.00±66.59
weight 970.00±71.27
(8.00)* (36.00) (6.00)*
(mg)
Pupal
860.00±47.03 1060.00±66.53 860.00±53.70
weight 820.00±66.31
(4.80)* (28.80) (4.80)*
(mg)
Shell
190.00±11.70 270.00±15.65 170.00±10.25
weight 150.00±14.81
(26.40) (79.20) (13.20)
(mg)
Shell ratio 18.09±0.95 18.79±0.93 16.50±1.01
15.46±1.34
(%) (17.01) (21.54) (6.73)*
Filament 721.07±51.04 857.14±69.11 690.15±55.14
463.31±33.54
length (m) (54.13) (82.70) (47.63)
N=30; Per cent deviation over control values in parentheses * not significant
Table 5: Economic traits of B.mori fed with MgSO4
Concentration (%)
Cocoon weight 1090.00±57.16 1130.00±52.47 1070.00±43.22
970.00±71.27
(mg) (12.00) (16.00) (10.00)
Pupal weight 890.00±40.27 920.00±33.15 880.00±50.75
820.00±66.31
(mg) (8.40) (12.00) (7.20)*
Shell weight 200.00±12.43 210.00±10.15 190.00±9.11
150.00±14.81
(mg) (33.00) (39.60) (26.40)
Shell ratio 18.30±1.17 18.50±1.02 17.75±0.82
15.46±1.34
(%) (18.37) (19.66) (14.82)
Filament length 792.30±68.21 819.20±79.50 651.06±47.53
463.31±33.54
(m) (69.08) (74.74) (39.42)
Table 6: Economic traits of B.mori fed with KCl2
Concentration (%)
Cocoon weight 1150.00±85.07 1350.00±76.08 1010.00±80.59
970.00±71.27
(mg) (18.00) (38.00) (4.00)*
Pupal weight 960.00±63.13 1130.00±52.49 920.00±43.77
820.00±66.31
(mg) (16.80) (37.20) (12.00)
Shell weight 210.00±11.95 240.00±12.24 190.00±12.65
150.00±14.81
(mg) (39.60) (59.40) (26.40)
Shell ratio 18.26±1.26 18.40±1.07 17.12±1.15
15.46±1.34
(%) (18.12) (19.02) (10.74)
Filament length 734.00±57.11 821.05±66.04 503.50±29.00
463.31±33.54
(m) (56.84) (75.12) (8.44)

Table 5 shows the economic traits of B.mori fed with MgSO4. Maximum increase was
observed with 4 per cent MgSO4 treated group. The control cocoon weight was 970.00±71.27
mg which was increased by 16.00 per cent. Shell weight was increased by 39.60 per cent and
the shell ratio also increased by 19.66 per cent, when compared to control. Filament length
(819.20±79.50 m) increased, when compared to control. Minimum cocoon weight, pupal
weight, shell weight, shell ratio and filament length (10.00, 7.20, 26.40, 14.82 and 39.42 per
cent) were observed when B.mori larvae fed with 6 per cent MgSO4. The economic
parameters of B.mori fed with KCl2 are presented in Table 6. The maximum cocoon weight,
pupal weight, shell weight, shell ratio and filament length was 1350.00±76.08 mg,
1130.00±52.49 mg, 240.00±12.24 mg, 18.40±1.07 % and 1.05±66.04m respectively, when
B.mori larvae fed with 4 per cent KCl2, when compared to 2 and 6 per cent treated groups
cocoon weight, pupal weight, shell weight, shell ratio and filament length (4.00, 12.00, 26.40,
10.74 and 8.44 per cent) were decreased, when B.mori larvae fed with 6 per cent KCl2.
DISCUSSION
Feeding trials conducted by several workers proved that the level of nutrients are vary
in B.mori have significant influence on growth and development and cocoon production. In
this present investigation, maximum protein content of silk gland was (18.47±1.43 mg/g)
observed in the middle silk gland when larvae fed with 1 per cent ascorbic acid compared to
control (10.20±1.48 mg/g). This study was supported by Thulasi and Sivaprasad (2013) and
who found out that the impact of ascorbic acid vis-à-vis lemon juice on the growth, protein
profile and economic parameters of silkworm.
In the present study, maximum silk gland ratio (47.25 per cent) was observed with 1
per cent folic acid. These findings are in accordance with Rahmathulla et al.(2007), who
reported that folic acid solution spraying on mulberry leaf and feeding to silkworm
significantly improved larval weight, silk gland weight and growth rate. Higher larval and
silk gland weight subsequently improved the economic parameters like cocoon weight, shell
weight and shell ratio of folic acid treated batches. The current findings are comparable with
the results of Das and Medda (1988), who stated that supplementation of mulberry leaves
with vitamin B12 could increase the synthesis of nucleic acids and protein in the silk gland of
silkworm. This result was supported by Nirwani et al. (1998). They found out that the oral
supplementation of riboflavin significantly increased 11 per cent silk gland weight in B.mori.

The silk conversion rate is an important factor to be considered while evaluating the
economic parameters of a silkworm. In this investigation, silkworm larvae supplemented
with ascorbic acid and folic acid the economic characters were increased. Maximum
increased was observed with 2 per cent folic acid. According to Sengupta et al. (1972) the
nutritional status of mulberry leaves can be improved by enriching them with extra nutrients,
such as, 0.5, 1, 1.5 per cent ascorbic acid and vitamin B complex to increase larval growth
and improve cocoon characteristics. Babu et al. (1992) observed that the first and second
instar larvae reared on 1.5% ascorbic acid enriched mulberry leaves resulted in higher silk
filament length, weight and denier value. Sarkar et al. (1995) reported that supplementation
of ascorbic acid to silkworm larva has increased the fecundity, cocoon yield and filament
length of B.mori. In the present study 1% ascorbic acid increased the shell ratio and cocoon
filament length of silkworm.
The current findings, while substantiating the positive impact of economic traits,
when the larvae fed with 4 per cent MgSO4 and KCl2. These results corroborate the earlier
findings of Ito (1980), He reported that calcium, iron, magnesium, potassium, phosphorus
and zinc are essential mineral elements and must be present in silkworm diet for better silk
yield. These results agreed with Sarkar et al. (1995), who obtained a significantly increased
of the cocoon weight, shell weight and shell ratio. Horie et al. (1997) explained this
phenomenon by the stimulation of the metabolic activity of the silkworm due to Ca, Mg and
Fe, which resulted in shorter larval duration and increased pupation rate. Vishwanath et al.
(1997) attempted to inter-relate the supplementation of mulberry leaves with combination of
secondary or micronutrients on the rearing performance of the silkworm, B.mori. They found
the reduction in the larval duration and increased not only in the larval weight, but also in the
effective rearing rate, cocoon weight, shell weight as well as in the filament length as
compared to those characters in the untreated control. The nutritional sources effect not only
the growth and development of the silkworm larvae, but also its final silk product.
CONCLUSION
In the present study, the treatment of vitamins such as ascorbic acid and folic acid at
the concentration of 1 and 2 may have beneficial effects on the feed efficacy, growth, silk
gland ratio and also increased the quantity of silk production than control and the other hand
supplementation of minerals such as MgSO4 and KCl2 (4 per cent) showed significant
improvement in above parameters. So, this supplementation could be prescribed to the
farmers to get more quantity of silk.

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Babu, M., Swamy, M.T., Rao, P.K. and compounds on some economic traits
Rao, M.S. 1992. Effect of ascorbic and physiological characters of
acid-enriched mulberry leaves on silkworm Bombyx mori, M.Sc
rearing of Bombyx mori L. Indian Thesis,Isfahan University of
J.Seric., 31: 11-114. technology, Iran.
Bhoopathy, S. and Gunesegar, N. 1998. Etebari, K., Ebadi, R. and Matindoost, L.
Effect of potassium dichromate on the 2004. Effect of feeding mulberry
pre and post cocoon characters in a enriched leaves with ascorbic acid on
hybrid silkworm, Bombyx mori L. J. some biological, biochemical and
Exper. Zool., 1: 61-67. economical characteristics of silkworm
Chakraborthy, M.K. and Medda, A.K. Bombyx mori L. Int.J.Entomol., 8: 81-
1977. Effect of cobalt chloride on 877
silkworm, (Bombyx mori L.). Nistari Hamano, K.,1989. Effect of dietary
race. Sci. and Cult.,44: 231-233. pyridoxine content on diet efficiency
Das, S. and Medda, A. 1988. Effect of of silkworm larvae Bombyx mori L.
cyanocobalamine on protein and Biochem.J., 92: 333-337.
nucleic acid contents of ovary of Horie, Y., Watanabe, K. and Ito, T. 1997.
silkworm, Bombyx mori L. during Nutrition of the silkworm, B. mori
larval, pupal and adult stages of XVIII. Quantitative requirements for
development. Insect Sci. Appl., 9: 641 the potassium, phosphorus, magnesium
646. and zinc. Bull. Ser. Exp St., 22(2): 181-
Etebari K and M. Fazilati. 2003. Effect of 193.
feeding on mulberry supplementery Ito T. 1978. Silkworm Nutrition; in the
leaves with multi mineral in some Silkworm an important Laboratory
biological and biochemical Tool. Tazima, Y. (Ed), pp. 121157,
characteristics of silk worm (Bombyx Kodansha Ltd, Tokyo.
mori). J. Sci. and Technol. Agric. and Ito, T. 1980. Application of artificial diets
Natur. Resour. 7, 233-244. in sericulture, silkworm. Physiol. Div.,
Etebari K. 2002. Effect of enrichment of Seric. Expt. Stn.,14: 163-168.
mulberry leaves (morus alba)with
some vitamins and nitrogenous

Krishnaswami, S. 1978. New technology J. Appl. Sci. Environ. Manage., 11(4):
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16(12): 7-15. Sarkar, A.A., Haque, M.R., Rab, M.A. and
Laskar N and Datta M. 2000. Effect of Absar, N. 1995. Effect of feeding
alfalfa tonic and its organic in mulberry (Morus sp.) leaves
gradients on growth and development supplemented with different nutrients
of silkworm, Bombyx mori L. race to silkworm (Bombyx mori L.).
instar. Environ. Ecol.,18:591-596. Curr.Sci.,69(2): 185-188.
Locke, M. and Nichol, H. 1992. Iron Sengupta, K., Singh B.D. and Mustafij, C.
economy in insects. Transport, 1972. Nutrition of silkworm, Bombyx
metabolism and storage. Recent mori L. Studies on the enrichment of
advances in silkworm nutrition. mulberry leaf with various sugars,
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Narasimhamurthy, C.V. and Govindappa, vigorous growth of the worm and
S. 1988. Effect of cobalt on silkworm increased cocoon crop production.
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Nirwani, R. B. and Kaliwal, B. B. 1996. Synergetic effect of ascorbic acid and
Effect of ferrous and magnesium lemon juice on the growth and protein
Sulphate supplementation on some synthesis in the silkworm, Bombyx
metabolic substances of bivoltine mori and its influence on economic
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B.B. 1998. Supplementation of Shankar, M.A. 1997. Feeding of
riboflavin on economic parameters and mulberry leaves supplemented with
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influence of folic acid administration. wood cliffs, 718.

STUDIES ON AQUATIC PHYTODIVERSITY IN VAGAIKULAM POND,
AMBASAMUDRAM TALUK, TIRUNELVELI DISTRICT, TAMILNADU
1
Paul David Selson S*2Darwin Paul Edison S and3Amish Abragam D
1,2,
Assistant Professor ,3Associate Professor
PG and Research Department of Botany, St.John’s College, Palayamkottai, Tirunelveli.
Affiliated to Manonmaniam Sundaranar University, Tirunelveli - 627 012,Tamil Nadu, India
*Corresponding Author: darwinpauledison@gmail.com
ABSTRACT
Aim of the study was to document the aquatic macrophyte wealth of a perennial fresh water
wetland ecosystem called Vagaikulam pond through qualitative survey. The study on the
biodiversity of aquatic plants was carried out in selected pond, perennial in nature located in
Tirunelveli district of Tamil Nadu state, India has the area about ca. 15 ha surrounded by
Agricultural land and human settlement. Ambasamudram is located in the foothills of
Western Ghats of Tirunelveli, Tamil Nadu, and Southern India. The area receives good
rainfall during both the north-east and south-west monsoons. The location is situated at 8°42’
latitude and 77°28’ longitude at an altitude of 64.8 m above MSL. The mean annual rainfall
of the area during the study period was 652.4 mm.A total of 35 plant species including 34
Angiosperm and one Pteridophyte were observed and collected from the pond. The most
speciose families were Poaceae followed by Cyperaceae and Nymphaeaceae. Among five
morpho-ecologic groups, emergent anchored with 15 species dominated the pond followed by
floating and floating leaved anchored. Anthropogenic pressure is escalating around the pond.
Further qualitative and ecological assessments are needed to conserve this irreplaceable and
invaluable perennial ecosystem.
KEYWORDS: Aquatic macrophyte, Aquatic ecosystems and Vagaikulam Pond
INTRODUCTION
Aquatic ecosystems are important one which provides livelihoods for the millions of
people who live around them. Man depends ponds for most of his needs like fishing,
agriculture, irrigation, and other domestic purposes. Ponds are playing a very good role in
rain harvesting, storage of water and regulation of ground water level. So in order to maintain
the ground water level we must conserve ponds and pond habitat. They are also a enormously

important natural resource, providing direct benefits to human communities across the world.
Major aquatic plants are mostly valued for their food, fodder, medicine etc. They are also
key species in the provision of wetland ecosystem services, such as water filtration and
nutrient recycling. An aquatic plant is defined here as a plant that is physiologically bound to
water (a hydrophyte) or as a terrestrial plant whose photosynthetically active parts tolerate
long periods submerged, floating and Emergent anchored (Cook 1996). Aquatic plants
represent between 2,900 and 5,800 species (Chambers et al., 2008, Vié et al., 2008).
India has a rich aquatic flora, but due to various reasons such as increased urban
features including roads and buildings, habitat destruction, dumping the municipal waste and
sewage, many species have become very rare and some are on the verge of extinction.
Therefore, the present study makes an attempt to explore the existing diversity of aquatic
plants from Vagaikulam pond, Ambasamudram Taluk, Tirunelveli District, Tamil Nadu.

Study Area
The study on the biodiversity of aquatic plants was carried out in selected pond,
perennial in nature located in Tirunelveli district of Tamil Nadu state, India has the area about
ca. 15 ha surrounded by Agricultural land and human settlement. Ambasamudram is located
in the foothills of Western Ghats of Tirunelveli, Tamil Nadu, and Southern India. The area
receives good rainfall during both the north-east and south-west monsoons. The location is
situated at 8°42’ latitude and 77°28’ longitude at an altitude of 64.8 m above MSL. The mean
annual rainfall of the area during the study period was 652.4 mm.
The mean maximum and minimum temperatures of Tirunelveli district are 31 ºC and
32ºC respectively with relative average humidity 65%. The average annual rainfall is around
814.8millimetres with two thirds of the annual rainfall received during the north east
monsoon (September-December). Mean depth of water column is varying across the season;
in rainy season (September-December) the mean depth is ca. 3.5 m while in summer it drops
around ca. 1.0-1.5 m. High volume of water is available in October. So far no studies are
available on the biotic wealth of Vagaikulam. Qualitative floristic survey was made through
regular field visit during 2008-09 to record the aquatic macrophytes wealth of Vagaikulam.
Angiosperm and Pteridophyte macrophytes were observed and collected include submerged,
submerged anchored and floating leaved anchored, emergent anchored and free floating. The
collected plant specimens were identified and confirmed with regional floras (Gamble and

Fischer., 1935, Nair and Henry., 1983, Henry et al., 1987, Henry et al., 1989,Matthew et al.,
1991 and Cook., 1996) and regional checklist for hydrophytes (3). Binomial and author
citation of all collected hydrophytes were checked with International Plant Names Index
(http://www.ipni.org/ipni/plantnamesearchpage.do). Further the extraction of bio-resources
from the study area was carried out through personal interviews in the regional language
(Tamil). Specimens were cross-check in St.Xavier’s College Herbarium, palayamkottai.
Changes in mean depth of pond were noted in the month of October of every year from 2003-
2009.

A total of 35 species including 34 flowering plants and one Pteridophyte species
spread in 32 genera and 22 families were recorded. Present study, the first of its kind in this
pond showed the aquatic macrophyte wealth of Vagaikulam pond. The most speciose
families were Poaceae, Cyperaceae, and Polygalaceae Convolvulaceae, Hydrocharitaceae,
Lemnaceae, (3 each). Areaceae, Nymphaeaceae and Scrophulariaceae were represented by
three species each. Whereas, only one species each was recorded for Acanthaceae,
Alismataceae, Amaranthaceae, Aponogetonaceae, Najadaceae Asteraceae, Brassicaceae,
Ceratophyllaceae, Lythraceae, Mimosaceae, Pontederiaceae, Sphenocleaceae and Typhaceae.
Nahlik and Mitsch (2006) reported that tropical wetlands are dominated by floating aquatic
macrophytes. Though Vagaikulam is a sub tropical perennial pond it is dominated by
emergent anchored morpho-ecologic group with 35 species followed by emergent anchored
(15),floating (11), floating leaved anchored and submerged anchored (4). Species richness
appears to be influenced by seasonal variations. In rainy season (September-December) 35
species were available whereas in summer season (April-June) as many as 32 species were
recorded. Marsilea quadrifolia, Nasturtium indicum, Cyperus bulbosa, C. rotundus, Ludwigia
adscendens and L. perennis mostly grow during summer in low depth areas near the
embankment. Five species such as Lemna gibba, L. polyrrhiza, Wolffia globosa, Pistia
stratiotes, Eicchorna crassipes and Salvinia molesta showed the seasonal appearance.

Table 1: Binomial, family and morpho-ecologic group of collected aquatic macrophytes.
Sl.No Binomial Family Family Morpho-ecologic group
1 Alternanthera sessilis (L.) R.Br. ex Amaranthaceae Floating
DC.
2 Ammannia baccifera L. Lythraceae Emergent anchored
3 Aponogeton natans (L.) Engler Aponogetonaceae Floating leaved anchored
4 Azolla pinnata R.Br. Azollaceae Floating
5 Ceratophyllum demersum L. Ceratophyllaceae Floating
6 Cynodon dactylon (L.) Pers. Poaceae Emergent anchored
7 Cyperus bulbosus Vahl. Cyperaceae Emergent anchored
8 Cyperus rotundus L. Cyperaceae Emergent anchored
9 Dactyloctenium aegyptium (L.) Willd. Poaceae Emergent anchored
10 Echinochloa colona (L.) Link Poaceae Emergent anchored
11 Eclipta alba Hassk. Asteraceae Emergent anchored
12 Eichhornia crassipes (Mart.) Solms- Pontederiaceae Floating
Laub.
13 Hydrilla verticillata (L. f.) Royle. Hydrocharitaceae Submerge anchored
14 Hygrophila schulli (Hamilt.) Acanthaceae Emergent anchored
M.R.Almeida & S.M. Almeida
15 Ipomoea aquatica L. Convolvulaceae Floating leaved anchored
16 Ipomoea carnea Jacq. Convolvulaceae Emergent anchored
17 Isoetes coromandelina L.f. Isoetaceae Emergent anchored
18 Kylinga bulbosa P. Beavu. Cyperaceae Emergent anchored
19 Lemna gibba L. Lemnaceae Floating
20 Lemna polyrhiza L. Lemnaceae Floating
21 Marsilea quadrifolia L. Marsiliaceae Emergent anchored
22 Monochoria vaginalis (Burm. F.) Presl Araceae Floating
23 Najas indica (Willd.) Cham. Najadaceae Submerged anchored
24 Nelumbo nucifera Gaertn. Nymphaeaceae Floating leaved anchored
25 Neptunia prostrata (Lam.) Baill. Mimosaceae Floating
26 Nymphaea nouchali Burm. f. Nymphaeaceae Floating leaved anchored
27 Ottelia alismoides (L.) Pers. Hydrocharitaceae Submerged anchored
28 Pistia stratiotes L. Araceae Floating
29 Polygala javana DC. Polygalaceae Emergent anchored
30 Polygonum glabrum Willd. Convolvulaceae Emergent anchored
31 Salvinia molesta D.S.Mitch. Salviniaceae Floating
32 Striga angustifolia (D.Don) Saldanha Scrophulariaceae Emergent anchored
33 Typha angustifolia L. Typhaceae Emergent anchored
34 Vallisneria natans (Lour.) Hara Hydrocharitaceae Submerged anchored
35 Wolffia globosa (Roxb.) Hartog & Lemnaceae Floating
Vander Plas .
CONCLUSION
Aquatic angiosperms are played an important role in the health of a wetland
ecosystem which support huge number of other aquatic organisms. Freshwater ponds are
considered as most important wetland ecosystems sustain huge numbers of rare and

threatened aquatic plants. Ponds are freshwater habitats throughout the world, although the
amount of water in them constitutes only a minute fraction of the total freshwater resource on
earth. Water bodies are the integral part of human civilization. Both are having impact on
each other. Due to urbanization several ponds which were truly necessity of village – as a
village pond, converted into urban pond. Distinct utilization pattern has been observed for
such pond on transformation. Either the entire pond is facing impact of such change or partly
some area of pond having specific pressure of human activity. The water quality of small
water bodies, particularly in the urban areas of India are under the influence of growing
population and development.
Development results in migration of rural population to urban areas; it is easy for

these migrants to settle on the open areas nearby ponds, lakes, canals etc. were solid and fecal
waster has been dumped into water body. Added to these the other sources of water pollution
of small water bodies are agricultural runoff, industrial waste and garbage dumping etc.
These water increases pollutants in terms of nutrients, organic matter and toxic substances in
the water bodies and disturbs its ecosystem. With increase in anthropogenic meddling as a
consequence of disregard to the socio- cultural values of water reservoir, there is increase in
quality deterioration of their water. The quality of physical and chemical parameters serves as
a good index in providing a complete and reliable picture of the conditions prevailing in a
water body. Now a days the pond ecosystem degraded, this the right time to study the nature
of the pond and uses, we must conserve the freshwater ponds.
REFERENCE
Chambers, P.A., Lacoul, P., Murphy, K.J. Cook, C.D.K. 1996. Aquatic Plant Book.
and Thomaz, S.M. 2008. Global diversity SPB Academic Publishing,
of aquatic macrophytes in freshwater. Amsterdam/New York.
In: E.V. Balian, C. Lévêque, H. Segers and Gamble JS, Fischer CEC, Flora of
K. Martens (eds.), The freshwater Presidency of Madras. Vol.1-3, Adlard and
animal diversity assessment. Son Ltd., London.1921-1935; 1-2017.
Hydrobiologia 595: 9-29. Henry AN, Chitra V, Balakrishnan NP.
Cook CDK. Aquatic and Wetland Plants of Flora of Tamil Nadu, India. Series 1, Vol.
India.Oxford University Press. United 3, Botanical Survey of India,
Kingdom. 1996; 1-385. Southern Circle, Coimbatore. 1989; 1-171.

Henry AN, Kumari GR, Chitra V. Flora of Nair NC, Henry AN. Flora of Tamil Nadu,
Tamil Nadu, India. Series 1, Vol. 2, India.Series 1, Vol. 1, Botanical Survey of
Botanical Survey of India, Southern India, Southern Circle, Coimbatore. 1983;
Circle, Coimbatore. 1987; 1-258. 1-184.
Matthew KM. An Excursion Flora of Vié,J.-C., Hilton-Taylor. and Stuart, S.N.
Central Tamil Nadu. Tamil Nadu: 2008. Wildlife in a changing world: An
Thiruchirapalli, Rapinat Herbarium. analysis of the 2008 IUCN Red List of
1991; 1-682. Threatened Species. Gland, Switzerland,
IUCN.

STUDIES ON PHYTOPLANKTON DIVERSITY IN
KEELAIDAYANKULAM POND, AMBASAMUDRAM TALUK,
TIRUNELVELI DISTRICT
Darwin Paul Edison S*1, Paul David Selson S2 and Amish Abragam D3
1,2,
Assistant Professor, PG and Research Department of Botany, St.John’s College,
Palayamkottai, Tirunelveli.
3
Associate Professor, PG and Research Department of Botany, St.John’s College,
Palayamkottai, Tirunelveli.
Affiliated to Manonmaniam Sundaranar University, Tirunelveli - 627 012,Tamil Nadu, India
*Corresponding Author: darwinpauledison@gmail.com
ABSTRACT
Phytoplanktons are microscopic organisms that swim of drift in water. Phytoplankton plays
an important role on the faunal biodiversity of aquatic ecosystems. Phytoplankton Samples
were collected on each month from the sampling pond for a period of two years, June 2015 to
May 2017 in Keelaidayankulam Pond. A total of 30 Phytoplankton species were identified in
Keelaidayankulam, 13 species belonged to Chlorophyceae, 5 species to Bacillariophyceae, 5
species to Cyanophyceae and 7 species to Euglenophyceae. Simpson’s Index was ranged
from 0.5601 to 0.9 27, Shannon’s Index was ranged from 0.9898 to 2.8 1, Pielou’s Index
was ranged from 0.7 48 to 0.91 6 and Margalef’s Index was ranged from 1. 06 to 3.297
during the study period.
KEYWORDS: Phytoplankton Diversity, Keelaidayankulam Pond and Diversity Indices
INTRODUCTION
Aquatic ecosystem is the most diverse ecosystem in the world. The biota of an aquatic
ecosystem directly reflects the conditions existing in the environment in terms of the quality
and quantity of the biota. Aquatic organisms are especially important as they form the most
sensitive component of the ecosystem and signal environmental disturbances. Phytoplanktons
are microscopic organisms that swim of drift in water. Phytoplankton plays an important role
on the faunal biodiversity of aquatic ecosystems. Phytoplankton is the major primary

producers in many aquatic systems and is important food source for other organism.
Phytoplankton not only serves as food for aquatic animals, but also plays an important role in
maintaining the biological balance and quality of water. However, Phytoplankton species
composition, abundance and diversity are regulated by environmental factors like physico-
chemical properties of water, meteorological characteristics of the region and morphometric
and hydrographic features of the water body (Dahl and Wilson, 2000 and Sukumaran et al.,
2008).
Phytoplankton are very sensitive to the environment they live in any alteration in the
environment leads to the change in the phytoplankton communities in term of tolerance,
abundance, diversity and dominance in the habitat. Therefore, Phytoplankton population
observation may be used as a reliable tool for biomonitoring studies to assess the pollution
status of aquatic bodies (Mathivanan et al., 2008).
Phytoplankton is one of the initial biological components from which the energy is
transferred to higher organisms through food chain. Phytoplankton are the microscopic single
celled aquatic plants forming the prime component in the food chain of aquatic ecosystems.
In any aquatic environment, phytoplankton constitute the most important group for the
production of particulate material in the food web and also act as the first link in all marine
food chains. These microscopic unicellular plants form the base of the marine food web,
fueling all of the higher organisms with the products of their photosynthesis (Sallie, 1992).
Phytoplankton are the important components in the energy exchange processes of the
oceans. They reduce atmospheric carbon dioxide, and thus play a crucial role in controlling
climatic changes and global warming. Phytoplankton is food for a variety of predators and
fuel food webs. Phytoplankton species distribution shows wide spatio-temporal variations
due to the differential effect of hydrographical factors on individual species and they serve as
good indicators of water quality including pollution (Liu et al., 2004).
Plankton is one of the important components of any aquatic ecosystem and which is
obvious from the abundant occurrence of planktonivorous animals in the fresh water
ecosystems. Among plankton, phytoplanktons are the primary source of food in the marine
pelagic environment, initiating the food-chain which may culminate even in large mammals
(Waniek and Holliday, 2006).
Physico-chemical parameters, species composition and seasonal variation in
phytoplankton abundance have been studied in other regions of Indian (Saravanakumar et al.,
2008; Vengadesh Perumal et al., 2009).

Study area
Keelaidayankulam
Keelaidayankulam Pond is situated in Melaambasamudram village. It is the second
feeding tank from North Kodaimelalagian channel. The capacity of the tank is 0.055
Mm2,total catchment area is 0.0894 Mm2 and Annual storage capacity of the tank is 0.0336
Mm3. The average depth of the tank is 2.5 m (Fig. 1)
Figure : 1. Keelaidayankulam Pond

Collection
Phytoplankton Samples were collected on each month from the sampling ponds for a
period of two years, June 2015 to May 2017. The collections were made early in the morning
by using the standard plankton net (No. 25) with 30 cm mouth diameter and length of 1 m.
100 litre of surface water was filtered and the filtrate was put into clean labeled plastic
containers. The volume of the concentrate was adjusted to 25 ml and it was preserved
immediately with 4 % formalin or Lugol’s solution for further analysis.
Counting
From the collected and concentrated filtrate, 1 ml of the sample was taken; the
concentrate was shaken, in order to get an even distribution of phytoplankton for analysis.
The analysis was repeated for 10 times and computed. The counting was done in a Sedgwick-
Rafter counting cell. From this, the number of cells per litre was calculated and the percent
compositions of various groups of phytoplankton were computed.
Identification
The collected Phytoplankton were identified by using Standard literatures. (Smith,
1951; Desikachary, 1959; Edmondson, 1959; Fritsch, 1971 and Anand , 1980).

Diversity Indices
Different phytoplankton species were subjected to diversity analysis using diversity

indices like Shannon-Wiener diversity index (Shannon-Wiener, 1949), Simpson’s index
(Simpson, 1949), Margalef’s index (Margalef, 1958) and Pielou Evenness index (Pielou,
1966).
Shannon-Wiener diversity index
Shannon-WienerDiversity index H = -H Pi In Pi
Where Pi = S/N
S = Number of individuals of one species
N = Total number of all individuals in the sample
In = logarithm to base e
Simpson’s diversity index
(a) Simpson’s index of dominance
D = Σ = ni (ni – 1) / N (N-1)
Where, ni = the total number of individuals of a particular species.
N = The total number of individuals of all species
(b) Simpson’s index of diversity
1-D
Where, D = Simpson’s index of dominance
Margalef’s Index
Margalef’s index was used as a simple measure of species richness (Margalef, 1958).
Margalef’s Index = (S -1)/ In N
S = total number of species
N = total number of individuals in the sample.
In = natural logarithm.
Pielou’s Index
For calculating the evenness of species, the Pielou’s Evenness Index (e) was
used Pielou, 1966).
e = H / In S
H = Shannon - Wiener diversity index
S = total number of species in the sample

RESULTS
SPECIES COMPOSITION
CHLOROPHYCEAE
List of Chlorophyceae members identified during the study period. Actinastrum
hantzschii, Ankistrodesmus falcatus, Ankistrodesmus spiralis, Chlorella vulgaris, Cosmarium
granatum, Desmidium swartzii, Oedogonium sp, Scenedesmus acuminatus, Scenedesmus
armatus, Scenedesmus maximus, Scenedesmus quadricauda, Spirogyra indica, Volvox sp
(Table: 1).
BACILLARIOPHYCEAE
List of Bacillariophyceae members identified during the study period. Fragilaria sp,
Navicula cincta, Amphora sp, Melosira sp, Gomphonema clevei. (Table: 1).
CYANOPHYCEAE
List of cyanophyceae members identified during the study period. Oscillatoria limosa,
Oscillatoria princeps, Nostoc sp, Merismopedia sp, Pseudoanabaena sp (Table: 1.).
EUGLENOPHYCEAE
List of Euglenophyceae members identified during the study period. Trachelomonas
armata var. steinii, Trachelomonas armata, Trachelomonas volvocinopsis var. volocinopsis,
Trachelomonas curta var. punctata, Phacus platalea, Euglena Caudata, Euglena limnophila
(Table: 1).
DISTRIBUTION OF PHYTOPLANKTON SPECIES
A total of 30 Phytoplankton species were identified in Keelaidayankulam, 13 species
belonged to Chlorophyceae, 5 species to Bacillariophyceae, 5 species to Cyanophyceae and
7 species to Euglenophyceae (Table: 1).
DIVERSITY INDICES
Diversity indices are used to estimate the population status in an ecosystem.
Simpson’s dominance Index, Shannon’s diversity Index, Pielou’s Index and Margalef’s Index
were analysed. In Keelaidayankulam, Simpson’s Index was ranged from 0.5601 to 0.9 27,
Shannon’s Index was ranged from 0.9898 to 2.8 1, Pielou’s Index was ranged from 0.7 48 to
0.91 6 and Margalef’s Index was ranged from 1. 06 to .297 (Table: 2).

Table:1List of the Phytoplankton species recorded in the sampling pond
S.No Phytoplankton Species Name
CHLOROPHYCEAE
1 Actinastrum hantzschii
2 Ankistrodesmus falcatus
3 Ankistrodesmus spiralis
4 Chlorella vulgaris
5 Cosmarium granatum
6 Desmidium swartzii
7 Oedogonium sp
8 Scenedesmus acuminatus
9 Scenedesmus armatus
10 Scenedesmus maximus
11 Scenedesmus quadricauda
12 Spirogyra indica
13 Volvox sp
BACILLARIOPHYCEAE
1 Fragilaria sp
2 Navicula cincta
3 Amphora sp
4 Melosira sp
5 Gomphonema clevei
CYANOPHYCEAE
1 Oscillatoria limosa
2 Oscillatoria princeps
3 Nostoc sp
4 Merismopedia sp
5 Pseudoanabaena sp
EUGLENOPHYCEAE
1 Trachelomonas armata var. steinii
2 Trachelomonas armata
3 Trachelomonas volvocinopsis var. volocinopsis
4 Trachelomonas curta var. punctata
5 Phacus platalea
6 Euglena Caudata
7 Euglena limnophila

Table: 2. Diversity Indices of Phytoplankton species during the study period in
Keelaidayankulam
Months Simpson’s Index Shannon’s Index Pielou’s Index Margalef’s Index

Jun.15 0.5815 0.9989 0.8351 1.206
Jul.15 0.5601 0.9894 0.871 1.762
Agu.15 0.8634 2.151 0.9015 1.721
Sep.15 0.8720 2.186 0.8151 2.627
Oct.15 0.9131 2.700 0.821 3.001
Nov.15 0.9111 2.689 0.8501 3.271
Dec.15 0.9157 2.718 0.7348 2.921
Jan.16 0.9327 2.831 0.854 3.297
Feb.16 0.9110 2.561 0.7801 2.7
Mar.16 0.8811 2.516 0.7801 2.401
Apr.16 0.8902 2.411 0.9204 1.962
May.16 0.8116 2.587 0.8431 3.111
Jun.16 0.7118 1.38 0.8403 3.214
Jul.16 0.7666 1.48 0.7816 3.09
Aug.16 0.8131 2.006 0.9136 1.306
Sep.16 0.8813 2.211 0.8271 0.955
Oct.16 0.927 2.77 0.861 1.921
Nov.16 0.932 2.87 0.904 3.216
Dec.16 0.9018 2.61 0.8191 3.276
Jan.17 0.9257 2.673 0.721 3.16
Feb.17 0.9215 2.816 0.7811 3.339
Mar.17 0.8691 2.161 0.8321 3.165
Apr.17 0.8681 2.168 0.721 2.016
May.17 0.8301 1.951 0.777 1.516
SEASONAL VARIATION
KEELAIDAYANKULAM
During the study period, the phytoplankton dominance in Keelaidayankulam was
ascended as Chlorophyceae (20%) < Euglenophyceae (23%) < Cyanophyceae (27%) <
Bacillariophyceae (30%) in Southwest monsoon, ascended as Euglenophyceae (18%) <
Cyanophyceae (20%) < Chlorophyceae (22%) < Bacillariophyceae ( 40%) in Northeast
monsoon, ascended as Bacillariophyceae ( 10%) < Cyanophyceae (12%) < Euglenophyceae
(18%) < Chlorophyceae ( 60%) in Post monsoon, ascended as 12% in Cyanophyceae (12%) <
Bacillariophyceae ( 20%) < Euglenophyceae ( 28%) < Chlorophyceae ( 40%) in Summer (Table:
3).

Table 3: Seasonalwise distribution order of Phytoplankton in Keelaidayankulam during
the study period (2015-2017).
Percentage of occurrence
Phytoplankton Group
2015 - 2017
SWM NEM PM SUM
Chlorophyceae 20 22 60 40
Bacillariophyceae 30 40 10 20
Cyanophyceae 27 20 12 12
Euglenophyceae 23 18 18 28
DISCUSSION
Phytoplankton acts as an important primary producers in any fresh water aquatic
ecosystem, it also an indicator of aquatic pollution. Phytoplankton is microscopic unicellular free
floating organism or filamentous multicellular organism which movement is more or less
dependent on water currents (Roy et al., 2016). Phytoplankton are considered as a important
living components to maintain equilibrium among living and non living components in any
aquatic ecosystem (Pandeyet al., 2004).
Bacillariophyceae considered as a major and important group of Phytoplankton among
micro algae. The presence of Bacillariophyceae like Fragilaria sp, Navicula Cincta, Synedra sp,
Gomphonema sp, Cymbella sp in the present study indivate the sampling ponds are organically
polluted. Cymbella sp and Navicula sp present in the sampling ponds indicates highly the ponds
are organically polluted (Nissa and Bhat, 2016).
Chlorophyceaespecies like Ankistrodemus sp, Chlorella sp, Desmidium sp, Oedogonium
sp, Scenedesmus sp and Spirogyra sp are present in all sampling ponds during the study period.
Similar findings was observed by Nissa and Bhar (2016) in Nigeen Lake. The distribution of
Chlorophyceae was highly influenced by temperature, phosphate and Nitrogen (Ganaiet al., 2010
; Nissa and Bhat , 2016).
Cyanophyceae member like Oscillatoria sp and Nostoc sp are present in all sampling
ponds. Merismopedia sp and Pseudoanabaena sp were present only in Melaidayankulam,
Keelaidayankulam, Periyakulam, Vagaikulam and Puthukulam during the study period.

Abundance of Cyanophyceae indicate the water body is eutrophic in nature due to the mixing of
household waste to the fresh water bodies (Nissa and Bhat, 2016).
Euglenophyceae members like Trachelomonas sp, Phacus sp and Euglena sp are present
all the sampling ponds during the study period. Trachelomonas armata var. steinii present only
in Keelaidaynkulam, Thirunankulam, Kalkandukulam, Periyakulam and Sumaithangikulam
during the study period Phacus sp and Eugelna spas a indicator species in pond ecosystem which
tell the pond ecosystem rich in organic pollution. Similar study was observed by Nissa and Bhat,
(2016) in Nigeen Lake.
ACKNOWLEDGEMENT
The first author is thanks to Dr.C.P.Balakrishnan, Professor and Head, Department of
Botany, Aditanar college of Arts and Science for identification of Phytoplankton species.
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TO COMPARATIVE ANALYSIS AND IMPACT OF VERMICOMPOST
ON VIGNA RADIATA L.(MUNG BEAN) VIGNA MUNGO (BLACK GRAM)
AND VIGNA UNGUICULATA( CHICK PEA).
*P. Mathiarasi1 and Dr. D. Amish Abragam2

1
Research Scholar, PG and Research Department of Botany, St. John’s College, Palayamkottai -
627002
2
Associate Professor of Botany, PG and Research Department of Botany, St. John’s College,
Palayamkottai -627002
ABSTRACT
Sustainable agriculture is one in which the goal is permanence, achieved through the utilization
of renewable resources. This leads to development of concept of organic natural farming. There
is an increasing awareness about organic farming in view of the energy shortage, food safety and
environmental concerns arising out of conventional farming. Organic farming involves
harnessing of soil organism to process to animal and plant residues and to produce slow release
of nutrients as needed by the crops. Earthworms can be called as biological indicators of soil
fertility for millions of years before the green revolution; these silent machines have been
performing a marvellous function of ploughing and fertilizing soils. Earthworms play a key role
in soil biology by serving as versatile bioreactor and converts organic wastes into valuable
orgnic manure. Vermicompost is arich source of vitamins, hormones, enzymes, macro and
micronutrients which when applied to plants help in efficient growth. The present investigation
deals with effects of vermicompost on germination effiency, growth, yield and quality of Vigna
radiata L.(Mung bean) Vigna Mungo (Black gram) and Vigna. Unguiculata( chick pea)were
observed in the 15 days regular intervals. The growth and yield performance of Vigna radiata.L
Vigna and Vigna. Unguiculata in vermicompost are significantly higher than the control.
KEY WORDS: Earth worms, Organic farming, Vermicompost, Vigna radiata L.(Mung bean)
Vigna Mungo (Black gram) and Vigna. unguiculata( chick pea).

INTRODUCTION
India generates 25 million tonnes of municipal solid wastes, 320 million tonnes of
agricultural wastes, 210 million tonnes of cattle manure and 3.3 million tonnes of poultry
manure. Such varied types of solid wastes can be converted into useful resources by subjecting
them to any type of bioconversion option such as biogas production, mushroom cultivation,
vermicomposting technology, etc, of all these types, incorporation of earthworms for recycling
organic wastes offers a most viable rural appropriate technology because the earthworm
consumes almost 100% of their weight as wet organic wastes. Earthworms are called friend of
formers or natural plough by Darwin (1981).Earthworms can be called as biological indicators of
soil fertility for millions of years before the green revolution; these silent machines have been
performing a marvellous function of ploughing and fertilizing soils. Darwin (1981) stated that
earthworms prepare the ground in an excellent manner for the growth of fibrous rooted plants
and for seedlings of all kinds. Among the soil organisms which contribute to soil fertility
earthworms are the most important. When earth worms are available in soils they alter soil
porosity, increase in soil air volume from 8% to 30% and always promote plant growth,.
(Wollny, 1890).
By adopting vermiculture biotechnology, we can convert any type of organic wastes into
useful manure. Composting is one of the biological treatments of solid waste which is an aerobic
process that converts organic wastes into humus. Vermicomposting is one of the eco-friendly
technologies for waste management. Vermicomposting cleans the environment and provides
remunerative organic manure. Vermicompost is 100% organic safe non-toxic and odour free. It
helps plants to grow faster and stronger. Earthworm can have a significant effect on soil fertility
parameters, because of their ability to modify their environment (Hauster et al., 1994). The
Worms feed on organic material, break it down and then excrete it a worm castings or
vermicomost. The castings are in the form of tiny pellets which are coated with a gel. This
crumb-like structure helps improve soil drainage and aeration. The organic matter also undergoes
chemical changes in the process. This makes the nutrients more readily accessible to plant roots
but in a form that is slowly released when required by the plants. Studies also significant
pathogen reduction in organic matter that has been through the vermicomposting process. The

vermicomposting acts like a buffer for plants where soil pH levels are too high or low making
soil nutrients available again to the plant. The castings are much higher in bacteria, organic
material and available nitrogen, calcium, magnesium, phosphorus and potassium than soil itself.
Vermicompost is biologically active and will continue to condition soils up to 4 years.
The present investigation deals with effect of vermicompost on Vigna radiata L. (Green
gram). Vigna mungo L Hepper,(Black gram) Vigna unguiculata (L) Walp (Chick pea). Pulses
play a vital role in providing balanced protein component in the diet of the people in developing
countries. Green gram, black gram are the third most important pulse crop in India. Mung bean
cover an area of 3 million hectares which is about 12% of the total area under pulses in India
with a production of 1.31 million tones and productivity of 425 kg/ha. Nutritionalists believe that
even non-vegetarians stand to benefits by including in their diet, a reasonable quantity of pulses
substitute. The role of pulses in the supply of high quality dietary protein especially by
complementing the stable cereals has been legendary. They also grow in a very short period. So
pulses are chosen for this present study. Some growth parameters such as shoot length, root
length, intermodal length, leaf area index, length of pod, number of seeds, weight of pod and
weight of seeds and some biochemical parameters like chlorophyll content, protein and sugar
content were analysed.
REVIEW OF LITERATURE
Kale and Bano (1986) stated that vermiculture can considerably reduce the use of other
fertilizers besides improving soil fertility. Bhawalkar (1989) reported that vermiculture a
promising source of biofertilizer. Sultan Ismail (1993) and Guna Thilagaraj (1994) stated that
vermiculture biotechnology is a man aspect of biotechnology for clearing up environment with
cost effective management technology contribution in the field of vermicompost. Edwards and
Burrows (1988) Albanell et al., (1988) stated that Vermicompost contain nutrients in forms that
are readily taken up by plants such as nitrates, exchangeable phosphorous, soluble potassium,
calcium and magnesium, and have less soluble salts, greater cation exchange capacity and
increased humic acid contents respectively. Vinceslas-Akpa and Loquet (1997) reported that the
Vermicompost product had a lower organic carbon and higher nitrogen ratio, which indicates
that the Vermicompost products were most suitable for soil amendment use.

Atlavinyte and Daciulyte (1969) reported the Vermi compost promote the accumulation
of vitaim B12 in the soil. Jadhav (1996) reported a reduction of 50% of the recommended dose of
nitrogen was supplemented by the use of Vermicompost.
Hopkins (1995) reported that Vermicompost increase the plant growth and development
as well as crop quality significantly.Krisnamoorthy and Vijrabhiah (1986) reported that
Vermicompost contain biologically active substances such as plant growth regulators. Shi-Wei
and Fuzhen (1991) stated that the Vermicompost contain large surface area for retention of
nutrients. Orozco et.al., (1996) reported the forms are readily taken by the plants.
Vermicompost helps the plant to fight soil – borne plant disease. Choui et al., (2002)
pointed out the applications of Vermicomposts have been reported to suppress plant fungi such
as Phytophthora, Fusarium and Plasmodiophora in tomatoes and cabbage, Pythium, Rhizootonia
in cucumber and radish, Verticilli in strawberries. Addabdo (1995) stated that the various forms
of organic matter amendments could often suppress plant parasitic nematode populations.
Edward and Bohlen (1996) stated that the nursery plants grown in the biologically enhanced
Vermicompost may have increased resistance to pests and pathogen.
Basker et. al., (1993) pointed out the studies carried out under field conditions indicated
that the castings of earthworms contained 2-3 times more available potassium than the
surrounding soil. Ellio et. al., (1990) found that earthworm castings generally have a higher
ammonium concentration and water – holding capacity than bulk soil samples and they
constitute sites of high denitrification potential.
Many authors have reported that Vermicompost enchance the plant growth in field soils
and green house media has been attributed to a variety of factors including physicochemical
properties. Chan and Griffiths (1998); Edwards and Burrows (1988); Wilson and Carlile (1989);
Mba (1998); Buckerfield and Webster (1998) reported the use of Vermicompost to soil or green
house container media. Edwards and Neuhauser (1988) reported increased plant growth in
potting – media enhanced with Vermicompost derived from animal manures. Subler et. al.,
(1998) pointed out the potential use of Vermicompost in the agricultural and horticultural
industries.

Plant Materials:
Vigna radiata Linn, Vigna mungo L Hepper, Vigna unguiculata (L) Walp. was taken for
investigation, the details of which is given below.
Systematic position
Vigna radiata Linn. Vigna mungo L Hepper, Vigna unguiculata (L) Walp are comes
under the
Family - Fabaceae
Sub – family - Papilionaceae
Tamil names - Patchai Payaru, karuppu ulunthu, Thattaipayaru respectively.
Collection of seeds:
Good quality healthy Seeds of Vigna radiata L., Vigna mungo L Hepper, Vigna
unguiculata (L) Walp.were purchased from agricultural extension centre,Annanager, Tirunelveli,
Tamilnadu.
Collection of Vermicompost:
Vermicompost provides the balanced nutrient to plants. It was collected from Sri
Paramakalyanki center for Environmental Sciences, Manonmanium Sundaranar University,
Alwarkurichi, Tirunelveli, Tamilnadu.
Experimental set up:-
For my investigation, the experiments were carried out the equal sized pots in the
medium designed with the following permutations.
C - Control (sand without any treatment)
V - Vermi compost (5kg sand+ 2kg vermicompost)
FYM - Farm Yard Manure (5kg sand+2kg Farmyard manure)
CF - Chemical Fertilizer (5kg sand+ 2kg chemical fertilizer)
Selection and Treatment of seeds:-
The good healthy certified seeds of the pulse crops seeds were washed with deionised
water and surface sterilized with 0.1% mercuric chloride solution to keep off from spores of
fungi and was sown in pots.
Analysing the morphological and biochemical parameters of the plants

Plants cultivated in the pots were watered regularly.The following morphological characteristic
of the plant were observed in every fifteen days interval. For biochemical analysis the samples
were collected in fully matured healthy experimental plants.
The following morphological characters are studied:-
1. Germination Percentage (No. of seeds germinated/ Total No. Of seeds sowed)
2. Shoot length (cm)
3. Root length (cm)
4. Inter nodal length (cm)
5. Leaf area index (sq.cm) (By using leaf area meter)
6. Length of pod (cm)
7. Number of seeds / per pod
8. Weight of pod (gm) / per pot
9. Weight of seeds (gm) /per pot
The following biochemical parameters are analysed
1. Protein (Lowry’s method,1951)
2. Starch (Lugols Iodine method)
In the present study, the impact of Vermicompost on different pulses variety namely
Vigna radiata L., Vigna mungo L Hepper, Vigna unguiculata (L) Walp and several growth and
yield parameters were investigated in control, all experimental samples.
I. Growth Parameters
1. Germination Percentage
The percentage of the germination of Green gram, Black gram and Chickpea was
observed on 10th day from sowing of seeds and presented in Table – 1
In this present study the germination percentage was observed all pulses in the control,
Vermicompost treated soil, Farm Yard treated soil (FYM) and chemical fertilizer treated soil. On
the 10th day the germination percentage of all grams were observed and tabulated.

Table – 1 Germination Percentage
Sl.No Sample Green Black Chick pea
gram Gram
1. Control 58 60 54
2 Vermicompost 87 85 78
3. Farmyard 84 82 73
manure
4. Chemical 81 83 73
fertilizer
Mc Coll et.al.,(1982) have observed the similar results of seed germination and plant nutrient
absorption capacity of wheat in the application of earthwarm humic matter on loamy soil.
2. Shoot Length :
The shoot length of the all pulses grown in pots with different samples are given in Table
2 .1 to 2.3
Table – 2.1 Shoot Length of Green gram (cm)
Sl.No Sample Interva ls
th
15 30th 45th day
day day
1. Control 19.7 28.7 39.6
2. Vermicopost 23.5 30.8 42.1
3. Farmyard manure 18.5 29 40.0
4. Chemicalfertilizer 21.5 26.1 39.7
Table – 2.2 Shoot Length of black gram(cm)

Sl.No Sample Intervals
15th day 30th day 45th day
1. Control 17.5 26.7 34.6
2. Vermicopost 20.5 27..8 40.1
3. Farmyard manure 18.5 28 39.0

Table – 231 Shoot Length of Chick pea(cm)
15th day 30th day 45th day
1. Control 21.7 32.7 45.6

2. Vermicopost 23.5 36..8 48.1
3. Farmyard manure 18.5 30.2 40.0
The shoot length was observed in every 15th day interval. The shoot length
ofexperimental pulses were higher in vermicompost treated soil than the other experimental
samples. Similar results are obtained in the Applications of casts showed significant increase in
the length and weight of the shoot and root systems of the Sorghum plant(Reddy et al., 1994).
3. Root Length:
The root length of the Green gram, Black gram, and Chick pea were recorded and
presented in Table – 3.1 to 3.3.
Table – 3.1 Root Length of Green gram (cm)
Sl.No Sample Root Length of Green gram
th
15 day 30 day 45th day
th
60th day 75th day
1. Control 3.8 5.9 10.9 16.2 17.9
2. Vermi 8.2 13.8 18.3 31.5 39.0
compost
3. Farmyard 7.7 12.5 16.1 26.3 38.0
manure
4. Chemical 7.6 12.0 15.2 27.1 36.0
fertilizer
Table – 3.2 Root Length of Black gram (cm)

Sl.No Sample Root Length of Black gram
th
th
60th day 75th day
1. Control 2.7 7.9 11.9 19.2 19.6
2. Vermi 6.2 13.8 17.2 27.5 38.0
compost
3. Farmyard 6.7 11.5 14.1 24.7 32.9
manure

4. Chemical 5.6 10.0 12.5 23.4 34.2
fertilizer
Table – 3.3 Root Length of Chick pea(cm)

Sl.No Sample Root Length of Chick pea
15 day 30th day 45th day
th
60th day
1. Control 4.8 9.9 14.9 45.2
2. Vermi 8.2 14.8 19.3 69.5
compost
3. Farmyard 8.7 12.5 17.1 56.8
manure
4. Chemical 6.8 11.0 16.2 47.4
fertilizer
The length of the shoot and root has increased steadily during the growth of plants. The
maximum shoot length and root length was recorded in vermicompost sample over control and
other experimental samples. Nijawan and kanwar (1952) have observed similar results of
increased root length than the control in the application of earthworm compost to wheat and
many other crop plants. A similar effect in Salvia and Aster grown in the plots was observed by
Grappelli et al., (1985). Increase in crop growth due to transport of minerals and other
compounds from deep down to the surface soil by the earthworms were found by Sharma (1986).
Grappelli et al., (1985) concluded the enhancement of root initiation, root elongation, root
biomass and rooting percentage by vermicompost.
4. Internodal Length
The Internodal length of Green gram in each treatment is listed in Table -4
Table – 4.1 Internodal Length of Green gram (cm)
Sl.No Sample Internodal Length of Green gram
(cm)
th th
1. Control 2.1 5.9 9.7
2. Vermi Compost 4.1 9.1 12.6
3. Farm Yard Manure 3.8 7.0 11.6
4. Chemical Fertilizer 3.5 6.6 10.7

Table – 4.2 Internodal Length of Black gram (cm)
Sl.No Sample Internodal Length of Black gram
(cm)
th th
1. Control 2.7 3.7 8.65
2. Vermi Compost 4.91 8.1 13.62
Table – 4.3 Internodal Length of Chick pea (cm)

Sl.No Sample Internodal Length of Chick pea
(cm)
th th
1. Control 3.2 4.3 6.8
2. Vermi Compost 4.6 9.7 12.8
In this work,all sample were thoroughly analysed in every 15 th, 30th and 45th day intervals.
The Internodal length was higher in vermicompost treated soil plants than the control, and other
samples. Reddy et al., (1994) found that the plant height and biomass of sorghum were
significantly higher when applied with earthworm casts and soil mixture than soil alone.
5. Leaf area index:-
The leaf area index of the treated plants grown in pots with different samples are given in
Table -5.1 to 5.3
Table – 5.1 Leaf Area Index of Green Gram (cm)
th th
15 30 45th day 60th 75th
day day day day
1. Control 3.63 9.06 15.37 17.8 19.2
2. Vermicompost 15.86 21.81 45.11 49 59
3. Farmyard 9.43 17.96 31.39 38.5 57.32
manure
4. Chemical 5.38 12.13 20.68 37.3 56.85
fertilizer

Table – 5.2 Leaf Area Index of black Gram
th th
15 30 45th day 60th 75th
day day day day
1. Control 2.56 7.56 17.38 16 16.2
2. Vermicompost 12.81 20.86 38.13 48.01 48.95
3. Farmyard 7.40 16.45 35.12 38.05 45.32
manure
4. Chemical 6.31 14.11 22.65 36.3 46.25
fertilizer
Table – 5.3 Leaf Area Index of Chick pea

15th 30th 45th day 60th 75th
day day day day
1. Control 1.63 2.06 4.57 5.12 5,92
2. Vermicompost 5.82 6.09 8.11 7.95 8.8
3. Farmyard 4.03 7.36 7.39 7.34 8.32
manure
4. Chemical 5.14 5.13 6.68 7.3 7.85
fertilizer
Among the experimental samples the vermin compost treated sample had highest leaf
area index. Leaf being the major site of photosynthesis, the increased surface area of
photosynthesis, the increased surface area of leaves and its longevity might be attributed to the
enhancement of plant growth and development during its ontogeny by endogenous growth
hormonal systems (Parvatham, 1990).
6. Pod Length:
The pod length of the all grams in all the treatments are given in Table.6
Table – 6 Pod Length of Green Gram
Sl.No Sample Green Black Chick pea
gram gram
1. Control 3.8 2.9 7.3
2. Vermicompost 8.2 3.9 13.0
3. Farmyard 6.3 3.7 12.7
manure
4. Chemical 7.1 3.3 12.2
fertilizer

In this present study, the pod length of Green, black gram and chick pea in the control, VM,
FYM and CF were measured on the 60th day. The pod length was quitedifferent respectively. The
pod length was highest in the plants grown in vermicompost treated soil. Hoppkins (1949) stated
that the increase in yield of crop plants was attributed to the release of beneficial chemicals from
the bodies of earthworms.
7. Number of seeds (Per Pod):
The number of seeds per pod in each treatment is listed in Table.7
Table – 7 Numbers of Seeds (Per Pod)
Sl.No Sample Green Black Chick
gram gram pea
1. Control 7 3 12
2. Vermicompost 11 8 18
3. Farmyard 9 4 15
manure
4. Chemical 8 4 13
fertilizer
In this work, the number of seeds (per pod) found in the control, VM, FYM and CF was
carefully counted. The vermicompost treated soil plants have more number of seeds than the
other experimental samples. Edwards and Bates (1992) found that earthworms significantly
increase the number, growth rate and yield of green gram.
8. Weight of Pod (Per Pot) :
The weight of pod per pot in all the treatments are given in Table.8
Table – 8 Weight of Pod (per pot) gm
gram gram Pea
1. Control 15 10 37
manure
fertilizer
The vermicompost treated soil plants have increased weight of pod. Kale (1998)
recommended the optimum amount of vermicompost without chemical fertilizers for some crops
for optimum yield as 15 tonnes for tomato, 10 tonnes for bringal and carrot, 8 tonnes for radish

and coriander, 12 tonnes for bhendi and 3 tonnes for cowpea per acre. The increased yield found
by Neilson (1965) may be due to the presence of plant growth promoting compounds elaborated
by earthworms and their castings.
9. Weight of seeds (per pot):
The weight of seeds per pot in each treatment is given in Table -9
Table – 9 Weight of seeds (per pot in gm)
gram gram Pea
1. Control 10 16 24
manure
fertilizer
The weight of seeds was more in the vermicompost treated soil plants. Increase in yield
upto 36% to 48% due to the application of vermicompost over control has been reported by
Gunjal and Nikam (1992).
II. Biochemical Parameters :-

1.Protein Content :
The protein content of the all grams in each treatment on 60 th day is listed in table 11.
Table – 10 Protein Content of 60thday.(µg/ ml)
Sl.No Sample Protein Content on 60th day
Green Black Chick
gram gram pea
1. Control 28.8 32.6 29.3
2. Vermi 72.5 68.6 60.3
Compost
3. Farmyard 69.8 54.3 49.7
manure
4. Chemical 70.1 47.3 45.3
fertilizer
The vermicompost treated soil plants had maximum protein content than the other
experimental samples. Bapat et.al., (1986) pointed out increased amount of S and P application

in green gram leads to increased protein content while S alone increase the amount of
methionine, cysteine contents in green gram grain. Very rich source of S and P present in
vermicompost.
2. Starch Content :-
The starch content of the grams in all the treatments is given in Table 12
Table – 11Starch Content on 75th day.(µg/ml)
Sl.No Sample Starch Content on 75thday
Green Black Chick
gram gram pea
1. Control 62.0 42.5 52.7
2. Vermicompost 89.1 71.02 82.02
3. Farmyard 82.0 54.7 66.4
manure
4. Chemical 81.0 43.5 54.3
fertilizer
In VM samples, the maximum starch content was recorded on 75 th day. The control plants
had minimum starch content on the same day. Bhagat et. al., (1995) studied the effect of
vermicompost and their growth regulators on yield and quality attributes mainly the sugar
content of green gram.Vermicompost contain more amount of nutrients such as nitrogen,
sulphur, potassium, phosphorous, calcium, magnesium etc.
CONCLUSION
The present investigation reveal that pulses grow much faster in vermicompost treated soil
than the control and other experimental samples. Seed germination percentages, shoot, root,
intermodal and pod length, leaf area, number of seeds, weight of pod and seeds seem to be
promoted in Green gram during the growth period in vermicompost. Control plants had
minimumgrowth in all experiments during all the stages of growth. Then the above results
suggest that the vermicompost treated plants had higher chlorophyll, protein and starch content
when compared to the control and other experimental samples.
FUTURE SCOPE
The yield of green gram, black gram and chick pea weres limited chiefly due to plant
nutritional problem, which is the ubiquitous shortage of total and available nitrogen to the plants

especially during seedling establishment. Growth,yield and quality of variety of pulses could be
improved by application of organing manures to encourage the farmers.
Extension and demonstration programme need to provide encouragement to utilize
material with lower C:N ratios as added organic manures especially vermicompost to derive the
maximam benefits. Increased Mung bean, black gram, and chick pea production in the country
can serve a very useful purpose in this direction.
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GROWTH, ECONOMIC FEATURES AND FECUNDITY OF SILKWORM
BOMBYXMORI UNDER THE INFLUENCE OF VITAMIN B- COMPLEX
P. Kumutha1 and R. Kanaga priya2

1
Head and Associate Professor, Department of Zoology,Govindammal Aditanar College for
Women, Tiruchendur.
2
Assistant Professor, Department of Zoology,Govindammal Aditanar College for Women,
Tiruchendur
ABSTRACT
Silk worms are monophagus silk production has direct relationship with larval growth on
mulberry. One of the alternative ways of important of larval feeding is enrichment of mulberry
leaves with supplementary nutrients such as B- complex vitamins. Final instar larvae of Bombyx
mori fed mulberry leaves (MR2) supplemented with B-complex vitamins. Silk worm exerted
significant improvement in the parameter studied such as larval weight, cocoon weight, shell
weight, shell Ratio percentage, silk gland weight and fecundity at 0.5% over the control batches
and showed declined performance at higher concentrations. Cocoon weight larva was increased
by 12.1%, 16.1% and 17.2% with the treatments 0.05%, 0.1% and 0.5% respectively over the
control. It might be due to the presence of increased amount of protein and other nutrients in
mulberry leaves which contributed to the extra growth of the silk worm, there by increasing the
weights of larvae and silk gland and improving the cocoon characteristics of Bombyx mori.
KEYWORDS: Vitamin B-complex, Bombyx mori, mulberry.
INTRODUCTION
Nutritional element of mulberry leaves determine the growth and development of the
larvae and cocoon production Sridevi et al., (2005).The Bombyx mori is essentially
monophagous and survives solely on mulberry leaves (morus sp), which play an important role
in the nutrition of the silkworm, and inturn cocoon and silk production, Nagaraju, (2002).
Fortification of mulberry leaves by using supplementary nutrient and feeding to the silkworms is

a useful modern technique to increase the economic value of cocoon Kumaraj (1972). The
supplementation of extra nutrients along with mulberry leaves results higher yield because the
production of superior quality and quantity of silk depends mainly on nutritional status and
healthiness of the larva. Nutritional back ground of the larval stage significantly influences the
status of the resulting larva, pupa and adult. [Takano and Arai, 1978]. In recent years, many
attempts have been made to improve the quality and quantity of silk through enhancing the
leaves with nutrients, spraying with antibiotics, hormones, plant products and Amino acids and
vitamins. Vitamins of B-complex group and certain essential sugars, proteins, aminoacids,
minerals etc are responsible for proper growth and development of the silkworm.(Faruki,1998).
Generally, vitamins present in the mulberry leaves satisfy minimum needs of silkworm but the
amount of vitamins present in mulberry leaves varies on the basis of environmental conditions
usage of fertilizers in field and mulberry varieties and other field practices. Sengupta et al.,
(1972) show that Bombyx mori requirs specific essential sugars, amino acids, proteins and
vitamins for its normal growth, survival and also for silk gland growth. Supplementation with B-
complex increased the resistance against poor environmental conditions and increased body
weight in silkworm [Das and Medda, 1998] The effect of vitamin supplementation on the growth
of Bombyx mori have been investigated by many researchers.[Rajabi etal., 2006; Faruki 2005;
Etabari and Matindoost, 2005.,Frahma and Hueea et al., 2007; Suprakash, E and Bai, 2012]. B-
Vitamin is important for healthy growth of human foetus and also reported that vitamin is
necessary for nucleic acid biosynthesis in insects. Hung and Li., 2004 nutritional interactions
exist between vitamin B and other groups of vitamin B. Niacin moiety acts as an electron
aceptor in many biological redox reaction [Swend seid and Jacob; 1994].
The present study was under taken to evaluate the influence of synthetic vitamin B-
complex in fifth instar biovoltine hybrid L×CSR2 Silkworm on growth rate, silk gland weight
and subsequent production of quality of cocoons and fecundity.
MATERIALS AND METHOD
Hybrid race of Bombyx mori L×CSR2 was used in the present investigation. Rearing was
carried out as per the method advocated by Rajan & Himantharaj (2005).The silkworm larvae
were reared on normal mulberry leaf without any treatment till the completion of 4 th instar.
Freshly ecdysed 5th instar larvae of L×CSR2 was classified in to 6 groups including control. Five

different concentrations were prepared in distilled water of synthetic vitamin B complex viz,
0.05%, 0.1%, 0.5%, 1% and 1.5% sprayed on mulberry leaves for feeding. Treatments were
given daily once from 1 st day of 5th instar till oneset of spinning. Each treatment was replicated
thrice with 50 larvae, in each replication. Control larvae also maintained. On 3 rd and 6th day of
supplementation, the larvae were randomly selected length and weight were measured.
Sericin and fibroin content
Individual cocoon were taken in a weighting crucible to which 20 ml of 0.5 percent KOH
was added and allowed to remain soaked for 6 hours. The protein sericin was removed by
washing in boiling distilled water twice leaving behind the other protein filament fibroin. Then
the crucible containing fibroin was oven dried at 900C for 24 hours. The weight of fibroin and
sericin was determined by the formula.
Sericin content (g) = Initial weight of the shell – weight of the shell after alkali treatment.
Fibroin content (g) = weight of the shell – sericin content.
Silkworm growth
Enriched mulberry leaves with additives led to an obvious improvement in several
measures, elevating larval weight, silkgland weight with cocoon characters and fecundity.The
larval growth rate of silkworm Bombyx mori in the control and experimental groups receiving
MR2 variety of mulbery leaves socked in various, concentration of vitamin B-complex (0.05%,
0.1%, 0.5%, 1% and 1.5%) is shown in the Table-1.
Table - 1 Effect of vitamin – B complex on the larval weight of silkworm
Bombyxmori(%change over control is given in parenthesis)(N=50)
Concentration of V instar (weight V instar (weight
% change over % change over
Vitamin B- in gram ) in gram )
control control
Complex in % (on 3rd day) (on 6th day)
Control 2.391 ±0.004 - 3.504 ± 0.007 -
0.05 2.46 ± 0.02 -2.89 3.796 ± 0.74 -8.3
0.1 2.482 ± 0.062 -3.81 3.83± 0.021 -6.468
0.5 2.601 ± 0.0212 -8.78 3.880 ± 0.005 -10.73
1.0 2.194 ± 0.002 -8.24 3.327 ± 0.001 5.05
1.5 2.172 ± 0.04 9.16 3.251 ± 0.064 7.22
Non significent [F=161. 45, 161.45; P>0.05]

Among the five different concentrations 0.05%, 0.1%, 0.5%, 1% and 1.5% tested as vitamin B-
complex to 5th instar larva, three concentrations 0.05%, 0.1% and 0.5% exhibited significant
improvement in the parameter studied. The enhancement of larval growth by nutritional
additives, mulberry varieties, may be due to the action of vitamin C and B as co-enzymes in
aminoacid metabolism and antioxidant agents, which may increase amino acid concentrations in
larval tissues, leading to improvements in productivity. This hypothesis consider with that of
Suprakash and Pal (2002) found that vitamin B – complex significantly improved growth and
development with beneficial effects on the economic characteristics of the cocoon. Multivitamin
supplements increased larval weight at 2.5%. (Kayvan Etabari and Leila Matindoost, 2005).
Adminstration of folic acid increases larval weight. (Raj etal., 2002) and thiamine increases
larval weight. (Nirwani and Kaliwal 1996 ). Ascorbic acid increases larval weight.
Silk gland weight
The silk gland weight of Bombyx mori larvae fed mulberry leaves supplemented with B-
Complex was studied at different concentrations. The higher silk gland weight (1.090 g/larva)
was observed on feeding silkworm with mulberry leaves subjected to treatment 0.5% followed
by 0.1% (1.014 g/larva) and 0.05% (1.012 g/larva) and 1% (0.869 g/ larva) and 1.5% (0.842
g/larva) Table 2 and 3.
Table - 2 Effect of vitamin –B complex on the larval length of silkworm Bombyx

mori(%change over control is given in parenthesis)(N=50)
V instar
Concentration of
V instar (length in cm) % change over (length in % change over
vaitamin B-
(on 3th day) control cm) (on control
Complex in %
6thday)
Control 5.2 - 6.2 ± 0.003 -
0.05 5.6 ± 0.032 -7.69 6.3 ± 0.42 -1.695
0.1 5.7 ± 0.401 -9.62 6.4 ± 0.332 -5.08
0.5 5.8 ± 0.254 -11.54 6.5 ± 0.241 -6.78
1.0 5.3 ± 0.06 -1.92 6.0 ± 0.071 -1.695
1.5 5.1 ± 0.07 1.92 5.7 ± 0.023 3.319
Non significant (F=161.45,161.45; P>0.05)

Table 3 Correlation co-efficient between length and weight relationship and weight and
silkgland weight relationship of silkworm larvae supplemented with B-complex vitamin.
V instrar V instar V-instar Weight of

Concentration
(weight in (length in (weight in the
of vitamin B- Correlation Correlation
gram) cm) gram) silkgland
complex in %
(6th day) (6th day) (6th day) (gram)
Control 3.504 6.2 3.504 0.975
0.05 3.796 6.3 3.796 1.012
0.1 3.83 6.4 0.838 3.83 1.014
0.958
0.5 3.880 6.5 3.880 1.090
1 3.327 6.0 3.327 0.869
1.5 3.251 5.7 3.251 0.842
Economic features
Among the five difficult concentrations i.e, 0.05%, 0.1%, 0.5%, 1% and 1.5% of vitamin
B-complex tested on 5th instar larvae, three concentration viz, 0.05%, 0.1% and 0.5% exhibited
significant improvement in the parameters studied. This findings supported by Suprakash and
Pal., (2003). They found that vitamin B-complex significantly improved economic
characteristics of cocoon. Sarkor et al., (1995) who reported that supplementation to silkworm
with ascorbic acid (1%) and vitamin B-complex (0.5%) improved cocoon yield and silk filament
quality.
The result was also supported by Rajabi et al., 2006., Etabari, 2002 reported that cocoon
weight improved by the vitamin folic acid. Increase in cocoon weight and filament characters
might be due to increased protein conversion efficiency of the silk gland as a result of increased
availability of vitamins (Table4).

Table - 4 Economic features of silkworm larvae fed with vitamin B-complex supplemented
mulberry leaves. (%change over control is given in parenthesis)(N=50)
Single cocoon character
Concentration
Sericin
of Vitamin Cocoon Pupal Shell Fibroin
Shell % ratio content
complex in % weight (g) weight (g) weight (g) content (g)
(g)
1.552 ± 1.281 ± 0.271 ± 17.46 ± 022 ±

Control 0.05 ± 0.020
0.0072 0.08 0.0086 0.065 0.036
1.740 ± 1.352 ± 0.378 ± 21.72 ± 0.23 ±
0.051 ± 0.04
0.05 0.0083 0.11 0.0071 0.073 0.027
(-2)
(-12.1) (-5.543) (-39.48) (-24.39) (-4.55)
1.802 ± 1.459 ± 0.343 ± 19.03 ± 0.234 ± 0.052 ±
0.1 0.0083 0.007 0.006 0.091 0.038 0.006
(-16.1) (-13.895) (-26.57) (-8.99) (-6.36) (-4)
1.819 ± 1.465 ± 0.344 ± 18.91 ± 0.235 ± 0.526 ±
0.5 0.009 0.005 0.06 0.107 0.046 0.001
(-17.2) (-14.36) (-26.9) (-8.30) (-6.36) (-9.52)
1.323 ± 1.117 ± 0.216 ± 0.214 ± 0.412 ±
16.326 ±
1.0 0.075 0.0083 0.038 0.029 0.003
0.043 (6.49)
(14.76) (12.8) (20.29) (2.27) (-724)
1.204 ± 0.992 ± 0.212 ± 17.60 ± 0.201 ± 0.348 ±
1.5 0.006 0.008 0.008 0.099 0.042 0.024
(22.4) (22.56) (21.77) (-0.8018) (8.636) (-596)
F=19,18.513; P>0.05
Silkworm Reproduction
A number of researcher works on the effect of vitamin enriched food on the reproduction of
Bombyx mori females (Saha and Khan,1999). 484 is the maximum recorded number of eggs in
the adults which were given 0.5% treatment during the larval stages. The highest concentration
i.e 0.64% had deleterious effect on all parameters. Rajabi et al., (2006) reported that Riboflavin
supplementation did not show significant difference in fecundity among the treatments and the
control. Faruki, (2005) reported that pyrol in various concentration from the third instar reduced
the fecundity. Many researchers reported vitamin at higher concentrations reduce fecundity and
fertility of silk worm (Faruki, 2005; Etabari and Matindoost, 2005; Rajabi et al., 2006).

Table - 5 Effect of vitamin B-complex supplementation on fecundity and hatchablityof Bombyx
mori female
Number of egg laid PRC percent Hatchability
Concentration in %
per mean SD reproduction percentage %
Control 403 ± 12 - 78
0.05 434 ± 8 -7.69 79
0.1 462 ± 11 -14.64 83
0.5 484 ± 9 -20.09 85
1 362 ± 6 10.17 70
1.5 316 ± 2 21.59 68
[F=161.45, 161.45 P>0.05]
CONCLUSION
According to the over all results of this study, The mulberry variety MR2 is highly
recommended for rearing silkworm in India. Fortification of vitamin B- complex orally during
5th instar silkworm larvae is beneficial, improving rearing and the quality of the silk filament and
fecundity at lower concentration i.e. 0.5% and was showing declining performance at higher
concentrations. Also, the supplementation with 0.5% vitamin B-complex significantly improved
larval weight, silk gland weight, cocoon characteristics of Bombyx mori. Cocoon weight larva
was increased by 12.1%, 16.1% and 17.2% with the treatments 0.05%, 0.1% and 0.5%
respectively over the control. It might be due to the presence of increased amount of protein and
other nutrients in mulberry leaves which contributed to the extra growth of the silk worm, there
by increasing the weights of larvae and silk gland and improving the cocoon characteristics of
Bombyx mori. Their exist a positive correlation (r = 0.958) between silk gland weight and larval
weight in the fifth instar larvae fed on mulberry leaves supplemented with vitamin B-complex.
Das and Medda., (1998) observed that the rate of increase in silk gland weight in proportionate to
the rate of increase in the body weight of silkworm. The present study also confirm these result.
This study would like to highlight the need and more comprehensive studies on this subject to
find better silk worm management.

REFERENCES
Das S, Medda A.Effect of cyanocobalamine the silk worm, Bombyx mori L.

on protein and nucleic acid contents of Bangladesh J. Zool. 24: 195-203, 1999.
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646, 1998. feeding silkworms. Indian J. seric. 11:
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biological characters in silkworm “Asia value on the basis of feeding and cocoon
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19.

ANALYSIS AND DETECTION OF BREAST CANCER MAMMOGRAM USING
MULTI-LAYER NEURAL NETWORK
G. Sam Singh1, R. Selva Priya2, K. Sabana Asmin2

1M. Tech, Assistant Professor, Dept. of Electronics and Communication Engineering ,
Dr.Sivanthi Aditanar college of Engineering., Tiruchendur, India

2B.E, Student, Dept. of Electronics and Communication Engineering, Dr.Sivanthi Aditanar
college of Engineering., Tiruchendur, India
ABSTRACT
Breast cancer is the leading cause of death in women all over the world. Over the years the help
of technology such as data mining and machine learning which can substantially improve
the diagnosis accuracy. However, standard and accurate classification is still poses a great
challenge for researchers. Machine learning techniques are currently instrumental in detection
and classification due to their adequate prediction performance .This paper formulates the
various pre-processing stages that are needed to be feed for the back propagation network, which
includes acquisition of image, extracting features from the mammograms, selecting more
optimal features from it and followed by classification using back propagation neural network
algorithm which can identify the appropriate class of mammogram thereby identifying the
cancerous cells.
KEYWORDS: Adaptive Median, Back Propagation, Feature Extraction, K-Means, Support
vector Machine.
INTRODUCTION
Cancer is not a single disease but a wide range of different diseases of which there
were hundreds of types. Thus, breast cancer refers to the erratic growth of cells that
originate in the breast tissues and nodules. . The group of these cells formed as a lump is
called tumours. Tumors can be malignant or benign. Malignant tumors penetrate deep into
nodes and destroy healthy among women between 40 and 60. It rarely occurs in men. Early
diagnosis and sustained treatment were the biggest hurdles and main indication of person
survival. To provide a better assistant for robust and reliable classification of breast tumour,

Back Propagation algorithm can be a powerful tool for such distributed diagnosis. This provides
an accurate result, whether the tumour is benign or malignant.
PROPOSED SYSTEM
The proposed method comprises four stages: pre-processing, segmentation, and feature
extraction and classification stage. The pre-processing stage involves techniques that include
noise removal , enhancement and segmentation process is done by K-means clustering. The
feature extraction stage entails extraction of features like entropy, standard deviation, texture,
Euclidean Distance, variance, tumour area, shape, concavity etc. Lastly, the classification stage
involves the use of artificial neural network by back propagation to classify breast tissue into
normal and abnormal.
BLOCK DIAGRAM:
Input Pre-Processing Segmentation
Breast (Active Median (K-Means)
Image Filter)
Feature Extraction
(Texture,
Classification Smoothness, Mean,
(SVM) Skewness, Standard
deviation, Euclidian
distance)
Back Propagation
Output
Pattern Recognition
Cancerous
Validating and Training

the Network
Non cancerous
Testing the network
View

METHODOLOGY
STEP 1: Input raw Breast Ultrasound screened images.
STEP 2: Pre-process using Adaptive Median Filter
STEP 3: Segment using K-Means Clustering on the ROI.
STEP 4: Extract features like entropy, Standard Deviation, Euclidean Distance, Variance,
Tumour area, Shape etc.
STEP 5: Classification of cancer stages using Back propagation Algorithm.
PRE-PROCESSING:
The primary tasks of pre-processing step is obtaining visually informative images and ease the
subsequent image processing and automated evaluation steps [10]. In this paper, the main focus
is on adaptive median filter.
A. Adaptive Median Filtering
The adaptive median filtering is an advanced method compared with standard median filtering.
The Adaptive Median Filter performs spatial processing and it will determine which pixels in an
image have been affected by noise. It compares each pixel in the image to its surrounding
neighbour pixels. The size of the neighbourhood is adjustable [1]. A pixel that is different from a
majority of its neighbours, as well as being not structurally aligned with those pixels to which it
is similar, is labelled as impulse noise. These noise pixels are then replaced by the median pixel
value of the pixels in the neighbourhood that have passed the noise labelling test.
Fig 1: Filtered image output using adaptive median filter
SEGMENTATION:
Image segmentation is a technique for dividing and discrediting an image into a number of
segments in order to modify the depiction of an image into more significant and easier one to

examine [4]. During the evaluation of mammogram images, it is necessary to delete background
from breast profile.
A. K-means clustering
Clustering is a method to divide a set of data into specific number of groups. K-Means
clustering is a popular method.. In K-Means clustering, It classifies a given set of data into k
number of disjoint cluster. The algorithm consists of two separate phases. It calculates the k
centroid in first phase and in the second phase it takes each point to the cluster which has
nearest centroid from the respective data point. There are different methods to define the
distance of the nearest centroid and one of the most used methods is Euclidean distance [9].
Then it recalculatethe new centroid of each cluster and based on that a new Euclidean distance
is calculated between each center and each data point. It assigns the points in the cluster which
have minimum Euclidean distance. Each cluster is defined by its member objects and centroid
of the image. The centroid for each cluster is defined as the point to which the sum of
distances from all the objects in that cluster is minimized. So K -means is an iterative
algorithm in which it minimizes the sum of distances from each object to its cluster centroid,
over allclusters [6].
The algorithm for k-meansclustering is following as:
1. Initialize number of cluster k andcentre for the image.
2. Calculate the Euclidean distance d, between the centre and each pixel of an image
using the relation givenbelow.
d=p(x,y)–ck
3. Assign all pixels of an image to the nearest centre based on distanced.
4. Recalculate new position of the centre after each pixel is assigned.
5. Repeat the process until it satisfies the requirement of given tolerance or errorvalue.
6. Reshape the clustered pixels intoimage.

Fig 2: K-Means output
FEATURE EXTRACTION
Feature extraction a kind of dimensionality decrease that productively speaks to
intriguing parts of a picture as a conservative component vector. This methodology is valuable
when picture sizes are huge and a decreased component portrayal is required to rapidly total
undertakings [7]. Feature identification, include extraction, and coordinating are regularly
consolidated to take care of normal PC vision issues.
A. Mean:
Mean is described by adding all pixel values of an image divided by total no of pixels in an
image.
∑ −1 ∑ −1
M= 1 ( , )
∗ =0 =0
B. Standard deviation:
It is the measure of probability distribution of an observed population and can serve as a measure
of homogeneity.
−1 ∑ −1
SD=�1 ∑ ( , )
∗ =0 =0
C. Skewness:
Skewness can be quantified to define the extent to which a distribution differs from normal
distribution.
1 ∑( ( , )− )
S (x)
k =
∗ 3
D. Euclidian distance:
The Euclidian distance is the straight line distance between two pixels.
( −
D(a, b) =� )2 +( − )2

Fig 3: Feature extracted image output
CLASSIFICATION:
The features or features subset are used by classifiers to classify images into normal and
abnormal. The classification technique consists of the training phase and the testing phase. The
training phase involves the analysis of features present in a known data by processing which
precedes classification. The testing phase validate the input and provide the required output.
A. Support vector machine classifier (SVM):
SVM is a method which works accurately and efficiently with high dimensionality feature
spaces. The original idea of the SVM is to construct a hyper plane as the decision surface in such
a way that the margin of separation between the positive and negative samples is maximized in
an appropriate feature space [2].Kernel functions are employed to perform the nonlinear
mapping, which computes the inner product matrix, the so-called kernel matrix, For kernel-based
methods, the kernel matrix acts as a bottleneck. All information are extracted from the Kernel
matrix.
B. Back propagation algorithm
It is a supervised multi layer perception based learning method and generalization of delta rule.
They can be used to find a pattern within the dataset to recognize the cancer automatically [9].
They works on self-learn logic principle as they require a database of inputs with desired output,
making up a training set and update their weights accordingly.
This algorithm performs four different operations
 Feed Forward computation
 Back Propagation to the output layer
 Back Propagation to the hidden layer

 Weight update
The training procedure for Back Propagation algorithm includes Feed forward and
Back Propagation in two parts. During feed forward procedure feature values from the input
layer propagates to the hidden layer and finally activation values of output node are generated.
During Back Propagation procedure, the errors of the output node can be obtained through
computing the difference between actual result and expected result [5]. Then the error signal
from the output layer propagates backwards through the network and updates the each node’s
weight. The algorithm is stopped when the value of error function is sufficiently small. This is
the rough and basic formula of Back Propagation algorithm.
Fig: Picture of back propagation network
Fig: Back propagation training window

The Back propagation algorithm detects whether it is cancerous or not.

Fig: Non –cancerous image output
Fig: Cancerous image output

If cancerous tumour is detected then further process can be done to identify the shape, size and
the exact location of the tumour.
Fig: Final output image(Cancerous)

CONFUSION MATRIX:
One measure of how well the neural network has fit the data is the confusion plot [3]. It returns a
confusion matrix plot for the target and output data in targets and outputs, respectively. On the
confusion matrix plot, the rows correspond to the predicted class (Output Class), and the
columns show the true class (Target Class). The diagonal cells show for how many of the
examples the trained network correctly estimates the classes of observations. That is, it shows
what percentage of the true and predicted classes match. The off diagonal cells show where the
classifier has made mistakes.
A..Representation of confusion Matrix:
Predicted Predicted
class 1 class 2
Actual class TP FN

1
Actual class FP TN
2
• True positive (TP): correct positive prediction

• False positive (FP): incorrect positive prediction
• True negative (TN): correct negative prediction
• False negative (FN): incorrect negative prediction.
Further, using this confusion matrix we can determine the performance of the system.
Fig: Confusion matrix

B. IMPORTANT MEASURES CALCULATED:
The main measures are given by,
SENSITIVITY
It measures the proportion of actual positives that are correctly identified
Sensitivity=TP/(TP+FN )
SPECIFICITY
It measures the proportion of actual negatives that are correctly identified
Specificity=TN/(TN+FP)
ACCURACY
The accuracy of a test is its ability to differentiate the patient and healthy cases
correctly.
Accuracy=TP+TN/(TP+TN+FP+FN )

Fig : Values of output measures calculated
CONCLUSION:
In this various levels of pre-processing techniques were studied and the stages of Breast images
which includes detection, pre-processing and classified using Back Propagation Algorithm, Also
to check its malignancy using MATLAB v2017. The values were also formulated from the
various parameters in detection of malignant tumour. It clearly demonstrates the effectiveness of
Back Propagation algorithm in the classification of breast tumours. Also this paper has the
reference on the parameters of processing the tumour. Further work can be incorporated by using
confusion matrix to find the accuracy level in detection of a malignant tumour.
REFERENCES
[1] Kesari Vermaa , Bikesh Kumar Singh, Neural Network” International
A.S. Thokec Zelst, “An Enhancement Journal of Latest Trends in
in Adaptive Median filter for Edge Engineering and Technology
Preservation”, Procedia Computer (IJLTET) , Vol 7 issue 2 July 2016.
Science 48 , Elseiver 2015. [4] R.C. Gonzalez , Woods R.E ,Digital
[2] V.Vijaya Lakshmi, G.Krishnaveni,” Image Processing, third ed , Pretice
Performance Assessment by using Hall, Englewood cliffs,NJ,2008.
SVM and ANN for Breast cancer [5] Paulin et al F,” Classification of Breast
Mammography image classification “, cancer by comparing Back
International Journal of Engineering propagation training algorithms”
Technology Science and Research International journal on Computer
IJETSR, Volume 4, Issue 9 science and Engineering, Vol. 3 No. 1
September 2017 Jan 2011.
[3] Kalpana Kaushik, Anil Arora, “Breast [6] S. M. Aqil Burney, Humera Tariq, K-
Cancer Diagnosis using Artificial Means Cluster Analysis for Image

Segmentation, International Journal Management(UCTM), Vol 3,Issue
of Computer Applications (0975 – 1,2016.
8887) Volume 96– No.4, June 2014 . [9] I.Salama, Gouda , M. B. Abdelhalim,
[7] Prannoy Giri and Saravanakumar,” and Magdy Abd-elghany Zeid.”Breast
Breast cancer detection using image Cancer Diagnosis on Three Different
processing techniques”, Orient Datasets using
J.Comp. Science &Tech, An Multiclassifiers”,Breast Cancer
International open Free access (WDBC) 32.569 (2012).
Journal, Vol 10,391-399,2017. [10]R.Ramani, N. Suthanthira Vanitha, and
[8] Pankaj Kumar Sharma ,”Implementation S. Valarmathy. ”The PreProcessing
of Cancer Detection Based on Back Techniques for Breast Cancer
Propagation” Vihar Univ, Detection in Mammography
International Journal of Converging Images.”International Journal of
Technologies and Image, Graphics & Signal Processing
5.5 (2013).

ISOLATION AND CHARACTERIZATION OF BIOACTIVE COMPOUNDS FROM
FUSINUS NICOBARICUS (ROEDING, 1798)
P.Subavathy* and R.D.Thilaga
PG and Research Department of Zoology, St.Mary’s College (Autonomous), Thoothukudi

Affiliated to Manonmaniam Sundaranar University, Abishekapatti, Tirunelveli - 627012,
Tamilnadu, India
*Corresponding Author: subavathy23@gmail.com
ABSTRACT
The marine environment is a huge source for discovering many novel drugs. The marine
environment comprises of complex ecosystem an extremely diverse reservoir of life, with a
plethora of organisms and many of these organisms are known to possess bioactive compounds
as a common means of defense. Apart from the food that is derived from the marine
environment, a wide variety of bioactive substances is being isolated and characterized several
with great promise for the treatment of human disease. The present study has been carried out to
characterize the bioactive component present in Fusinus nicobaricus through GC-MS analysis.
The GC-MS analysis of methanol extract confirmed the presence of 13 active compounds.
Several compounds were confirmed in the methanolic extract of gastropod with the prominent
peak of oleic acid (20.29%). These compounds might be responsible for different
pharmacological actions.
KEYWORDS: Fusinus nicobaricus,methanol, GC-MS analysis, bioactive compounds
INTRODUCTION
The seas and oceans, which cover 70% of the world’s surface, are one of the man’s great
hopes for future food supplies (Jerome and Williams, 1979). There are about 30 million species
of living organisms estimated on earth. Marine bioresource is known to be one of the richest
among all the living ecosystems. There is a rich marine biota in different parts of the ocean, from

the surface to the deepest part and from the estuarine region to the offshore region. Ocean waters
can support life from the microscopic bacteria to the gigantic whale (Shanmugam, 2008).
Among the various phyla represented in the marine environment, the phylum Mollusca is
the second largest phylum in the animal kingdom next to Arthropoda (Abbott, 1954). The
molluscs in total constitute a natural resource of sizable magnitude in many parts of the world.
About 28 different species of bivalves and 65 species of gastropods are of very great importance
under these categories (Ramjee and Naganathan, 2008). The study of marine organisms for their
bioactive potential and importance in the marine ecosystem has accelerated in recent years along
with the growing recognition of their importance in human life (Nazar et al., 2009). A number of
biologically active compounds with varying degrees of action such as antimicrobial, antioxidant,
anticancer, antileukemic, antiviral, antiproliferative, cytotoxic, antibiotic and antifouling
properties have so far been isolated from marine sources (Villa and Gernwick, 2010).
Molluscs are soft-bodied animals and the term ‘mollusca’ was derived from the latin
word ‘Mollis’ which means ‘Soft’. Among them gastropods are the largest and most certainly the
well-known class of all the molluscs. About one - third of the drugs being used now were
discovered from nature, the balance are synthetic. Today the number of active products
discovered from land resources is declining and this has prompted the scientists to look for
marine natural resources. So the present study has been carried out to isolate and characterize the
bioactive compounds from Fusinus nicobaricus and to evaluate their pharmacological potential.
Collection of experimental organism
Specimens of Fusinus nicobaricus used in the present study were collected from
Thoothukudi coast (8°35′ - 9°25′ N; 78°08′ - 79°30′E) along Gulf of Mannar region. Specimens
of Fusinus nicobaricus were collected during low tides from the sea in their natural habitat by
divers and from trawl nets used for crab fishing. They were brought to the laboratory and
maintained under laboratory conditions for further observations. In the present study whole body
tissue extract of Fusinus nicobaricus were used for the GC-MS analysis. The freshly collected
samples were cleaned and washed with fresh sea water to remove all impurities. The shells were
removed and the tissues were then dried in hot air oven at 56ºC for 48 hours, powdered and used
for further studies.

GC – MS analysis
GC-MS analysis was carried out on a GC Clarus 500 Perkin Elmer System comprising a
AOC 20i auto sampler and gas chromatography interfaced to a mass spectrophotometer (GC-
MS) instrument employing the following conditions such as Column elite – 5MS fused silica
capillary column (30 x 0.25mm ID x 0.25µm df, composed of 5% Diphenyl/95% Diphenyl Poly
Siloxane), operating in electron impact mode at 70eV: Helium (99.999%) was used as a carrier
gas at constant flow of 1ml/min and an injection volume 3µl (split ratio of 10:1) injector
temperature 250ºC. The oven temperature was programmed from 110ºC (isothermal for 2min),
with an increase of 10°C/min to 200°C, then 5ºC/min to 280ºC. Mass spectra were taken at
70eV; a scan interval of 0.5s and fragments from 45 to 450 a.
Identification of compounds
Interpretation on mass spectrum was conducted using the database of National Institute of
Standard Technology (NIST Ver.21), WILEY 8 and FAME having more than 62,000 patterns.
The unknown components found in the body tissues of Fusinus nicobaricus were matched with
the spectrum of the known components stored in NIST, WILEY and FAME, the MS library and
predicted from Duke’s Ethno Botanical Database.
RESULT
GC-MS chromatogram of the methanolic extract of Fusinus nicobaricus indicates the
presence of 13 chemical constituents. On comparison of the mass spectra of the constituents with
the NIST library the 13 chemical constituents were characterized and identified (Table.1). The
chemical compounds identified with their retention time (RT), molecular formula and
concentration (peak area %). GC-MS analysis from the experimental organism Fusinus
nicobaricus revealed 13 compounds that could be identified as Butanedial, Undecanal, 2-methyl-
Benzoic acid, 4-ethoxy-,ethyl ester, Pentadecanoic acid, 14-methyl, methyl ester, 1,2-
Benzene dicarboxylic acid, butyl octyl ester, Hexadecanoic acid, ethyl ester, n-
Hexadecanoicacid, 9-Octadecenamide, oleic acid, Cyclopropanedodecanoic acid, 2-octyl methyl
ester, Arachidonic acid methyl ester, 1-Heptatriacotanol, Octadecanoic acid, 3-hydroxy methyl
ester (Fig.1). Among the compounds characterized present, the prominent peak was found in

oleic acid (20.29%). The various chemical compounds which contributes to the pharmacological
activity of the gastropod Fusinus nicobaricus.
Table 1. Activity of components identified in the methanol fraction of Fusinus nicobaricus

by GC-MS
No RT Name of Molecular MW Peak Compound **Activity
compound formula Area Nature
1. 2.66 Butanedial C4H6O2 86 7.41 Aldehyde Antimicrobial,
Compound Anti-inflammatory
2. 3.01 Undecanal, 2- C12H24O 184 2.85 Aldehyde Antimicrobial
methyl- compound
3. 8.22 Benzoic acid, C11H14O3 194 4.24 Aromatic Antimicrobial
4 –ethoxy-, ethyl ester Preservative
ester.
4. 12.69 Pentadecanoic C17H34O2 270 1.85 Fatty acid No activity reported
acid, 14-methyl-, ester
methyl ester
5. 13.13 1,2- C20H30O4 334 3.70 Plasticizer Antimicrobial,
Benzenedicarbox compound Antifouling
ylic acid, butyl
octyl ester
Antioxidant,
Hypocholesterolemic,
6. 13.53 Hexadecanoic C18H32O2 284 2.70 Palmitic Nematicide, Pesticide,
acid, ethyl ester acid ester Lubricant,
Antiandrogenic,
Flavor, Hemolytic 5-
Alpha reductase
inhibitor
Antioxidant,
7. 15.33 n – Hexadecanoic C16H32O2 256 2.78 Palmitic Nematicide, Pesticide,
acid acid Lubricant,
Antiandrogenic,
Flavor, Hemolytic 5-
Alpha reductase
inhibitor
Anti-inflammatory,
Antiandrogenic,
8. 16.32 9– C18H35NO 281 8.80 Oleic acid Cancer preventive
Octadecenamide, amide Dermatitigenic,

(Z)- [Oleic acid Hypocholesterolemic,
amine] 5-Alpha reducatase
inhibitor,
Anemiagenic,
Insectifuge
Anti-inflammatory,
Antiandrogenic,
9. 17.94 Oleic Acid C18H35O2 282 20.29 Oleic acid Cancer preventive,
Dermatitigenic,
5-Alpha reductase.
10. 19.63 Cyclopropanedod C24H46O2 366 16.13 Ester No activity reported
canoic acid, 2- compound
octyl-, methyl
ester
11. 20.41 Arachidonic acid C21H34O2 318 7.87 Omega -3 Anticancer,
methyl ester fatty acid Anticholesterol
compound
12. 21.05 1- C37H76O 536 10.73 Aliphatic Antimicrobial
Heptatriacotanol alcoholic
compound
13. 22.47 Octadecanoic C19H38O3 314 10.65 Fatty acid No activity reported
acid, 3-hydroxy-, ester
methyl ester compound
Fig.1 GC-MS spectra of some compounds present in methanolic extract of

Fusinus nicobaricus
Butanedial n-Hexadecanoic acid

Oleic acid
DISCUSSION
Marine molluscan extracts are usually complex mixtures of bioactive molecules mainly
proteins, peptides and sterols. Many studies have been carried out to find out the bioactive
compounds from marine sources. The present study based on various analysis revealed the
presence of alkaloid, ketone, amino - ester, nitrogen, aldehyde, alcoholic, amide and steroid
compounds (Table.1). The GC – MS study of Fusinus nicobaricus reveals the probable bioactive
compounds such as Butanedial, Undecanal, 2-methyl-, Benzoic acid, 4-ethoxy, ethyl ester,
Pentadecanoic acid, 14-methyl -, methyl ester, 1,2 –Benzenedicarboxylic acid, butyl octyl ester,
Hexadecanoic acid, ethyl ester, n-Hexadecanoicacid, 9-Octadecenamide(Z)-(oleic acid amide),
Oleic acid, Cyclopropanedodecanoic acid, 2-octyl-, methyl ester, Arachidonic acid methyl ester,
1 – Heptatriacotanol, Octadecanoic acid, 3 – hydroxy-, methyl ester which could be responsible
for antimicrobial, anti-inflammatory, antioxidant, antifouling, hypocholesterolemic, nematicide,
pesticide, lubricant, antiandrogenic, cancer preventive, dermatitigenic and anticholesterol
activities.
The present findings are in agreement with Emiliano Manzo et al., (2007) who reported
that two novel triterpenoids, aplysoils A and B. β Etzionin a tyrosin derived compound exhibited
antibacterial activity against Bacillus subtilis (Lindquist and Fenical, 1990). Brominated indoles
6-bromo 2-methylthio indolin-3-one extracted from Australian muricid Dicathdis orbita has been
identified as anticancer drug indole derivatives of 6, 6 ′dibromoindigo have also shown
antimicrobial activity (Benkendorff et al., 2001).
The large diversity of exotic plants in rainforests has provided sources for some of these
clinically active compounds, but many have come from the sea (Rayl, 1999) and from marine
invertebrates such as sponges, tunicates, bryozoans and molluscs (Haefner, 2003). From the

result of the present study it is understood that, the rich diversity of marine biota with unique
physiological adaptations exerted or provided by organisms to the harsh marine environment,
affords a fruitful source for the discovery of life saving drugs.
CONCLUSION
World’s oceans could play paramount role in supplying life saving drugs in future.
Although substantial progress has been made in identifying novel drugs from the marine sources,
great endeavours are still needed to explore these molecules for clinical applications without
altering or disturbing the biodiversity on marine organism. The potential of marine molluscs as a
source of biologically active products is largely unexplored in India. Hence a broad based
screening of marine molluscs for bioactive compounds is necessary.
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(Part I). Polyplacophora and Immunopharmacol. Immunotoxicol,
32: 228-237.

GREEN SYNTHESIS OF SILVER NANOPARTICLES USING ATROCARPUS
HETEROPHYLLUS LEAVES AND THEIR SUPPLEMENTARY EFFECT ON
SILKWORM BIOLOGY
P.Kumutha1 and M. Vidya Kalaivani2

1
Associate Professor, Department of Zoology, Govindammal Aditanar College for Women,
Tiruchendur.
2
Assistant Professor, Department of Zoology, Govindammal Aditanar College for Women,
Tiruchendur.
ABSTRACT
Silkworms are monophagous. Silk production has direct relationship with larval growth.
Larval growth can be improved by supplementary nutrients such as nanoparticles. Nanoparticles
of 14.5 nm were prepared by ecofriendly bioreduction method using Atrocarpus heterophyllus
leaves. Final instar larvas of Bombyx mori were supplemented with 25%, 50%, and 75% of
nanoparticles through feed. 25% of silver nanoparticles stimulate growth and silk gland weight,
75% of silver nanoparticles enhanced economic features such as pupal weight, shell weight and
cocoon weight and worms fed with 100% nanoparticles showed maximum fecundity during their
adult stages. Thus silk industry can be improved by nanotechnology.
KEY WORDS: Nanoparticles, Bombyx mori, Ecofriendly.
INTRODUCTION:
The silkworm, Bombyx mori, is a monophagous lepidopteran insect; the silkworm rearing
is a traditional industry in Asia. The economic importance of silk production extensively
increases the physiological study of this species. Mulberry leaves are suitable as food for
silkworms, it contain several chemical constituents such as water (80%), proteins (27%) and
carbohydrates (11%), other extracts, mineral matters, vitamins etc. (Koul, 1989). But this is not
enough to increase the larval growth and cocoon quality as well as quantity. The
supplementation of extra nutrients along with mulberry leaves results higher yield because the
production of superior quality and quantity of silk depends mainly on the nutritional status and

healthiness of a larva. Nutritional background of the larval stage significantly influences the
status of the resulting larva, pupa and adult (Takano and Arai, 1978). Recently, much research
has been done on the diet supplementation of mulberry leaves with antibiotics, hormones, plant
products, amino acids and vitamins to fed silkworms.
Fortification of mulberry leaves by using supplementary nutrient and feeding to the
silkworms is a useful contemporary technique to increase the economic value of cocoon
(Kamaraj, 1972, Etebari 2002; Etebari and Fazilati, 2003). When silver nanoparticles were
administrated to the silkworms there is a shift in nitrogen metabolism in favour of increasing
body weight, increased output of silk, cocoon characters, fecundity and hatchability. The
syntheses of silver nanoparticles using physical and chemical methods are extremely expensive
and also involve the use of toxic, hazardous chemicals, which may pose potential environmental
and biological risks. Now a day’s silver nanoparticles are mainly synthesized with plants. Silver
ions were synthesized from silver nitrate along with different source of plant extract like
Geranium (Pelargonium gravelolens), Cinnamomum camphora leaf extract (Sankar et al., 2003,
Huang et al., 2007) and bark power of cinnamon zeylanicum tree. Gold nano triangle and
spherical nano silver particles synthesized by Aloevera leaf (Chandran et al., 2006). It is an
unavoidable fact that the silver nanoparticles synthesized have to be handled by humans and
must be in available rates for their effective utilization thus, there is a need for an experimentally
and economically feasible way to synthesize these nanoparticles by biological method, which
solve this entire problem.
MATERIALS AND METHODS:
Silkworm rearing
The eggs of silkworm Bombyx mori popular Indian (CSR XL) race were collected from
V.M.Chathram at Tirunelveli, Tamilnadu, India. The eggs were placed at ambient temperature of
25 ±2o c and relative humidity of 70 to 80% in an incubator for hatching. After hatching, the
larvae were reared in the laboratory following the procedure of Krishna swamy method (1978).
Preparation of nanoparticles:
Silver nanoparticles were synthesized by the bio-reduction method using the extracts of
Atrocarpus heterophyllus leaves and silver nitrate solution (0.1M). In the method, the plant
extract containing flavonoids and other phenolic compounds act as reducing as well as

stabilizing agents and a pale red colored colloidal solution was obtained indicating the formation
of silver nanoparticles. Silver nanoparticles exhibit a pale red colour in aqueous solution due to
excitation of surface plasmon vibrations in silver nanoparticles (Sarkar et al., 2010. Schultz et
al., 2000, Rai et al., 2009). The UV – visible absorption spectrum reveals the formation of silver
nanoparticles by the interaction of the extract of Atrocarpus heterophyllus leaves with silver
nitrate by showing surface plasmon absorption maxima at 418nm and from XRD, the mean
particle size is found to be 14.5 nm.
Feeding pattern
After the fourth moult, the fifth instar larvae were divided into 5 batches of 25 worms
each. The first batch was given normal feeding 5 times a day and treated as control. The second,
third, fourth and fifth batches were treated as experimental samples and the larvae reared under
these categories were fed with mulberry leaves fortified with 25%, 50%, 75% and 100% silver
nanoparticles respectively. Treatments were given daily once from I stday of 5th instar till the
onset of spinning. Each treatment was replicated thrice control larvae were also maintained.
Weight measurement
On 2nd to 5th day of supplementation the larvae were randomly selected length and weight
were measured. When the larvae stopped eating and emptied their gut, 10 larvae was randomly
selected anesthetized with ether and dissected to remove the silk gland and washed with Ringer
solution. The fresh weight was estimated gravimetrically. Five days after spinning the cocoon
were harvested cocoon weight, shell weight and pupal weight were measured. After hatching,
adults were allowed to pair after oviposition fecundity and hatchability were found out.
Study reveals the enriched mulberry leaves with additives led to an obvious improvement
in several measures elevating larval weight, length, silk gland weight and cocoon characters and
fecundity. The larval growth rate of silkworm Bombyx mori in the control (First group) and
experimental (2nd to 5th) groups, receiving mulberry leaves socked in various concentration of
Silver nanoparticles (25%, 50%, 75% and 100%) is shown in Table 1,2. The mean larval weight
was plotted against mean larval length and regression line was fitted by methods of least squares
(Table 3). From statistical analysis the correlation coefficient was found to be positive. The

supplementation of silver nanoparticles to silkworm larvae in low concentration (25%) gave best
results regarding weight and length.
The silk gland weight of Bombyx mori larvae fed mulberry leaves supplemented
with silver nanoparticles was studied at different concentrations The results have shown higher
silk gland weight on feeding silkworm larvae with 25% nano particles (table: 1, 2). The mean
larval weight was plotted against silk gland weight and the regression line was fitted by method
of least squares (Table 3) and the correlations coefficient was found to be positive.
The economic features measured (Table 4) initially and after experiment cocoon
parameters were varied in different concentration. Among the four concentrations i.e. 25%, 50%,
75% and 100% of silver nanoparticles treated to 5th instar larvae, three concentrations viz, 25%,
50% and 75% exhibited significant improvement in the parameter studied. In these observations
larvae treated with 25% 50% and 75% silver nanoparticles produced cocoon with high cocoon
weight, shell weight and pupal weight than groups which were given 100% , suggests that silver
nanoparticles which were stimulated silkworm nutrient intake up to 75% than at 100% and
control.
Silver nanoparticles have wide range of application in science, as catalyst (Schultz et al.,
2000), antimicrobial and therapeutics (Raj et al., 2009, Crooks et al., 2001) microelectronics
(Glittin et al., 2000), medical devices and health care products (Lands down et al., 2006) in
textile fabrics, food storage containers and in home appliances (wang et al., 2006; Marambio –
Jones et al., 2010). In the present study it enhanced the growth of silkworm, silk gland weight,
economic features and fecundity when it fed to 5th instar silk worm larvae. The nutritive value of
mulberry leaves depends on various agro climatic factors and any deficiency of nutrients in
leaves affect silk synthesis by the silkworm. Nutritional management directly influences the
quality and quantity of silk production (Hiware, 2006, Vyjayanthi and Subramanian 2002). In the
present study, the larval and cocoon characters significantly increased in some groups. Silver
nanoparticles exhibit certain growth stimulant activity and can be used to increase the silk in
commercial silk worm rearing with reference to sericulture industry (Balasundaram et al., 2012).
Grasserie, a polyorganotrophic disease caused by Bombyx mori nucleopolyhedro virus accounts
for lethal infection to fifth instar silkworm larvae. It was found that nanoparticles induced
morphological transformation of BmNPv polyhedral and could reduce the infectivity of BmNPv

both in cell line and in silkworm larva. Govindaraju et al.,(2010) confirmed the anti-microbial
activity of Solanus torvum mediated silver nanoparticles against pathogenic bacteriae and fungi
of silkworm Bombyx mori such as Pseudomonas aeruginosa, Staphylococcus aureus, Aspergillus
flavus and Aspergillus niger and Linga Rao and Savithramma (2011) recorded antibacterial
activity of nano-based leaf extract of Svensoniahyderabadensis against Aspergillus niger,
Fusarium oxysporum, Curvulari alanata and Rhizopus arrhizus.
Table – 1 Effect of Silver Nanoparticles on the larval weight of Silkworm Bombyx mori
(% change over control is given in parenthesis)
Concentration V instar % change V % V % V %

of (weight overcontrol instar change instar change instar change
Silvernitrate in gram) (weight over (weight over (weight over
nanoparticle on II in control in control in control
day gram) gram) gram)
on III on IV on V
day day day
Control 2.221 ± - 2.699 ± - 2.899 ± - 2.915 ± -
0.124 0.344 0.0075 0.034
25% 2.707 ± -21.88 3.319 ± -22.97 3.452 ± -19.07 3.532 ± -21.16
0.013 0.028 0.012 0.047
50% 2.412 -8.59 3.224 ± -19.45 3.346 ± -15.41 2.470 15.26
±0.006 0.241 0.0132 ± 0.024
75% 2.394 ± -7.78 3.119 ± -15.56 3.031 ± -4.55 2.324 ± 20.27
0.014 0.024 0.0148 0.028
100% 2.129± 4.14 2.616 ± 3.07 2.668 ± 7.99 2.061 ± 29.29
0.004 0.007 0.002 0.0241
Non-significant (F=9.276, 215.71; p>0.05)
Table – 2 Effect of Silver Nano particles on the larval length of Silkworm Bombyx mori
(% change over control is given in parenthesis)
Concentration V % change V % V % V instar %

of Silver instar overcontrol instar change instar change (length change
nanoparticle (length (length over (length over in cm) over
in cm) in cm) control in cm) control on V control
on II on III on IV day
day day day
Control 4.8 ± - 4.9 ± - 5.1 ± - 5.2 ± -
0.002 0.81 0.132 0.242
25% 5.0 ± -4.16 5.2 ± -6.12 5.3 ± -3.92 5.5 ± -5.66
0.004 0.204 0.014 0.013

50% 4.7 ± 2.08 4.8 ± 2.04 5.2 ± -1.96 5.4 ± 1.88
0.1042 0.208 0.164 0.037
75% 4.2 ± 12.5 4.7 ± 4.08 5.0 ± 1.96 5.3 ± 3.77
0.128 0.004 0.137 0.309
100% 4.0 ± 16.6 4.6 ± 6.12 4.8 ± 5.88 4.7 ± 11.32
0.054 0.119 0.124 0.274
Non-significant (F = 9.276, 215.71; p>0.05)
Table - 3 Correlation co-efficient between length and weight relationship and weight and
silk gland weight relationship of V instar (V day) silkworm larvae supplemented with silver
nanoparticles
V instar V instar Weight of

Concentration V instar
(length in (weight in the silk
of Silver (weight in Correlation Correlation
cm) Vth gram) V gland
nanoparticles gram) Vth
day thday (gram)
in % day
Control 2.915 5.3 2.915 0.70
25% 3.532 5.6 3.532 0.84
50% 2.470 5.2 0.94 2.470 0.69 0.96
75% 2.324 5.1 2.324 0.61
100% 2.061 4.7 2.061 0.54
Table 4 Economic features of silkworm larvae fed with silver nanoparticles Supplemented
mulberry leaves (% change over control is given in parenthesis)
Concentration % % %
Cocoon Pupal weight Shell weight
of silver nano Change Change Change
weight (g) (g) (g)
particles in % Over Over Over
Control Control Control
Control 1.142 ± 0.010 - 0.987 ± 0.042 - 0.155 ± 0.062 -
1.497 ± -31.08 -23.10 0.282 ± -81.93
25% 1.215 ± 0.021
0.1005 0.0403
50% 1.767 ± 0.167 -54.72 1.397 ± 0.031 -41.54 0.37 ± 0.04 -138.7
75 2.112 ±0.182 -84.93 1.732 ± 0.094 -75.48 0.523 ± 0.034 -237.41
100 1.37 ± 0.090 -19.96 0.847 ± 0.024 14.18 0.38 ± 0.027 -145.16
Non significant (F =19.00, 199.50; p>0.05)
Table - 5 Effect of silver nanoparticles supplementation on Fecundity and hatchability of

Bombyx mori female
Number of egg laid PRC percent Hatchability

Concentration in %
per mean SD reproduction percentage %
Control 610 ± 10 - 64%
25% 634 ± 12 -3.93 65%

50% 648 ± 14 -6.22 70%
75% 651 ± 20 -6.72 74%
100% 769 ± 23 -26.06 80%
Non – significant (F = 161.45, 161.45; p>0.05)
In silkworms silver nano particles are used for disease treatment, disease prevention,
disease control and health maintenance or growth promotion. Ponrajganesh prabhu et al., (2011)
evaluated that food ingestion and digestibility and growth in larval stages are stimulated by
Silver nanoparticles when it fed to larval stages of silkworm Bombyx mori. In the present study,
it has been observed that silkworm fed by 25% Silver nanoparticles have enhanced the larval
length and weight and silk gland weight. This work is corroborated with Ganesh prabu et al.,
2012; Ponrajganesh prabu et al.,(2011) Thilagavathi et al.,(2013) suggested that this
improvement in larval weight and length related to enhancement of feeding activity and
metabolic pathway in supplemented larvae. Similar findings were also observed in the present
study.
Studies on economic features reveals, the larva fed with 75% silver nanoparticles showed
maximum cocoon weight, shell weight and pupal weight. Regarding fecundity and hatchability,
the Vth instar larvae fed with 100% silver nanoparticles (during their adult stage) laid more eggs
compared to other concentrations (25%, 50%, and 75%). It may be due to delayed expression of
silver nanoparticles during the later stages of development (or) later stages are much receptive to
the stimulatory action of silver nanoparticles and also may be due to its antimicrobial activity,
silkworms are allowed to grow in the disease free environment.
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nanoparticles and their antibacterial

ANTIMICROBIAL ACTIVITY & SELECTIVITY STUDIES OF POLY (O-TOLUIDINE)
ZR (IV) TUNGSTOIODO PHOSPHATE NANOCOMPOSITE ION-EXCHANGER
M.Vijayakumari
Assistant professor, Dept. of Chemistry, Dr.Sivanthi Aditanar College of Engineering,
Tiruchendur, Tamil Nadu, India.
Corresponding Author:vimalavijaya87@gmail.com
ABSTRACT
Nano composite material is growing very rapidly. “Conducting Composites” means
incorporation of the Conducting polymer into a suitable matrix.Nano composites are
combination of two or more nano particles in some suitabletechnique, Organic polymeric part of
the composite provide mechanical and chemical stability where as inorganic part support the ion
– exchange behaviour, thermal stability and also increase the electrical conductivity. Thus, the
synthesis of polymeric/inorganic composites has received a great deal of attention because it
provided new materials with special mechanical, chemical, electrochemical and optical as well as
magnetic properties.Few such excellent ion – exchange materials have been developed and
successfully being used in chromatographic techniques. It was therefore considered to synthesize
such hybrid ion–exchangers with a good ion–exchange capacity, high stability, reproducibility
and selectivity for heavy metal ions, indicating that they are useful in environmental
applications. An organic- inorganic nano composite ion-exchanger, poly(o-toluidine) Zr(IV)
tungstoiodo phosphate was synthesized via sol-gel mixing of poly(o-toluidine) into the matrices
of inorganic precipitate of Zr(IV)tungstoiodophosphate.Theantimicrobial activity and selectivity
of metal ions were also carried out.
KEYWORDS: tungstoiodophosphate, polymer, nano composite
INTODUCTION
Nano composites are a special class of materials originating from suitable combination of
two or more nano particles in some suitabletechniqueOne of the best ways is the formation of
conducting composites. “Conducting Composites” means incorporation of the Conducting

polymer into a suitable matrix. The matrix may be insulating or conducting. Conducting
composites are having attractive properties The conducting polymer based nano composites have
shown great potential in gas sensing[1].One of the composites contains a conducting polymer
[2-3]
(Polypyrrole, Polyanilineetc) and metal oxides . Nano composite contains polymer and
inorganic ion – exchange material prepared by sol – gel method has shown conducting behaviour
[4-5]
and excellent ion – exchange properties with electrochemical application.
Organic polymeric part of the composite provide mechanical and chemical stability
where as inorganic part support the ion – exchange behaviour, thermal stability and also increase
the electrical conductivity. Such a modified composite materials can be applied as
electrochemically switchable ion – exchanger [6-7] for water treatment, especially water softening.
This ion – exchanger can be regenerated without chemical additives or water electrolysis. The
synthesis of hybrid ion-exchangers with controlled functionality and hydrophobicity could open
new avenues for organometallic chemistry, catalysis, organic host-guest chemistry,analytical
chemistry[8-10] ,hydrometallurgy, antibiotic purification and separation of radioactive isotopes,
and find large scale application in water treatment and pollution control .Thus, organic-inorganic
hybrid materials are expected to provide many possibilities as new composite materials that
exhibit very different properties from their original components (organic polymer and inorganic
materials), especially in the case of molecular level hybrids. Thus, the synthesis of
polymeric/inorganic composites has received a great deal of attention because it provided new
materials with special mechanical, chemical, electrochemical and optical as well as magnetic
properties [11-14].
Few such excellent ion – exchange materials have been developed and successfully being
[15-17]
used in chromatographic techniques . It was therefore considered to synthesize such hybrid
ion–exchangers with a good ion–exchange capacity, high stability, reproducibility and selectivity
for heavy metal ions, indicating that they are useful in environmental applications.In the present
work, the selectivity of metal ion studies of organic-inorganic nano composite ion-exchanger by
incorporation of organic polymer ie; poly(o-toluidine) and inorganic ion-exchanger ie;
Zirconium (IV) tungstoiodophosphate was analysed.

Synthesis of organic polymer-poly(o-toluidine)
Poly(o-toluidine)emeraldine salt can easily be synthesised by the oxidative

polymerisation of o-toluidine in aqueous acidic media by using KPS oxidant.The organic
polymer, poly(o-toluidine) was prepared by mixing 0.46g of Potassium perdisulfate (KPS)
(0.25M) to 0.2 M (o-toluidine) in 1M HCl with continuous stirring by a magnetic stirrer for 20
minutes to form an emulsion of POT / Hydrochloric acid; a green – colored precipitate was
obtained and it was kept for 24 hrs at 00C in the refrigerator. The emeraldine salt was washed
with organic solvents (methanol &acetone) to remove oligomers. The POT salt is dried under
vaccum for 24 hrs to obtain green-black powder of POT – Hydrochloric acid emeraldine salt.
Synthesis of inorganic precipitate-Zr(IV)tungstoiodo phosphate
ZTIP prepared by mixing a 0.1M zirconium oxychloride in 4M HCl (at the flow rate at
0.5 ml min-1 )to an aqueous solution of Potassium iodate and 1M ortho phosphoric acid (H 3PO4)
in 1:1:3:1 volume ratios by shaking of the mixture and maintaining the pH = with constant
stirring was done using a magnetic stirrer at room temperature.Now the white-colored gel was
obtained and it was filtered off, washed thoroughly with demineralised water to remove excess
acid and the material was dried in an electric oven at 60ºC.
Preparation of poly(o-toluidine) Zr(IV)tungstoiodo phosphate
The composite ion exchanger was prepared by the sol-gel mixing of poly(o-toluidine), an
organic conducting polymer, into the inorganic precipitate of
zirconium(IV)tungstoiodophosphate. In this process, when the gels of poly(o-toluidine) were
added to the white inorganic precipitate of zirconium (IV) tunstoiodophosphate with a constant
stirring for 7 hrs. The resultant mixture were turned slowly into a greenish black colored
slurries. The resultant greenish black-colored slurries were kept for 24 hrs at room
temperature.Now the poly(o-toluidine) based composite gels were filtered off, washed
thoroughly with DMW to remove excess acid and any adhering trace of KPS. The product was
dried in an air oven.

SELECTIVITY (sorption) STUDIES
The distribution behaviour of metal ions plays an important role in the determination of
selectivity of the material. In certain practical applications, equilibrium is most conveniently
expressed in terms of distribution co-efficient of the counter ions.
The distribution coefficient (Kd values) of various metal ions an poly(o-toluidine Zr(IV)
tungstoiodophosphate was determined by batch method in various solvent system. Various
200mg of the composite cation exchanger in the H+ form were taken in flasks with 20ml of
different metal nitrate solutions Pb(NO3)2, Cu(NO3)2, CO(NO3)2 in the required medium and kept
for 24 hour with continuous shaking. The metal ions in the solution before and after equilibrium
were determined by titrating against standard 0.25M solution of EDTA. The distribution
quantity is given by the ratio of amount of metal ion in the exchanger phase and in the solution
phase. In other word, the distribution coefficient is the measure of a fractional uptake of metal
ions competing for H+ ions from a solution by an ion-exchange material and hence
mathematically can be calculated using formula given as,
m moles of metal ions / g of ion exchanger
Kd (ml g-1) =
m. mole of metal ions / ml of solution
(I-F) V
Kd = ml g-1
F M
where
I - initial amount of metal ion in the aqueous phase (mol dm-3)
F - Final amount of metal ion in aqueous phase (mol dm-3)
V - Volume of the metal ion (ml)
M - amount of cation exchanger(g)
Table- 1 Kd values of some metal ions on poly(o-toluidine) Zr(IV)

tungstoiodophosphate column in different solvent systems
Solvents Pb2+ Cu2+ Co2+

DMW 334 100 30
10% ethanol 244 112 34
10% acetone 367 150 18
1M H2SO4 89 64 42

In order to find out the potentiality of this composite material in the separation of metal ions,
distribution studies for three metal ions were performed in four solvent systems. The distribution
studies showed that Kdvalues varied with the nature and composition of contacting solvents. It
was also observed from the sorption studies (Kdvalues) showed that the composite has a
maximum selectivity towards Pb2+ because lead was highly adsorbed in all solvents, while
remaining metal ions were poorly adsorbed. Pb2+ is a major pollutant in the environment that
can be removed easily by the poly(o-toluidine) Zr(IV)tungstoiodophosphate.
ANTIMICROBIAL ACTIVITY STUDIES

The antimicrobial activity studies of the Poly – o – toluidine and POT/ ZrTIP hybrid ion
exchanger were carried out against two bacteria namely Escherichia coli and proteus vulgaris.
The test solutions of these two samples were prepared in NMP solvent.
A Standardized filter paper disc – agar diffusion procedure, known as the Kirby – Bauer
method, is frequently used to determine the drug susceptibility of micro – organisms isolated
from infectious process. In this procedure, filter paper discs of uniform size are impregnated
with specified concentrations of different antibiotics and then placed on the surface of an agar
plate that has been seeded with the organism to the tested. The medium of choice is Mueller –
Hinton agar, with a PH of 7.2-7.4, which is poured into plates to a uniform depth of 5mm and
refrigerated on solidification. Prior to use, the plates are transferred to an incubator at 370C for
10-20 mins to dry off the moisture that develops on the agar surface. The plates are then heavily
innoculated with a standardized innoculum.
Following incubation the plates are examined for the presence of growth inhibition,
which is indicated by a clear zone surrounding each disc. The susceptibility of an organism to a
drug is determined by the size of this zone.
A measurement of the diameter of the zone of inhibition is millimeters is made and its
size is compared to that contained in a standardized chart. Based on this comparison the test
organism is determined to be resistant, intermediate, or susceptible to the antibiotic.
Materials :
i) Culture:
0.85% saline suspensions of bacteria are adjusted to an O.D. of 0.1 at 600 mμ.

ii) Media :
Per designated student group; seven Mueller – Hinton agar plates.
iii) Antimicrobial – sensitivity discs.
Penicillin – G – 10 Mg; Streptomycin – 10 Mg; tetracycline – 30 Mg; chloramphenicol – 30 Mg;
gentamicin – 10 Mg; vancomycin – 30 Mg; and sulfanilamide – 300Mg.
iv) Equipment
Sensi disc dispensers of forceps, Bunsen burner, sterile cotton swabs, glass ware marking pencil
and millimeters rules.
Procedure.
1) Place agar plates right side up in an incubator heated to 37oC for 10-20 mins with the covers
adjusted so that the plates are slightly opened.
2) Label the covers of each of the plates with the name of the test organism to be inoculated.
3) Using sterile technique, innoculate all agar plates with their respective test organisms.
4) Allow all culture plates to dry for about 5minutes.
5) Using the sensi – disc dispenser over the other surface and pressing the plunger, depositing the
discs simultaneously onto the agar surface.
6) Gently press each disc down with the wooden end of a cotton swab or sterile forceps to ensure
that the discs adhere to the surface of the agar. Do not press the discs into the agar.
7) Incubate all plate cultures in an inverted position for 24-48 hrs at 37oC.The antimicrobial
activity study results are summarized in the Table.8
Zone of inhibition in (mm)
S.No. Organisms Media
Control POT POT/ZrTIP
1. Escherichia coli Muller Histon 18 17 18
2. Proteus vulgaris Agar 20 19 16
From the table, Poly- o- toluidine and POT / ZrTIPnano composite ion –
exchanger showed higher antimicrobial activity against Escherichia coli and proteus vulgaris and
an inhibition zones were formed due to their antimicrobial activity.

In the present study, new and novel organic – inorganic electrically conducting nano
composite ion exchanger was chemically prepared by sol- gel mixing of organic conducting
Polymers like Poly (o- toluidine) into the matrix of inorganic ion – exchangers ( ie) Zirconium
(IV) tungstoiodophosphate.
The mechanism for the formation of conducting Polymeric – inorganic nanocomposite
ion – exchanger.
i) Poly (o- toluidine) gel was prepared by oxidation coupling using K2S2O8 in acidic
medium.
Y Y Y
+ +.
2-
4 NH3 +5S O
28 2 NH NH
+ 12H+ + 10 SO 2-
4
ii) The binding of Poly ( o- toluidine ) into the matrix of Zr (IV) tungstoiodo phosphate
is possible due to ionic interaction between the radical cation of Poly ( o- toluidine)
and anionic group Zr(IV) tungstoido phosphate.
Y Y Y Y
+.
- +.
NH NH NH NH
X + -
X
ZZrTIP C onducting polymer C onducting polymeric- inorganic

ion-exchnager
Where,
Y = CH3 o – Toluidine

The distribution studies for three metal ions were performed in four solvent system. The K d
values shows the maximum selectivity towards Pb2+, because Pb2+ has highly adsorbed in all
solvents while remaining metal ion were poorly adsorbed.
The distribution studies for three metal ions were performed in four solvent system.In the
present work Poly (o – toluidine) Zirconium (IV) tungstoiodo phosphate is newly synthesised
organic- inorganic nanocomposite material developed that possessed all such characteristics.
The Kd values shows the maximum selectivity towards Pb2+, because Pb2+ has highly adsorbed in
all solvents while remaining metal ion were poorly adsorbed and is highly selective for lead, a
hazardous toxic metal in the environment.The material can be used in making Pb (11) ion
selective membrane electrode. Antimicrobial activity study shows the organic Polymer and
nanocomposite ion exchanger are effective against E. Coli &Proteius vulgaris.
CONCLUSIONS
Conducting organic–inorganic nanocomposite ion–exchange materials were synthesised
by sol- gel mixing of organic conducting polymers into the inorganic phosphate. The analytical
importance of the material was deduced from Kd values for different metal ions in different
solvents. The material had high affinity for Pb2+ ion which is a major pollutingmetal in the
environment. The material can be used in making Pb (11) ion selective membrane electrode.and
also antimicrobial activity study shows the organic Polymer and nanocomposite ion exchanger
are effective against E. Coli &Proteius vulgaris.
Acknowledgement
We would like to acknowledge thesupport of Dr.Sivanthiaditanar College of Engineering,
Tiruchendur.
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INTERACTION OF MONOTERPENES OF ESSENTIAL OIL WITH Β-
CYCLODEXTRIN: ANTIMICROBIAL AND ANTIOXIDANT APPLICATION
A. Antony Muthu Prabhu

Department of PG Chemistry, Aditanar College of Arts and Science, Virapandianpatnam,
Tiruchendur 628 216, Tamilnadu, India
Corresponding Author: antonyphdchem@yahoo.com,
ABSTRACT
Essential oils are more complex and comprise the number of volatile and natural bioactive
compounds often used in various food industries and in therapeutic. The major compounds of
essential oil represent a potential anti-oxidant, anti-microbial and anti-fungal activity through
various mechanisms. Essential oils are playing a key role in various biological activities
including food, cosmetic and therapeutic field associated with human health. The interaction of
monoterpenes of essential oil with β-cyclodextrin was investigated theoretically and also
applicable for these inclusion complexes to antimicrobial and antioxidant activity. The
encapsulated essential oil can be used in diverse applications in the food, cosmetic and
agriculture sectors and improvement of antimicrobial, antioxidant activity.
KEYWORDS:Essential oil, β-cyclodextrin, inclusion complex, biological activity
INTRODUCTION
Essential oils are aromatic and volatile liquid naturally present in plant parts such as
flowers, seeds leaves, peel, bark, stems, roots and whole plants [1]. It has been widely used in
various countries as medicine, perfumery, and cosmetic and as food preservatives. Initially it was
used as medicine in nineteenth century due to their aroma and flavor. Till date there are 3000
essential oils were identified and about 300 are used commercially in the flavor and fragrances
market [2]. Basically, essential oils are secondary metabolites and play a key role in plant
defense mechanism hence it has various medicinal properties including antimicrobial activity [3].
Since then, essential oil and their Phyto-constituents have been shown wide range of antibacterial
activity [4], also have reported its antiparasitic [5], antiviral [6] and antifungal activity.

Chemical composition of essential oil
Essential oils are very complex nature it composed of natural mixture of about 20-60
components at quite different concentrations. Basically, two or three major components at fairly
concentration of about 20-70% compared to other constituents which are present in trace
amounts. For example, Origanum compactum have carvacrol (30%) and thymols (27%) are the
major chemical components and linalool is the major components in Coriandrum sativum and
other essential oil composition like α and β a- and b-thuyone (57%) and camphor (24%) of the
Artemisia herba-alba are present in high concentration. These major components are responsible
for the various biological activities of essential oils. Generally, these major components are
classified as two major groups of distinct biosynthetically origin [7]. The mainly it composed of
terpenes and terpenoids and the other of aromatic and aliphatic constituents, all characterized by
low molecular weight. Essential oils are mixture of over 300 different compounds majorly it
consists of volatile compounds with low molecular weight about below 300. The volatile
compounds are majorly categorized into various chemical class such aldehydes, ketones, alcohol,
amines, amides, phenols and mainly terpenes.
Terpenes
The basic structures of terpenes are 5-carbone base units known as isoprene. The
biosynthesis of the terpenes consists of three important step in the first step isopentenyl
diphosphate (IPP) precursor was synthesized and in the second step there is a repetitive addition
of IPPs to form the prenyl-diphosphate precursor of the various classes of terpenes, in the third
step a slight modification of the allylic prenyl-diphosphate by terpene specific synthetases to
form the terpene skeleton and finally, it undergo redox reaction for secondary enzymatic
modification of the skeleton which attributed the functional properties to the different terpenes.
Monoterpenes (C10) and sesquiterpenes (C15) are the major type in addition to that other types
such as hemiterpenes (C5), diterpenes (C20), triterpenes (C30) and tetraterpenes (C40) also exist.

(a) (b) (c) (d) (e)
Figure 1. T he(f)mol ecular ge omet

(g) ry of maj or(h) (j) ellol, (b) α-
(i) es sential oil, (a) Citron
mo noterpenes of
Pinene, (c) Camphor, (d) E-β-Ocimene, (e) Geraniol, (f) Limonene, (g) Linalool, (h) Myrcine, (i)
Nerol and (j) Terpineol.
(a) (b) (c) (d) (e)
Figure (f) (g) etry of ma jor (h)

2. T he molecular geom (i) tial oil, (a) C itrone
mon oterpenes o f essen (j) llol – β-
CD, (b) α-Pinene – β-CD, (c) Camphor – β-CD, (d) E-β-Ocimene – β-CD, (e) Geraniol – β-CD,
(f) Limonene – β-CD, (g) Linalool – β-CD, (h) Myrcine – β-CD, (i) Nerol – β-CD and (j)
Terpineol – β-CD.

β-Cyclodextrin
Cyclodextrins (CDs) are cyclic oligosaccharides consisting of 6, 7, and 8 units of 1,4-
linked glucose units, and are named alpha (α)-, beta (β)-, and gamma (γ)-cyclodextrins,
respectively. These macromolecules, which can be spatially represented as a torus with wide and
narrow openings corresponding to secondary and primary hydroxyl groups respectively, can
encapsulate a large variety of compounds due to the hydrophobic character of their internal
cavity [8]. Although the depth of the cavities for the three CDs is the same (~0.78 nm), their
cavity diameters are ~0.57, 0.78, and 0.95 nm respectively. Due to the unique chemical structure
of CDs, the inner side of the cavity is hydrophobic and the outer side is hydrophilic. The
hydrophobic nature of the CD cavities facilitates the ability of CDs to act as host for both
nonpolar and polar guests, which include small molecules as well as polymers [9]. Once the
inclusion compound is formed, the stability of the guest molecules increases due to the binding
forces (van der Waals attractions, hydrogen bonding, hydrophobic interactions, etc.) between the
host (CDs) and guest molecules [10]. The CDs also have several advantages in other areas, such
as the food, cosmetics industries, and agro chemistry [11], especially owing to their capacity to
protect the guest molecules against oxidation, light-induced reaction, and loss by evaporation.
Additionally, they usually enhance the aqueous solubility of poorly soluble or even insoluble
compounds such as organic and drug molecules [12].
Cyclodextrins are crystalline compounds comprised of torus-like macro-rings of
glucopyranose units, β-cyclodextrin (β-CD), for example, having seven glucose units. Various
3D structures result from joining the single molecules that differ in their chemical and physical
properties (size, functionality or solubility), many of which are very interesting from a
pharmaceutical point of view. Their particular molecular structure means that, these molecules
have a chemical tendency to form inclusion complexes with other ionic species. Such complexes
are of great interest for practical application and in fundamental research, because, when suitably
manipulated, they provide information on non-covalent intermolecular forces.
Antimicrobial activity
Essential oils and their constituents play a key role in antimicrobial activity. Due to the
hydrophobicity nature essential oils are significantly move across the lipids of the cell membrane

of bacteria and disrupting the cell wall structure and making them to more permeable [13]. This
membrane permeability change leads the leakage of ions and other cellular materials [14]. The
greater loss of cellular content and ions lead the cell death. Essential oils show both single and
multiple target activity. The major components of essential oil such as such as carvacrol,
eugenol, and thymol, have important antibacterial activities which disrupting the cytoplasmic
membrane, the driving force of protons, electron flow, activity transport and coagulation of cell
contents. Generally essential oils are more active in gram-positive bacteria compared to gram
negative bacteria due to the presence of peptidoglycan layer lies outside of outer membrane.
Antioxidant activity
Several studies have been reported that essential oils have prominent antioxidant activity.
The antioxidant potential of essential oils is mainly depending on its chemical composition.
Mostly the essential oils are having oxygenated monoterpenes such as alcohols (Achillea
filipendulina), aldehydes (Galagania fragrantissima), ketones (Anethum graveolens, Artemisia
rutifolia, Hyssopus seravschanicus, Mentha longifolia, and Ziziphora clinopodioides), and esters
(Salvia sclarea) as major chemical composition. Thymol and carvacrol, are important
monoterpenes present in several types of essential extracted from Origanum tyttanthum, Mentha
longifolia and Thymus serpyllus are play a key role in the antioxidant properties of several other
essential oils [15]. Antioxidant activity of essential oils was measured by different kinds of test
like 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical inhibition and inhibition of lipid peroxidation
by measuring the index of thiobarbituric acid reactive substances (TBARs).
CONCLUSION
In the present study, the inclusion of monoterpenes of essential oil with β-cyclodextrin
was achieved. These studies have addressed the physical and chemical conditions of the
encapsulation reactions, employed several types of essential oils and characterized the
microcapsules as to their ability to release encapsulated active principles. The essential oils
studies with cyclodextrin encapsulation processes have been highly varied. These inclusion
complexes have been studied with antimicrobial and antioxidant capacities.

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GREEN APPROACH TO CORROSION INHIBITION OF MILD STEEL
USING EMBLICA OFFICINALIS (AMLA SEED) IN ACID MEDIUM
R. Rajkumar1* and C. Kavitha2

1
Assistant Professor, PG Department of Chemistry, Aditanar College of Arts and Science,
Tiruchendur - 628 216, Tamilnadu, India
2
Assistant Professor, Department of Chemistry and Research centre, Aditanar College of Arts
and Science, Tiruchendur - 628 216, Tamilnadu, India
*Corresponding Author: rajkumarr96@ymail.com
ABSTRACT
The inhibitive performance of alcoholic extract of Emblica Officinalis (Amla seed) towards the
corrosion of Mild steel in 1N Hydrochloric acid medium has been studied by weight loss
measurement with various periods of contact and temperature. The present study revealed that
the percentage of inhibition efficiency is enhanced with the increase of inhibitor concentration
and decreased with the rise in immersion time. The temperature studies reflect that the
adsorption of inhibitor on metal surface takes place via physisorption. The calculated values of
∆Gads suggested that the adsorption may be spontaneous process.It was found that the adsorption
of Amla inhibitor follows Langmuir adsorption isotherm. The results obtained showed that the
extract of Emblica Officinalis (Amla seed) could serve as an effective inhibitor of the corrosion
of mild steel in hydrochloric acid media.
KEYWORDS: Mild steel, Corrosion inhibition, Amla seed, Weight loss, Adsorption isotherm
INTRODUCTION:
Acid solutions are commonly used for removal of undesirable scale and rust in metal
finishing industries, cleaning of boilers, and heat exchangers. Among these, hydrochloric acid is
one of the most widely used agents in the process of acid pickling. Use of inhibitors is one of the
most practical methods for protection against corrosion especially in acid solutions to prevent
unexpected metal dissolution and acid consumption [1]. A mild steel corrosion phenomenon has
become important particularly in acidic media because of the increased industrial applications of

acid solutions [2]. The known hazardous effect of most synthetic corrosion inhibitors is the
motivation for the use of some natural products. The use of chemical inhibitors has been limited
because of the environmental threat, recently, due to environmental regulations. Plant extracts
have become important because they are environmentally acceptable, inexpensive, readily
available and renewable sources of materials, and ecologically acceptable [3]. Plant products are
organic in nature and some of the constituents including tannins, organic and amino acids,
alkaloids, and pigments are known to exhibit inhibiting action. Moreover, they can be extracted
by simple procedures with low cost [4, 5].
Seeds are of great concern for corrosion inhibition studies. Tobacco (Nicotiana), black
pepper (Piper nigrum), castor seeds oil (Ricinus communis), acacia gum, and lignin can be good
inhibitors for steel in acid medium. Papaya, Poinciana pulcherrima, Fedegoso (Cassia
occidentalis), and Datura (Datura stramonmium) seeds are efficient corrosion inhibitors for steel
[6–8]. In our present investigation consists of the inhibitive and adsorption properties of Amla
seed extract on the corrosion of Mild steel in 1N Hydrochloric acid have been observed at
various concentrations of inhibitor using mass loss measurements at different time duration and
temperature. However, the Amla seed extract have never been used as a corrosion inhibitor in
acidic medium.
Specimen preparation:
Mild steel of thickness 1.4mm was obtained locally and was mechanically cut into
coupons of 5×1×0.14cm. A small hole was drilled at one end of the coupons for easy hooking.
The coupons were degreasedin absolute ethanol, dried in acetone and stored in a desiccator.
Preparation of Amla Seed Extract:
The amla seed were collected, shade dried and powdered. The selected materials are
dried in shade so as to enrich the active principles in them, by reducing their moisture content.
The stock solution of the inhibitor materials was prepared by refluxing 5g dry powder with 100
ml of 1N Hydrochloric acid for 3 hours. The refluxed solution was allowed to stand overnight
and filtered through ordinary filter paper. The residue was repeatedly washed with small amounts
of 1N HCl and the filtrate made up to 100ml. From this stock solution, different concentrations

of inhibitor solutions ranging from 100 to 500 ppm were diluted. The chemical used were of
analytical grade and double distilled water for their preparation.
Weight loss measurements:
In mass loss measurement, mild steel specimen is immersed exactly in 50ml of the test
solution in the presence and absence of the inhibitor. The specimens were withdrawn from the
test solutions after an hour at the temperature range of 303K to 333K and after 24 to 120hrs at
room temperature. From this observed data, the corrosion rate (mmpy), percentage inhibition
efficiency (%I.E) and surface coverage (θ) was calculated using the following formula
87.6 X w
Corrosion Rate (mmpy) = --------- (1)
DAT
Where mmpy = millimeter per year, w = weight loss (g), D = Density (g/cm3), A = Area of
specimen (cm2), T = time in hours
W1  W2
%I.E. = X 100 --------------- (2)
W1
W1  W2
= ---------------- (3)
W1
Where W1 and W2 are the weight loss in the absence and presence of the bio-inhibitor
respectively.
(i). Effect of Time:
The corrosion parameters of mild steel containing various concentration of AS extract
with different exposure time in 1N HCl. It reveals that the loss of mass increased (0.1236g to
0.3114g) with increase of exposure time (24 hours to 120 hours) in the absence of inhibitor
concentration. But in the presence of AS extract, the corrosion rate was significantly reduced
from 0.0114 to 0.0032 mmpy for 24 hours and 0.0057 to 0.0019 mmpy for 120 hours
respectively. The maximum inhibition efficiency of AS extract was achieved at 500ppm of
inhibitor concentration and the I.E (%) is 73.30%. The values of AS extract are shown in the
figure 1.
This was mainly due to the blocking effect of surface by film formation which reduces
the corrosion rate by the attack of acid environment. This achievement exhibited by the

heteroatoms likes nitrogen, oxygen and olefinic bonds present in the inhibitor. The observed
surface coverage were almost greater at 500 ppm was mainly due to the formation of insoluble
stable complex between the ion in the metal surface and the heteroatom present in the phyto
constituent of the extract.
Figure 1: Variation of Inhibition efficiency with concentration of AS extract on Mild

steel in different exposure time
(ii). Effect of Temperature:
Effect of Temperature on the inhibitory action of the inhibitor was determined by weight
loss method at various concentrations of AS extract at 303K to 333K and the plots are shown in
the figure 2. From the data revealed that the temperature increased from 303K to 333K, the
corrosion rate increase while the I.E (%) and surface coverage (θ) decrease. This suggests that
some desorption of some of the adsorbed inhibitor from the metal surface at higher temperature.
This gives a clue that the mechanism of adsorption of the inhibitor may be mainly due to
physisorption, because the physisorption which is due to weak vander waal’s force disappears at
elevated temperature. Thus as the temperature increases, the number of adsorbed molecules
decreases, leading to a decrease in the inhibition efficiency.

Figure 2: Variation of Inhibition Efficiency with concentration of AS extract on Mild
steel in different temperature
Adsorption Isotherm
The values of adsorption parameters deduced from Langumir, Tempkin and Freundlich
isotherms are recorded in table 1. From the tables, the degree of linearity (R 2) was also closed to
unity indicating strong adherence of the adsorption of AS extract on the surface of the mild steel.
The equilibrium constant of adsorption of AS extract on the surface of the mild steel are related
to the free energy of adsorption (∆G°ads) according to equation,
∆G°ads = -2.303 RT log (55.5 k)
Where R → Gas Constant, T → Temperature, K → Equilibrium constant of adsorption, 55.5 →
molar heat of adsorption of water. Values of K obtained from intercept of Langmuir, Tempkin
and Freundlich isotherm were used to compute for ∆Gads according to equation and the result in
table 1. From the result ∆Gads values were found to be negative and less than the threshold values
of -40KJmol-1 required for chemical adsorption hence the adsorption of AS extract on the surface
of mild steel is spontaneous and follows physical adsorption mechanism [9].

Table 1: Langmuir, Tempkin and Freundlich adsorption parameters for the adsorption of
AS extract on Mild steel in 1N HCl
Adsorption Temperature ∆Gads
Slope Kads R2
Isotherms (K) (KJmol-1)
303 1.1451 76.111 0.9998 -21.035
313 1.2233 93.539 0.9996 -22.265
Langmuir
323 1.1646 147.253 0.9997 -24.196
333 1.1893 174.153 0.9990 -25.409
303 0.1927 0.3591 0.9940 -07.539
313 0.2300 0.3596 0.9969 -07.791
Tempkin
323 0.2463 0.3547 0.9969 -08.003
333 0.2205 0.3289 0.9969 -08.042
303 1.2016 0.2577 0.9964 -06.703
313 1.2517 0.2234 0.9969 -06.552
Freundlich
323 1.2292 0.2239 0.9969 -06.768
333 1.1476 0.2482 0.9969 -07.262

Figure 3: Langmuir Adsorption Isotherm of AS extract on Mild steel in 1N HCl
Figure 4: Tempkin Adsorption Isotherm of AS extract on Mild steel in 1N HCl
Figure 5: Freundlich Adsorption Isotherm of AS extract on Mild steel in 1N HCl
CONCLUSION
Amla seed extract has shown excellent inhibition performance for mild steel in 1N HCl
solution. The corrosion rate of mild steel increases with increasing the concentration of HCl and
the inhibition efficiency increased with the increase of inhibitor concentration of AS extract.
The maximum inhibition efficiency was achieved at 89.23%. The inhibition efficiency gradually
decreased with increasing the immersion time (i.e.) 73.30% to 68.59% for 24hrs and 120hrs
respectively. Also the inhibition efficiency of AS extract decreases as the temperature increases,
which is due to the destability of the formed protective film on metal surface. The adsorption of
AS extract on the surface of the mild steel followed Langmuir adsorption isotherms.

ACKNOWLEDGEMENT
The authors sincerely thanks to the management of Aditanar College of Arts and Science,
Tiruchendur, for providing all the lab facilities.
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SEAWEED EXTRACT OF RED ALGAE AS ORGANIC FERTILIZER AND
THEIR IMPACT ON GROWTH PARAMETERS, BIOMASS
PRODUCTIVITY AND ANTIOXIDANT OF AMARANTHUS TRISTIS L.
T. Kumareswari1, *, S. Maria Victorial Rani2,

1
Department of Botany, Kamaraj College, Thoothukudi, Tamil Nadu, India
2
Department of Botany, St. Mary’s College (Autonomous), Thoothukudi, Tamil Nadu, India,
Affiliated to Manonmaniam Sundaranar University, Abishekapatti, Tirunelveli – 627012, Tamil
Nadu, India.
*Corresponding Author: jkumareswari@yahoo.com
ABSTRACT
Seaweed extracts are used as organic fertilizer in agriculture and horticulture particularly to
increase plant growth and productivity. In this study, we investigated the effect of seaweed
extracts (1% SWEs) made from red algae such as Gracilaria verrucosa, Hypnea musciformis and
Kappaphycus alvaerezii, on growth parameters, biomass productivity and antioxidant of green
leafy vegetable, Amaranthus tristis L. Soil application of extract of Kappaphycus alvaerezii
increased the shoot length (96.7% ), stem circumference (97.2%), root length (94.2%) more than
control. SWE of K. alvaerezii and H. musciformis showed better impact on fresh and dry
biomass accumulation. Extracts of H. musciformis increased total phenol (22.8%), total
flavonoid (18.5%), vitamin C (24%) and carotenoid (34.4%) in relation to control. However,
SWEs of all the seaweeds enhanced antioxidants at different degrees with respect to control. The
results of the present study showed that seaweed extract of three seaweeds enhanced well the
growth, biomass and antioxidant of Amaranthus tristis L.
KEY WORDS: seaweed extracts, organic fertilizer, soil application, biomass accumulation,
antioxidant
INTRODUCTION
Green leafy vegetables are important sources of minerals, fibre and vitamins, which
provides essential nutrients for the human health. Increased consumption of vegetables

significantly reduces the incidence of chronic diseases, such as cancer, cardiovascular diseases
and other aging-related pathologies (Prakash et al., 2012). About 9,000 macro algae species are
classified into three main groups depending on the pigmentation including brown, green and red
algae. Seaweeds among the algae are used in the agriculture (Hong etal., 2007). They are bio-
degradable, non-toxic, non-polluting and non-hazardous to human, animals and birds
(Dhargalkar and Pereira, 2005). Seaweeds are considered to be a rich source of antioxidants
(Indu and Sreenivasan, 2013). The presence of antioxidant substances in seaweeds is found to be
an endogenous defense mechanism as a protection against oxidative stress due to extreme
environmental conditions (Aguilera et al., 2002). The antioxidant activity of red seaweed
extracts correlated with their polyphenol content (Duan etal., 2006). Polyphenols form a large
group of phytochemicals with excellent antioxidant properties and play an important role as free
radical scavengers required in the maintenance of “redox homeostasis” responsible for various
degenerative diseases (Prakash et al., 2012). At present, wide spread requirement for
environment friendly agriculture for the production of quality and healthy food to nourish the
increasing population is in high demand feasibility and efficacy of organic sources, not only
renovating soil productivity but also increasing crop productivity. In this scenario, seaweeds are
considered as a potential element which is available in plenty, easily accessible. Seaweed
extracts can be manufactured in liquid or powder form and used as fertilizers for all type of
crops, grasses and trees (Shruthi Nagaral et al., 2018). Soil application of seaweed can rectify
micronutrient deficiency and remarkably increase soil porosity, structure and water holding
capacity, soil organic matter. They bring about aggregation of soil particles which are
responsible for crumb structure associated with soil aeration (Ouedraogo et al., 2001; Zodape et
al., 2010). This study aims to evaluate the effect of soil application of extract of Gracilaria
verrucosa,(Huds.)Papenfuss,Hypnea musciformis (Wulfen)J.V.Lamouroux and Kappaphycus
alvaerezii (Dody) Dody ex P.C.Silva)on growth, biomass and antioxidant parameters of green
leafy vegetable, Amaranthus tristis L.
Collection of seaweed
Gracilaria verrucosa,(Huds.)Papenfuss,Hypnea musciformis (Wulfen)J.V.Lamouroux
and Kappaphycus alvaerezii (Dody) Dody ex P.C.Silva) were collected during low tide, at Hare

Island, Thoothukudi from November 2016 to February 2017. The sample was washed thoroughly
with seawater followed by fresh water to remove sand particles and macroscopic epiphytes. After
draining, the seaweed was shade-dried, powdered, sieved and used for the preparation of
seaweed concentrate.
Preparation of seaweed extract for soil application
Seaweed extract (SWE) was prepared by adopting the standard method (Rama Rao,
1990) with certain modifications. About 20g dried seaweed powder with 200ml distilled water
was heated to 60oC and maintained at the temperature for 24 hr in a hot air oven. The extract was
filtered and then centrifuged at 10000 rpm to remove suspended impurities. The filtrate was
stored in air tight bottles at 4oC (100% seaweed concentrate) for further use.
Experimental design
A pot culture experiment was conducted during February to April 2017 at Plant Research
Centre, St. Mary’s College Campus, Thoothukudi, Tamil Nadu, India. The pots were filled with
3kg of garden soil. 50 seeds of A. tristis were sown in each pot. After the emergence of
seedlings, they were thinned to ten plants per pot and allowed to grow for a period of 30 days.
Weeding and watering were done at regular intervals throughout the experimental period. 1%
SWE was applied in soil (along with 100ml of distilled water in the ratio of 1: 10) after
expansion of first leaf and was continued for twenty days. Enough replicates were maintained.
Analysis
Chemicals of analytical grade were used for all the analyses. Plants from each treatment
were randomly drawn to study the growth and antioxidant parameters. Growth contributing
parameters such as shoot and root length, number of leaves/plant, leaf area, above ground and
below ground biomass were measured. Root length and shoot length were measured using ruler
and recorded in centimeter. Leaf area was measured by using graph paper method as described
by Santra et al., (1989). Vitamin C (Baker and Frank, 1968), total phenolic content (Duan et al.,
2006), total flavonoids (Zhishen et al., 1999) were also recorded. Leaves harvested from 30 days
old plants were used for all analyses.
Data collected in this study was analysed by using Microsoft Excel 2007. One way
ANOVA was used to compare differences in the means of vitamin C, total phenolic content, and

total flavonoids of control and treated plants. A significant difference was considered at the level
of p<0.05.
Table 1. Effect of soil application of SWE on growth parameters of Amaranthus tristis L.
Treatments Stem
Shoot length Root length Leaf area Number of
(SWE) circumference
(cm) (cm) (cm2) leaves/plant
(mm)
Control 5.25±0.77 5.19±0.55 1.82±0.02 3.75±0.17 4.4±0.39
G.verrucosa 9.39±0.69* 8.45±0.77* 2.98±0.05* 5.45±0.48* 7.0±0.82*
H.musciformis 10.55±0.79* 9.72±0.82* 3.15±0.01* 7.50±0.79* 7.5±0.55*
K.alvaerezii 12.25±0.81* 10.08±0.95* 3.59±0.03* 6.75±0.56* 7.2±0.42*
Values are the mean of ten replicates ± standard deviation. All the variables were recorded on 30
days old plants. Control= Plants were irrigated with water. SWE= Seaweed extract formulated
from G. verrucosa, H. musciformis and K. alvaerezii (1%) were applied as soil application. Note:
*significant at 5% level of probability.
Table 2.Effect of soil application of SWE on biomass productivity of Amaranthus tristis L.

Treatments Above ground Below ground Below ground
Above ground dry
(SWE) fresh fresh dry
biomass (g)
biomass (g) biomass (g) biomass (g)
Control 0.745±0.075 0.107±0.001 0.102±0.005 0.020±0.002
G.verrucosa 2.036±0.179* 0.202±0.021* 0.215±0.023* 0.031±0.003*
H.musciformis 2.068±0.324* 0.294±0.014* 0.381±0.025* 0.070±0.001*
K.alvaerezii 2.075±0.107* 0.301±0.011* 0.388±0.017* 0.076±0.002*
Values are the mean of ten replicates ±standard deviation. All the variables were recorded on 30
days old plants. Control= Plants were irrigated with water. SWE= Seaweed extract formulated
from G. verrucosa, H. musciformis and K. alvaerezii (1%) were applied as soil
application . Note: * significant at 5% level of probability
Table 3. Effect of soil application of SWE on antioxidant in leaves of Amaranthus tristis L.

Treatments Total phenolic
Total flavonoid Vitamin C
(SWE) content Carotenoid
(mg QEs g-1 (mg AAEs g-1
(mg GAEs g-1 (mg g-1 FW)
FW) FW)
FW)
Control 2.58±0.72 35.56±0.38 3.84±0.27 51.71±0.39
G.verrucosa 3.02±0.27* 38.10±0.52* 4.55±0.40* 58.81±0.75*
H.musciformis 3.17±0.39* 42.15±0.75* 4.78±0.26* 69.50±1.16*
K.alvaerezii 3.15±0.48* 41.12±0.36* 4.47±0.47* 60.29±0.49*

Values are the mean of three replicates ±standard deviation. Control=Plants were irrigated with
water. SWE= Seaweed extract formulated from G. verrucosa, H. musciformis and Kappaphycus
alvaerezii (1%) were applied as soil application. GAEs = Gallic Acid Equivalents. QEs=
Quercetin acid Equivalents. AAEs=Ascorbic Acid Equivalents. *significant at 5% level of
probability.
Effect of SWE on growth parameters

Among the three seaweeds, extract of K. alvaerezii significantly increased the shoot
length and was 96.7% more than the control. Extracts of H. musciformis (66%), G. verrucosa
(60.9%) also notably enhanced the shoot length. Moreover, all the seaweeds exerted highly
significant positive impact on shoot growth. Similar results were reported by Abou El-Yazied
and Mady, (2012), Ganapathy selvam et al. (2013). Extracts of K. alvaerezii and H. musciformis
boosted the radial growth of stem by 97.2% and 73% respectively in relation to control.Seaweed
concentrates contain growth promoting hormones like IAA and IBA and cytokinins (Khan et al.,
2009) that possibly have stimulated cambial activity, cell division and cell elongation, resulting
into increased stem length and diameter.The stem of A. tristis is consumed as vegetable like
other vegetables by most of the vegetarian in Tamil Nadu and increase in diameter further add up
the consumable value. Extract ofK. alvaerezii significantly boosted the root growth by 94.2%
respectively in correspondence with control. However, seaweed extract of H. musciformis (85%)
and G. verrucosa (62%) also promoted the root growth moderately in relation to control. There
were numerous reports about the role of SWE on growth of plants. SWE application induced
resistance and improved growth and yield responses under different conditions (Beckett and van
Standen 1989, Erulan et al., 2009). Extract of H. musciformis promoted the leaf formation
comparatively higher than all other SWEs. This is in accordance with earlier reports (Kumari
et al., 2011; Anisimov et al., 2013).
Effect of SWE on biomass productivity
Young tender leaves of A. tristis are more popular as green leafy vegetable and
production of more leaf biomass in a cost effective environmentally healthy means adds to food
security. Soil application of SWE influenced more in promoting biomass productivity. SWE of
K. alvaerezii and H. musciformis as soil application exerted higher positive impact on fresh and

dry biomass accumulation (Table 2). The result of the present study is in agreement with others
(Kumar and Sahoo, 2011; El-Din, 2015)).Liquid seaweed application in soil results in
improvement of plant growth and soil texture. Improved soil structure is invariably associated
with better aeration enhanced nitrogen fixation of soil micro organisms and raised proliferation
of soil organism. Capillary action is also increased and as a result, root system of plant is
stimulated into further growth. The SWE application appears to bring about mobilization of
certain trace elements in soil where these tend to be in unavailable form (Zodape, 2001).
Effect of SWE on antioxidants
Soil application of H. musciformis increased total phenol (22.8%), total flavonoid
(18.5%), vitamin C (24%) and carotenoid (34.4%) in relation to control. However, SWEs of all
the seaweeds enhanced antioxidants at different degrees with respect to control. Flavonoids and
phenolic acids, the largest class of plant phenolics, are biosynthetically derived from the acetate
and shikimate pathways and from phenyl alanine or tyrosine (Dewick, 2009). These
phytochemicals were found to have excellent antioxidant activity both in vitro and in vivo
investigations. Moreover, they are known to interact with other physiological antioxidants like
ascorbate, tocopherol and synergetically amplify their biological effects (Michalak, 2006).
Furthermore vitamin C, vitamin E, flavonoids and phenolic compounds have been reported to
exert pro-oxidant properties at high concentrations (Marcio Carocho and Isabel Ferreira, 2013).
In the present work, SWE stimulated the production of various antioxidants investigated,
however to the level below their proxidant concentrations. So, seaweed grown A. tristis could be
effectively utilized as green leafy vegetable without side effects.Plant derived antioxidants, such
as flavonoids, phenolic compounds, carotenoids, chlorophyll, vitamin C have multiple biological
effects, as they combat oxidative stress in the body by maintaining a balance between oxidants
and antioxidants (Rochfort and Panozzo, 2007). Therefore it is pertinent to conclude that
seaweed grown vegetables would serve as an alternative resource to alleviate malnutrition and to
add health benefits in the current scenario.
CONCLUSION
From this study, it is concluded that root growth and shoot growth, biomass and
antioxidants are stimulated by seaweed application, however with species specific differences.
Also, soil application of these SWEs is labour free and more profitable in organic agriculture.

ACKNOWLEDGEMENT
The author is grateful to Dr. S. Maria Victorial Rani, Former HOD of Botany and Dean
of Research, St. Mary’s College (Autonomous), and Thoothukudi for giving valuable
suggestions and constant support to complete this research work.
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SOCIO-ECONOMIC AND DEMOGRAPHIC CORRELATES AND RATE OF
CONSANGUINITY IN CERTAIN COMMUNITIES OF KANYAKUMARI DISTRICT
K. Hemarana1*, Jayashree KV2 and Elango T3

1
Research Scholar [Reg.No.8173], Centre for Marine Science and Technology,
Rajakkamangalam, ManonmaniamSundaranar University, Abhishekapatti, Tirunelveli, Tamil
Nadu, India – 627 012
2
Associate Professor, Sree Ayyappa College for Women, Nagercoil, Manonmaniam Sundaranar
University, Abishekapatti, Tirunelveli – 627 012
3
Professor, Amrita Sai Institute of Science and Technology, Paritala, Andhra Pradesh – 521180
*Corresponding Author: mhemarana@gmail.com
ABSTRACT
Consanguineous unions are blood related marriages whose rate varies by region and religion
influenced by so many socio-economic and demographic factors. Education, occupational status,
family income, marital age and marital year were some of the socio-economic and demographic
factors chosen to study their influence over consanguinity among ten communities in
Kanyakumari district.Inverse relationship of high education and low rate of consanguinity was
noted among Pallar, Paravan, Sambavar (both sexes) and wives of Chakkalar and Kanikkar. But
the opposite of low education and high rate was noticed among the husbands of Sambavar,
Chetty, Kattunayakan and wives of Bharatar, Sambavar, Chetty, and Kanikkar. Highly
significant association was found between consanguinity and occupation among Pallars,
Thandans and Kattunayakans (wives) and Bharatars (husbands) (1%). No clear association
between consanguinity and family income could be predicted except among Sambavars,
Bharatars, Chakkalars and Kammalars, where Sambavars showed negative association and the
rest positive association. Highly significant association between two was observed among
Kanikkars (1%) and less among Chakkalars and Kammalars (5%). It was interesting to note that
women (Pallar, 66.2%, Sambavar, 46%, Chetty, 62.3% and Kattunayakan, 76.5%) and men
(Pallar, 86.7% and Bharatar, 100%) who married below 20 years were more consanguineous.

The percentage of consanguinity was observed to be higher for marriages which were performed
before 1950 (100% among Bharatars, Thandans, Chakkalars and Chetty-76.9%), 1951-1960
(Pallars, kammalars and Kattunayakans- 74.5, 77.1 and 100% respectively) and 1961-1970
(Paravan and Kanikkar, 91.7 and 58.5 respectively. The study shows the influence of education,
occupation, family income and marital age in the rate of consanguineous unions. Alsodecline in
the rate of consanguineous unions was found since 1950.
KEY WORDS: consanguineous, factors, socio-economic, demographic, significance
INTRODUCTION
The rate of consanguinity varies by region, religion etc., from time to time. It is found
more in Muslim countries especially in Middle East. It records 70% in Pakistan (Ullahet al.,
2017) and 23 to 78% in Iran (Shavaziet al., 2008).Decline in the rate of consanguinity has been
recorded in Western world, Europe, USA, Lebanon, Morocco, Saudi Arabia, Kuwait, Israel,
Arab, Jordan and Palestine territories (Khlat, 1985, Lamdouar, 1994, Al-Abdul Kareem and
Ballal, 1998, Radovanovicet al., 1999, Jaberet al., 2000, Zlotogoraet al., 2002). Studies
underlying the prevalence of consanguinity throughout the world highlighted the influence of
certain factors like religion or civil laws on its rate. Research works on the factors which
influence consanguinity were carried out world-wide including India. Many studies indicated
social status in terms of educational level had strong influence on the rate of consanguinity.
Negative correlation between the rate of consanguinity and literary status i.e., lower the level of
literacy, higher the rate of consanguinity and vice-versa has been observed by Hussain and
Bittles (1998) and Mathew et al., 2006, whereas strongly positive association was observed
among immigrant Tamil Brahmin group residing in the Palakkad district of Kerala
(Jyothilekshmi, 2015). Similar to education, occupational status also has controlling effect on the
rate of consanguinity.
Positive association between consanguinity and occupational status was reported in the
Muslim groups of different countries (Bittles and Hussain, 2000) and many forward and
backward communities of Kerala (Mathew and Jyothilekshmi, 2017) whereas highly significant
negative correlation with consanguinity was found in many tribal groups like Mudugars, Irulars,
Karuvazhipulaya, Kurumbapulaya and Muthuvans. Family income is also an important factor in

influencing consanguineous marriages which was found to be negative among many scheduled
caste and most tribal communities of Kerala state (Mathew and Jyothilekshmi, 2017).
Descending trend of consanguinity was evident from older to younger generation among many
Scheduled castes and tribal groups in Brazil, Japan etc (Imaizumi, 1986, Krishnamoorthy and
Audinarayana, 2001, Assaf and Khawaja, 2009). Likewise, fairly high rates of consanguinity was
constructed among low age at marriage groups in most of the Middle East and Asian countries
including South India (Hussain and Bittles, 2004, Mathew et al., 2006, Islam, 2018, Mathew and
Jyothilekshmi, 2017). Kanyakumari is the Southern-most tip of Tamil Nadu and India where ten
communities were chosen to study the socioeconomic and demographic influence on
consanguinity.
The data regarding socio-economic and demographic factors of consanguinity was
collected among ten communities chosen from Kanyakumari district by direct door to door visit.
The ten communities include Pallar, Bharatar, Paravan, Thandan and Sambavars from SC,
Chetty, Chakkalar and Kammalar from BC and Kanikkar and Kattunayakan from ST category.
Totally 6922 data was collected. The three socio-economic factors studied include education,
occupational status and family income and demographic factors include year of birth, year of
marriage and spousal age at marriage of spouses. The educational status was classified in to
three, low (illiterate and up to primary), middle (secondary, higher secondary and technical) and
high (university level), occupational status in to three, un-employed, low (labourers, skilled
workers, marginal farmers and class 1V Government employees), middle (school teachers, para
professionals and class 11 and 111 Government employees) and high (executives, professionals,
landlords, businessman and class 1 Government employees) and family income in to low (below
250000), lower middle (25 to 75000), upper middle (75 to 2 lakhs) and high (above 2 lakhs).
Marital age of the spouses were categorized in to six classes such as age below 20, 20-24, 25-29,
30-34, 35-39 and 40 and above and marital year such as before 1950, 1950-59, 1960-69, 1970-
79, 1980-89, 1990-99, 2000-2009 and 2010 and above. Chi-square analysis was carried out to
study the significance between socio-economic factors and consanguinity.

RESULTS
Frequency of consanguinity by the level of educational status of both spouses is
furnished in Table 1.From the table it is evident that consanguineous marriage was contracted by
spouses in all the three categories but with varying percentage. No clear-cut relationship was
observed in the population selected for study between educational level and consanguinity
among different castes. An inverse relationship of high education and low rate was noted among
some castes like Pallar, Paravan, Sambavar (both sexes) and wives of Chakkalar and Kanikkar.
But the opposite of low education and high rate was noticed among the husbands of Sambavar,
Chetty, Kattunayakan and wives of Bharatar, Sambavar, Chetty, and Kanikkar. The
consanguinity rate among husbands of Pallar, Paravan, and Kammalar and wives of Pallar,
Paravan, Chakkalar and Kammalar was noticed to be high among middle category. It was
observed that among husbands and wives of Thandan the consanguinity rate showed a positive
correlation ie. Highest rate of consanguinity among highly educated category. Highly significant
association was observed among Kammalars and less significance among Paravans and wives of
Thandans.
Table 1 Frequency of consanguinity by the level of education for different castes
% of consanguinity - Education
Husband Wife
Caste
Low Middle High Low Middle High
Pallar 52.9 60.8 29.3 56.8 58.6 35.3
Bharatar 54.1 62 64.1 69.5 47.6 63
Paravan (H*, W*) 59 72.2 65.2 64.3 72.5 50
Thandan (W*) 55 51 91.4 51.2 57 65.2
Sambavar 45.4 38.5 36.6 46.9 39.4 24.7
Chetty 63.2 59 62 65.8 59.4 60.1
Chakkalar 56 57.4 61.5 56.7 61.3 44.4
Kammalar(H**, W**) 59.9 62.5 40.4 58.1 64.4 51
Kanikkar 48.4 45.5 53.3 49.2 45.7 30
Kattunayakan 76.6 64.1 66.7 75.6 64.4 75
**Significant at 0.01 level, *Significant at 0.05 level

Influence of occupational status on consanguinity is shown in Table 2 which
evidence consanguineous unions among all the four categories in varied percentage. In majority
of the castes, except Thandan, negative association was observed between consanguinity and
occupation (i.e. un-employed and low category employed men contracted consanguineous
marriages in high percentage), whereas the trend was reverse (positive) with highly employed
category men of Thandan. Among women the rate of consanguinity was found to be higher in
the middle category of Bharatar, Paravan, Thandan and Kammalar. Highly significant
association was found between consanguinity and occupation among Pallars, Thandans and
Kattunayakans (wives) and Bharatars (husbands) (1%). Less significant association was found
among Thandans and Chakkalars (husbands) and Kanikkars (wives).
Table 2 Frequency of consanguinity by the level of occupation for different castes
% of consanguinity - Occupation
Unemployed Low Middle High

Caste
H W H W H W H
Pallar (W**) 35.3 53.9 57.4 59.1 46.5 22.2 0
Bharatar(H**) 33.3 63.3 61.1 35.2 59 82.4 0
Paravan 87.5 65.7 65.7 68.8 57.1 70 0
Thandan(H*, W**) 70.5 53.6 54 57.1 59 86.2 100
Sambavar 0 44.2 43.6 28.1 25.7 0 0
Chetty 63.6 61 59.6 51.4 60.4 58.3 55.6
Chakkalar(H*) 0 56.7 56.9 100 64.2 75 0
Kammalar 71.4 61.2 60.8 48.6 24 68 0
Kanikkar (W*) 100 48.8 45.3 41.5 20 0 0
Kattunayakan (W**) 77.8 77 70.2 55.3 44.4 0 0
The frequency of consanguinity based on family income is shown in Table 3.
Consanguineous marriages were contracted by spouses of all castes irrespective of their family
income but with varying rates. Therefore, no clear association between consanguinity and family
income could be predicted except among Sambavars, Bharatars, Chakkalars and Kammalars,

where Sambavars showed negative association and the rest positive association. Highly
significant association between two was observed among Kanikkars (1%) and less among
Chakkalars and Kammalars (5%).
Table 3 Frequency of consanguinity by the level of family income for different castes
% of consanguinity - Family Income

Caste
LIG LMIG UMIG HIG
Pallar 54 73.1 73.9 16.7

Bharatar 61.1 44.7 56.9 62.5
Paravan 65.9 57.1 73.1 0
Thandan 54.6 58.5 64 61.9
Sambavar 46 23.4 31.3 0
Chetty 60.4 60.9 60.3 56.5
Chakkalar* 57.1 25 66.1 100
Kammalar* 62 54.4 46.3 69.7
Kanikkar** 46.9 75 10 0
Kattunayakan 69.2 87.5 50 0

Marital age and its influence on consanguinity is furnished in the Table 4. From the
table it is evident that the percentage of consanguinity is observed in all age groups. Among men
negative association (ie. lower age with high percentage of consanguinity) between
consanguinity and marital age was observed among Pallars and Kammalars whereas others
showed positive association (ie. higher age with higher consanguinity). Women among Pallar,
Paravan, Thandan, Sambavar, Chetty and Kattunayakan showed negative association and others
positive association. It was interesting to note that women (Pallar, 66.2%, Sambavar, 46%,
Chetty, 62.3% and Kattunayakan, 76.5%) and men (Pallar, 86.7% and Bharatar, 100%) who
married below 20 years were more consanguineous. Likewise, men (Paravan, 76.5%, Thandan,
65.2%, Chakkalar, 78.9%, Sambavar, 65.5% and Kanikkar, 75%) and Women (Bharatar, 100%,
Thandan, 64.3%, kammalar, 100% and Chakkalar, 100%) who married above 30 years were also
found to be highly consanguineous. Highly significant association between marital age and

consanguinity was observed among husbands of Bharatar, Thandan and Sambavar (1%) and less
significance among wives of Bharatar and Thandan and husbands of Kanikkar (5%).
Table 4 Frequency of consanguinity by the level of marital age for different castes
% of consanguinity – Marital age
Husband Wife
Caste 20 – 25 - 30 – 35 - 20 - 25 - 30 - 35 –
<20 ≥40 <20
24 29 34 39 24 29 34 39
Pallar 86.7 57.7 56.2 47.6 52.2 6.3 66.2 54.8 38 30.8 8.3
Bharatar
100 71.2 57.6 50 40 33.3 68 53.6 54.7 100 66.7
(H**,W*)
Paravan 0 69.1 68.2 60.3 76.5 60 61.6 70.8 62.6 45.5 66.7
Thandan
33.3 62 55.1 46.2 65.2 44.4 60.8 49.8 51.2 64.3 0
(H**W*)
Sambavar
29 46.5 41 32.8 12.5 48.1 46 43 24.6 0 0
(H**)
Chetty 50 57.4 61.9 60.3 57.1 0 62.3 59.8 56.5 50 0
Chakkalar 0 53.2 59.9 45.5 78.9 50 56.3 57.9 56.7 33.3 100
Kammalar 0 65.5 58.5 61.2 46.7 0 65.1 58.5 59.4 100 0
Kanikkar
50 46.8 43.5 50.4 66.7 75 43.2 51.6 48.3 50 0
(H*)
Kattunayakan 50 76.7 62.7 66.7 50 66.7 76.5 66.1 56.3 50 0
The rate of consanguinity in various year groups were presented in Table 5. Analysis
of the data revealed that the percentage of consanguinity was observed to be higher for marriages
which were performed before 1950 (100% among Bharatars, Thandans, Chakkalars and Chetty-
76.9%), 1951-1960 (Pallars, kammalars and Kattunayakans- 74.5, 77.1 and 100% respectively)
and 1961-1970 (Paravan and Kanikkar, 91.7 and 58.5 respectively). In the case of Sambavars,
marriages made after 2010 were found to be at higher rate (53.7) than the early years. Highly
significant association was observed between consanguinity and year of marriage among

Thandans, Sambavars and Kanikkars whereas less significant association was observed among
Paravans, Chakkalars and Kammalars.
Table 5 Frequency of consanguinity by the level of marital year for different castes
% of consanguinity – Marital year

1951- 1961- 1971- 1981- 1991- 2001-
Caste ≤1950 >2010
1960 1970 1980 1990 2000 2010
Pallar 66.7 74.5 60.8 54.1 63.9 53.3 43 58.8
Bharatar 100 70 62.5 61.4 61.2 53.6 56.7 66.7
Paravan* 0 75 91.7 55 64.2 71.4 60.6 60
Thandan** 100 66.7 68.6 51.2 57.5 57.2 48.3 46.6
Sambavar** 28.6 41.7 39.6 41.5 28.5 39 48.5 53.7
Chetty 76.9 60 66.7 59.6 58 51.8 61.5 76.3
Chakkalar* 100 80 71.1 57.1 47.4 57.6 60.3 38.5
Kammalar* 57.9 77.1 63.2 57.9 60.3 65.5 58.2 45
Kanikkar** 0 31.3 58.5 40.5 40.9 46.4 54.7 41.5
Kattunayakan 75 100 80 72.7 87 51.3 68.4 75
DISCUSSION
Generally, consanguinity is influenced by geographical, demographical, religious,
cultural and socio-economic factors (Hussain, 1999; Bittles, 2001; Saadat, 2007). These factors
may vary from region to region, population to population and caste to caste. Mostly spouse
selection depends on socio-economic and literacy levels. The highest rate of related unions is
related to low economic status, low education and residence in rural areas (Jurdi and Saxena,
2003; Alperet al., 2004; Bener and Hussain, 2006). Relatively high rates of consanguinity occur
in more traditional and rural areas and among the illiterate and poor societies (Bittles, 1998).
In Europe and the New World, low rate of consanguinity is negatively associated with
urbanization, industrialization, literacy and church ban whereas high rate occurs in traditional
rural areas among illiterates and poor (Bittles, 1998). Among racial isolates, endogamous castes
and tribal groups, factors like ethnicity, tribal and caste bonds play a major role. In most of the
Christian countries church ban decreases the rate of consanguinity while in many Muslim

countries related unions are encouraged for religious reasons (Tadmouriet al., 2009; Schulz,
2017). Among Hindus in South India, close kin unions are practiced more as part of their social
custom and tradition. Marital distance is also a major factor in influencing consanguinity where
the people are in isolated and rural areas. Arranged marriage system still prevalent in South
Asian countries like India and Pakistan favours related mate selection. Close kin alliances are
also favoured by joint family system because of the faith to have optimum environment for
compatibility between the bride groom and her in-laws, greater protection for new life, mutual
knowledge and greater stability system. Economic factor is also considered as a primary and
dominant factor in influencing the rate of consanguinity.
Socio-economic variables such as educational level, occupational status and income of the
family, together with geographic determinants like residential area and marital distance and
demographic variables like marital age, marital year and spousal age difference have influencing
role in consanguinity. These factors are used to stratify the social classes of a population in
general. This type of stratification allows 1) definition to control variables while inferring the
effects of consanguineous unions on public health parameters 2) discussion of social profile of
the study population to be addressed by the public health programme 3) providing useful
information about the problems of related unions. Consanguineous unions are reported fairly
high in less educated families especially from rural habitations (Riazet al., 2016). The role of
different factors in influencing the rate of consanguinity among the selected castes for the study
is analysed.
Educational level is a reflection of the social status of an individual/population.
Consanguinity studies carried out in diverse population groups world over proved its strong
influence on mate selection. In almost all studies, except few, the literacy level is negatively
correlated with the rate of consanguinity (Joseph and Mathew, 2002; Vasanthakumari, 2003;
Mathew et al., 2006; Kerkeniet al., 2006; Lekshmi and Sudhakaran, 2012; Cicekliogluet al.,
2012; Jyothilekshmi, 2015; Bhasin and Kapoor, 2015; Charmode, 2016; Khan and Mazhar,
2018) showing that the lower the literacy level the higher is the consanguinity rate and vice
versa. Men with higher education are more exposed to modern ideas about independent mate
selection and consequently may view arranged marriage as undesirable. Educated men may also
have greater opportunities for interaction with women who are non-relatives than do men with

less education (Thakeb, 1985). The same is also true with educated women. Studies carried out
world across documented evidences of negative correlation with the rate of consanguinity
(Sureenderet al., 1998; Hussain and Bittles, 1999; Mathew et al., 2006). In majority of the cases,
education of women was found to be an important issue of responding against or for to the
compulsion and pressure for consanguineous unions. In the present study, the literary status of
the spouses was categorized into a) illiterate b) low (up to primary level), c) medium (upper
primary and high school level) and d) high (university level). The data (Table 1) showed decline
in the consanguinity rate correlated with educational status of men in Pallar, Sambavar, Chetty,
Kammalar and Kattunayakan castes and women in all castes except Thandan.
Occupational status also plays a vital role in influencing consanguinity (Bhasin and
Kapoor, 2015). In the present study the occupational status was categorized into unemployed,
low, middle and high. Highly employed were very meagre in number hence neglected from the
study. The percentage of consanguinity was high among unemployed and low occupational
classes of men. Positive (Liascovichet al., 2001) as well as negative association (Kerkeniet al.,
2006; Joseph, 2001; Jyothilekshmi, 2015) was reported between the rate of consanguinity and
occupational status in many studies. In most cases occupational status has association with
educational level which together play important role in marriage transaction. Caste wise analysis
shows higher percentage of consanguinity among the un-employed and low occupational status
except women of Bharatar and Thandan (Table 2). Similar report of consanguinity was obtained
in Pakistan among lower category and blue-collar jobs (Kamal et al., 2015).
Among women the rate of consanguinity was high among middle educated. This may be
due to the financial independence and decision of working women to make choice of husband
(Kamal et al., 2015). It is plausible that working women have greater exposure to or contact with
unrelated men than do women who do not work. Pingle (1983) has reported that low
occupational mobility accounted for the very high incidence of related marriages in tribal groups.
Frequency of consanguinity was observed to be very low (below 15%) in high education class in
all the three communities (SC, ST, BC) studied by Vasanthakumari (2003) from the same region.
Employed men/women in many cases are unable to find professionally compatible mates among
his/her close-kins, and as a result they are liable to make mate selection with unrelated spouses.

Among the socio-economic determinants, the most important at the household level
is income or wealth. Family income is a primary measure of the financial status of the family. It
is considered as a valid indicator of the social status of a family. Household income indeed
represents the current flow of economic resources to the family and, as such, is a good measure
of the family's capacity to purchase health through food, medical services, and household
amenities (Tekce and Shorter, 1984). Negative association between consanguinity and family
income was reported in many studies (Jyothilekshmi, 2015).
The families chosen for the study were classified into four categories such as low,
lower middle, upper middle and high based on the annual income. The rate of consanguinity
based on family income is shown in Table 3. In the present study, Pallars, Paravans and
Thandans, recorded high rate of consanguinity among upper middle income grade, Bharatars,
Chakkalars and Kammalars among high income grade, Chetties, Kanikkars and Kattunayakans
among lower middle income grade and Sambavars among low income grade. 100%
consanguinity was observed in high income grade Chakkalars. While considering the whole data
irrespective of castes, family income showed highly significant negative association with
consanguinity. Among consanguineous couples higher percentage of consanguinity was recorded
among low income grade. The reason for this may be due to the possibilities to reduce hidden
uncertainities in health and financial issues, to reduce dowry and bridal payments (Bittles, 2001;
Mathew et al., 2006).
Premarital negotiations in connection with financial matters of marriage can be
sometimes made in less cost (Hamamy, 2012). Non-consanguineous households were more
prosperous than consanguineous families (Hussain and Bittles, 1998). Family income,
occupation and education status are inter-dependent factors and have implications on the life
style, social activities etc. Usually it may be difficult for a person of high financial position to get
a suitable compatible partner from among the same kindred. Hence, such people try to make
suitable match selection from outside the kinship boundary, thus reducing the frequency of
consanguinity in such category.
Relation of consanguinity with various demographic factors was effectively used to
interpret the ascending or descending temporal trend of consanguinity. Year of marriage, spousal
age at marriage and spousal age difference were the demographic factors analysed for the study.

Age at marriage is an important demographic factor that determines the marital life and fertility
span of couples (Bittles and Black, 2010). Low aged close-kin alliances are arranged by parents
which have been the practice in most Middle East and Asian countries including South India
(Hussain and Bittles, 2004; Mathew et al., 2006; Islam, 2018). The influence of marital age of
spouses on frequency of consanguinity was carried out by grouping the marriage age into six
from below 20 years to above 40 years and the results are furnished in Table 4. Earlier marriage
of spouses leads to a longer reproductive period combined with reduced family planning
measures and results in population growth with greater genetic risk. Age at marriage is greatly
influenced by literacy status of spouses especially women (Hussain and Bittles, 1999). Literacy
can obviously elevate the age at marriage in different ways. The time required for a desired
course of education can delay the age at marriage as is evident in the present study (Joseph,
2001).
A woman who marries relatively late may be able to avoid having her marriage
arranged. This should increase her chances of marrying a non-relative. Assessment of the factors
which encourage early marriage is necessary to minimize the problems due to consanguinity
among inbreeding communities. Few studies with spousal age at marriage had been carried out
in Kerala (Joseph, 2001) and Karnataka (Hussain and Bittles, 1999). The present study shows
highly significant positive association between the marital age of spouses and consanguinity.
Declining trend with increasing age of men except Thandans, Chakkalars and Kanikkars and
women except Bharatars, Thandans, Chakkalars and Kammalars was noticed in the present study
(Table 4).
There are so many factors responsible for lower marital age among consanguineous
couples. Lower educational level, prior marriage planning and availability of suitable partner are
some factors to consider (Sivaramet.al. 1995; Hussain and Bittles, 1999). In the present study
also most of the consanguineous women are less educated and had lower age at marriage.
Positive association between literacy and spousal age at marriage especially among women was
reported by, Pillai and Mathew (1995) and Hussain and Bittles (1998). Raj et al. (2010) and
Nasrullahet al. (2013) reported disadvantageous situation due to lower marital age linked with
higher fertility and negative effects. Similar situation was observed in the highly consanguineous
Kattunayakans of the present study.

One of the demographic factors which influence consanguinity is the year of
marriage. It helps to understand the direction of temporal trend of the phenomenon of
consanguinity. Recent studies evidenced decline in the rate of consanguinity since 1960 in many
countries (Fusteret al., 2001; Sindhu, 2000). The reasons spotted for this include migration of
populations, civilization, urbanization, secular family and changing socio-economic condition of
the population through time (Krishnamoorthy and Audinarayana, 2001; Ullahet al., 2018).
Small- sized families result in lesser number of children which consequently reduces the number
of eligible cousins. Some countries like Japan, Arabian and India have persisted rate of
consanguinity due to maintenance of their social and traditional customs. Present study
registered high rate of consanguinity before 1950 among five castes like Pallar, Bharatar,
Thandan, chetty and chakkalar. The percentage of consanguinity was higher among Kammalar
and Kattunayakan during 1951 and 1960 (Table 5). Considering the whole data together also
suggests highly significant association between year of marriage and consanguinity.Before the
year 1950, Thandan, Chakkalar and Bharatar castes recorded 100% consanguinity but after 2010
the record declined to 46.6%, 38.5% and 66.7% respectively. In Japan studies by year of
marriage have indicated a clear downward trend of consanguinity rate down the decades (Lebel
and Opitz, 1983).
A consistently declining trend in consanguinity from older to younger generation is
not evident in the study of Mudaliars (Lekshmi and Sudhakharan, 2012) suggesting their strong
belief in following Dravidian culture, together with socio-economic and educational
backwardness acting hand in hand may be the reason for the high prevalence of consanguinity
among them. This contradicts the general trends prevailing in most areas of the world, where
remarkable decline in the rate of inbreeding has been reported from older to younger generations
(Sanghvi, 1966; Prothro and Diab, 1974; Bittles, 1994; Richard and Rao, 1994; Jyothilekshmi,
2015). The rate of decline of consanguinity may be due to laws banning consanguinity,
awareness, demographic constraints and socio-economic development. Mathias et al., (2000)
reported steady decline in consanguinity in Virginia. Reduction in the size of the family may also
be the reason for the decline in the rate of consanguinity.

CONCLUSION
From this study it is evident that the rate of consanguinity in Kanyakumari district is
in decline which may be due to the increased secular family system and awareness about the
genetic abnormalities connected therewith. Education, occupational status, family income,
marital age and marital year have some positive and negative influence on the rate of
consanguinity.
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ECOLOGY OF TUTICORIN AND VEMBAR GROUP OF ISLANDS, GULF
OF MANNAR
M.Sheeba1 and T.Mohanraj2
1
Dept of Zoology, PSGR Krishnammal College for Women, Coimbatore.
2
Dept of Zoology, Aditanar College of Arts & Science, Tiruchendur.
ABSTRACT
The present study deals with the assessment of various ecological parameters such as
temperature, pH, salinity, dissolved oxygen, light penetration depth (LPD), calcium carbonate,
organic matter and sand-silt-clay composition of the Tuticorin and Vembar group of Islands. The
water and sediment samples were collected from the seven islands namely: Vaan, Kasuvar,
Kariyachalli, Vilanguchalli, Upputhanni, Puluvinichalli and Nallathanni islands, during July
2009 and February 2010. The samples were then subjected to several physical and chemical
investigations to know the ecological status of the study areas. Statistical analysis like Pearson
Correlation matrix was performed using the collected data to identify the relationship between
the studied parameters.
KEY WORDS: parameter, temperature, salinity, sediment,Pearson correlation
INTRODUCTION
The Gulf of Mannar Biosphere Reserve is one of the world’s richest coastal regions in
Asia, from the marine biodiversity perspective; as it harbours over 3,600 species of flora and
fauna. It comprises 21 islands, enclosed by estuaries, mudflats, beaches, coral reefs, salt
marshes, mangroves, along with the marine floral communities such as algae and sea grasses.
These 21 small islands in the Gulf of Mannar region cover a total area of nearly 2.41 square
miles (6.23 sq km). They are bordered by astonishing species of coral reefs that are often
referred as ‘Underwater Tropical Rain Forest’, endowed with a habitat for unique marine
biodiversity. The islands serve as a nesting ground for marine turtles, and home to a variety of
migratory bird species. All the 21 islands are located between 8o 47’ N; 79o 14’ E and are

clustered into four groups namely: Tuticorin (4 Islands), Vembar (3 Islands), Kilakarai (7
Islands) and Mandapam (7 Islands) group.
Nearly 60 percent of the world population live near the coastal zone and seem to be
dependent directly or indirectly on its resources. The coastal environment of Gulf of Mannar is
typically dynamic with many cyclic and random processes owing to a variety of resources and
habitats that are influenced by seasonal patterns. Factors including urbanization, population
growth, industrial expansion, foreign trade and unsustainable resource utilization are known to
pose serious threats to the survival of unique life support system of the Gulf of Mannar
Biosphere Reserve.
In recent years global climate change has imposed a remarkable transformation over
several environmental variables. The marine environment as a complex ecosystem has been
influenced by a variety of physical, chemical and biological processes. Hence study of the
environmental parameters would serve as a useful tool for the ecological assessment and
monitoring of different coastal systems (Sankar et al., 2010 and Saravanakumar et al., 2008). In
general, factors like salinity, temperature, Dissolved Oxygen together with sediment
characteristics have greater influence on the distribution of marine organisms (Jayaraj et al.,
2007). Moreover, assessment of the ecological parameters is very essential since the quality of
coastal water resources rely mainly on the physico-chemical status and the source of the
pollution loads (Reddi et al., 1993). In temperate regions, the chemical component especially the
nutrient salts and the physical factors including pH and temperature operate as limiting factors in
the distribution and abundance of marine flora and fauna. A study on hydrography of the east
coast by Ramamurthy (1953, 1953a) and investigation by Malupillay (1962) on physico-
chemical conditions of Gulf of Mannar, reveal the importance of ecological studies of the marine
ecosystem. Most of the marine life forms could withstand a wide range of environmental
conditions,whereas some are very sensitive to the slight alterations in their surroundings(Raibole
and Singh, 2011). Hence, knowledge on the shifting pattern of physical and chemical factors of
sea water is very crucial for obtaining a scientific view of the existing and the upcoming threats
to the marine life.

Study area
Forty-eight surface sediment and bottom water samples were collected manually with the
help of Scuba divers from Tuticorin. Two sets of samples were collected for the study; one
during July 2009 and another set during February 2010. All underwater photographs were taken
using a CANON Powershot digital SLR camera (Model: S45). The samples were collected from
the vicinity of seven islands namely: Vaan Island (08° 49' N; 78° 11' E), Kasuvar Island (08° 49'
N; 78° 14' E), Vilanguchalli Island (08° 56' N; 78° 16' E), Kariyachalli Island (08° 57' N; 78° 15'
E), Upputhanni Island (09° 05' N; 78° 29' E), Puluvinichalli Island (09° 06' N; 78° 32' E) and
Nallathanni Island (09o 10' N; 78° 57' E). The former four islands are named as Tuticorin Group,
while the latter three as the Vembar Group of coral islands (Fig. 1).
Water Sample
Water samples were preserved by adding a few drops of chloroform (Newcombe et al.,
1939). The Bottom water temperature (BWT) and pH were measured on board, the former using
a thermometer, and the latter using a pH meter. Light penetration depth (LPD) was determined
using a black and white Secchi disk. Salinity was estimated using the standard titration method
and equation proposed by Knudsen (1901), while the dissolved oxygen (DO) content was
determined UV-spectrophotometrically (Duval et al., 1974).
Sediment Sample
The sediment samples were stored in a mixture of one part of buffered formalin in nine
parts of water (4% solution) with a pinch of CaCl2 to achieve neutrality (Walker et al., 1974).
From the sediment sample Calcium carbonate and organic matter was estimated adopting
methodology after Loring and Rantala (1992) and Gaudette et al. (1974), respectively. Sand, silt
and clay percentages were evaluated through sieving and pipette procedures, the latter in
accordance with Krumbein and Pettijohn (1938).
RESULTS
Water and sediment sampling were made at different stations around the Tuticorin and
Vembar group of islands to record various ecological parameters. The collected samples were
placed in sterile polythene covers and brought to the laboratory immediately for further analysis.

All parameters with the mean value of the data and the significant variations between the study
areas have been depicted in Table 1&2.
Water depth (m)

The average water depth evidenced during the study varied between 5.42m to 10.67m.
The lowest range of water depth was recorded at Upputhanni Island (1.0m – 12.1m) and highest
depth range at Vilanguchalli Island (3.8m to 16.2m). A similar trend was observed during both
the surveillance period.
pH
pH of water is vital for the biotic communities as most of the marine flora and fauna
endure a narrow range of pH, from slightly acidic to alkaline condition (Goher MEM, 2002).
The pH of the water samples remained alkaline throughout the study period. During July 2009
and February 2010, pH value ranged from 8.17 to 8.4 and 8.15 to 8.45 respectively (Fig. 2a).
The maximum pH was reported in the Kaswar Island and the lowest at the Nallathanni Island.
The minimum value was recorded post monsoon, which could be attributed to the influx of rain
water.
Water Temperature (°C)

The bottom water temperature of the chosen study areas fluctuated between 25.98 oC and
29.94 oC. The highest temperature was recorded at the Upputhanni Island during June 2009 (Fig.
2b). It is possibly indispensable to note that the average bottom-water temperature for the
shallowest station had the highest temperature range.
Light Penetration Depth (m)

Sea water selectively scatters and absorbs certain wavelengths of visible light up to a
particular depth; this light penetration depth (LPD) was estimated at all the study areas. The LPD
was seen the maximum at Vilanguchalli island with a value of 2.88m, whereas the lowest LPD
was recorded from the Upputhanni island i.e. 0.95m (Fig. 2c).

Salinity (ppt)
Salinity is one of the fundamental water quality parameter that has an influence over the
distribution of marine biota. Generally the salinity range of sea water is estimated at 5‰ or
35ppt. In the present study highest salinity was observed at Kariyachalli Island with an estimate
of 5.57‰. The samples collected from Nallathanni Island during July 2009, showed a lower
salinity range of 4.89‰, but post monsoon analysis i.e February 2010 study evidenced much
lesser value of .1‰ at the Vilanguchalli Island (Fig. 2d).
Dissolved Oxygen (ml/L)

Higher the value of dissolved oxygen greater is the incidence of aquatic life. The amount
of dissolved oxygen around the Tuticorin and Vembar group of islands ranged between 4.03 to
4.46mg/L and 3.93 to 4.47mg/L during July 2009 and February 2010 samplings respectively
(Fig. 2e). The highest value was recorded post monsoon due to the turbulence of water, thus
facilitating the diffusion of atmospheric oxygen resulting in an increased solubility of oxygen in
water.
Organic matter (%)

Usually, all natural waters contain organic matters (OM), which are the resultants of
biogenic material and biological excretion products. In this study the mean organic matter varied
between 0.38 and 1.32 mg/l obtained from Nallathanni and Vaan islands, respectively. The
organic carbon that is deposited in marine sediments form a vital component of the global
carbon cycle. The OM could be considered as an estimator of the processes occurring within the
water stratum.
Calcium carbonate (CaCO3)

In the present study deposition of CaCO3sediments in the ocean was dominated by the
input of calcium primarily from the corals, molluscs as well as the foraminifera and
Coccolithophores (algae). As the organisms die their calcareous shells get accumulated at the sea
floor along with the sea sediments. During the sediment analysis high CaCo 3 concentration
(28.32%) was recorded at Vaan Island, followed by the Kaswar and Vilanguchalli islands.

Soil texture (%)
The sediment texture of the selected stations showed presence of sand, silt and clay in
varying proportions. All the islands were primarily sandy in nature. During both the survey
period Upputhanni Island comprised the maximum silt composition. Similarly, the sediment
collected from Kaswar Island showed relatively higher percentage of clay (1.02%) in
comparison to the other islands.
The correlation matrix was performed to demarcate the relationship between the studied
ecological parameters of the Tuticorin and Vembar group of islands. The Pearson correlationand
their significant values are shown in Tables 3&4.
DISCUSSION
Changes in the ecology of marine environment have gained importance in the biological
research around the globe; since itis considered as one of the prime factors determining the
survival and productivity of the flora and fauna. Increased level of pollutants in water has
steadily altered the physico-chemical and biological parameters, thus affecting the normal and
healthy oceanic habitat.Hence the present study was undertaken with the objective to investigate
the important ecological parameters(water depth, temperature, Light penetration depth, pH,
salinity,dissolved oxygen, organic matter and soil texture) of the Tuticorin and Vembar group of
islands, Gulf of Mannar.
During the study a wide range of depth variations were recorded around the seven coral
islands that seem to have a considerable effect over the physical and chemical properties of
water. The prevailing ecological condition of the coastal waters has been influenced greatly
bythe arrival of monsoon (Marichamy et al. 1985). A report by Ramamirtham and Jayaramau
(1960) illustrated fall of temperature values in the Arabian Sea, with the onset of monsoon. In
the present study also a similar phenomenon was observed during July 2009 (pre-monsoon)
sampling, in whicha lower temperature range was recorded between 25.98 oC - 28.42oC, around
the Nallathanni and Kaswar islands respectively. During both the surveys pH remained alkaline
at all stations with minimum values observed during July 2009; however the pH range showed
slight variations between the study areas. The sequential change in pH is apparently due to
change in the Primary production, respiration by organisms and mineralization and

decomposition of organic matter in the sediments(Zingde and Desai 1987). The highest pH of
8.45 recorded during the present study might be the indicative of elevated rate of illumination
and subsequent photosynthesis, as said by Anjana and Kanhere (1995). Rajasegar (2003) stated
that seasonal fluctuations in pH values of seawater is mainly attributed to factors like elimination
of CO2 through bicarbonate degradation, freshwater influx, dilution and low primary
productivity. In addition, pH might also change as a result of temperature-salinity changes and
biological activity (Srilatha et al., 2012). The light penetration depth (LPD) was recorded the
maximum during the post-monsoon season, which wasgenerally due to the monsoonalrains
coupled with low intensity of solar radiation.The increased concentration of dissolved organic
matter, suspended sedimentsand turbulence in the water could also possibly result in a higher
LPD value.
One of the major ecological factors responsible for the distribution of biotic elements in
the marine ecosystem is salinity (Manivasagan, 2009).Its variation caused by dilution and
evaporationis most liable to influence the species diversity and distribution pattern
(Chandramohan and Sreenivas, 1998). Fluctuation in salinity would definitely affect the
biological characteristics of the marine environment and is primarily dependent on rainfall, land
water runoff, the rate of evaporation of water and water current.During the present study,there
was no wide salinity variation observed between the stations.A maximum salinity of 35.57ppt
observed at the Kariyachalli Island during July 2009, reveal the higher rate of evaporation
prevailing during summer and alsothe oceanic water current that probably brings in high salt
concentration to the coastal seawater. A significant reduction in salinity value observed during
the post-monsoon survey was mainly due to the rainfall and input of freshwater. The salinity
profile of the seven coral islands of Gulf of Mannar is well comparable with the earlier works of
Thangaraj et al. (1979), Sivakumar (1982), Nair et al. (1983) and Chandran and Ramamoorthi
(1984) in the coastal waters of Bay of Bengal.
In the present study, the dissolved oxygen (DO) concentration was low at most of the
stations during the July sampling and was relatively high during post-monsoon (February 2010).
A similar observation has been made by Varadharajan and Soundarapandian (2014), which
recorded a higher value of DO during the month of February at the sampling sites of

Muthupettai. Temperature is one of the major factors controlling oxygen saturation in any water
body (Hung 1972). The oxygen solubility in water is inversely proportional to temperature
(Carpenter 1996). The low DO concentration observed during July may be due to the higher
range of salinity and temperature values, coupled with biological processes such as consumption
by the organisms for respiration and active decomposition of organic matter during the pre-
monsoon (summer) season.The observed high DO values might be due to the combined effect of
higher wind velocity and the subsequent freshwater mixing (Vijayakumar et al., 2000; Mitra,
1990).
The distribution of calcium carbonate and organic matter are chiefly controlled by the
availability of coral rubbles and distribution of sea grass in the study areas.CaCO3generally
recognized as dilutor of trace metal concentrationenters the marine environment through
terrestrial run off and organisms in water column (Sundararajan and Srinivasalu, 2010). During
the sediment sample analysis maximum concentration of CaCO3 (28.32%) was recorded at the
Vaan island during the July 2009 survey; However it reduced at all the seven islands during the
subsequent (post-monsoon) sampling. Similarly, the occurrence of sediment organic matter
(OM)is assumed to be in one of the chief factors influencing the community structure and
metabolism of the benthos (Mills, 1975; Graf et al., 1983). It plays a key role in the mobility and
availability of trace elements in soil and sediments (Kirk, 2004).The result indicated high level of
organic matter recorded at the Vaan Island; which shows thepresence and incorporation of
increased level of organic materials into the study area. Similarly, lower concentration
estimatedat most of the sampling locations was due to thelesser-availability of organic matter
sources including the sea grasses.
The benthic landscape of any marine environment is influenced by the
distributionalpattern of its sediment particles (Swift, 1970). Sediments in aquatic systems serve
as sinks and sources of contaminants because of theirvariable physical and chemical properties
(Rainey et al., 2003; Marchandet al., 2006). The relative percentageof Sand, Silt and clay content
of all the study areas has been presented in Fig. 3. A study by Jonathan (2004), reported that the
surface and core sediments off Tuticorin have high content of sand in the continental shelf
region. Likewise, all the seven islands are predominantly sandy in nature with silt and clay
contents in minor proportions.

Fig. 1: Map of the study area
Table 1: Ecological parameters of Thoothukudi and Vembar group of islands during July 2009
Ecological Vaan Kasuvar Kariychalli Vilanguchalli Upputhanni Puluvinichalli Nallathanni

Parameters Island Island Island Island Island Island Island
WATER
Depth (m) 6.08 7.7 8.23 10.67 5.42 7.36 6.92
pH 8.21 8.4 8.17 8.17 8.24 8.4 8.35
BWT (° C) 28.45 29.75 29.76 29.87 29.94 29.5 29.25

LPD (m) 1.19 1.22 1.18 1.27 0.95 0.97 1.06
Salinity (‰) 35.25 35.27 35.57 35.36 35.25 35.04 34.89
DO (ml/l) 4.04 4.05 4.05 4.03 4.46 4.28 4.25
SEDIMENT
Organic 1.32 0.51 0.63 0.64 0.58 0.40 0.38
matter (%)
CaCO3 (%) 28.32 25.35 24.77 25.52 24.42 21.28 21.01
Sand content 80.75 81.12 86.98 78.5 79.82 80.97 81.4
(%)
Silt content 19.46 17.85 12.52 20.96 21.73 19.52 19.35
(%)
Clay content 0.35 1.02 0.47 0.53 0.4 0.47 0.4
(%)

Table 2: Ecological parameters of Thoothukudi and Vembar group of islands during February
2010
Ecological Vaan Kasuvar Kariychalli Vilanguchalli Upputhanni Puluvinichalli Nallathanni

prameters Island Island Island Island Island Island Island
WATER
Depth (m) 6.08 7.7 8.23 10.67 5.42 7.36 6.92
pH 8.31 8.45 8.3 8.2 8.18 8.23 8.15

BWT (° C) 27.67 28.42 27.88 28.08 28.35 26.32 25.98
LPD (m) 2.74 2.87 2.78 2.88 2.23 2.25 2.64
Salinity (‰) 33.25 33.07 33.16 33.1 33.16 33.15 33.2
DO (ml/l) 4.23 4.47 4.4 4.4 3.93 4.14 4.1
SEDIMENT
Organic 0.65 0.51 0.52 0.59 0.69 1.25 1.12
matter (%)
CaCO3 (%) 24.07 23.5 21.17 22.57 21.44 24.26 23.78
Sand content 82.4 84.07 81.47 77.97 77.83 80.03 79.8
(%)
Silt content 17.04 15.47 18.07 21.52 21.92 19.58 19.45
(%)
Clay content 0.53 0.45 0.45 0.5 0.24 0.37 0.32
(%)
CONCLUSION
Monitoring the ecology of coastal waters provide a significant contribution towards the
knowledge onmarine environment. The analysis carried out during the study revealed the
ecological (Temperature, pH, dissolved O2, light penetration depth, Salinity, Organic matter,
and sediment analysis) status of the Tuticorin and Vembar group of islandslying in the Gulf of
Mannar region. The hydrological parameters exhibited distinct variations during both the study
period. A complete database of the physico-chemical characteristics of the seven coral islands
was made. Thedata contains comprehensive information that subsequently helps understand and
monitor thehealth of the ecologically sensitive habitats and implement effective management
andconservation practices. This baseline information on the ecological status could
fundamentally forma support for the futuristic studies on various aspects of this marine
ecosystem.

Depth pH BWT LPD Salinity DO OM CaCo3 Sand Silt Clay
Depth 1
pH -0.26538 1
BWT 0.402168 -0.00261 1
LPD 0.658673 -0.39952 -0.09417 1
Salinity 0.092173 -0.13274 -0.03896 0.146891 1
DO -0.21312 0.136158 -0.50246 0.490372 -0.29299 1
OM -0.21263 -0.53969 -0.71454 0.396485 -0.22603 0.613353 1
CaCo3 0.030954 -0.59249 -0.31586 0.635378 -0.29324 0.696809 0.856908 1
Sand -0.03347 -0.16409 0.035502 0.078712 0.358114 -0.10134 -0.05182 -0.06606 1
Silt -0.17464 0.159757 -0.0648 -0.32281 -0.31204 -0.03965 0.024744 -0.04928 -0.95897 1
Clay 0.284769 0.470156 0.380607 0.413762 -0.18812 0.499828 -0.29737 0.103711 -0.03458 -0.17026 1
Table 3: Pearson correlation matrix (July 2009)
Table 4: Pearson correlation matrix (Feb 2010)
depth pH BWT LPD salinity DO OM CaCo3 sand silt clay

Depth 1
pH 0.038616 1
BWT 0.134065 0.493734 1
LPD 0.593238 0.495462 0.328699 1
Salinity -0.58897 -0.35645 -0.42821 -0.228 1
DO 0.713686 0.690254 0.374116 0.857754 -0.51136 1
OM -0.41646 -0.65245 -0.77979 -0.81359 0.30633 -0.78903 1
CaCo3
-0.11709 0.131665 -0.57829 0.003868 0.209819 -0.00621 0.37379 1
Sand -0.1399 0.898596 0.1375 0.477927 -0.02619 0.566497 -0.45783 0.354915 1
Silt 0.125102 -0.87638 -0.09917 -0.50572 0.003013 -0.57147 0.449901 -0.38068 -0.99792 1
Clay 0.533886 0.541257 0.269777 0.777468 -0.03893 0.806849 -0.70491 0.230725 0.504062 -0.51602 1

Fig. 2: Variation in Ecological parameters recorded around the Tuticorin and Vembar group of
islands during July 2009 and February 2010 – (a) pH, (b) Bottom Water Temperature, (c) Light
Penetration Depth, (d) Salinity and (e) Dissolved Oxygen.
a b
8.5 31
8.4 30
BWT (0C)
29
8.3 28
pH
8.2 27
26
8.1 Jul-09 25 Jul-09
8 24
Feb-10 Feb-10
c d
36
3.5 35.5
3 35
Salinity ‰
2.5 34.5
LPD (m)
2 34
1.5 33.5
1 33
Jul-09 32.5 Jul-09
0.5
32
0 Feb-10 Feb-10
31.5
e
4.6
4.5
4.4
DO (mg/L)
4.3
4.2
4.1
4
3.9
3.8 Jul-09
3.7
3.6 Feb-10

Fig. 3: Variation in soil texture recorded around the Tuticorin and Vembar group of islands
100%
90%
80%
70%
Soil texture (%)
60%
50%
40% Clay
30% Silt
20% Sand
10%
0%
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30. 245-268.

OBSERVATION ON CAPTURED MARINE FISHES OF MANAPADU
COAST
A. Vanmathi1 and T. Mohanraj2

1
Department of Zoology, Govindammal Aditanar College for Women, Tiruchendur, Tamil Nadu
2
Department of Zoology, Aditanar College of Arts & Science, Tiruchendur, Tamil Nadu
ABSTRACT
The present study the marine fishery sector of Manapad has been continues to be one of the
major source of fish .The primary data on the various crafts and gears used in fishing practices.
To identify the fishes at the fore most taxa using FAO identification sheets. The result showed
that the most dominant fish families were Carangidae, Serranidae, Lutjanidae, Lethirinidae,
Scombridae. Other fish families were Clupidae, Cynoglossidae. The maximum occurrence of
species recorded from February followed by April. To calculate the ANOVA to know the
significance between families and species.
KEYWORDS:Serranidae, FAO,Tiruchendur, Terapontidae,
INTRODUCTION
Mariculture has been contributing around 30.3% of the global aquaculture production by
quantity and 29.2% of the total value. Finfish culture in the sea is expanding rapidly with an
average annual rate of 9.3% from 1990 to 2010 (Gopalakumar, 2014).Currently, the total is of
the order of about 2.3 million tones. This production has the estimated value of about Rs. 10,400
cores at the landing centre price and about Rs. 17,800 crores at the retail level. During the last
year, the exports of marine fishery products earned about Rs. 5,300 cores foreign exchange
(Srinath, 2012).
Manapad area (8.3765° N, 78.0563° E) fish landing centre is located in Tiruchendur taluk
of Thoothukudi district. The fishery data were collected from the study area for about four
months, i.e. From January 2017 to April 2017. The types of gears operating in the study area was
undertaken during the first week of the study period by visiting the landing site and by

questioning the fisher folks on the operating, usage and the composition of catches of these
gears. The landed fish species were photographed at the landing centre. The identification was
done using FAO identification sheets, Book of Smith’s fishes and fish base. The fishes were
classified up to species level. The commonly used gears for fishing at Manapad includes:
(hook& line, gillnet, sardine gillnet, bottom set gillnet, seine net). Sardines, salmon, cod are the
fishes mainly caught by gillnet. Mid water trawl is used to catch pelagic fishes such as
anchovies, shrimp, tuna and mackerel.
RESULT
The present research recorded with various species of bony fishes from the study area
for the period of January- April 2017. The identified fishes are presented in table 1. Totally 76
species of bony fishes that belongs to 30 families has identified. Of which 9 species belongs to
carangidae, 8 species belongs to the family serranidae, 7species belonged to the family
scombridae. And 6 species of the family Lethirinidae, 5 species each to Sciaenidae and
Lutjanidae, 4species each to Clupidae and cynoglossidae, 2 Species each to Scaridae,
Sphyraenidae, Chirocentridae has also recorded. A single species each to Synodontidae,
Balistidae, Pomacanthidae, Barbourisiidae, Parascorpidae, Ariidae, Mullidae, Haemullidae,
Pricanthidae, Rachycentridae Siganidae, Plotosidae, Terapontidae, Leiognathidae, Menidae and
Psttodidae also recorded during this study.
Table 1: Shows list of fish species recorded during the study period (January – April 2017)
S.No Family Common name Scientific name
1 Ariidae Spotted sea cat fish Arius maculates

2 Balistidae Masked trigger fish Sufflamen fraenatus
3 Barbourisiidae Valvet whale fish Barbourisia rufa
Indian oil sardine Sardinella longiceps
Clupidae Chacunda gizzard shad Anodontostoma chacunda
4
Gold strip sardinella Sardinella gibbosa
White sardine Escualosa thoracata
Cynoglossus macrolepidotus
Cynoglossidae Natal tongue sole Cynoglossus acaudatus
5
Tongue fish Paraplagusis bilineata
Bengal tongue sole Cynoglossus cynoglossus
Dorap wolf herring Chirocentrus dorab
6 Chriocentridae Wolf herring Chirocentrus nudus

Gold fusilier Caesio caerulaurea
7
Caesionidae Caesio cunning
Malabar trivial Carangoides malabricus
Shirmp scad Alipes djedaba
Razorbelly scad Alepes kleini
8 Carangidae Rorpedo scad Megalaspis cordyla
Bigeye scad Selar crumenophthulmus
Bigeye travelly Caranx sexfasciatus
Golden trevelly Gnathanodon speciosus
Gangetic anchovy Thryssa mystax
9 Engraulidae Stolephorus insularis Stolephorus insularis
10 Haemullidae Plectorhinchus Plectorhincus sp
Dory snapper Lutjanus fulviflamma
Blue stripe snapper Lutjanus kasmira
11 Lutjanidae Mangrove red snapper Lutjanus argentimaculatus
Five lined snapper Lutjanus quinquilineatus
Lutjanus sp
Thumb print emperor Lethirinus harak
Lethirinidae Emperor fish Lethirinus lentjan
Sweet lip snapper Lethirinus miniatus
12
Spangled emperor Lethirinus nebulosus
Lethirinidae Emperor Lethirinus ornatus
Lethirinus sp
13 Leiognathidae Bony fish orange Leiognathus bindus
14 Malabar grouper Epinephelus malabaricus
Orange spotted Epinephelus suilius
Serranidae grouper
Spiny cheek grouper Epinephelus diacanthus
Brown spotted grouper Epinephelus chlorostigma
Greasy grouper Epinephelus tauvina
Epinephelus sp
Yellow-edged lyre tail Variola louti
Indian mackerel Rastrelliger kanagurta
King mackerel Scomberomorus cavala
Indo-Pacific king Scomberomorus guttatus
Scombridae mackerel
15
Narrow-barred Spanish Scomberomus commerson
mackerel
Mackerel tuna Euthynnus affinis
Yellowfin tuna Thunnus albacores
16 Synodontidae Greater lizardfish Saurida tumbil
17 Scaridae Tri-color Parrotfish Scarus tricolor

Daisy parrotfish Scarus sordidus
Sphyraenidae Pickhandle barracuda Sphyraena jello
18
European barracuda Sphyraena sphyraena
Tigertooth croaker Otolithes ruber
Karut croaker Johnius carutta
19 Sciaenidae Lesser tiger tooth Otolithes cuvieri
croaker
Solider croaker Nibea soldado
Siganidae Golden spotted rabbit Siganus chrysospilos
20
fish
21 Mullidae Indian goatfish Parupeneus indicus
22 Menidae Moon fish Mene maculate
Nemipterus sp.
23 Nemipteridae Japanese threadfin Nemipterus japonicas
bream
24 Pomacanthidae Ray-finned fish Pomacanthus semicirculatus
25 Parascorpididae Jutjaw Parascorpis typus
26 Priacanthidae Moon tail bullseye Priacanthus hamrur
27 Plotosidae Eel tail catfish Plotosus canius
Psttodidae Spiny turbots Psettodes crumei
28
29 Rachycentridae Cobia Rachycentron canadum

Terapontidae Tiger bass Terapon jarbua
30
Table 2: Shows the number of fish families and species recorded during month wise survey
Month Survey Number of Number of

number family species
January 1 7 16
2 10 21
3 8 13
4 15 26
February 1 11 24
2 17 32
3 16 27
4 9 18
March 1 12 28
2 18 39
3 22 37
4 19 32
April 1 21 33

2 15 22
3 17 29
4 18 25
Table 3: One way ANOVA calculated between families and species reported from the study
area
Source of SS Df MS F P-value F crit

Variation
Between Groups 4371.125 1 4371.125 10.41674 0.017967 5.987378
Within Groups 2517.75 6 419.625
Total 6888.875 7
A high number of species diversity was observed in the month March. The same is very
low in the month of January (Figure1).The fishes were captured of using hook& line , gillnet,
sardine gillnet, bottom set gillnet, seine net. The majority of the fishes were captured in the gill
net& hook line in the study area (figure2).
There are totally 124 fishing vessels at manapad. The one way analysis of variance
(ANOVA) computed from the collected data (table3) for a significance level of 5% shows p<
0.05 i.e. The p-value is 0.017, therefore, the null hypothesis is not accepted. This means that
there is statistically significant difference between the groups.
DISCUSSION
They are effectively captured on hook and line, in gill nets and baited basket traps.
Highest catches are obtained in Maharashtra and Karnataka in the fourth quarter, in Kerala and
Tamil Nadu in third quarter and in Andhra Pradesh in second quarter. (James, 1991).
In total landings, major pelagic finfish groups were those of oil sardine (14.4%),
carangidas (6.2%), ribbonfish (5%), mackerel (5.5%), Bombay duck (5.3%), lesser sardines
(3%), anchovies (4.7%), seer fish (1.8%), Hilsa (1.7%) and tunas (1.7%) of the total production.
The Indian mackerel showed signs of recovery from the progressive decline in catches
experienced. Since 2001 as the catches in 2005 were 1.25 lakh tones (ICAR). The percentage
contributions of other important pelagic fishes along this coast are anchovies and white baits
(9%), ribbon fish (9%) and 9ther clupeids (4%). Small, coastal water fishes belong to the three
genera, Leiognathus, Secutor and Gazza. They occur together and contribute to major

commercial catches in the states of Tamil Nadu, Andhra Pradesh and Kerala. Heavy landings are
obtained along the southeast-coast, around Mandapam. Peak catches are obtained in the third
quarter in Tamil Nadu and Kerala and in the first and second quarters in Andhra Pradesh (James,
1991).The percentage contributions of other important pelagic fishes along this coast are
anchovies and white baits (9%), ribbon fish (9%) and 9ther clupeids (4%). An increasing trend is
seen in the catches of lesser sardines, anchovies and white baits on the east coast of India
(Dharma raja and Varughese Jacob, 1972).
The result showed that the most dominant fish families were Carangidae, Serranidae, Lutjanidae,
Lethirinidae, Scombridae. Other fish families were Clupidae, Cynoglossidae. The maximum
occurrence of species recorded from February followed by April.
REFERENCE
Abdussamad, E. M. Pillai, N. G. K fisheries research institute cochin-1 1
Mohamed Kasim, H. Habeeb Mohamed, Indian council of agricultural research.
O. M. M. J. Jeyabalan, K. (2010). Aswathy, N., Narayanakumar, R. And Somy
(Central Marine Fisheries Research kuriakose. (2014). “Economic
Institute, Kochi - 682018, Kerala, sustainability of marine fisheries in
(India)). Fishery, biology and population india: a total factor productivity
characteristics of the Indian mackerel, approach”. Journal of aquatic biology an
Rastrelliger kanagurta (Cuvier) fisheries vol. 2/no. 2/2014/pp. 69 to 74.
exploited along the Tuticorin coast. Claire Hornby, Brajgeet Bhathal, Daniel
Indian Journal of Fisheries 57(1) p. 17- Pauly and Dirkzeller. (2015).
21. Reconstruction of India’s marine fish
Alagaraja, kurup, srinath & balakrishnan. catch from 1950-2010, University of
Analysis of marine fish landings in British Columbia, Canada.Coasts.
India. A new approach central marine David, E. (2007). Biodiversity and fisheries
fisheries research institute P. B. No. management opportunities in the
1912, cochin.682 018, India, Indian Mekong river basin, Programme Co-
council of agricultural research.Annual ordinator, Fisheries Programme Mekong
report for (1970). ICAR central marine River Commission Secretariat C/o P.O.

Box 7980 Vientiane, Lao PDR. FAO. inshore waters of India.” Central marine
2016. The State of World Fisheries and fisheries research institute mandapam
Aquaculture 2016.Contributing to food regional centre, Mandapam Camp.
security and nutrition for all. Rome. 200 Mohanraj. T. and Prabhu, K. (2012). “Food
pp. Habits and Diet Composition of
FRAD, CMFRI (2015). Marine fish Demersal Marine Fishes from Gulf of
landings in india (2014). Technical Mannar, Southeast Coast of India”
report, CMFRI, kochi. Food and Advances in Biological Research 6 (4):
agriculture organization of the united 159-164.
nationsRome, 2011. Pillai, V. N and Menon, N.G.
Gopalakrishnan., CMFRI (2015). Annual (2000).”Marine fisheries research and
Report 2014-15. Central Marine management .” Central marine fisheries
Fisheries Research Institute, Cochin, research institute tatapuram .p.o.,
353. Cochin. 628014 Kerala, India. Srinath.
Gopalakrishnan.A. And Imelda Joseph M. (2012). Marine Fishery Statistical
(2015). Mariculture research towards a System in India - Issues and
sustainable blue revolution in India. Approaches, CMFRI, cochin.
ICAR-Central Marine Fisheries Mohanraj.T and K. Prabhu., (2012). “Food
Research Institute Post Box No. 1603, Habits and Diet Composition of
Ernakulam North P. O., Kochi - 682 Demersal Marine Fishes from Gulf of
018, Kerala, India. Mannar, Southeast Coast of India”
James ,P.S.B.R. (1991). “Exploited and Advances in Biological Research 6 (4):
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MORPHO-ANATOMICAL STUDIES OF RED ALGAE GRACILARIA
FERGUSONII J. AG
P. Jenifer1, CP Balakrishnan1,* AND S Chidambaram Pillai2

1
Department of Botany, Aditanar College of Arts and Science, Virapandianpatnam, Tiruchendur
- 628 216, Tamil Nadu, India
2
PG and Research Department of Botany, V.O. Chidambaram College, Tuticorin- 628 008,
Tamil Nadu, India
*
Corresponding Author:sharubala08@gmail.com
ABSTRACT
Seaweeds are less consumed natural resource but abundantly available in the vast coastal
areaand are rich source of nutrients that can be used as important medicinal drugare extensively
used for treating variety of ailments in various system of medicine. Present study was made on
theanatomical characters, organoleptic evaluation and fluorescence characters of Rhodophycean
algae Gracilaria fergusonii J. Ag. Collected algal specimen was investigated using different
microscopic techniques (Micro techniques, SEM and CLSM) for observed anatomical features of
cell constituents. In micro technique, a clear distinct cell wall mucilaginous region and medulla
regions were observed under 10μm of toluidine blue staining. Whereas, SEM analysis revealed
that the clear cut image of multilayered cell wall galactans and inter cellular matrix also
observed. In Energy dispersive atomic X-ray microanalysis (EDAX) shows that the presence of
macro and trace elements in the cell wall composition.
KEYWORDS: Gracilaria fergusonii, Microtechniques, SEM, CLSM, EDAX
INTRODUCTION
Seaweeds are a commercially important, marine living renewable resource.
Commercially available varieties of marine macroalgae are commonly referred to as ‘Seaweeds’.
Macroalgae are classified as red algae (Rhodophyta), brown algae (Phaeophyta) or green algae
(Chlorophyta), depending on pigmentation, morphological and anatomical characters
(Manivannan et al., 2009). Cell wall polysaccharides occur in the structural elements of certain

seaweeds. In marine algae polysaccharide may constitute up to 70% of dry matter of some red
algae. Usually ester sulfate is associated with galactans are the polymer of α- L- and /or α-D or ß
– D- galactopyranosyl units in Rhodophyceae. Agarophytic Rhodophyceae member of
Gracilaria species are commonly available in the shallow coastal region of study area of
Manapad, Tamil Nadu, India. Cell wall polysaccharides and other cell constituents of Gracilaria
have immense bioactive substances for various ailments. The cell wall polysaccharide and
glycoprotein components found in plants have been well characterized structurally. The present
study was deals with the anatomy of red algae G. fergusonii using different microscopic
techniques.
Collection and preparation of seaweeds
Seaweed G. fergusoniiJ. Agardh was collected from the intertidal rocks of Manapad coast
of Tamil Nadu, India (8.3775°N; 78.0522°E) at low tide. Specimen was washed thoroughly in
seawater to remove extraneous matter such as epiphytes and sand. After collection, fresh samples
were taken into plastic jar and brought back to the laboratory immediately. Samples were
washed by tape water for several times, then gently brushed and rinsed with distilled water.
Fresh specimens were used for anatomical studies. The collected specimen was authenticated
and preserved in the form of herbaria. The voucher specimen (ACBHR33) was lodged in the
Department of Botany, Aditanar College of Arts and Science, Tiruchendur, Tamil Nadu.
Samples were washed by tape water for several times, then gently brushed and rinsed with
distilled water and then dried at room temperature. The dried seaweed powder was stored in
refrigerator for further uses.
Morpho-anatomical study
Macroscopic feature of fresh specimens were analyzed by using the keys given by
Umamaheswara Rao (1970) and (1987). In micro technique method, the fresh specimen was cut
into small pieces and fixed in FAA (Formalin: Acetic Acid: Alcohol) for 24 hours. After fixation
the specimen was washed thoroughly in distilled water simultaneously dehydrated using alcohol
and xylol for remove excess of water. Dehydrated specimen was embedded in paraffin wax
(Merck Product, 550C Melting Point) after infiltration (at 550C in Hot air oven) and sectioned
using Rotary Microtome to the thickness of 5 to 10 µm (Sass, 1940). Sectioned materials were

stained by using different standard stains via Toluidine Blue by O’ Brien et al., (1964) method
and Eosin Blue, Safranin and Crystal Violet by Johansen, (1940) method under downward and
upward series using alcohol and xylol at 1 minute interval. Stained specimens were mounted
using DPX (Distrene Plasticizer Xylene); is the most commonly used non fluorescent mountant
that preserve most routine stains and dries rapidly Culling, (1963). The mounted sections were
taken photomicrographs by using compound binocular microscope (Olympus CH20i) with built
in analogue camera and the help of Adobe Photoshop version 11.0 of computer.
Confocal Laser Scanning Microscopic (CLSM) study
The fresh unstained specimens were used for this study. The images were obtained by
LSM 710 Carl Zeiss CLSM instrument equipped with Brightfield and DIC imaging microscope
system. The specimen was excited with a range of 488 nm argon laser line at 25mW. For
fluorescent of cell constituents, green and red filters were used. Autofluorescent of
photosynthetic pigments were avoided by using wavelength ranging between 491 nm to 581nm.
Images were detected by LSM 710 Carl Zeiss CLSM equipped with 4 channels Zeiss Meta
(QUASAR) detector and processed with Zeiss ZEN 2009 Windows Vista software.
Scanning Electron Microscopic (SEM) and EDAX-Ray microanalysis
3 mm size of specimen fixed in 3% glutaraldehyde and 0.1 ml of phosphate buffer. After
fixation, the specimen was dehydrated through a graded series of alcohol for 5 minutes in each.
The dehydrated sectioned specimen examined under TESCAN-VEGA3-LMU-USA
instrument.The quantification of elemental analysis to identify the weight percentage of major
and minor elements presents in the samples was done using BRUKER Energy dispersive X-ray
Microanalysis (EDAX). The cross sectional area of the specimens was taken and the elements
were also identified.
Thallus of G. fergusoniiis cylindrical or flattened, branched stem, dark greenish red with
monopodial central axis consist bunches of dichotomous bulbous short braches at the apex and
are attached to the substratum by a sole organ or lack of attachment (Fig.1a). In micro technique,
a clear distinct cell wall region and medulla regions were observed under 10μm magnification of
Toluidine blue staining. The cell wall matrix appeared in dark purple in colour compare than

medulla regions (Fig. 1b). The sulphated polysaccharides of agarophytes and carrageenophytes
were metachromatic with toluidine blue O (Panikkar et al., 1991).
Confocal microscope is a powerful tool that creates sharp images of a specimen and
better contrast than that of a conventional microscope (Diaspro, 2002 and Hibbs, 2004). The cell
wall galactans possess highly electronegative charge density from their sulfated esters which
allow them electrostatic interactions with specific protein triggering in consequence their
biological effect. Here the images revealed that the cell wall has a more fluorescence than inter
cellular matrix (Fig. 1c and d). Due to the reason is some of the minerals from seawater may
absorb by osmotic as direct flow by ionic form Pillai, (1956). On other hand water soluble
minerals Ca, Mg and K and ionic salt Na have emit fluorescence. Evidence has been given by
Chapman (1950) showed that agar and algin exist in the form of Ca and Mg compounds. SEM
analysis revealed that the clear cut image of multilayered cell wall galactans and inter cellular
matrix of G. fergusonii. The insoluble galactans along with crystals were clearly showed under
higher magnification (Fig. 1 e and f). Morphology and anatomy of marine macro algae as early
studied under electron microscope by Clayton and Ashburnur (1990) and Balakrishnan et al.,
(2013).

Figure 1: Gracilaria fergusonii thallus (a) Entire view (b) 10μm magnification of
microscope view (c and d) CLSM view (e and f) SEM view
EDAX-Ray microanalysis
The results of the X-ray microanalysis show the presence of different chemical elements
in the cell wall of G. fergusonii(Fig.2).

cps/eV
K
S C O Na Mg Si S K Ca
2 Ca
0
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0
keV
Figure 2: Elemental composition under EDAX- ray microanalysis of G. fergusonii

There are eight elements such as carbon, oxide, sulphur, calcium, magnesium, sodium,
potassium, silica are present in this cell wall of the thallus. Thus the eight elements are
distribution from higher to lower rank are as following order: C>O>S>Ca>Mg>Na>K>Si.
Weights of these eight elements are given as % values. They are 43.45% of carbon, 48.25% of
oxide, 4.69% of sulphur, 2.70% of calcium, 0.60% of magnesium, 0.15% of sodium, 0.10% of
potassium and 0.06% silica respectively (Table -1). All seaweeds contain large amounts of both
macro-minerals (Ca, Mg, Na, P and K) and trace elements (Zn, I and Mn) (Matanjun et al., 2009
and Polat and Ozogul, 2009). Seaweeds generally contain 8–40% of minerals, and the essential
minerals and trace elements needed for human nutrition are present in seaweeds (Mabeau and
Fleurence 1993).
Table 1: Shows the chemical elements present in the G. fergusonii thallus

El AN Series unn. Unn. C norm. C Atom. C Error (1
[wt.%] [wt.%] [wt.%] [at.%] Sigma)
[wt.%]
C 6 K- series 43.45 43.45 52.56 5.57

O 8 K-series 48.25 48.25 43.82 6.11
S 16 K-series 4.69 4.69 2.12 0.20
Ca 20 K-series 2.70 2.70 0.98 0.11
Mg 12 K-series 0.60 0.60 0.36 0.06
Na 11 K-series 0.15 0.15 0.09 0.04
K 19 K-series 0.10 0.10 0.04 0.03
Si 14 K-series 0.06 0.06 0.03 0.03
Total 100.00 100.00 100.00

ACKNOWLEDGEMENT
The authors gratefully acknowledge the University Grants Commission (UGC), New
Delhi, India, for the financial assistance of this project (Ref. No. 42-935/2013) under MRP
scheme. The authors also thanks to Dr. N. Thajuddin, Professor and Head, Dept of Microbiology,
Bharathidasan University, Tirchy for providing CLSM facility to carry out the anatomical
studies.
REFERENCE
Balakrishnan, C.P., P. Jenifer and M. Hibbs, A.R 2004, (Ed). Confocal
Esakkilingam 2013, Algal Microscopy for Biologists, New York:
documentation and Phytochemical kluwer Press.
studies of red algae Gracilaria corticata Johansen, D.A 1940, Plant Microtechnique.
of Manapad Coast, Tamil Nadu. Journal 1st Edn, McGraw -Hill Book Company,
of Pharmacognosy and Phytochemistry., New York and London, 182-203.
2(4): 193-197. Mabeau, S., Fleurence J., 1993, Seaweed in
Chapman, V.J 1950, Seaweeds and their food products: Biochemical and
uses. The Camelot Press Ltd., Methuen nutritional aspects, Trends Food Sci.
and Co. Ltd., London and Sothampton. 2 Tech., 4, 103–107.
nd ed. 63-85. Manivannan, K, Thirumaran G, Karthikai
Clayton, M.N and C.M. Ashburnur 1990, Devi G, Anantharaman P,
The anatomy and ultrastructure of Balasubramanian T 2009, Proximate
conducting channels in Ascoseira composition of different groups of
mirabilis (Ascosetirales, Pheophyceae). seaweeds from Vedalai coastal waters
Bot. Mar., 30: 63-70. (Gulf of Mannar): southeast coast of
Culling, C.F.A 1963, Handbook of India, Middle-East, J. Sci. Res., 4 (2):
Histopathological Techniques, 2nd ed. 72–77.
Butterworths, London. Matanjun, P., Mohamed S., Mustapha N.M.,
Diaspro, A 2002, (Ed). Confocal and two - Muhammad K., 2009, Nutrient content
Photon Microscopy: Foundations, of tropical edible seaweeds, Eucheuma
Applications, and Advances. New York: cottonii, Caulerpa lentillifera and
Wiley-Liss.

Sargassum polycystum, J. Appl. Phycol., Polat, S., Ozogul Y., 2009, Fatty acid,
21 (1), 1–6. mineral and proximate composition of
O'Brien, TP, N. Feder, M.E. McNull 1964, some seaweed from the northeastern
Polychromatic staining of plant cell Mediterranean coast, Ital. J. Food Sci.,
walls by Toluidine blue-O, 21, 317–324.
Protoplasma., 59: 364-373. Sass, JE 1940.Elements of Botanical
Panikkar, M.V.N, P. Ampli and V.D. Microtechnique. McGraw Hill Book Co,
Chauhan 1991, Histochemical New York.
localization of polysaccharides in the Umamaheswara Rao, M 1987, Key for
tissue of three Indian marine algae. identification of economically important
Phykos., 30 (1&2): 61-66. seaweeds. Bull. Cent. Mar. Fish. Res.
Pillai, V.K 1956, Chemical studies on Indian Inst., 41: 19-25.
seaweeds. I: Mineral constituents. Umamaheswara Rao, M 1970. The
Proceeding of Indian Academic Science, economic seaweed of India. Bull. Cent.
B 45: 3-29. Mar. Fish. Res. Inst. 20: 1-68.

PHARMACOGNOSTIC STANDARDIZATION AND UV SPECTRUM
ANALYSIS OF MARINE RED ALGAE GRACILARIA SPECIES OF
MANAPAD COAST, TAMIL NADU
P Jenifer1, CP balakrishnan1,* and S Chidambaram Pillai2

1
Department of Botany, Aditanar College of Arts and Science, Virapandianpatnam, Tiruchendur
- 628 216, Tamil Nadu, India
2
PG and Research Department of Botany, V.O. Chidambaram College, Tuticorin- 628 008,
Tamil Nadu, India
*
Corresponding Author: sharubala08@gmail.com
ABSTRACT
The present study was made on organoleptic evaluation, fluorescence characters and UV
spectrum analysis of marine red algae Gracilaria species of Manapad Coast, Tamil Nadu. The
quality and purity of algal drug standardized by methods such as organoleptic, fluorescence and
UV spectrum analysis using with different solvent and reagents were recorded. The results of
UV spectrum profile showed different bands that were confirmed the presence of phenols,
organic acids, lipids and related compounds. Pharmacognostic evaluation of this study helps to
identify the phytoconstituents of the marine macro algae Gracilaria species.
KEYWORDS: Gracilaria species, organoleptic, fluorescence, UV spectrum
INTRODUCTION
In the field of pharmaceutical research, the analytical investigation of bulk drug
materials, intermediates, drug products, drug formulations, impurities and degradation products,
and biological samples containing the drugs and their metabolites is very important.
Pharmacognosy is a simple and reliable tool, by which complete information of the crude drug
can be obtained (Padmavathi et al., 2011). The process of standardization can be achieved by
stepwise pharmacognostic studies that help in identification and authentication of a plant
material (Thomas et al., 2008). Correct identification and quality assurance of the raw material
are the important prerequisites in herbal therapy to ensure its quality, efficacy and safety (Nayak

and Patel, 2010). The marine algae have been closely associated with human life and are being
exhaustively used in numerous ways as a source of food, feed, fertilizer, medicine and chiefly for
economically important phycocolloide (Darcy- Vrillon, 1993). The present research work deals
with Pharmacognostic standardization and UV spectrum analysis of marine red algae Gracilaria
species of Manapad Coast, Tamil Nadu.
Marine red algae Gracilaria species such as Gracilaria fergusonii, G. foliifera and G.
edulis were collected from Manapad coast of Tamil Nadu, India (8.3775°N; 78.0522°E) at low
tide. Specimen was washed thoroughly in seawater to remove extraneous matter such as
epiphytes and sand. After collection, fresh samples were taken into plastic jar and brought back
to the laboratory immediately. Samples were washed by tape water for several times, then gently
brushed and rinsed with distilled water and then dried at room temperature. The dried seaweed
powder was stored in refrigerator for further uses.
Organoleptic evaluation
The organoleptic characteristics of algal powdered sample their appearance and colour in
day light, nature, color, odour, shape (under microscopic), taste and texture etc were also studies.
Organoleptic evaluation of seaweed powder was carried out according to the methods of Kokate
et al., (2005) and Kumar et al., (2012).
Fluorescence analysis
Fluorescence analysis is a fundamental parameter for the first line standardization of
crude drug. Fluorescence characteristic of the seaweed powder was treated with different solvent
and reagents (Chloroform, Petroleum ether, benzene, acetone, distilled H2O, xylene, HCL,
HNO3, H2SO4, NaOH, Acetic acid, FeCl3, Ammonia solution, K2Cr2O7, iodine solution,) and
colour changes was observed under ordinary/visible light and ultra violet light by using
HORIZONTAL LAMINAR FLOW-TECHNICO LAB FURNITURE AND SYSTEMS (MM
WG.SP 0.82) instrument. Fluorescence of analysis of the seaweed powder were carried out
according to the methods of Chase and Pratt (1949), Kokoshi et al., (1958) and Kumar et al.,
(2012).

UV Spectrum Analysis
The methanolic algal extract of Gracilaria species was examined under UV spectral
analysis. The extract was centrifuged at 3000 rpm for 10 min and filtered through Whatmann No.
1 filter paper by using high pressure vacuum pump. The sample is diluted to 1:10 with the same
solvent. The methanolic extracts were scanned in the wavelength ranging from 200-300 nm
using UV-Spectrophotometer and the characteristic peaks were detected.
The nature of coarse powder of the seaweed Gracilaria species were evaluated on
organoleptic characters. The observations for nature, odour, taste, texture, colour and shape were
noted for different species of Gracilaria and are tabulated (Table -1). Organoleptic study forms
an important part of powder analysis and is a technique for the qualitative evaluation based on
the study of morphological and sensory profiles of whole drugs (Kokate et al., 2007).
Fluorescence study is an essential parameter for first line standardization of crude drug. The
results of Fluorescence studies of dried powder algae Gracilaria species are treating with various
solvent and reagents under visible and UV light has been recorded in Table -2. Fluorescence is
the phenomenon exhibited by various chemical constituents present in the plant material. Some
constituents show fluorescence in the visible range in daylight.
Table 1: Organoleptic evaluation of marine red algae Gracilaria species
S. Observation
Characters
No. G. fergusonii G. foliifera G. edulis
1 Nature Coarse powder Coarse powder Coarse powder
2 Odour Characteristic Characteristic Characteristic
3 Taste Tasteless Tasteless Tasteless
Texture
4 (microscopic Rough Rough Rough
observation)
Colour
Pink with light Reddish pink with Yellowish pink/ dark
5 (microscopic
green green red
observation)
Shape Ellipsoidal/ Ellipsoidal/ Triangles/
Ellipsoidal/
6 (microscopic Pentagonal/ Rhombus/ Pyramid/ rounded/
Triangles/ curved
observation) Rounded/ Curved Curved
Hence, some crude drugs are often assessed qualitatively in this way and it is an important
parameter of Pharmacognostical evaluation (Kumar et al., 2012 and Gupta et al., 2006). The

fluorescent method is adequately sensitive and enables the precise and accurate determination of
the analyzed over a satisfactory concentration range without several time consuming dilution
steps prior to analysis of pharmaceutical samples (Pimenta et al., 2006).

Table 2: Fluorescence analysis of powered drug of marine red algae Gracilaria species
G. fergusonii G. foliifera G. edulis
S. Powdered drug Under
Under ordinary / Under UV Under ordinary Under UV
No with Reagent ordinary / Under UV light
visible light light / visible light light
visible light
Greenish Greenish
1 Powder as such Brown Dark green Light green Light green
brown brown
Powder + Conc. Yellowish Yellowish
2 Light brown Dark brown Greenish brown Dark green
HCL green brown
Powder + 50% Reddish Yellowish Yellowish Yellowish
3 Brownish pink Yellowish green
HCL black green brown green
Powder + Conc. Yellowish Yellowish
4 Yellowish green Yellowish green Pale yellow Pale yellow
HNO3 brown green
Powder +50 % Yellowish
5 Yellowish green Pale yellow Yellowish brown Pale yellow Pale yellow
HNO3 brown
Dark
Powder + Conc. Greenish
6 Greenish red greenish Greenish red Greenish black Brown
H2SO4 black
brown
Powder + 50% Greenish Greenish
7 Brown Dark Brown Greenish brown Brown
H2SO4 brown brown
Powder + 1N
Greenish Greenish
8 NaOH Brown Dark green Greenish brown Brown
brown brown
(Aqueous)
Powder +1N Dark reddish Greenish
9 Brownish white Brown Dark brown Greenish brown
NaOH (Alcohol) brown brown

Powder + 10%
Yellowish Yellowish
10 Glacial Acetic Pale brown Brown Blackish brown Pale brown
brown brown
acid
Powder + 50% Brownish Dark yellowish Yellowish
11 Brownish black Black Brown
FeCl3 yellow brown brown
Powder +
Yellowish Yellowish
12 Ammonia Brownish yellow Green Dark green Green
brown brown
solution
Powder +10 % Yellowish Blackish Reddish
13 Yellowish brown yellowish brown Reddish brown
K2Cr2O7 brown brown brown
Powder + 5% Blackish Yellowish
14 Pale brown Black yellowish brown Black
Iodine solution yellow brown
Powder + Greenish Blackish
15 Brown Black Black Black
Xylene brown yellow
Powder + Yellowish Blackish
16 Brown Black Black Brown
Chloroform black green
Powder + Yellowish
17 Brown Black Black Blackish brown Dark brown
Petroleum ether brown
Powder Yellowish Yellowish
18 Brown Black Blackish brown Dark brown
+Benzene brown brown
Powder + Blackish light
19 Brown Black Brown Black Black
Acetone green
Powder + Yellowish Greenish
20 Brown Brown Greenish brown Pale brown
Distilled water brown brown

G. fergusonii G. foliifera
0.45 0.40
0.38
229
0.40
0.36
253 259
absorbance
absorbance
0.34
266
0.35
0.32
0.30
0.30
0.28
0.25 0.26
200 220 240 260 280 300 200 220 240 260 280 300
wave length nm wave length nm
0.21
0.20
0.19
230 255 271
0.18
absorbance
0.17
0.16 210
0.15
0.14
0.13
200 220 240 260 280

Wave length nm
G. edulis
Fig 3: Ultra violet spectrum of methanol extract of Gracilaria species
The UV spectroscopic analysis of the methanolic extract of Gracilaria species were

studied at a wavelength range from 200 to 300 nm. Band was recorded with
absorbance values (Fig 1a-c). The result of UV spectrum profile confirms the
presence of phenols, organic acids, lipids and related compounds in the methanolic
extract of different Gracilaria species.
CONCLUSION
It can be concluded that the standardization of Gracilaria species using
parameter such as organoleptic, fluorescence and UV spectrum analysis plays an
important role in the determination of quality and purity of the drug. It will also
helpful to carry out further research and revalidation of its use in traditional system of
medicine.
ACKNOWLEDGEMENT
The authors are gratefully acknowledges the University Grants Commission
(UGC), New Delhi for the financial assistance of this project (Ref. No. 42-935/2013)
under MRP scheme.

REFERENCE
Chase, CR. and RF. Pratt 1949, radiation. J. Amer. Pharm. Assn.,
Fluorescence of powdered 38(10): 715-717.
vegetable drugs with particular Kumar, D, Gupta J, Kumar S, Arya R,
reference to the development of Kumar T, Gupta A 2012,
systems of identification, J. Pharmacognostic evaluation of
American Pharm. Assoc. 38: 324- Cayratia trifolia (Linn.) leaf. Asian
333. Pac J Trop Biomed., 2(1): 6-10.
Darcy- Villon, B 1993, Nutritional Nayak, BS, Patel KN 2010,
aspect of the developing use of Pharmacognostic studies of the
marine macro algae for the human Jatropha curcas leaves, Int J
food industry. Int.J. food sci. Nutr. Pharm tech Res., 2(1): 140-3.
44:S23-S35. Padmavathi, D, Susheela L, Bharathi
Gupta, MK, Sharma PK, Ansari SH, RV 2011, Pharmacognostical
Lagarkha R 2006, evaluation of Barringtonia
Pharmacognostical evaluation of acutangula leaf, Int J Ayurveda
Grewia asiatica fruits, Int J Plant Res., 2(1): 37-41.
Science., 1(2): 249-251. Pimenta, AM, Montenegio MC, Ara
Kokate, CK 2005, Practical Ujo AN, Mart'inez JC 2006,
pharmacognosy, New Delhi: Application of sequential injections
Vallabh Prakashan p. 107, 108, analysis. J. Pharm. Biomed. Anul.,
115-120, 122-123. 40: 16-34.
Kokate, CK, Purohit AP, Gokhale SB Thomas, S, Patil DA, Patil AG,
2007, Pharmacognosy. Nirali Chandra N 2008, Pharmacognostic
Prakashan; 38th edition, Pune. evaluation and physicochemical
Kokoshi, CJ, Kokoshi RJ 1958 Sharma analysis of Averrhoa carambola L.
M. Fluorescence of powdered fruit. J Herb Med Tox., 2(2): 51-4.
vegetable drugs in ultra violet

STUDIES OF PHYSICOCHEMICAL AND BIOCHEMICAL COMPOSITION
OF MARINE RED ALGAE GRACILARIA FERGUSONII J. AG
P Jenifer1, CP Balakrishnan1,* and S Chidambaram Pillai2

1
Department of Botany, Aditanar College of Arts and Science, Virapandianpatnam,
Tiruchendur - 628 216, Tamil Nadu, India
2
PG and Research Department of Botany, V.O. Chidambaram College, Tuticorin- 628
008, Tamil Nadu, India
*
Corresponding Author: sharubala08@gmail.com
ABSTRACT
The present study deals with physicochemical and biochemical analysis
of Rhodophycean algae Gracilaria fergusonii J. Ag was collected from Manapad
coast, Tamil Nadu, India. Collected algal specimen was investigated proximate and
chemical composition were using standard procedure. The moisture content, total ash
value, acid insoluble and water soluble ash value was 24.6±0.324%, 36.61±0.69%,
24.24±0.19%, and 26.69±0.02% were respectively. The solvent extractive value and
other biochemical composition were also recorded.
KEYWORDS: Gracilaria fergusonii, physicochemical, biochemical, vitamins,
pigments
INTRODUCTION
Seaweeds are reported to be rich in soluble dietary fibres, proteins, minerals,
vitamins, antioxidants, phytochemicals and polyunsaturated fatty acids, with a low
caloric value (Mohamed et al., 2012). Commercially available varieties of marine
macroalgae are commonly referred to as ‘seaweeds’. Macroalgae are classified as red
algae (Rhodophyta), brown algae (Phaeophyta) or green algae (Chlorophyta),
depending on pigmentation, morphological and anatomical characters (Manivannan et
al., 2009). Red and brown algae are mainly used as human food sources (Dawczynski
et al., 2007). In marine algae polysaccharide may constitute up to 70% of dry matter
of some red algae. Seaweeds are known as valuable sources of protein, elements,
dietary fibers, vitamins, essential amino acids and essential fatty acids. Moreover,
seaweeds also contain potential bio-active compounds which exhibit antibacterial,

antiviral and antifungal properties (Marinho-Soriano et al., 2006). Agarophytic
Rhodophyceae member of Gracilaria species are commonly available in the shallow
coastal region of study area of Manapad, Tamil Nadu, India.The present study was
deals with the physicochemical and biochemical composition of red algae G.
fergusonii.
Seaweed G. fergusoniiJ. Agardh was collected from the intertidal rocks of
Manapad coast of Tamil Nadu, India (8.3775°N; 78.0522°E) at low tide. Specimen
was washed thoroughly in seawater to remove extraneous matter such as epiphytes
and sand. After collection, fresh samples were taken into plastic jar and brought back
to the laboratory immediately. Samples were washed by tape water for several times,
then gently brushed and rinsed with distilled water and then dried at room
temperature. The dried seaweed powder was stored in refrigerator for further uses.
Physicochemical analysis
The dried powdered sample was used to physicochemical analysis.
Physicochemical parameters such as total ash value, acid insoluble ash value, water
soluble ash value, different solvent (petroleum ether, benzene, chloroform, acetone
and methanol) extractive values, and moisture content were determined as per method
described Anonymous (1996) and crude fibre were determined (Sadasivam and
Manickam, 1992).
Quantitative estimation
The dried powder sample of G. fergusonii was used to biochemical analysis
such as estimation of carbohydrate, protein, amino acid, lipids, total free
phenols,tannins, organic carbon, vitamin A, B3, C, and pigment analysis such as
estimation of anthocyanin, ß carotene,chlorophyll a, b, total chlorophyll and
carotenoids were carried out using standard methods. The total carbohydrate was
estimated by anthrone method (Sheifter et al., 1950). Protein was estimated by
Lowry’s method (Lowry et al., 1951). Amino acid was estimated by ninhydrin
method as suggested by Rosen (1957). Lipids were estimated by Bligh and Dyer
(1959). Other biochemical’s like estimations of total free phenols by Folin-
Ciocalteu’s method (Sadasivam and Manickam, 1992), tannin was vanillin-HCl
method by Burns (1971), Vitamin A (Eitenmillar and Landen, 1998), Vitamin B3 and

C (Sadasivam and Manickam, 1992)was carried out, Organic carbon by Walkley and
Black (1943), and photosynthetic pigments like chlorophyll a, b, total chlorophyll and
carotenoids by Arnon (1949) as modified by Harborne (1973) method, anthocyanins
(Vivekanadan, 1991), and ß Carotene by AACC (1995) methods are followed.
Physicochemical analysis
The physicochemical parameters are mainly used in judging the purity and
quality of the drug. In this study, Quantification of physicochemical parameters such
as moisture content, total ash value, acid insoluble ash value, water soluble ash value,
different solvent (petroleum ether, benzene, chloroform, acetone and methanol)
extractive value and crude fibre were showed in Table 1. The moisture content, total
ash value, acid insoluble and water soluble ash value of G. fergusonii was
24.6±0.324%, 36.61±0.69%, 24.24±0.19%, and 26.69±0.02% were respectively. Ash
analysis showed high mineral contents that might be the result of the absorption of
inorganic salt from seawater or of the association of cations with algal
polysaccharides (Lahaye, 1991). Then the solvent extractive value like petroleum
ether, benzene, chloroform, acetone, methanol and water was 0.40±0.01%,
0.55±0.02%, 0.25±0.02%, 0.27±0.008%, 1.66±0.05% and 4.24±0.03% respectively.
Extractive value is useful for the evaluation of a crude drug as it gives idea about the
nature of chemical constituents present in it and is useful for estimation of chemical
constituents, soluble in that particular solvent used for extraction (Lincy and Mathew,
2011).
The physicochemical results suggest that the seaweed have high extractive
value in methanol and water extract than that of other extract. Seaweeds were rich in
dietary fiber (>50% dry weight), particularly in the soluble form (Mabeau and
Fleurence, 1993). The crude fiber content of G. fergusonii was 87.8±1.485%
recorded. The consumption of dietary fibers and plant cell walls containing such fiber
components protects human organisms against a number of chronic diseases e.g.,
colon cancer (Guidel and Goni, 2002).
Biochemical Analysis
The nutritional value of a food depends on its chemical composition such as
carbohydrates, proteins, lipids, sugars, and also the minerals present in them.
Knowledge of the chemical composition of marine macro algae is important for the

nutritional value and evaluation of potential sources of protein, carbohydrate and lipid
for commercial use or for possible human consumption (Chapman and Chapman,
1980). Biochemical such as carbohydrate, protein, amino acid, lipid, total free
phenols,tannins, organic carbon, vitamin A, B3, C, pigment analysis such as
estimation of anthocyanin, ß Carotene,chlorophyll a, b, total chlorophyll and
carotenoids were determined present in Table 2 and 3. Carbohydrates are the most
abundant substances in most seaweeds, since they occur in cell wall (ex.: agar,
cellulose) and as storage products (ex.: starch, laminaran). The dry weight of primary
metabolites carbohydrate contain 0.160±0.014mg/g, protein was 0.62±0.03 mg/g,
amino acid was 0.257±0.0015 mg/g and lipid was 0.373±0.003 mg/g were present in
red algae G. fergusonii. Carbohydrate is one of the important components for
metabolism and it supplies the energy needed for respiration and other most important
processes. Edible seaweeds contain significant quantities of protein, lipids, minerals
and vitamins (Norziah and Ching, 2002). The secondary metabolites like total free
phenols andtannins were 3.08±0.022 and 0.473±0.029 mg/g respectively. The
organic carbon content of G. fergusonii was 2.331±0.016mg/g recorded. Whereas
vitamins like A, B3, and C content via 54.31±0.27, 50.81± 0.89 and 3.790± 0.431
were respectively.
The photosynthetic pigments like chlorophyll a, chlorophyll b, total
chlorophyll, carotinoids, fresh weight content of red algae G. fergusonii was
0.088±0.008 mg/g, 0.127±0.011 mg/g, 0.219±0.016 mg/g, 56.33±1.08mg/g, and dry
weight content of anthocyanin and ß Carotene was 0.0133±0.002 mg/g, and
0.851±0.009 mg/g were respectively. Many studies have been performed worldwide
to characterize algae species according to their chemical composition (Norziah and
Ching, 2002, Abdallah et al., 2006 and Kamenarska et al., 2002). Marine resources
offer important bioactive molecules that have advantages on the human body. They
can be applied in many fields such as the drug, cosmetic, and food industries.
Functional foods can easily be developed from marine products since they are widely
available and they have the ability to prevent certain diseases and cure some illnesses.

Table 1: The weight percentage of moisture content, ash values, different
solvents soluble extractive values and crude fibre of red algae G. fergusonii
Sample Gracilaria fergusonii
Moisture content% 24.6±0.324
Total ash % 36.61±0.69
Acid insoluble ash % 24.24±0.19
Water soluble ash % 26.69±0.02
Petroleum ether % 0.40±0.01
Benzene % 0.55±0.02
Chloroform % 0.25±0.02
Acetone % 0.27±0.008
Methanol % 1.66±0.05
Water % 4.24±0.03
Crude fibre % (dry wt.) 87.8±1.485
The results are expressed as mean value ± standard deviation
Table 2: Estimation of biochemical compositions of powdered drug G. fergusonii
(dry wt)
Sample G. fergusonii
Carbohydrate (mg/g dry wt) 0.160±0.014
Protein (mg/g dry wt) 0.62±0.03
Amino Acid (mg/g dry wt) 0.257±0.0015
Lipid (mg/g dry wt) 0.373±0.003
Total free Phenols (mg/g dry wt) 3.08±0.022
Tannins (mg/g dry wt) 0.473±0.029
Organic Carbon % (dry wt) 2.331±0.016
Vitamins A (mg/100g dry wt.) 54.31±0.27
Vitamins B3 (mg/100g dry wt.) 50.81±0.89
Vitamins C (mg/100g dry wt.) 3.790±0.431
Table 3: Estimation of photosynthetic pigments analysis of G. fergusonii
Sample G. fergusonii
Chlorophyll a (mg g-1 fr. wt) 0.088±0.008
Chlorophyll b (mg g-1 fr. wt) 0.127±0.011
Total Chlorophyll (mg g-1 fr. wt) 0.219±0.016
-1
Carotinoids (mg g fr. wt.) 56.33±1.08
Anthocyanin (mg/g dry wt) 0.0133±0.002
ß Carotene (mg/g dry wt) 0.851±0.009

ACKNOWLEDGEMENT
The authors are gratefully acknowledges the University Grants Commission
(UGC), New Delhi, India, for the financial assistance of this project (Ref. No. 42-
935/2013) under MRP scheme.
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A COMPARISON OF THE EFFECTS OF COBALOXIMES ON
ANTIBACTERIAL AND ANTIFUNGAL PROPERTIES
M. Amutha selvi a *, C. Balachanderb, A. Dayalanc, S. Lingathuraid AND S.

Ignacimuthub
a
Department of Chemistry, Avinashilingam Institute for Home Science and Higher
Education for Women University, Coimbatore, Tamil Nadu, India
b
Entomology Research Institute, Loyola College, Chennai, Tamil Nadu, India
c
PG & Research Department of Chemistry, Loyola College, Chennai, Tamil Nadu,
India
d
Thiruchedur, Tamil Nadu, India
ABSTRACT
Cobaloximes were prepared using dimethylglyoxime as equatorial network and
pyridine derivatives as axial ligands and characterized by the spectroscopic
techniques and were also screened against microorganism for antimicrobial activity
with Streptomycin as standard. The cobaloximes containing iodine were found to be
active against most of the microbes.
KEYWORDS: Cobaloximes, antimicrobial activity of cobalt (III) complexes,

Halocobaloximes
INTRODUCTION
Organocobaloximes, originally proposed as models of Vitamin B12 nearly
three decades ago, have been studied extensively and reviewed (Bresciani Pahor et al.,
1985). In the B12 field, a lot of protein-free corinoids or alkyl cobalt complexes
(Randaccio, 1989 and Calligaris, 1972) have been synthesized as models of the
cofactor due to the flexibility of their equatorial oxime ligands is quite similar to that
of corrin in natural cofactor. The difficulty of obtaining large amounts of pure natural
products, high molecular weights, extreme molecular complexity and limited
solubility make the elucidation of the chemical behavior of the metal ion difficulty
when compounds of biological origin are studied directly.

Model studies in chemical research are to understand the biological roles of
metal ions. Complexes with dimethylglyoxime (dmgH2) as equatorial ligand undergo
the most extensive investigations of models of Vitamin B12. (Calligaris, 1972).
Cobalt(III) Complexes are an important class of coordination compound not only
because of them being interesting B12 models and oxygen carriers, but also due to
their interesting magnetic properties, spectroscopic characteristics and the interaction
of metal-to ligand bonds. Co(III) is also found in certain cobalt-porphyrin containing
proteins (Hatchikian, 1981). A number of examples of stable Co(III) complexes have
also been reported. Although polydentate ligands with N, O and S donor atoms in the
coordination sphere of cobalt are the most common ligands used to stabilize Co 3+ ion.
Herein we describe the synthesis and characterization of the novel Co(III) complexes
as the potential candidates for antimicrobial activity against standard.

Cobalt(II) chloride, dimethyl glyoxime, various substituted pyridines
(purchased from SD Research Laboratory), Cobalt (II) Bromide (purchased from
sigma Aldrich) were used for the preparations of the complexes. Dimethyl sulpoxide
was dried over calcium hydride and distilled at reduced pressure. The distilled
solvent, stored under molecular sieves, was used for the study of the antimicrobial
activity.
Preparation of the cobaloximes
Preparation of choloro and bromo complexes
The green dihalo cobalt complex: H[Co(dmgH) 2Cl2], H[Co(dmgH)2Br2], was
prepared as reported in the literature (Costa, et al, 1967., Jothi, et al, 2008). About
0.005 moles of the above complex, in ethanol, was stirred for 10 minutes with 0.005
moles of the ligands (viz., 4-hydroxy pyridine, 4-methyl pyridine, 4-N,N’-
dimethylamino pyridine) in different lots. The reaction mixtures were refluxed for
about 2-3 hrs at 40oC and allowed to settle for 1 hr, after which the brown colored
products were collected in a glass sintered crucible, washed successively with ethanol,
ether and finally dried in vacuum desiccators.
Preparation of iodo complexes
The green colored dichloro complex, [Co(dmg)(dmgH2)Cl2], (0.01 mole) was
placed in 75 ml of water taken in a conical flask. The slurry was stirred for 10 min,

mixed with 0.01 mole of potassium iodide. The reaction mixture was stirred for about
an hour at 60ºC. The green color turned into brown color solution indicating the
formation of aquoiodocomplex (Tucker et al., 2004)
To 0.01 mole of the light brown colored aquoiodocomplex, viz.,
[Co(dmgH)2(H2O)I], in 50 ml water 0.005 mole of ligands in different lots. The
reaction mixtures were stirred for about an hour at 60°C on a water bath. The light
brown color turned into dark brown. It was allowed to stand for an hour and filtered
through sintered crucible. The dark brown color complex formed was washed with
alcohol, ether and dried over vacuum desiccators (Yield: 80%).
The prepared complexes are 1. [Co(dmgH)2(HP)Cl] 2. [Co(dmgH)2(HP)Br]
3.[Co(dmgH)2(HP)I] 4.[Co(dmgH)2(MP)Cl] 5. [Co(dmgH)2(MP)Br] 6.
[Co(dmgH)2(MP)I]
7. [Co(dmgH)2(DMAP)Cl] 8. [Co(dmgH)2(DMAP)Br] 9. [Co(dmgH)2(DMAP)I]
Microbial organisms
The following bacteria and fungi were used for the experiment. Bacteria:
Salmonella paratyphi-B, Klebsiella pneumoniae MTCC 109, Proteus vulgaris MTCC
1771, Shigella flexneri MTCC 1457, Micrococcus luteus, Enterobacter aerogenes
MTCC 111, Staphylococcus aureus MTCC 96, Staphylococcus aureus (MRSA-
methicillin resistant). The reference cultures were obtained from Institute of Microbial
Technology (IMTECH), Chandigarh, India -160 036; fungi: Candida albicans MTCC
227, Malassesia pachydermatis,Trichophyton mentagrophytes 66/01, Trichophyton
rubrum 57/01 and Aspergillus flavus. All the other cultures were obtained from the
Department of Microbiology, Christian Medical College, Vellore, Tamil Nadu, India.
Antimicrobial assay
Antibacterial and antifungal activities were carried out using disc-diffusion
method (Murray et al., 1995). Petri plates were prepared with 20 ml of sterile Mueller
Hinton agar (MHA) (Hi-media, Mumbai). The test cultures were swabbed on the top
of the solidified media and allowed to dry for 10 min and a specific amount of crude
extract at three different concentrations (1, 2 and 3 mg/disc) and the compound at 250
µg/disc was added to each disc separately. The loaded discs were placed on the
surface of the medium and left for 30 min at room temperature for compound

diffusion. Negative control was prepared using respective solvents. Streptomycin
(10µg/disc) was used as positive control against bacteria. Ketoconazole was used as
positive control for fungi. The plates were incubated for 24 h at 37°C for bacteria and
for 48 h at 28°C for fungi. Zones of inhibition were recorded in millimetres and the
experiment was repeated twice.
Minimum Inhibitory Concentration (MIC)

Minimum inhibitory concentration studies of the isolated compound were
performed according to the standard reference methods for bacteria (Duraipandiyan
and Ignacimuthu, 2009), for filamentous fungi (CLSI, 2008) and yeasts
(NCCLS/CLSI, 2002). The required concentrations (1000 µg/ml, 500 µg/ml,
250µg/ml, 125µg/ml, 62.5 µg/ml, 31.25µg/ml and 15.62µg/ml) of the compound were
dissolved in DMSO (2%), diluted to give serial two-fold dilutions that were added to
each medium in 96 well plates.
An inoculum of 100 from each well was inoculated. The antifungal agents
Ketoconazole for fungi and Streptomycin for bacteria were included in the assays as
positive controls. For fungi, the plates were incubated for 48 to 72 hours at 28°C and
for bacteria the plates were incubated for 24 h at 37°C. The MIC for fungi was
defined as the lowest extract concentration, showing no visible fungal growth after
incubation time. 5 µl of tested broth was placed on the sterile MHA plates for bacteria
and incubated at respective temperature. The MIC for bacteria was determined as the
lowest concentration of the compound inhibiting the visual growth of the test cultures
on the agar plate.

Spectral Studies
The IR spectra were obtained using PERKIN ELMER FTIR
spectrophotometer using KBr pellets
FT-IR spectra
All the complexes showed of a peak in the range of 490-510 cm–1 indicates the
formation of Co-Naxial bonds. The C=N stretching characteristic of oxime in its
complexes was observed around 1400 cm-1 and the intra molecular hydrogen bonded
–OH around 3400 cm-1 (Figure 1).

Figure 1 FT-IR Spectrum of the complex [Co(dH) 2(DMAP)Cl]
A sharp peak around 1240 cm-1 may be attributed to the –N-O stretching of
dimethylglyoximate. The –C=N- and -N-O- stretching vibrations of dmgH2 are
shifted to lesser frequencies (from 1500 cm-1 to 1385 cm-1) on complexation
(Mojumdar et al., 1998).
Antimicrobial activity
All the complexes exhibited moderate zone of inhibition towards tested

bacteria. All the nine complexes showed high zone of inhibition against
staphylococcus aureus compare to the standard. The complex 1, 3 and 9 alone showed
activity against Salmonella paratyphi-B, and all other complexes showed nil activity.
The complexes 5 and 6 showed nil activity against Proteus vulgaris (Plate 1). The
complex 9, which is iodo cobaloxime containing 4- N,N’dimethylaminopyridine as
axial ligand showed good zone of inhibition towards all the bacteria (Table 1).
Among all the complexes the complexes containing 4-N’dimethylaminopyridine as
axial ligand showed good activity when compare to 4-Hydroxypyridine and 4-
Methylpyridine.
Bacterial growth inhibitions were identified with MIC values. Based on our
results were expressed as µg/ml (Table 2). The MIC studies revealed that, most of the
complexes showed activity greater the concentration of control. The MIC values for
were in the range of 125 to 250 µg/ml for the complex of 7 and in the range of 125 to
500 µg/ml for complex 6. The results of our study showed that complex 7 had higher
antimicrobial effects. Complex 7 showed effective inhibition against Shigella flexneri,
Micrococcus luteus, Staphylococcus aureus and Staphylococcus aureus (MRSA)
bacterial strains at very minimum concentration (125 µg/ml).

Table -1 Antimicrobial activity of complexes (Zone of inhibition in mm)
(1µg/disc)
[Co(dmgH)2(DMAP)C
[Co(dmgH)2(DMAP)I]
[Co(dmgH)2(DMAP)B
[Co(dmgH)2(MP)Br]
[Co(dmgH)2(MP)Cl]
[Co(dmgH)2(HP)Br]
[Co(dmgH)2(HP)Cl]
[Co(dmgH)2(MP)I]
[Co(dmgH)2(HP)I]
Organism
C
Bacteria
Salmonella paratyphi-B 10 - 13 - - - - - 12 18
Klebsiella pneumonia 13 13 10 16 13 14 13 12 15 20
Proteus vulgaris 12 10 8 18 - - 14 8 17 30
Shigella flexneri 16 18 15 17 16 16 18 18 12 30
Micrococcus luteus 13 15 17 15 17 18 17 18 19 26
Enterobacter aerogenes 16 15 15 14 12 13 15 15 16 22
Staphylococcus aureus 18 16 16 19 19 17 18 18 18 14
Staphylococcus aureus 21 16 18 19 15 15 18 17 25 30
MRSA
Fungi C
Candida albicans 20 23 22 21 18 24 23 23 20 28
Malassesia pachydermatis 19 13 14 13 15 16 10 15 23 26
Trichophyton mentagrophytes - - - - - - - - - 26
Trichophyton rubrum - - - - - - - - - 28
Aspergillus flavus - - - - - - - - - 30
C- control: Streptomycin - standard antibacterial agent; Ketoconazole - standard
antifungal agent.
All the complexes showed good zone of inhibition towards fungi Candida albicans
and Malassesia pachydermatis (Plate 2).
Plate 1 Zone of inhibition of [Co(dH)2(B)(X)] complexes against bacteria

Plate 2 Zone of inhibition of [Co(dH) 2(B)(X)] complexes against Malassesia
pachydermatis
The iodocobaloximes showed good zone of inhibition compare to choloro and bromo
complexes. All the complexes showed nil activity against Trichophyton
mentagrophytes, Trichophyton rubrum, Aspergillus flavus.
This is the first report of this complex against antibacterial activity with
greater MIC values. Remaining other complexes viz., 1, 2, 4, 5, 6 and 9 showed
minimal activity against bacterial strains. Complex 3 and 8 showed very least activity,
because it ranges about 125 to 1000 µg/ml values. All the experiments were compared
with respective standard antibacterial drugs.
In the case of antifungal growth inhibition were assessed and evaluated
effective complexes. Schreier et alalso investigated the effects of iodine derivatives
on microbial cells and found that it affects the structure and functions of enzymes and
cell proteins and damages bacterial cell function by blocking hydrogen bonding and
altering the membrane structure.Only two microbes (Candida albicans and
Malassesia pachydermatis) were tested against for the complexes. The results showed
that ranges about 62.5 to 500 µg/ml. Very low concentration was observed (62.5
µg/ml) 1, 2, 3, 6, 7 and 8 complexes against Candida albicans. Remaining complexes
were showed at 125 µg/ml level. Complex 1 was very effective towards M.
pachydermatis at 125 µg/ml and remaining complexes exhibited that ranges from 250
to 500 µg/ml.

Table-2 Minimum inhibitory concentration of complexes
[Co(dmgH)2(DMAP)I]
[Co(dmgH)2(DMAP)
[Co(dmgH)2(DMAP)
[Co(dmgH)2(MP)Br]
[Co(dmgH)2(MP)Cl]
[Co(dmgH)2(HP)Br]
[Co(dmgH)2(HP)Cl]
[Co(dmgH)2(MP)I]
[Co(dmgH)2(HP)I]
Organism
C
Cl]
Bacteria
Salmonella 125 - 125 - - - - - 250 6.25
paratyphi-B
Klebsiella 250 250 500 250 250 250 250 500 125 6.25
pneumoniae
Proteus vulgaris 250 500 1000 125 - - 250 1000 500 6.25
Shigella flexneri 500 125 500 500 500 250 125 125 250 <0.78
Micrococcus luteus 250 500 125 250 500 125 125 125 125 >100
Enterobacter 250 500 500 250 500 250 250 250 125 <0.78
aerogenes
Staphylococcus 500 250 250 125 125 500 125 125 500 <0.78
aureus
Staphylococcus 500 250 250 125 250 250 125 500 250 <0.78
aureus (MRSA)
Fungi C
Candida albicans 62.5 62.5 62.5 125 125 62.5 62.5 62.5 125 25
Malassesia 125 250 250 250 250 500 500 250 250 12.5
pachydermatis
C- control; (–) no activity; Synthesis compound; Ketoconazole (standard antifungal
agent):
Streptomycin (standard antibacterial agent)
CONCLUSION
Hence, it may be concluded that cobaloximes will show antimicrobial activity
depending on the nature of the axial ligands and halogens. Commonly
iodocobaloximes shows good activity compare to choloro and bromo complexes. The
antimicrobial action mode of Iodine’s is not fully understood, but it is believed to be
associated with its ability to rapidly penetrate the cell wall of microorganisms.These
multiple modes of action ensure the rapid death of microbes and help to prevent the
development of bacterial resistance.

ACKNOWLEDGEMENT
The authors are thankful to Principal, Loyola College, for providing the
necessary facilities.
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ANTIFEEDANT AND TOXIC PROPERTIES OF CALOTROPIS
PROCERA (AITON) W.T. AITON EXTRACT ON HELICOVERPA
ARMIGERA HUBNER
Bakavathiappan GA1,*, Baskaran S2and Lingathurai S3

1
Department of Zoology, Saiva Bhanu Kshatriya College, Aruppukottai, Tamil Nadu,
India
2
PG Department of Zoology, Ayya Nadar Janaki Ammal College, Sivakasi, Tamil
Nadu, India
3
*Corresponding Author: ga.bakavathiappan@gmail.com
ABSTRACT
The antifeedant and larvicidal activities were studied with different solvent extracts of Calotropis
procera such as, hexane, chloroform, ethyl acetate, methanol, acetone and ethanol
against third instar larvae of Helicoverpa armigera by leaf dip method. All the
extracts prepared in different concentrations such as, 0.625%, 1.25%, 2.5% and 5%
level. Antifeedant activity was directly proportional due to the concentration of the
extracts. In Helicoverpa armigera, highest antifeedant activity was observed in
chloroform extract at 5% concentration. The experimental results showed that highest
larval mortality was observed in chloroform extract. Antifeedant effect in insect is one
of the major parameters to assess the efficacy of crop protections. All the results were
clearly indicated chloroform extract of C. procera possesses many useful properties to
control insect pests.
KEYWORDS: Antifeedant activity,bioassay, Calotropis procera,pest
control,Helicoverpa armigera.
INTRODUCTION
India is basically an agriculture based country and more than 80% of Indian
population depends on it. Agricultural productivity influences the Indian economy.
Insect pests are known to cause significant damage to crops and affect agricultural
productivity (Summarwar, 2013). Among the insect herbivores, 10,000 species are

sporadic pests and 1000 species are serious pests in the world. Losses due to insect
herbivores have been estimated about 10-20% for major crops are a significant factor
in limiting food production (Ignacimuthu, 2005). Insects developed resistance against
most of the modern classes of synthetic insecticides like DDT, organophosphate,
pyrethroids and carbamates (Neoliya et al., 2007). The misuse and excessive use of
synthetic insecticides may cause some undesirable effects, not only to the agricultural
ecosystem but also to human health due to insecticide residue in food. Interest in
plants with insecticidal properties has been on the increase recently around the world
today, either singly in integrated pest management or in conjunction with synthetic
pesticide (Adewole, 2013). Antifeedant is defined as any substance that reduces
consumption (feeding) by an insect. Many antifeedants do not kill pests immediately;
suppress insect feeding for several days and cause mortality after two weeks.
Antifeedants are considered as insecticides. Antifeedant principles may cause
insect mortality in lepidopteron pests due to a combination of starvation and contact
toxicity (Ignacimuthu and Vendan, 2008). Botanical pesticides are ecofriendly
economic, target specific and easily biodegradable (Ignacimuthu, 2004). Botanical
pesticides are derived from plants. They degrade rapidly and therefore are considered
safer to the environment than the common synthetic chemicals. Hence, the present
investigation was conducted to study the antifeedant activity of different solvent
extracts of Calotropisprocera leaves against third instar larvae ofHelicoverpa
armigera.
Helicoverpa armigera is a pest belonging to the order Lepidoptera and the
family Noctuidae. This moth is a major pest because the larvae can feed on a wide
range of economically important crops including cotton, corn, tomato, legumes and
tobacco. Calotropis procera, is a popular medicinal plant distributed in arid to semi-
arid regions. Botanical pesticides are eco-friendly, economic, target-specific and
easily biodegradable.

Extraction of plant materials
Calotropis procera (Apocynceae) leaves were collectedin and around Sivakasi
Virdhunagar District, Tamil Nadu, India and washed with tap water. The plant leaves
were shade dried and powered in a domestic grinder and stored in refrigerator for

further use. The powder was extracted with hexane, chloroform, ethyl acetate,
methanol, acetone and ethanol in a Soxhlet apparatus separately.
Collection and rearing of Helicoverpa armigera

Eggs of H. armigerawere collected from cotton fields in and around Sivakasi,
Virdhunagar District, Tamil Nadu, India and cultured on tender cotton leaves at
laboratory conditions 12-hrs photoperiod 28±2°C and 65-75% R.H. Laboratory
emerged adults were maintained in 5% sugar solution. Eggs were maintained in the
laboratory conditions. Laboratory emerged third instarH. armigera larvae were used
for this experiment.
Concentrations and antifeedant bioassay
The crude extracts were tested at four different concentrations viz., 0.625,
1.25, 2.5 and 5%. The antifeedant activity wastested by no-choice method (Bentley et
al., 1984).H. armigerafed with crude extract coated cotton leaf discs (4 cm
indiameter). The experiment was performed in a 9 cm diameter petridish containing
moist filter paper to avoid early drying of the leaf disc. Four hours pre-starved H.
armigeralarvae were introduced to treated and control leaf disc taken in the petridish.
Leaf area consumed by the larva in control and treatments was determined after 24
hrs using a leaf area meter (Delta-T Devices, Serial No. 15736 F 96, UK). Antifeedant
activity was calculated by the modified formula of Bentley etal. (1984).
Insecticidal bioassay
The insecticidal activity of crude extracts was also tested at four different
concentrations viz., 0.625, 1.25, 2.5 and 5 %. The treatments were given cotton leaf
discs by the same procedure as described in the antifeedant bioassay test. After 24 h
treatment period the larvae were reared on fresh untreated leaves. The mortality in
treated and control groups was recorded by the method of Abbott (1925) and LC50
and LC90 values were determined using probit analysis (Finney, 1971).
Table 1 shows the antifeedant activity of H. armigera treated with crude

extract of six solvents of C. procera. Highest antifeedant activity of 72.02 %was
observed in chloroform extract at 5% concentration. Next to chloroform

extract,acetone extract shows increased percent antifeedant activity such as 13.17
(0.625%), 24.57 (1.25%), 47.11 (2.5%) and 69.17 (5%).
Antifeedant activity was increased from 17.21 (0.625%) to 51.79 (5%) in
hexane extract. Moderate amount of percent antifeedant activity was observed in
ethanol extract 34.61(5%) and in methanol extract as 31.06 (5%). Very low
antifeedant activity was observed in ethyl acetate extract. Maximum percent
antifeedant activities of 72.02, 69.17, 51.79, 34.61, 31.06 and 24.08 were found in
chloroform > acetone > hexane > ethanol > methanol > ethyl acetate respectively at
5% concentration. The antifeedant activity was directly proportional to the
concentration of the extract (Table 1).
Table 1. Antifeedant activity (%) of crude leaf extracts of Calotropis procera
against third instar larvae of H. armigera
Solvent Concentration of leaf extracts of Calotropis procera (%)

extract
0.625 1.25 2.50 5
Hexane 17.21 ± 4.90de 23.79 ± 2.12cd 27.20 ± 3.24c 51.79 ±5.14b
Chloroform 20.41 ± 1.12d 21.76 ± 5.74 d 62.52 ± 4.70ab 72.02 ±5.86a
Ethyl acetate 04.91 ± 0.89g 07.76 ± 0.72fg 19.95 ± 2.02d 24.08 ± 2.46cd
Methanol 10.61 ± 1.56f 16.63 ± 1.36de 24.38 ± 2.99cd 31.06 ± 1.56bc
Acetone 13.17 ± 2.50ef 24.57 ± 3.67cd 47.11 ± 5.62b 69.17 ± 1.16a
Ethanol 13.96 ± 2.71ef 15.71 ± 2.60e 24.15 ± 2.08cd 34.61 ± 2.29bc
Within columns, mean ± SD followed by the same letter do not differ significantly
using Tukey’s test, P ≤ 0.05.
The LC50 values of six solvent extracts such as, hexane, chloroform, ethyl
acetate, methanol, acetone and ethanol were 8.590, 6.128, 14.151, 13.193, 14.556 and
8.748(%) respectively against H. armigera. In these solvent extracts, the highest toxic
effect was recorded with chloroform extract at 5% concentration with the LC 50 value
of 6.128% followed by hexane extract (8.590%) in H.armigera. Irrespective of the
concentration and the solvents used for extraction, the insecticidal activity has been
varied (Table 2). Baskar et al. (2009) showed the LC50 and LC90 values of 2.46% and
5.41% respectively in hexane extract of Atalantia monophylla leaf against third instar
larvae of H. armigera.

Table 2. Effect of different solvent extracts of Calotropis procera on the larval
mortality (%) of third instar larvae of H. armigera
Solvent Concentration of leaf extracts of Calotropis procera (%)

extract
0.625 1.25 2.50 5
Hexane 4 11 21 37
Chloroform 0 12 29 42
Ethyl acetate 4 17 21 29
Methanol 0 8 8 25
Acetone 4 12 17 29
Ethanol 8 17 21 37
Sahayaraj et al. (2008) reported similar findings in LC50 values of S. litura

with Pedalium murex. In higher concentration of P. murex (above 0.4%) died at the
early period of the treatment. But those animals which fed with lesser concentration
(below 0.2%) failed to complete the moulting and died either in the larvae or in
pupae.
In the present study toxic effects was observed in all the solvent extracts of C.
procera against the larvae of H. armigera. The present investigation is in accordance
with the earlier findings of Kavitha et al. 2009 showed the larval mortality of H.
armigera against neem derivatives. The percent larval mortality varied from 45.83
(0.5%), 79.16 (2.0%) in neem oil and 29.16(1.0%), 70.83(5%) in neem cake extract.
The insect endocrine system is an excellent target for insecticides; because small
changes in the endocrine balance can be greatly amplified resulting in a reduced
ability to compete in the environment or death.
In H.armigera, among the tested extracts, highest larval mortality was
observed in the chloroform extract with increasing concentrations from 0.625%(0),
1.25%(12), 2.50%(21) and 5%(42) whereas in other extracts the larvicidal activity
was less than the chloroform extract (Table 3).

Table 3. Toxic effect (%) of different solvent leaf extracts of Calotropis procera
treated against third instar larvae of H. armigera
Chi square
Solvent extract LC50 LC90 Slope ± SE value (χ2)
Hexane 8.590 74.004 1.37 ± 0.23 2.383*
Chloroform 6.128 26.327 2.02 ± 0.27 4.949*
Ethyl acetate 14.151 88.286 1.09 ± 0.23 2.988*
Methanol 13.193 78.120 1.65 ± 0.31 5.920*
Acetone 14.556 72.810 1.19 ± 0.24 0.921*
Ethanol 8.748 90.898 1.90 ± 0.31 1.428*
Units LC50 and LC90 = % / w, applied for 96h, a95% lower and upper fiducial limits
are shown in parenthesis.
Vendan et al. (2009) studied the larvicidal effects of Hydnocarpus alpina

against the larvae of H. armigera. Ethyl acetate extract killed maximum number of
larvae and the mortality was recorded as 77.78 percent at 5 percent concentrations.
Even though the larvae consumed very little quantity of treated leaf, they were
affected by the toxin present in the plant extract.
Lingathurai et al. (2008) studied the antifeedant effects of Morinda tinctoria
against the third instar larvae of S. litura. Chloroform extract showed the highest
antifeedant activity (33.66%) followed by ethyl acetate (31.38%) and hexane
(28.40%) at 5% concentration. The antifeedant activity was directly proportional to
the concentration of the extract. Vendan et al.(2008) showed the antifeedant activity
of chloroform extract of Clerodendron phlomidis (75.14%), ethyl acetate extract of
Mundulea sericea (71.78%) , Hexaneextract of Citrullus colocynthis
(67.36%),chloroform extract of Artemisia vulgaris (61.47%)andhexane extract of
Argemone mexicana (55.69%) against third instar larvae of H. armigera. C.procera
showed potent antifeedant and larvicidal activities against H.armigera. This could be
used for the development of newpesticide formulations for the control of this serious
polyphagous pest.

REFERENCES
Abbott,W.S., 1925. A method of feeding deterrents for spruce

computing the effectiveness of budworm
an insecticide, J.Econ. Choristoneurafumiferana
Entomol.,18 : 265-276. (Lepidoptera: Tortricidae),
Adewole,A., Oderinde,A., Bankole, Annals. Entomol. Soc. ofAmer.,
O., Faparusi, F. and R.T. 77 : 393-397.
Oyede,2013. Larvacidial Finney,D.J., 1971. Probit analysis,
activities of three plant extracts Third Edition., Cambridge
of common wire weed (Sida University Press, Cambridge.
Acuta), Catnip (Nepeta Ignacimuthu, S., 2004. Green
Cataria) and pesticides for insect pest
Neem(Azadirachta Indica) management, Curr.Sci., 86 :
against the larva of mosquito 1059.
(Anopheles Gambiae), Igancimuthu, S., 2005. Biodiversity
Academia Journal of Medicinal and insect pest management.
Plants1 (2): 37-40. Curr. Sci., 88 (10) : 1543-1544.
Baskar, K., Kingsley,S., Vendan,S.E., Ignacimuthu,S. and S.E.Vendan, 2008.
Paulraj,M.G., Botanical pesticides in insect
Duraipandiyan,V. and pest management. Ins. Pest
S.Ignacimuthu, 2009. Mgmt. & Envl. Safety (Suppl),
Antifeedant, larvicidal and 4 (1) : 141-154.
pupicidal activities of Atalantia Kavitha, E., Kingsley,S., Revathi,N.
monophylla (L.) against and M.Sathivel, 2009.
Helicoverpa armigera Insecticidal activity of neem
(Hubner) derivatives against okra fruit
(Lepidoptera:Noctuidae), borer Helicoverpa armigera,
Chemosphere, 75 : 355-359. Inter.J. Agric.Sci.,5 (2) : 528-
Bentley,M.D., Leonard,D.E., 530.
Stoddard,W.F. and Lingathurai,S., Vendan, S.E., Paulraj,
L.H.ZalKow, 1984. M.G. and S. Ignacimuthu,
Pyrrolizidine alkaloids as larval 2008. Antifeedant and growth

inhibiting effects of Morinda Indian Journal of Fundamental
tinctoria Roxb, against and Applied Life Sciences.
Spodoptera litura (Fab.) Vendan,S.E., Baskar,K.,
(Lepidoptera: Noctuidae). In : Lingathurai,S., Ramar,M.,
Recent Trends in Insect Pest Paulraj,M.G. and S.
Management (Eds.) Ignacimuthu, 2008. Effects of
Ignacimuthu, S. and S.Jayaraj, plant extracts on larval
Elite Publishing House Pvt. development of Helicoverpa
Ltd, New Delhi. 58-63. armigera, In : Recent Trends
Neoliya,N.K., Singh, D. and in Insect Pest Management,
R.S.Sangwan, 2007. (Eds.) S.Ignacimuthu and
Azadirachtin-based insecticides S.Jayaraj, Elite Publishing
induce alteration in House Pvt. Ltd., New Delhi,
Helicoverpa armigera head 215-219.
polypeptides, Curr.Sci., 92 : 94- Vendan,S.E., Baskar,K., Paulraj,M.G.
99. and S.Ignacimuthu, 2009.
Sahayaraj, K., Venkateshwari, M. and Antifeedant and larvicidal
R. Balasubramanian, 2008b. effects of Hydnocarpus alpina
Insecticidal and antifeedant extracts against the larvae of
effect of Pedalium murex Helicoverpa armigera.In
(Linn) root and on Spodoptera :Ecofriendly Insect Pest
litura (Fab.) (Lepidoptera: Management, (Eds.)
Summarwar,S. and J.Pandy, S.Ignachimuthu and
2013.Effect of plant extract B.V.David, Elite Publishing
Azadirachta indica on feeding House Pvt. Ltd., New Delhi,
behaviour of Spodoptera litura, 210-216.

BIOMONITORING OF AIR POLLUTION AT MILLERPURAM, JUNCTION,
PALAYAMKOTTAI ROAD, THOOTHUKUDI BY ANALYSING SELECTED
PLANTS
M. Isakkiyammal
Department of Botany, A.P.C.Mahalaxmi College for Women, Thoothukudi, Tamil

Nadu, India
Corresponding Author: skeikannan@gmail.com
ABSTRACT
The present study was concluded to biomoniter the air pollution by studying some
morphological and biochemical parameter of selected plants at Millerpuram Junction.
Palayamkottai Road, Thoothukudi. The selected plants showed increased pH,
decreased ascorbic acid content, decreased total chlorophyll and reduction of controls
plants. To protect the plant species the authorsuggests installation of pollution control
equipment in vehicles and industries,use of lead free petrol by adding
substitutes,effective legislative control measures and good traffic management.
KEYWORDS: Air pollution, Biomonitoring, Indicator plants, Suggestions.
INTRODUCTION
Thoothukudi, which is termed as “SOUTHERN GATE WAY OF INDIA” and

also as PEARL CITY is at the south east cost at 8.45 N latand 78.9 Elong. Having a
colourful past. Thoothukudi is a port city situated in the gulf of Manner about
125KM.north of Cape Commorinand 650KM south of Chennai. The bay formed by
the Hare Island,Devils point and the main land and gives ample protection to
thelightens from monosoonic weather. The port hinterland of Thoothukudi extend to
the districts of Madurai, Kanyakumari,Thirunelveli,Ramnad and also southern parts
of Tiruchirappali and encompasses an area of about 41,600 Sq.Km. and a population
of over 10 million, This hinterland is a fast growing industrial belt of South India.
Thoothukudi is a Municipal Corporation andranks 6th town in Tamilnaduwith an
extent of 13.47 sq.km.

Millerpuram Junction at Palayamkottai road Thoothukudi Municipal
Corporation
Thoothukudi is served by three main roads which radiate Madurai,

Tiruchendur, Tirunelveli. It is also the terminal of broad gauge railway line which
connects Thoothukudi with southern railway board gauge system. The
PALAYAMLOTTAI ROAD is between 3rd mile and old bus stand.And Thoothukudi
is having more petrol and diesel operated vehicles are plying through this road.
Increasing urbanisation and high density traffic have created environmental

vehicular pollution problems. The degradation of air quality causes much worry to
the city dwellers. The sounds we here in the streets which offend our ears, emotions,
and our general physiology come only partly from industry. Much of thenoise meets
our urgent personal and business needs (Nagiet al.1999).
Air pollution is responsible for innumerable problems like ozone depletion,

greenhouse effect, acid deposition, dieses outbreaks , physicochemical changes of soil
etc. (Khan and Chalkoo, 2005), which consequently have serious effects on the biotic
and abiotic components of the environment. Regional impacts of air pollution on
different plant species is one of the major ecological issues, as they are continuously
and directly exposed to the changing environment as compared to the animal
population. Several studies have been carried out to highlight the impact of air
pollution, vehicular pollution on the morphological, anatomical, physiological and
biochemical aspects of different plants (Chauhan et al.,2004)
Using plants as indicator of air pollution is the possibility of synergistic action

of pollutants. The ambient environment of the urban are may be contaminated with
several pollutants and the plants growing there would be exposed not only to one but
to many pollutants and their different conditions It is possible to estimate the overall
effect of a large number of pollutants as total pollution by measuring changes in the
plants (Tiwari et al.,1993) Hence the plants are considered to be efficient bio-
indicators which take note of many changes in the environment and will soon become

valuable records of contaminations and very convenient for environmental
monitoring.
The present work is an attempt to bio monitor the air pollution by studying
some morphological and biochemical parameters of selected plants at
MillerpuramJunction, Palayamkottai Road, Thoothukudi. Ten plants were assessed at
this site for tolerance index to establish the air pollution level.
Description of the selected Plants
The selected flora of study area listed (Table 1) and their description is as
follows (Gamble, 1921).
Table.1: Plants selected at Millerpuram Junction, Thoothukudi.
S.NO Plant Family

1 Acalypha indica Euphorbiaceae
2 Boerhavvia diffusa Nyctaginaceae
3 Calotropis gigantea Asclepiadaceae
4 Clerodendrum inerme Verbenaceae
5 Coccinia indica Cucurbitaceae
6 Jatropha curcas Euphorbiaceae
7 Passiflora foetida Passifloraceae
8 Prosopisspocigera Leguminaceae
9 Tephrosia purpurea Fabaceae
10 Tridax procumbens Compositae
1. Acalypha indica. Linn. (Euphorbiaceae)
It is an erect, herbaceous, annual growing to height of 0.75 to 2 feet or more,

with many spreading of ascending ranches.
2. Boerhaavia diffusa, Linn.(Nyctaginaceae)
It is a diffusely branched perennial with ascending or prostrate habit. Stem is

slender, swollen at nodes, sub - glabrous and often pinkish.
3. Calotropis gigantea ,R.,Br.(Asclepiadaceae)

A large much – branched milky shrub,3-6 feet high and covered all over with
very soft cottony tomentum. Stem is erect, stout and round.
4. Clerodendrum inerme, Linn. (Verbinaceae)
Trees and shrubs, sometimes straggling or climbing. Leaves obovate or

elliptic, even orbicularobtuse or emarginated, opposite or rarely ternate, up to 2 in
long 1-5 in board.
5. CocccinaindicaWand A
A prostrate are climbing herb, branches are glabrous grooved with simple
tendrils.
6. Jatropha curcas, Linn (Euphorbiaceae)
It is an erect robust shrub 3-4 feet height and thrive best in sandy loams stem
is round, thick and contains transparent juice7.Passiflorafoetida,Linn (Pssifloraceae).
It is a twining plant with tendrils. Leaves are simple and palmately lobed, usually
glandular.
8. Prosopisspicigera,Linn (Leguminosae).
It is a small to moderate – sized deciduous tree with many thorns on board

bases, the leaflets distant, linear – along, about five inches long.
9. Tephrosiapurpurea, Pers.(Fabaceae)
It is a freely branching perennial under shrub. Stem is woody at base, round

and pubescent on your parts that become glabrous with age.
10. Tridax procumbns,Linn.(Compositae)
This is another example of an introduced that has become naturalised in the

presidency and it is now a part of the flora.
The site Millerpuram junction was selected along Thoothukudi-Palayamkottai

Road in Thoothukudi Municipal Corporation to study the traffic density. Ten

common plants species were selected and leaves were collected in the site. Leaves
were sampled were mixed separately to get a composite sample, which was analysed
for different parameters. The same plants collected from unpolluted areas were taken
as controls for comparison.
1. pH (Mir et al., 2008)
2. Estimation of biomass/ relative water content (Dutta and Mookerjee, 1981)
3.Estimation of Ascorbic acid (Sadasivam and Manickam, 1992)
4.Estimation of cholorophyll (Arnon, 1949)
5. Air pollution tolerance index (APTI) (Kousar et al., 1998)
6.Estimation of carotenoids (Ridely, 1982):
The present study is to evaluate the effect of air pollution on the morphology
and selected biochemical parameters of the selected plants at Millerpuram Junction,
Palayamlottai Road in Thoothukudi Municipal Corporation.
Table.2: Air Pollution Tolerance Index of the selected plant at Millerpuram

junction, Thoothukudi.
S.No Name of the plant pH W.C A.A T.C APTI Response

1 Aclypha indica 5.64 3.41 11.21 9.49 1.73 S
2 Boerhavia diffusa 6.42 2.81 40.47 5.65 0.56 MS
3 Calotropis gigantea 5.89 3.11 7.54 5.98 0.92 MS
4 Clerodendrum 5.96 2.76 8.67 7.63 1.20 S
inerme
5 Coccinia indica 5.98 2.82 8.65 7.92 1.23 S
6 Jatropha curcas 5.88 2.58 6.64 7.85 0.93 MS
7 Passiflora foetida 5.99 3.02 12.56 9.52 1.97 S
8 Prosopis foetida 5.67 3.12 22.21 7.52 2.96 S
9 Tephrosia purpurea 5.69 3.67 16.17 6.32 1.98 S

10 Tridax procumbans 5.99 3.51 20.14 5.36 2.32 S
pH-pH of leaf extract, W.C –Relative water content, A.A- Ascorbic acid, T.C -Total
chlorophyll , APTI Air pollution tolerance index.
pH:
All the selected plants showed more or less increased pH than the control plant
leaf samples. Table. 2) Similar reports were reported by Kousar et al., (1998) in
Warangal city, Andhra Pradesh.
Biomass /relative water content:
All the selected plants showed decreased biomass than the controls at the site.
The increased traffic density decreased the biomass content. Air pollution due to
industries and auto – exhaust are the cause for this decrease in biomass.
Ascorbic acid content:
All the ten plants species showed decreased ascorbic acid content than the
controls at the site (Table.2). The increased traffic intensity decreased the ascorbic
acid content gradually. Alquainy, (2007) reported increased ascorbic acid / vitamin–
C production and saline tolerance in Vicia faba and Pisum sativum during the
germination of at seedling growing under saline stress conditions.
Chlorophyll pigment concentration:
The total pigment concentration at the selected site decreased with increasing
traffic density than the controls in all the ten selected plants (Table.2). Similar results
were obtained by Mir et al., (2008) in Lucknow city in Alstonia scholaris results,
Ficus religiosa, Nerium odorum and Polyalithia longifolia leaves.
Air pollution tolerance index (APTI)
From table 2, it was evident that the selected plant species showed the degree
of tolerance index against air pollution. Our study revealed that, in the selected site,
Acalyphaindica, Clerodendrum inerme, Coccinia indica, Passiflora foetida, Prosopis
spicigera, Tephrosia Purpurea and Tridax procumbens are sensitive to air
pollution.Boerhavvia diffusa, Calotropis gigantea and Jatropa curcas are more

sensitive to air pollution (Table 2). This is attributed due to high air and vehicular
pollution.
Morphological observation:
The selected plant species generally did not show any visible symptoms of
injury. The exhaust and dust from automobiles and industries settled the selected
plant species and give them dull look. This may be attributed to the coating of the
dust and exhaust fumes due to which the original shone and brightness of the leaves is
lost. Thus the air pollution causes both visible and invisible injuries to the plants. No
other external visible symptoms of injury were observed. Similar observations were
reported by Salgare andlyer (1991) in some common roadside weeds.
SUGGESTIONS/ RECOMMENDATIONS
As Thoothukudi is a fast growing industrial and port city of southern Tamil

Nadu, there are more industrial and automobiles are plying. Auto exhaust pollutants
are carbon monoxide (CO), Oxides of nitrogen (NOx),Lead (Pb),Sulphurdioxide
(SO2), Ozone (O3), and smoke industrial pollutants are SO2, fly ash, dust particles,
toxic gases etc. These are the major threats to the plants, animals and human beings.
In the light of the data collected the following suggestions are recommended for the
management of air pollution;
 Design of the fuel efficient/ changed fuel engines (using gas, electricity, solar
power battery etc.)
 Installation of pollution controlequipmentin vehicles and industries.
 Use of lead free petrol by adding substitutes.
 Improved refining of fuel. Sulphur content of diesel may be reduced to 0.25%.
Benzene content may be reduced to 30%.
 Good traffic management.
 Encouragement of public transport system.
 Control by legislation.
 Wide roads, good road maintenance.
 Air pollution trees may provide a natural sink for air pollutants and they may
be grown on large scale.

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isolated chloroplasts. Narain, K. and Yuns, (2008).
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Effects of fumigation of SO2 avenue plants.Roll.Res. 27(1)
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Response of some common Tiwari, S and Bansal, S. (1993).
plants to air pollution, Expected performance indices
India.J.Applied& pure Boil. 20 ofsome planted plants of
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M.A.Sinngarachyarya
,(1999).IJEP.19 (7). 488-492.

AMINO ACID CONTENT OF EARTHWORM EISENIA FETIDA
CULTURED ON AQUATIC WEED WATER HYACINTH
T. Sakthika
Dept. of Zoology, A.P.C.Mahalaxmi College for Women, Thoothukudi, Tamil Nadu,
India
Corresponding Author: sakthikasaravanan@gmail.com
ABSTRACT
Earthworms were cultured on Aquatic weed waterhyacinth for 60 days. Essential and
Non essential aminoacids were analysed from the earthworm extract and compared
with the worms cultured on garden soil. Highest level of essential amino acid in the
experimental worm was leucine (8.6g / 100g), followed by Lysine (7.3g / 100g). The
non-essential amino acid in the experimental earthworm was dominated by glutamic
acid (7.80g / 100g), followed by Aspartic acid (7.80g / 100g).The present
investigation proves that the conversion of aquatic weed biomass into worm biomass
is an effective eco-friendly technology for not only producing aminoacid enriched
protein but also for managing the rapid growth of aquatic weeds.
KEY WORDS: Waterhyacinth, Eisenia fetida, Vermiculture, Aminoacid,
INTRODUCTION
Amino acid utilized as proteins are primary constituents of structural and
protective tissues, including skin, feathers, bone, ligaments, as well as muscles and
organs. The highest essential amino acid of earthworm was dominated by histidine
(0.63% of dry matter basis) and earthworm meal was dominated by isoleucine (1.98%
of dry matter basis). Histidine was essential for protein synthesis, involved in
carnitine and haemoglobin synthesis. It was effective in allergic disease and tension of
the autonomic nervous system, meanwhile isoleucine was involved in protein
synthesis, energy production and muscle building (Padmavathiet al 2017).
Tram et al. (2005) reported that the highest essential amino acid composition
of P. excavates was leucine (3.47% and 0.76% of dry matter basis respectively). The
non essential amino acid of earthworm and earthworm meal was dominated by
glutamic acid (1.52% and 3.60% of dry matter basis respectively). Glutamic acid was

involved in protein synthesis and as source of energy for cells lining the intestine.
Facilitates immune function and improve anti-inflammatory effects, aids in
preventing and healing of peptic ulcer and ulcerative colitis.
The high essential amino acid composition of earthworm meal would produce
great result when added in animal feedstuff ratio. By protein content rich with amino
acids, the earthworm meal presents around 98% of absorption by animal organism
due the balance between vitamins and amino acids. (Julendra, 2003), showed that
earthworm meal of Lumbricus rubellus had 65.63% crude protein content (Damayanti
et al., 2008), earthworm meal of Lumbricus terestris contained 32.60% protein and
earthworm meal of Perionyx excavatus contained 57.2% crude protein and had
complete amino acid (Tram et al., 2005) Earthworm L. rubellus contained ‘lumbricin
I’ which had antibacterial activity, included in peptide group which contained 62
amino acids (Salzet et al., 2006).
Earthworms could serve as a source of Essential Aminoacids especially
Lysine which is limiting in many basic food stuffs (Albarran, 1996). The content of
Lysine in earthworm flour is significant representing the daily requirement for
children between the age of 2 and 5 years. Earthworm meal was shown to have an
aminoacid composition very similar to that of fishmeal and potentially superior to
meat meal and the protein was shown to contain such essential aminoacids such as,
phenyl alanine leucine lysine, metionine and valine (Gabriel and Dedeke, 2010).
According to Gajalakshmi et al (2001), water hyacinth could be converted to compost
by earth worms. The high protein content makes the water hyacinth a potential feed
for live stock such as cows, goats, sheep and chickens. Abdelhamid and Gobu (1991),
after chemical analysis on water hyacinths collected from a canal and a ditch reported
them are having 9.5% DM, and in the DM ,74.3% organic matter ,19% Crude protein,
and 18.9% Crude fiber. Poddar et al (1991), reported the chemical composition of
water hyacinth as 83.6% organic matter, 16.3% Crude protein and 16.4% Crude fiber
(on DM basis). Aboud et al (2005) reported that water hyacinth could provide large
quantities of nutritious feed as was a potential source of ruminant nutrition.
Water hyacinth (Eichhornia crassipes) is a fast growing perennial aquatic
plant found in wetlands and which prefers nutrient-enriched water (Wilson et al.,
2005). It can cause infestations over large areas of water surfaces and leads to series
of problems such as decrease of biodiversity, blockage of rivers and drainage systems,

depletion of dissolved oxygen, alterations in water chemistry, environmental
pollution, decreased fish population, restricting access to fishing sites and loss of
fishing equipment (Malik, 2007). An alternate source for waterhyacinth management
is vermicomposting.
The objective of the present study is to determine the amino acid content of
the earthworms Eisenia foetida cultured on Aquatic weed waterhyacinth which is
generated as a solid waste in the Tamirabarani River, near Eral using
vermitechnology.
Earthworms and Substrates
Earthworms, Eisenia fetida were obtained from a culture bank maintained in
DCW Ltd, Arumuganeri, Thoothukudi District. Aquatic weed Eichornia sps was
taken from Thamirabarani River, near Eral. Garden soil was used as control. Cow
dung was obtained from a local farmyard.
Experimental design
The Water hyacinth was dried in air, cut into small pieces and mixed with cow
dung (nutrient mixture) on dry weight basis in a ratio of 5:1 for the experiment. This
mixture was pre decomposed for 15 days to make it palatable for the earthworms.
The compost was prepared in wooden box of 3 feet breadth and 2 feet height.
A thin layer of 1.5 cm thick sterilized soil is filled at the bottom as the supporting
material for vermicomposting. Partially decomposed cow dung was placed over the
soil layer. The experiment was setup by taking 5kg nutrient mixture (on dry weight
basis) in each wooden box and no extra feeds were provided during the study. Fifty
earthworms, Eisenia foetida were released over the mixture. The compost of mixture
was covered with paddy straw. Two vermibeds were prepared for control and
Waterhyacinth. Three replications were setup for statistical analysis of the results.
Vermicomposting was conducted under laboratory conditions in darkness at an
average ToC at 25oC and a substrate moisture content of 70 -75%. The experiment
was conducted for 60 days after releasing the earthworms. The aminoacid content of
the earthworm was analysed after 60days.
Preparation of Earthworm Extract: The earthworm was washed in running tap
water to remove dirt if any over the body surface and submerged in warm water (40
°C) for 2 hours to allow its gut-soils to be excreted out. Later on, the earthworms were

washed thoroughly with distilled water and then kept in a hot air oven at 45 °C for 15
days to get it completely dried.
Assay of total amino acids: Total amino acids content of protein free supernatants
were estimated by modified dinitrophenyl (DNP) derivatization method (Varley, et al
1980). In brief, the methanol extract (100 µl) of biological fluid or the reference
standard (equimolar mixture of glutamate:glycine) was made up to 250 µl with 80%
(v/v) methanol. Equal volume (250 µl) of borate buffer was added to each of the
tubes, followed by 0.5 ml of DNFB reagent. Tubes were incubated at 45°C for 30
minutes and allowed to attain room temperature. After adding 1 ml of 0.25 M HCl to
each of the tubes and mixing, the absorbancy was measured at 420 nm. Apart from
reagent blank and reference standards, some of the samples also carried a known
amount of glutamate:glycine mixture to assess the recovery of total amino acids
through different steps of assay procedure.
A total of seventeen (17) amino acids consisting of Nine (9) essential namely
Lysine, Histidine, Arginine, Threonine, Valine, Methionine, Isoleucine, Leucine and
Phenylalanine (Table.1) and (8) non-essential amino acids namely Aspartic acid,
Serine, Glutamic acid, Proline, Glycine, Alanine, Cystine and Tyrosine (Table. 2)
were recorded in this study through the high performance liquid chromatography.
Table 1: Essential amino acid contents (g per 100g) of earthworm tissue
Essential Aminoacid Control Worm Worms Increased
Cultured On percentage
Aquatic Weed
Lysine 4.5±0.1.7 7.3±0.66 62.22
Histidine 2.3±0.25 3.6±0.22 56.52
Arginine 4.1±0.98 6.2±0.76 51.21
Threonine 3.7±0.55 4.5±0.43 21.62
Valine 3.2±0.94 5.0±0.56 56.25
Methionine 1.8±0.45 2.2±0.20 22.22
Leucine 4.2±0.1.2 7.6±0.46 80.95
Isoleucine 2.7±0.70 4.6±0.32 70.37
Phenylalanine 2.5±0.70 3.3±0.32 32.00
Table 2: Non Essential amino acid contents (g per 100g) of earthworm tissue
Non Essential Control Worm Worms Cultured On Increased
Aminoacid Aquatic Weed percentage
Aspartic acid 5.30 ±0.90 6.68 ±1.45 26.03
Serine 2.40 ±0.60 3.03 ±0.54 26.20

Glutamic acid 4.40 ±1.00 7.80 ±0.78 77.27
Proline 2.55± 0.32 2.76 ±0.34 8.23
Glycine 0.64 ±0.23 0.75 ±0.12 17.18
Alanine 2.80± 0.34 3.70 ±0.36 32.12
Cystine 0.61 ±0.45 0.70± 0.12 9.37
Tyrosine 2.30 ±0.27 2.48 ±0.56 7.83
The earthworm meal contains around 98 per cent of absorption by animal
organism due the balance between vitamins and amino acids. The amino acid contents
of earthworm meal were varied depending on species and food source. Higher values
of all the essential amino acid contents in the Earthworm cultured on Waterhyacinth
compared to that of control in the present study was due to the high protein content of
water hyacinth. It is a potential feed for livestock such as cows, goats, sheep and
chickens. Abdelhamid and Gabr (1991) after chemical analysis on water hyacinths
collected from a canal and a ditch reported as having 9.5% DM, and in the DM 74.3%
organic matter 19% crude protein and 18.9% crude fiber. Aboud et al (2005) reported
that water hyacinth could provide large quantities of nutritious feed and was a
potential source for ruminant nutrition.
Highest level of essential amino acid in the experimental worm was leucine
(8.6g / 100g), followed by Lysine (7.3g / 100g). Similarly, Isoleucine, valine,
methionine, Histidine, Arginine, Threonine and Phenylalanine were increased to
70.37%, 56.25%, 22.22%, 56.52%, 51.21%, 21.62% and 32.00% respectively than the
control. Leucine works with the amino acid Isoleucine and Valine to repair muscle,
regulate blood sugar level and provides the body with energy. Lysine is important for
proper growth to convert fatty acids in to energy and reduce blood cholesterol level.
Methionine is a powerful antioxidant and critical for proper neurological,
immunological and gastrointestinal functions. Methionine also aids the proper
absorption of selenium and zinc and the removal of heavy metals, such as lead and
mercury. Histidine facilitates growth, the creation of blood cells, and tissue repair. It
also helps maintain the special protective covering over nerve cells, which is called
the myelin sheath (Yongqing Hou, et al2015). Histidine supplements in food help to
relieve symptoms of rheumatoid arthritis. It is also important to normal sexual
functioning.
The protein content in the earthworm powder has been used in agriculture for
soil fertility (Medina et al. 2003), in pharmaceutical as an anticancer, antibiotic, anti-

hyperglycemia, anti-diabetes, anti-hypertension and anti-hypotension (Paoletti et al.
2003).
The non-essential amino acid in the experimental earthworm was dominated
by glutamic acid (7.80g / 100g), followed by Aspartic acid (7.80g / 100g). Similarly,
Serine, Proline, Glycine, Alanine, Cysteine and Threonine were increased to 26.20%,
8.23%, 17.18%, 32.12%, 9.37% and 7.83% respectively than the control. Highest
level of glutamic acid among non-essential amino acids in earthworms was reported
in Perionyx excavatus (1.42g / 100g) and Lumbricus rubellus (1.52g / 100g),
(Istiqomah et al., 2009).
The results of the study conducted by Sakthika et al (2014) indicated that fish
fed with earthworm meal prepared from water hyacinth showed increased BWG and
SGR than the control group of fish Mystus montanus. Selvaraj, (2006) reported that
the number of adult worms was found to be 4,590 (more than three times increase
over the initial number of worms introduced) in the water hyacinth compost and each
worm weighed about 1.70gms in the water hyacinth compost. This evidence supports
the high nutritive content of earthworm cultured on waterhyacinth and there by the
reason of increasing concentration of Glutamic acid. It is involved in protein synthesis
and as source of energy for cells lining the intestine. It also facilitates immune
function and improves anti-inflammatory effects, aids in preventing and healing of
peptic ulcer and ulcerative colitis (Istiqomah et al., 2009).
It had long been assumed that NEAA are synthesized sufficiently in animals
and humans to meet the needs for maximal growth and optimal health (National
Research Council, 2012). However, no experimental data substantiate this assumption
(Wu et al 2013). Certain NEAA can regulate gene expression in animal cells, micro-
RNA biogenesis, and epigenetics (Wang et al 2012). For example, dietary glutamine
reduces intestinal expression of genes that promote oxidative stress and immune
activation Wang et al 2008). NEAA affect digestive and absorptive function of the
small Intestine. NEAA arginine, glutamine, and proline regulates immune responses,
including expression of T-cell receptors; lymphocyte proliferation; the production of
cytokines and antibodies; macrophage polarization (i.e. the population of M1 and M2
cells); killing of pathogens by NO, superoxide anion, and H 2O2 modulation of
intestinal microbiota and its function; and prevention of infectious disease (Tan et al
2009).

In managing organic wastes with earthworm, some researchers have been
carried out to determine the ability of certain species of earthworm to degrade organic
materials. Tripathi and Bhardwaj (2004) comparing Eisenia fetida and Lampito
Mauritii noted that rate of decomposition and mineralization were higher in E. fetida
and concluded that E. fetida was a better species for decomposition of kitchen manure
and cow dung. Balasubramanian and Kasturi (1995) noted that the degradation ability
of the earthworm is influenced by waste type. Using cow dung, water hyacinth and
biogas plant effluent, the efficiency of waste breakdown by earthworm was found to
be highest in cow dung followed by water hyacinth and then biogas plant effluent
(Balasubramanian and Kasturi, 1995).
CONCLUSION
Earthworm has high content of essential and non-essential amino acids. Water
hyacinth is a good source of earthworm feed. In the present study earthworm cultured
on water hyacinth showed higher content of aminoacids than earthworm cultured on
garden soil. This eco friendly economical method to produce aminoacid enriched
Earthworm protein is certainly a promising one to meet the dietary requirement as
well as a chance to utilize the aquatic weed to reduce its menace in the aquatic habitat,
more over the vermicompost is an alternate source of chemical fertilizer.
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STUDIES ON THE GROWTH PERFORMANCE OF AZOLLA
PINNATAR.BR. IN DIFFERENT WATER MEDIUM
Kamala Devi N*, Lakshmanan G and Mohammed Ariff Sehriff
Tiruchendur, Tamil Nadu
ABSTRACT
Different types of water medium (cow dung, vermicompost combination and

distilled) each with different nutrient medium were used to grow and evaluate the
growth performance of Azolla pinnata. A. pinnata were grown high with
vermicompost combined water. Because the increased in biomass when available
nutrients of this water medium. High sustainable growth rate (SGR) % were observed
in the vermicompost fed groups 3.5146% day and followed by cow dung 3.339% day
and control 1.807% day were observed. And it is a fast grower and it will enhance the
growth of fish and as well as ornamental fish in a juvenile stage to adult stage and it is
a very good food for poultry, cattle, duck, and white pigs etc. The SGR value is also
clearly shown in the growth performance of A. pinnata fed with different nutrients
source. Based on these analysis indicated that a combination of vermicompost
medium contents explained the observed increased in biomass of A. pinnata grown in
the different water sources.
KEY WORDSAzolla pinnata, vermicompost, cow dung, sustainable growth rate,

biofertilizer
INTRODUCTION
Azolla pinnata, a free-floating, nitrogen (N2) fixing aquatic fern, is an

established N2 biofertilizers for agriculturally important crops. Phosphorus (P) is the
most critical and limiting input for A. pinnata rice cultivation (Majumdar et al 1993).
A. pinnataabsorbs P from the flood water makes it available to the plant. Different
species of A. pinnata have different requirementsto efficient phosphorus scavenging
strain are needed to ensure an adequate phosphorus supply.A. pinnata is very rich in
proteins, essential amino acids, vitamins (A, B12, Beta carotene) and growth promoter
intermediaries and minerals (calcium, phosphorus, potassium, ferrous, copper,
magnesium). It is the most economic and efficient feed substitute for livestock.

A. pinnata is a floating fern in shallow water and dry weight basis is
constituted of 25.35% protein content, 10.15% mineral content and 7 – 19% a
combination of amino acids, bio-active substances and bio polymers are present. The
carbohydrate and oil content are very low and can be easily digestible by the
livestock, owing to its high protein and low lignin content. Also increase milk yield in
animal like cow up to 15-20% replaced by other fodder.
A. pinnata feeding does not affect the milk production, improves quality of
milk and health and longevity of livestock. The poultry bird improves the weight of
the broiler chicken and increases the egg production of layers. It can be fed to sheep,
goat, pig and rabbits as feed substitute 25 to 30% water also needs to be replaced with
fresh water once in 10 days to prevent nitrogen built up in the bed. Replacement of
water and soil should be followed by fresh inoculation of A. pinnata at least once in
six months.
Food is considered as an important ecological factor influencing the

population dynamics of fishes. Fish culture is induced primarily by the need for
increased protein supply. One of the essential prerequisites for the successful
management of fish culture programme is a comprehensive understanding of feeding
(Halver, 1972). The increase in cost and demand of feed protein from conventional
resources necessitates fish culturists of the developing countries to incorporate cheap
and locally available ingredients in fish feeds.
Our modern agriculture is heavily dependent upon chemical fertilizers for

increasing crop yield which slowly accumulated in the environment and pose a danger
to the activity of many other organisms including man. More so, one of the
constraints to high yields is the limited availability and high prices of N 2& P
fertilizers. It has therefore become imperative to seek alternative to the chemical
fertilizers through renewable organic sources will not disturb ecology. Photosynthetic
biofertilizers have drawn considerable attention to maintain the fertility status of soil.
The conversion of molecular nitrogen into organic from by cynobacteria is considered
today as one of the most direct method of utilization of solar energy and also
considered as an extremely low cost biofertilizers. Azolla – Anabaena symbiotic
system has proved to potential N source in water logged rice ecosystem which is most

effective when ploughed into the soil. The natural floating ‘nitrogen factory’ consists
of two plants-the water fern A. pinnata and blue green alga Anabaena azollae living
together in symbiotic association.
The use of A. pinnata with its ‘nif’ genes receives considerable interest as an
efficient biofertilizers. The algal symbiotic Anabaena is harboured in the ventral side
of the dorsal leaves of A. pinnata and remained present during all stages of frond
development. It is very sensitive to the presence or absence of suitable concentration
of nutrients and soil phosphorus. A. pinnata grows well in alkaline soils.
Recently the utilization of aquatic plants having high food value are used to
supplement fish food has taken a new dimension for producing the much required
animal protein at low cost (Lakshmanan et al., 1967).A. pinnata, which grows in
association with the blue green algae Anabaena azolla, is perhaps the most promising
from the point of view of ease of cultivation productivity and nutritive value
(Lumpkin and Plucknett 1982; van Hove lopez 1983). Fish require diets relatively
higher in protein than those of commercially cultured animals. Biochemical and
physiological changes in organisms influence the activities of some enzymes and
metabolic levels of some tissues a (Murty, 1986). Hence the present work is designed
to study the effect of A. pinnata as a fish feed on bioenergetics and biochemical
parameters.
The enormous studies were made on the potential of A. pinnata as a

biofertilizers, yet a very little attempt has so far been made on its productivity under
contaminated soils. Keeping in view, these soils as an important environmental stress,
the present investigation has been undertaken to utilize these soils as a cheap source
for cultivation of A. pinnata. So in the present study is mainly focusing on the growth
performance and biochemical composition of the different medium fed groups in the
laboratory condition.
The experiments were carried out at Research laboratory, Department of

Zoology, Aditanar College of Arts and Science, Tiruchendur, Tamil Nadu, India. The
collected A. pinnata along with native water were maintained in the plastic trough and

acclimatized to laboratory condition (29 ± 2ºC). Before the commencements of the
experiment the medium were prepared with 10 litre capacity fresh water with cow
dung (Experimental set up 1) followed by vermicompost (Experimental set up 2)
paddy field water and control distilled water with cow dung. Each experiments were
carried out in triplicates. A. pinnata were fed with vermicompost (500 g) medium and
control only water with the trough size length 42 cm, width 31 cm height 14 cm and
the water level is 10 litre and control vice versa. In each treatment we have provided
50g A. pinnata water level should was maintained throughout the experimental
duration of 28 days. And every week the weights of the samples were measured with
balance Dried A. pinnata was used as the feed supplement with other ingredients.
The following formula was calculate supplement growth rate,
SGR % day =Final log weight – Initial log weight / days X 100
In this present study the effect of A. pinnata as a protein supplement feed

evidenced by the positive growth performance were clearly noticed in vermicompost
fed groups (Fig 1) and followed by cow dung. The vermicompost shows the best

growth performance followed by other nutrients fed groups (Fig 2). The fig 2 which
shows the best SGR of A. pinnata fed with different nutrient medium in a week
interval. The earth worm is one of the eco friends to the agriculture farmer. It’s a best
tiller in the agric land and it will enhance the soil fertility to give a best yield in the
cultivate lands. And the mass culture of A. pinnata was also done In abroad country
like Thailand and the small scale were also done In the Andhra Pradesh. And its one
of the best feed for the duck, pigs and integrated farming area.
The best growth is due to the worm secret an enzyme that will move to the soil
and enriches the organic manure in addition to the natural sources. After a week the
increment of A. pinnata was shown in figure 1. The nitrogen component is playing a
major role in the growth of A. pinnata. It doubled the growth by a week and
simultaneously increases the growth rate at a week interval. As A. pinnata grows even
in the drainage system and the stagnant water so it’s very simple to culture the A.
pinnata in our environment with low cost nutrient medium.

In the present study the best SGR % day and were observed in the
vermicompost fed groups 3.5146% day and followed by cow dung 3.339% day and
control 1.807% day were observed (Fig 2). And it is a fast grower and it will enhance
the growth of fish and as well as ornamental fish in a juvenile stage to adult stage and
it is a very good food for poultry, cattle, duck, and white pigs etc. the SGR value is
also clearly shown in the growth performance of Azolla fed with different nutrients
source (fig 2). Then the initial and final weights of the A. pinnata are also showed in
Fig3. The pH and temperature is also play a major role in the A. pinnata culture. It’s a
best feed for cultivable fresh water fishes and very important one in the aquaculture
industry
CONCLUSION
In our present study revealed that the best growth performance in A. pinnata
was observed in the vermicomposting fed groups followed by cow dung and then
control. Thus we conclude in saying that vermicomposting is a best biofertilizers to
the agricultural field.

REFERENCES
Majumdar J, Rajagopal V, Shantaram tropical Agriculture, series No

MV. 1993. Rock phosphate is 15,230 p.
an effective phospate carrier for Van Hove, C and Lopez, Y. 1983.
Azolla. Int.Rice Res. Notes Fisiologia de Azolla. In:
18(1):40. Boletin Teenico, Universidad
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Boulder, Colorado. West view

SYNTHESIS OF SILVER NANO PARTICLES FROM EXPIRED
PARACETAMOL TABLETS
Kamala Devi N*, LakshmananG andKrishna sagarV.M
Department of Zoology and Research Centre, Aditanar college of Art and science,
ABSTRACT
The present investigation is a low cost green approach capable of producing verities
of nano materials from expired medicines. This approach consolidates the idea that
also expired medicines can play a part in nanotransformation. Also, this synthetic
method might be a step towards controlling the menace of pollution and open away
for the pharmaceutical companies to recycle their waste medicines or drugs by
synthesizing nanomaterials. Success of such a rapid timescale or synthesis of metallic
nanoparticles is an alternative to chemical synthesis protocols and low cost reluctant
for synthesizing iron nano particles. The various nano particles synthesized from
expired medicine also evaluated for their antibacterial activity and it showed a
positive result which proved to be an effective antibacterial drugs.
KEY WORDS: nano particle, paracetamol, antibacterial activity, zone of inhibition
INTRODUCTION
Nanotechnology in undoubtedly one of the most important technology of the

21st century. With an estimated $ 147 billion worth of so-called ‘nano-enabled’
commercial and consumer products sold in 2007 and a projected value of $ 3.1 trillion
by 2015 (LuxResearch 2009). Nanotechnology has been referred to as the next
industrial revolution (Schmidt, 2009). Nanotechnology shows great promise in
providing solutions to many of today’s problems in medicine, energy production, and
environmental sustainability. Governments and investors properties of nonmaterial
opportunities which exist within nearly every industrial and technological sector.
Nanotechnology, involving the creation of novel materials that function more

effectively and can perform new roles. Nanosized particles will therefore be

chemically more reactive per unit mass than their bulk sized counterparts. This
property makes the m particularly desirable for use within bio medical applications
where dissolution rate is enhanced (Gupta and Kompella, 2006) and within chemical
processes as catalysts.
Silver has been known for antibacterial activity since the ancient Greece
times. Currently, the investigation of this phenomenon has gained more attention due
to the increase of bacterial resistance of antibiotics, caused by their overuse. Silver
nanoparticles can be medicine to reduce infections in burn treatment, to prevent
bacteria colonization on dental materials, stainless steel materials, to eliminate
microorganisms on textile, or they can be used for water treatment (tomisc et al.,
2008).
Copper nanoparticles, due to their unique physical and chemical properties

and the low cost of preparation, have been of great interest recently. Copper
nanoparicles are exploited in wound dressings and socks to give them bio-cide
properties (Borkow et at., 2010) Furthermore, copper nanoparticle have potential
industrial use such as gas sensors, catalytic processes, high temperature
superconductors solar cells and so on (Li and Guo et al., 2007).
In green nanotechnology, different microorganisms produce inorganic

materials, either intracellular or extravellularly with properties similar to chemically
synthesized materials (Bauerlein et al., 2003). (Usha et al., 2010) reported a green
synthesis of copper oxide by Streptomyces sp. for development of antimicrobial
textiles which can be useful in hospitals to prevent or to minimize infection with
pathogenic bacteria (Usha et al., 2010).
Iron oxide nanoparticles have been widely researched for MRI, as they are
mainly superparamagenitic. There are several types of iron oxide nanoparticles,
namely maghemite,  - Fe2 O3, magenetite, Fe 3O4, and haematite,  - Fe2 O3,
among which magnetite, Fe3 O4, is very promising, because of its proven
biocompatibility (Gupta et al., 2005). For molecular imaging purposes,
superamagnetic iron oxide nanoparticles (SPIONS) need to be biocompatible, non-
oxic and magnetic. They also need to bind to a range of drugs, proteins, enzymes,
antibodies, or other molecular targets, there have been a number of approaches to the

production of SPIONS for use as MRI contrast agents, and each method produces
particles with different sizes and magnestisation parameters. The iron oxide
nanoparticles can also be coated with a surface player, usually of organiz materials,
that provides an interface between the core and the surrounding environment
(Rochelle et al., 2007). This surface layer can be used to direct the particles to a target
site.
Paracetamol or arcetaminophen (ACOP) is a widely used analgesic anti

pyretic drug. It is a suitable alternative when he patients are sensitive to aspirin (Wade
et al., 1979). It is used to reduce fever cough and cold, and reduce mild to moderate
pain, including instances of tension headache, migraine headache, muscular aches,
chronic pain, neuralgia, backache, joint pain, general pain and toothache (Koch Wese
et al., 1976). It is also useful in osteorathristis therapy (Brandtetal., 2003) and it is
sometimes used for management of cancer pain. Recent research suggests that
paracetomol may help to protect from charges leading to hardening of arteries that
cause cardiovascular disease (Toylar et al., 1998). It also remains the analgedic of
choice for people with asthma. There is also some evidence to suggest that
paracetamol may offer some protection against ovarian cancer (Cramer D.W et al.,
198) ACOP rapidly gets absorbed and distributed after oral administration and is
easily excreted in urine. Generally, paracetamol (PC) does not exhibit any harmful
side effects by hypersensitivity or overdoses in ew cases leads to the formation of
some liver and nephrotoxic metabolites (Patelet al., 1992). Because PC is being
increasingly used for therapeutic purposes, its determination and quality control are of
vital importance (Lourence et al., 2009). Disposal of paracetamol in an indiscriminate
manner cause serious impact on the environment.
Nanotechnology mainly deals with the fabrication of nanoparticles having

various shapes, sizes and managing their chemical and physical parameters for further
use in human benefits. Their growing applications in various fields like biosensors
(Wade et al., 1979) bioremediation of radioactive wastes (Koachweser et al., 1976),
functional electrical coating (Clissold et al., 1986), synthesis ofenzyme electrodes and
particularly in medicine such as delivery of antigen for vaccination (Brandtt et al.,
2003). Gene delivery for treatment of prevention of genetic disorder inspired the

scientists to develop environment friendly procedures for the synthesis of
nanoparticles and to avoid use of hazardous chemical, which are traditionally used.
To do this, different nanomaterial has been synthesized using expired

medicines. This study aims to provide a green protocol to synthesize different
nanomateials using a green chemistry approach. Green chemistry is the design,
development and implementation of chemical products and the process of reducing or
eliminating the use and generation of substances hazardous to human health and the
environmentally benign solvents, biodegradable polymers, and non- toxic chemicals.
The expired medicine which is considered merely as a waste could be used for the
synthesis of nanoparticles.The objectives of the present investigate environmental
friendly method of waste disposal. And synthesize various nanoparticles from expired
tablet and also test their antibacterial activity.
In the present investigation, an attempt was made to synthesize various

nanoparrticles such as silver, copper, cadmium and iron from the expires tablet,
Pracetamol.
Peparation of broth from expired drug (Paracetamol)
Four-month expired Paracetamol (500mg) tablets were dissolved slowly in

100ml sterile distilled water. To this was added 5 ml N/20 HCL to ensure a better
degree of dissolution through gentle heating over a steam bath of up to 40  C until a
light yellow color appeared in the conical flask. Then 25 ml of this source broth was
diluted 4 times by the addition of distilled water. This source solution (pH=3) was
used to synthesize nanomaterials.
Biosynthesis of Silver Nanoiparticles
AgNO3 of 1 mm was prepared by adding 0.015 g of Ag NO3 to 90 ml of

distilled water and used for the synthesis of silver nanoparticles. Then 10 ml of flower
exract was added into 90ml of prepared aqueous solution of 1 mm AgNO3 for
reduction into Ag + ions and kept in magnietic stirrer for 1 hour at room temperature.

Biosynthesis of Cadmium nanparticles
Twenty ml of 0.25(M) Cd Cl2 solution was purged with H2S and this was
heated for 10 min over a water bath in order to expel the surplus hydrogen sulfide.
This was charged to the broth solution shifting the pH to 4. Immediately, the CdS
appear fluffy and on progressive heating the pH further shifted to 5 leading to a
successful transformation as indicated by the yellowish / Orange color of the broth
and the deposition of the precipitate at the bottom.
Biosynthesis of Copper nanoparticles
Twenty mL of Fehling A (2% sodium carbonate in 01. N NaOH) and Fehling

B (0.5% copper sulphate solution in 1% sodium postassium tartarate) was boiled and
mixed with 20 mL of expired tablet broth. Red precipitate is formed; indicate the
presence of copper naniparticle.
Biosynthesis of Iron nanoparticles
10 ml of Ferric chloride solution (8.1 g of Ferric chloride was dissolved in 50

ml deionized water and stirred for 15 minutes) was mixed with 10 ml of the tablet
brother in a breaker. After 10 minutes, the color of the solution changed from brown a
black indicating the formation of iron oxides nanoparticles.
UV-VIS Spectra analysis
The reduction of nanoparticles was monitored by measuring the UV-Vis

spectrum of the reaction medium at 5 hours after diluting a small aliquot of the
sample in to distilled water. UV-Visible spectra of nano particles were recorded with
an Elico 1800 UV Spectrophotometer.
Antibacterial test for nanopartilces
Agar diffusion assay is used widely to determine the anti0 bacterial activity of
nanoparticles. Nutrient agar prepared was poured in the Petri dish. 24 h growing
bacterial culture were swabbed on it. Sterile discs obtained from Himedia
Laboratories Pvt ltd were used for antimicrobial studies. The sterile disc was dipped
in solution containing naniparticles and placed on the Petridis. Plain sterile disc was

used as control. The plates were then incubated at 37C for 24 h. The inhibition
diameter was measured.
RESULTS
Visual observation and UV- Vis spectral study
Formation and stability of AgNPs in sterile distilled water is confirmed using

UV –vis spectrophotometer in a range of wavelength from 380 to 1100 mm. As soon
as, the extract was mixed in aqueous solution of silver ion complex, the reduction of
pure Ag+ ions to AgO was monitored by measuring UV- vis spectrum of the reaction
media at regular intervals. UV-vis spectra were recorded as function of reaction time.
We observe that there is no peak showing no sign for the synthesis of silver
nanoparticles but after 5 min the surface Plasmon resonance of silver occur at 425 nm
(Fig 1` ) and steadily increasing with the time of reaction without much change in the
peak wavelength.

The reduction of iron ions into iron particles occurred after mixing ferric chloride
with tablets broth, followed by colour change in pH of solutions. As the tablet broth
was mixed in the aqueous solution of the iron ion complex, it started to change the
color due to reduction of iron ion, which may be the indication of formation iron
nanoparticles (Plate 1). The UV-visible spectrum of nano particle is shown in the
above figure. The two absorption peaks at wavelengths of 525 nm and 650 nm (Fig 2)
indicates the formation of nano iron particles.
DISCUSSION
In the present investigation, the expired tablet paracetamol is used as a raw

material for the synthesis of various nano particles such as silver, iron, copper and
cadmium.
Paracetamol is an antibiotic commonly used for any ailment. Man y

fluorinated compounds are currently widely used in the treatment of a variety of

diseases. These include antidepressants, anti-inflammatory agents, anti-malarial
drugs, anti- psychotics, antiviral agents, steroids and general anesthetics, (Dollery,
1999). The chemistry and medicinal chemistry of fluoroorganic compounds and drugs
have been reviewed (Resnati 1990). The inclusion of a fluorine atom in a drug
molecule can influence both the disposition of the drug and its interaction with its
pharmacological target. For example, the effects of fluorine substitution on the inter
and intra molecular forces that affect binding of ligands to cholinergic and adrenergic
receptors, and this introduce receptor subtype selectivity, are now well understood
(Bravo, 1992).
The replacement of a hydrogen atom or hydroxyl group by a fluorine atom is a

strategy being widely used in drug development of alters biological functions.
Although it is generally thought that fluorine for hydrogen substitution causes,
minimal static effects at receptor sites, the actual Van der Waals radius of fluorine
(1.47 A) lies between that of oxygen (1.57 A) and hydrogen (1.2 A). Despite the fact
that fluorine has a greater size than hydrogen, several studies have demonstrated that
it is reasonable to believe that hydrogen mimics and exerts only a minor steric
demand at receptor sites, at least for nano functional analogs (O’ Hagan, 1997).
In contrast with noble metals, such as silver and gold, the synthesis of copper
nanoparticles is much more challenging, since copper nanoparticles are fairly unstable
in aqueous solution. However, copper costs significantly less the silver and gold,
therefore, it is economically attractive. The additional of ferric chloride solution to
any extract containing carbohydrates as a major component which have aldehyde
group may cause the partial reduction of Fe3+ to form Fe3 O4. First, FeCl3, hydrolyzes
to form ferric hydroxide and releases H+ ions in the proper pH value and temperature.
After that, ferric hydroxide is partially reduced by the plant extract containing
carbohydrates (glucose) to form Fe3 O4., While aldehyde group is oxidized to
corresponding acide (Senthil and Ramesh, 2012)
Antimicrobial activity of colloid silver particles is influenced by the

dimensions of the particles. The extremely small size of nanopartivles results in the
particles having a large surfaced area relative to their volume. This allows them to
easily interact with other particles and increases their antibacterial efficiency. In

heteorneous catalysis, it is well known that the increasing proportion of surface atoms
with decreasing particles size makes small metal particles highly reactive. Silver has
been utilized as oxidation catalyst especially for production of ethylene oxide from
ethylene (Shiraish and Toshima 1999)
Cadmium nanoparticle showed maximum inhibition activity than order

nanoparticles. The Cd2+ accumulation culture showed a similar accumulation under
neutral and basic conditions (Volesky and Holan 1993) B. subtilis was resistant to
ambient conditions, such as pH, ionic strength and the presence of metal chelators or
complexing agents. Once Cd2+ was transported into the cytoplasam, I formed a
complex with the metellothionein (MT) protein. In the induction at the early and late
logarithmic phases, Cd2+ was accumulated. It was speculated that this was due to the
incorporation based on the polysaccharide composition of each particular organism,
and is highly variable among distinct genera and even strains from the same species.
Gram-positive cells has tricholic acids associated to the cell wall, whose phosphate
groups are key components for the uptake of metals (Beveridge 1989).
Furthermore, this suggests that nano silver, iron cadmium and particles could
be potentially effective to defend against bacterial of fungal attacks and could be
prepared even from and expired medicine which is considered as an in disposable
waste.
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http://www.nano.gov.

SYNTHESIS OF LEAD NANO PARTICLES FROM CRAB SHELLS
A.Kavitha*, N. kamalaDevi*, B. Megala and G.Lakshmanan*
ABSTRACT
Many scientists paid their attention towards the synthesis of silver and copper
nanoparticles. Hence, today more works have been done our synthesis of silver and
copper nanoparticle. But only dearth of knowledge is available for synthesis of lead
nanoparticles. So that in the present study we have also concentrated and done the
synthesis of lead nanoparticles from the crab shell. Green chemistry approach for
synthesis of lead nanoparticles has many advantages such as ease with which the
process can be scaled up, economic viability etc.
INTRODUCTION
Nanotechnology is the technology by which an atomic and molecular scale
level matter is material was skillfully managed as tool for various applications. It
deals with nanometer sized object. Nanoparticles are the fundamental building blocks
of nanotechnology. Nanoparticles synthesized from microparticles which are present
at nanoscale level in different parts of plants and animals. For example nanoparticles
present in root, stem, leaves, flowers and seeds of plants. Especially antennas,
cuticles, hair, feathers, and shells of animals.
Recently the animals are used to synthesis functional nanoparticles has been of
great interest. Synthesis of nanoparticles using biological entities has greatest
attention from the scientists throughout the world. It is due to their unique properties
such as size and shape depending optical, electrical and magnetic properties that have
antimicrobial application, biosensor materials, composite fibers, cryogenic
superconducting materials, cosmetic products and electronic components. For the
reasons of many applications in the field of synthesizing nanoparticles such as drug
delivery system, gene delivery system, destruction of tumor etc. The present study

was taken in synthesize of nanoparticles from crab shell. Though the crab shells are
harmful to the environment they may be transformed with valuable products.
The synthesis of lead nanoparticles by the reduction and thermolysis of
organo lead compounds in organic solvents and polymer matrices, the electrolytic
deposition from aqueous solutions, photolysis, radiolysis and the dispersion of molten
lead is organic or metallic media were the methods used. Green chemistry approach
towards the synthesis of lead nanoparticle has many advantages than other methods.
Green chemistry approach for synthesis of lead nanoparticles has many
advantages such as ease with which the process can be scaled up, economic viability
etc. Application of lead nanoparticles such as ecofriendly nanoparticle in the
electronic industry make this method potentially more relevant for the large scale
synthesis. Biological methods are highly effective hence in this project this method is
followed for the synthesis of lead nanoparticles.
Collection of crab shell:
Crab shells were collected from fish market, coastal areas from vast shore
fauna.Then cleaned thoroughly with distilled water in order to remove debris. It is
kept for few minutes. Then it powdered and sieved. The fine powder collected and
stored for further analysis.
Preparation of extract:
5g of crab shell powder was weighed and dissolved in 30ml of distilled water. It
is stirred vigoursly for few minutes then filtered thrice. The crude filtered extract of
crab shell was stored for further analysis.
Synthesis of lead nanoparticles:
Crab shell extract is taken in different concentration viz 1, 1.5, 2, 2.5,3, 3.5, 4, 4.5, 5
ml. It is then added to 5ml of 1Mm lead acetate for each concentration.Then it will be
incubated at room temperature for five hours. The appearance of mild colour changes
occurs.
Characterization of lead nanoparticles
Spectral analysis:
The reduction of lead ions was determined in a range of 250 – 500 nm using
UV- Vis spectrophotometer.

Morphological analysis:
Morphological analysis of lead nanoparticles was done by using Scaning
Electron Microscope (SEM). It was carried out in the Institute of Forest Genetics and
Tree, Coimbatore.
Plate: 6 Different concentration of crab shell extract for synthesis of lead

nanoparticles
Observation of white colour
RESULTS
UV-Vis Spectrum analysis of Lead Nanoparticles:

Lead nanoparticles synthesis was confirmed using UV-Vis absorption
spectroscopy. Small lead nanoparticles exhibit the absorption of visible
electromagnetic waves by the collective oscillation of conduction electrons at the
surface. The lead acetate is the reducing agent for the lead nanoparticles. The
reduction reaction take place for 5hours and watery solution turns white in colour.
Different concentrations of crab shell extract shows different ranges of peak value
between 264 – 393 nm the time takes from the reduction for lead nanoparticles is 5
hours. The observed peak broadening and noise were probably related to the effect of
nanosized particles and the presence of various crystalline biological macromolecules
in the crab shell extract. The results from UV-Vis of the sample was shown in the
figure 15-23.

Crab Shell Extract Lead Wave length
Concentration acetateSolution (UV-Vis)
(ml)
1ml 5 264
1.5ml 5 378
2ml 5 312
2.5ml 5 379
3ml 5 341
3.5ml 5 370
4ml 5 298
4.5ml 5 328
5ml 5 313
Fig 15: UV-Vis Absorbance Spectra of lead Fig 16: UV-Vis Absorbance Speectra of
nanoparticles using 1ml crab Shell extract nanoparticles using 1.5ml Crab shell extract
4.0
378 nm
3.5
1.5ml
3.5 3.0
3.0
265 nm 2.5
1 ml
Absorbance
2.5 2.0
2.0 1.5
Absorbance
1.5 1.0
1.0 0.5
0.5 0.0
250 300 350 400 450 500

0.0
Wavelength nm
-0.5
250 300 350 400 450 500
Wavelength nm
Fig : 17 UV- Vis Absorbance Spectra of lead Fig: 18 UV- Vis Absorbance Spectra of lead
nanoparticles using 2 ml of crab shellextract nanoparticles using 2.5 ml of crab shell extract
3.5
379 nm
3.0
2.5 ml
2.5
4.0
3.5 2.0
312nm
Absorbance
2ml
3.0 1.5
2.5 1.0
Absorbance
2.0 0.5
1.5
0.0
1.0 250 300 350 400 450 500

Wavelength nm
0.5
0.0
250 300 350 400 450 500

Wavelength nm
Fig :19UV- Vis Absorbance Spectra of lead Fig :20 UV-Vis Absorbance Spectra of lead
nanoparticles using 3 ml of crab shell extract nanoparticles using 3.5 ml crab shell extract
4.0
3.5 341 nm
3 ml
3.0
2.5
Absorbance
2.0
1.5
1.0
0.5
0.0
250 300 350 400 450 500

Wavelength nm
Fig :21 UV – Vis Absorbance Spectra of lead Fig : 22 UV – Vis Absorbance Spectra of lead
nanoparticle using 4ml crab shell extract Nanoparticle using 4.5 ml crab shell extract

4.0
3.5 298 nm
3.5 313 nm
3.0 4 ml 5ml
3.0
2.5
2.5
Absorbance
2.0
Absorbance
2.0
1.5
1.5
1.0
1.0
0.5
0.0 0.5
250 300 350 400 450 500 0.0

Wavelength nm
250 300 350 400 450 500
Wavelength nm
SEM analysis of the synthesized samples were performed in order to investigate the
morphology and distribution of Lead Nanoparticles. The SEM images are observed at
different magnification such as X 7,000, X 10,000, X 20,000, X 30,000 with 20kv.
All of them shows rod and irregular ball in shape. From the SEM images it is evident
that the morphology of lead nano particle is rod and irregular ball which is good
agreement with the shape of SPR band in the UV-Vis spectra. The SEM analysis of
lead nano particle is shown in the figure 24-27.
Fig:24 SEM Analysis of Lead Fig:25 SEM Analysis of Lead
Nanoparticles using Crab Shell Nanoparticles using Crab Shell Extract
Extract with X 7,000 with X 10,000

Fig:26 SEM Analysis of Lead Fig:27 SEM Analysis of Lead
Nanoparticles using Crab Shell Nanoparticles using Crab Shell Extract
Extract with X 20,000 with X 30,000
DISCUSSION
In the present study, formation of silver, copper, lead nanoparticles was
confirmed by UV-Vis spectra analysis and SEM analysis for the synthesis of various
metal nanoparticles, the colour changes followed by time measures for reductioin
reaction and absorbance ranges in nanometric scale predict the conformation of
nanoparticles in the sample.
UV-Analysis of lead nanoparticles:
In the current work the presence of lead nanoparticles was reported. The
reduction reaction takes for 5 hrs and watery solution turns white in colour. The
particle size ranges from 264- 393nm. It is found that crab shell extract is a good
source for the presence of lead nanoparticles
This could be compared to the present work of Pavani et al.,(2012). They
have reported the synthesis of lead nanoparticles using grapes skin extract. The
nanoparticles were assessed by UV-Vis spectroscopy with particle size range form
661 – 796 nm. They showed the phytochemical compounds such as flavones and
anthocyanins cause bioreduction of lead ions into lead nanoparticles.
SEM analysis of lead nanoparticles
From the crab shell extract the lead nanoparticles appeared in rod and irregular
ballshapped structure.Because of non – availability of relevant literature, this work is
not compared with others.

SUMMARY
The waste crab shells are used for synthesis of nanoparticles such as silver,
copper and lead.For silver nanoparticles, the time taken for reduction reaction is 48
hrs. Reddish brown colour developed from the extract after the introducing reducing
after silver nitrate.For copper nanoparticle the time taken for reduction reaction is
only 5 minutes. Black colour is developed during reduction reaction. It may be due to
the occurrence of copper metal is higher in the crab shell.For lead nanoparticles the
time taken for reduction reaction is 5 hrs.Significant amount of lead is also present in
the crab shell.Peak value for silver, copper and lead observed in UV
spectrophotometer.SEM Analysis have been done from the SEM analysis, it was
found that the crab shells are one of the main sources for the presence of silver,
copper and lead nanoparticles.
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SURVEY OF MARINE MOLLUSCS FROM TIRUCHENDUR
LOCATIONS OF THOOTHUKUDI COST, TAMIL NADU, INDIA
Kamala Devi N*, LakshmananG andMuthu lingamS
Department of Zoology and Research Centre, Aditanar college of Art and science,
ABSTRACT
The objective of this project is to provide standard survey that can be used by field
personnel to determine presence/absence for marine mollusc species, i.e., to
determine what marine mollusc species are present in a selected sea shore. This
methodology can be used to collecting information about what species are present. It
can also be used during general inventories of marine molluscan for distribution or
other efforts to determine distribution or habitat associations.
KEYWORDS: marine mollusc, distribution, soft body, consumption
INTRODUCTION
Mollusca are one of the most diverse groups of animals on the planet, with at
least 50,000 living species.It includes such familiar organisms as snails, octopuses,
squid, clams, scallops, oysters, and chitons.These belong to the groups of
monoplacophorans.Molluscs are soft bodies which typically have a “head” and a foot
region.Molluscs are extremely important members of many ecological
communities.Distribution from Terrestrial Mountain tops to the hot vent and cold
seeps of the deep sea, and range in size from 20- meter – long.These creatures have
been important to human’s thought history as a source of food, jewellary, tools and
even pets.They also have a very long and rich fossil record going back more than 550
million years. The study of molluscan diversity along Thoothukudi coast and Gulf of
Mannar is very limited.

STUDY AREA
The study area Punnakkayal coast is located between 8 º,39;09 N latitudes

780,09 and 22E longitudes. It is located at the middle of Thoothukudi and Tiruchendur
coast. The distance from Thoothukudi is 20 km and Tiruchendur is 25 km.

After collection the shells are washed with 10% hydrochloric Acid solution.
The remaining debris removed with the help of knife then washed with pure fresh
water dried in sunlight and then identified
IDENTIFICATION OF GASTROPODS
GASTROPODS FEATURES
Radial with two elongated teeth in each row, jaws are
absent; esophagus with a large poison gland; siphon and
CONUS
proboscis are well developed; shell fusifrom with an
elongated spine.

Shell in cypraea is oval, rounded on the top and flattened
beneath and consists of a large whorl. The shell opening is
CYPRAEA long and narrow, toothed on both sides and channeled at
each and .Their extremely smooth, highly polished and
variously colored shells make them very conspicuous and
are prized by the collectors.
EBURNA Shell fusiform with a pointed sprine and along; Nassa with
two slender, posterior appendages in the foot.
Long proboscis and siphon well developed redial with at

most three teeth in a row; Marginals absent; carnivores
MUREX highly predatory. Shell with prominent spire ornamental
with spines; eyes at the base of the tentacles, it produces a
purple due secreted by the anal gland.
Long proboscis and siphon welldeveloped; with at most

PTEROCEROUS three teeth in a rows; marginalsabsent, carnivorous highly
LAMBIS predateory. Shell with prominent spire ornamental with
spines.
Foot narrow, laterally compressed notadapted for creeping;
STROMBUS
progresses by a sort of leaping movements carrying its
(WING SHELL)
heavy shell and rolling along in a series of jumps in a most
singular and grotesque manner.
Helix can survive a water loss equal 1 to 50% of the body
HELIX weight and the sluglimax can survive an 80% loss. In the
shelled Helicide (helix) the vagina contains an oval dart
sac, which secretes the calcareous specula.

External shell stout, relatively solid able to accommodate
the animal during defensive retraction, consisting of up 8
whorls. Horny operculum present.Cephalic shield forming
ACETON a pair of antero-lateral lobes, and a pair of postero-lateral
flaps which may conceal part of the shell. Parapodial lobes
not developed. Posterier pallial lope in conspicuous;
mantle prolonged inside the shell to form a spiral caecum.
Gill present in mantle cavity. Large penis, non-retractile.
These gastropods are larger and bottom – dwelling. They
BUSYCON
feeds on bivalves, other gastropods, and echinoderms.
Most of the times, they bunied with In shallow ocean.
Holiotes inhabited in Inter tidal and shallow water rocks of
HALIOTES the ocean. The shells are designed for water resistance and
protection.
BIVALVES
BIVALVES FEATURES
The shell consists of two valves. Which are usually
symmetrical. The two valves articulate by a system of
CARDIUM
teeth and sockets to Forma hinge and are also tightened
by ligaments. The teeth in some cases are reduced or
reduced or may absent.
The outer surface of the valves is covered with
CARASSOSTREA foliaceous / aminae, which closely overlap. The left
MADRASENIS valve is much deeper than the right. The inner surfaces
PRESTON of the valves are smooth, glossy and deeper than that of
the edges and sometimes almost approximates to jet
black the shell attains a fairly large size.
MERTRIX This is a large clam with a thick shell and grows to a
MERETRLX length of about 75 mm. The periostacum is thin, delicate

and grey or straw colour on the posterodorsal margin of
the shell.
Shell large, elongate, sub trigonal with its anterior end
pointed arched and beak like dorsal margin slightly
concave in middle; posterior margin broadly rounded;
PERNA VIRDIS
ventral margin slightly concave. Surface of shell
strongly decussately striated ligament strong and
elongated. Shell surface covered by a firm horny, bright
green periostracum.
Pecton is commonly known as “scallop”. The unequal
PECTON
shell valves are beautifully sculptured and bear radiating
IRRADIAND
ribose striations. The hinge ligament is straight and
toothless.
Pinna like pen shells anchor themselves in sand These
mollusks place their anterior end downward, while their
elongate, wing –shaped valves extend above the
PINNA CARNEA
substratum. The foot and other anterior structures are
reduced.Most of the mantle edge in pinna is free from
the shell. It may also excel broken shell particles and
other debris.
Shell more or less trigonal, sometimes oblong,
SACCOSTRES
extremely hard and plaited plaits more or less angular,
CUCULLATA
generally small, lower value thick, overlapping at
(BORN)
margin, hooded under the margin, hinge elongated
frequently produced at the apex
The shell consists of two valves. The two valves
TELLINA
articulate by a system of teeth, and sockets to from a
hinge and are also tightened by ligaments.
Shell thick, tumid, irregularly oval or rhomboidal in
MELONGENA outline; umbones anterior bus sub terminal, with a
projecting, rounded, anterior, anterior margin; dorsal
margin markedly convex, ventral margin slightly

concave. Umbonal ridges well-developed. Young
specimen with numerous long, smooth, slender
periostracal spines, frequenly with attached detritus, lost
as the shell grows, represented only by a roughened area
at posterior end. Sculpture of fine concentric lines and
ridges, growth stages often clear (M. modiolus –drawin).
CEPHLOPODA
CEPHALOPODA FEATURES
The sepia is commonly found in shallow waters of our
coast. Sepia is active swimmer. It feeds an animal like
SEPIA crabs, pawns, small mollusks and marine worms. The
internal shell is present. The shells of dead animals may be
seen washore and they are called as ‘Cuttle bones’.
Nautilus or pearly nautilus is found in seawater in Indo

pacific region.The body is enclosed in a spirally coiled shell
which is divided into interval chambers by transverse septa.
NAUTILUS The animal lives in a last chamber which is the largets and
the body can be withdrawn for protection. The air fills all
the chambers expect the last. This made the shell buoyant
and enables the animal to swim easily.
Shells are small to medium-sized, circular or polygonal in

section. Longituditudinal rib lets are frequently lacking in
aduIt stage but present on the apical portion of the shell in
DENTALIUM
juvenile stages. Apical orifice generally with a v-shaped
notch on or near the concave side and generally bearally
bearing a solid plug with a central pipe or orifice, rarely
simple.

DISCUSSION
The molluscans resources offer an immense scope for increasing food

production.Cultivated employing cheap but effective methods.Small scale artificial
culture of pearls industry.Shellfish will also result in greater availability of quality
lime, much in demand for construction of human habitation.Harvesting processing
and marketing of the utilizable species, besides culturing some of them will provide
employment opportunities.The best know phylum in the invertebrate word is the
Molluscan lay persons, and scientists alike collect the shells of these, often beautiful
and fascinating animal.Today, with 80,000 living and 35,000 fossil species, the
molluscs, rant as the largest non-arthropod group. Molluscs have been used by human
though the centuries as tools, jewelry, and money.
Molluscs live in nearly all environments. They are numerous on the ocean
floor,where they range from great depths to coastal and intertidal zones. Pelagic and
plank tonic molluscs are also known. Freshwater bivalves and gastropods are well
established within the phylum is infrequent.The generalized characteristics on which
the unity of the phylum is based. Bilateral,coelomate, probably unsegmented
invertebrates Animals with a dorsal shell secreted by an underlying mantle, a visceral
hump and a ventral muscular foot. Animals with a complete digestive tract featuring a
radula and paried digestive diverticula. Animal with an open circulatory system,
respiratory gills within a mante cavity, and complete metanephridia. Animals whose
movement depends on muscular waves and an antagonistic relationship between the
muscles and a hemocoelic skeleton.
Typical pot stones with a trochophore and often veliger larva in the life
cycle.For this reason their fossils are indispensable to the science of Geology in
general and to the science of petroleum Geology in particular. Through an extensive
study of marine molusc fossile, the petroleum geologists are able to locate valuable
oil deposits which often lie several miles beneath the surface of the earth.

Plate: Identified molluscan species

REFERENCES
Rees, W.J. 1965. The aerial dispersal invertebrates from the United
of Mollusca. Proceedings of the States and Canada: mollusks,
Malacological Society of 2nd edition. American
London 36:269-282. Fisheries Society special pub.
Smith, D. G. 2000. Notes on the 26, Bethesda, MD
taxonomy of introduced Davis, G.M. 1967. The systematic
Bellamya relationship of Pomatiopsis
(Gastropoda:Viviparidae) lapidaria and Oncomelania
species in northeastern North hupensis formosana
America. Nautilus 114(2):31- (Prosobranchia: Hydrobiidae).
37 Malacologia 6: 1-143.
Turgeon, D.D., J.F. Quinn, Jr., A.E. Dindal, D. L. (ed.) 1990. Soil Biology
Bogan, E.V. Coan, F.G. Guide. J. Wiley, N.Y. 1359 pp.
Hochberg, W.G. Lyons, P. Duncan, N. 2006. Report on
Mikkelsen, R.J. Neves, C.F.E. mussel survey techniques and
Roper, G. Rosenberg, B. Roth, results for the Umpqua Basin,
A. Scheltema, F.G. Thompson, Douglas County, OR. Roseburg
M. Vecchione, & J.D. Bureau of Land Management,
Williams. 1998. Common and Roseburg.
scientific names of aquatic

COMMERCIALLY IMPORTANT MARINE MOLLUSCS FOR
HUMAN CONSUMPTION IN TUTICORIN, TAMIL NADU, INDIA
Kamala DeviN*, Lakshmanan G and Petchimuthuananth B
Tiruchendur, Tamilnadu, India
ABSTRACT
Increased human population needs more marine food resources. Marine habitat offers
a variety of food to the human beings. Scientists from different nations urged to
develop a conventional food from nonconventional food. Marine bivalves play a vital
role in human foods and food industries. Clams are the non-conventional food
sources but readily available at all seasons. This is non-conventional clams are
modified into conventional nature by adopting some cooking methods. A calm pickle
is tastier and contains high valuable energy. Ready to serve soup powder is also high
nutritive one. Chutney powder is also prepared easily and which provide good tested.
A significant number of native species that have aquaculture potential should be
studied for commercial culture practice as well as to conduct studies on their biology
on those species that are considered over-exploited and allow recovery of the species
in its habitat.
KEYWORDS: Molluscs; Commercial; Consumption; Human; Acapulco; México
INTRODUCTION
Large numbers of molluscan species are utilizable as a source of cheap but

nutrients food. Some members of the group produce beautiful pearls priced as high as
gems. Molluscan shells are put to varied uses: their exquisite from and durability of
texture, often coupled with pleasing color patterns have made some of the sea shells
covetable possessions of man from primitive to the present day civilizations. The
largest utilization of the molluscan shells is for burning them into lime to be used in
masonry constrictions and white washing of buildings. Molluscs have many other
uses too: for instance, some of the snails yield purple for dyeing cloth and the feather
shells of the Family pinnnidae yield silky byssus threads which could be woven into
socks or similar useful articles. As in agricultural farming, some of the species lend
themselves admirably for cultivation to offer good quality of hygienically clean food
for human consumption.
Molluscan species inhabiting the coastal waters, backwaters and estuaries are
economically of much greater importance than those found on land or in fresh water
habitats.
The main utilizable species come under three major taxonomic groups, namely,
1. Lamellibranchiata which includes mussels, oysters, clams, pearl oysters and

the windowpane oyster.
2. Gastropoda which includes snails, top shells and turban shells,
andCephalopoda which includes the Nautilus, squids, cuttlefish and the octopi.
In the following account the distribution of the molluscan resources and the
extent to which they are utilized at present in India are dealt with indicating the future
scope for their fuller utilization.
Mussels
Mytilus viridis, the green mussel belonging to the family Mytilidae, is a fast
growing bivalve attaining a length of over 13 cm. mussels adhering together from
thick encrustations over the submerged coastal rocks. The species is distributed
widely along the east and the west coasts, often extending into the estuaries and
backwaters. Mussel beds are extensive in kerala and Southern Mysore, but rather
sparse in Karwar, Goa and further north. On the east coasts its occurrence is much
restricted, being found to some extent around Madras and in the vicinities of the
Mahanadi estuary in Oriyya. Mussels are a favorite item of food many of the coastal
populations.In Spain, France, Portugal, Holland and other European countries mussels
are cultivated by collecting the young mussles called the seed mussels and rearing
them either on stakes fixed in shallow waters or suspending them in the surface
waters by means of floating rafts. Under such conditions they grow fast and thrive
well as they are not smothered by the bottom mud and are also prevented from attacks
by predatory organisms.

Oysters
The edible oysters of which several species occur belong to the Family
Ostreidae under the Lamellibranchiae. The Indian backwater Oyster, Crassostrea
madrasensis, has a wide distribution in all the east coast, west coast back waters and
estuaries.
Clams and other bivalves
The clams are burrowing and available in the bays, creeks, estuaries,
backwaters and the surf- beaten sandy shores. They are collected in considerable
quantities and used as food by the coastal people. There are no organized fisheries for
them and they never reach the markets except at a few places on the west coast.
Nevertheless, they support fisheries of much local importance. The “bay clam”,
Meretrix meretrix occurs extensively in most places on the west coasts and is
comparatively less abundant along the east coast. Meretrix casta, known as the
“backwater clam” is much more abundant on the east and west coast backwaters and
estuaries. Paphia malabarica, the false clam, Gafrarium tumidum, the cockle clam
and Katelysia opima, the inflated clam, are also common along both the coasts.
Other bivalves utilized for food belong to different families, viz., Arcidae,
represented by the ark shell, Arca granosa, in the muddy sand bays and creeks,
Donacidae by the wedge clams, Donax cuneatus and D. scortum of the surf-beaten
sandy shores, Cardiidae by the cockles, Cardium spp., from mud flats, Corbiculidae
by the black clam, Villorita cyprinoids from the West Coast backwaters, Solen spp.,
from the partly muddy low lying flats of the east and the west coasts. The culture
practices of clams are very limited. Most species exhibit rapid growth in culture. The
cultural techniques here are even much similar than in mussel culture. The only
requirement is to lay the seed on especially hardened beds. The bottom soil should not
be either too loose or too hard.
Donax species
Donax species are commonly available on Trichendur Coast. The peculiarity

of the Donax, is it occurs in all seasons. But in postmonsoon season, high density
observed on coastal regions. Hence, the postmonsoon period (December to February)

is very ideal for collection. The study of food quality and making conventional food
in Donax species is very limited or almost nil. Hence the present work is aimed to
collect and make them as pickle, chutney and ready to serve powder.
MATERIAL AND METHODS
Methods of preparation
The Clams were collected by manual from the sea shores of Trichendur. The
shells were removed and Flesh parts were washed in potable water to remove dirt and
pigments. The edible portions were boiled for 20 minutes to remove mucus and then
cut into thin round slices of about 1 mm thickness. The sliced meat was used for the
smoking purposes. The slices were blanched separately in 5 % boiling brine for 5
minutes. The blanched meat was then drained and spread on trays and air dried for 20
minutes to facilitate smoke penetration. A conventional vertical type kiln was used for
smoking the meat by burning sawdust. During smoking, samples were drawn every
15 minutes to observe the effect of smoking time on the quality of the meat. The good
quality smoked meat (5 % brine, 60 minutes) was collected from the kiln, dried in
electrical meat drier and each pack in separate polythene bags. Samples were drawn
bimonthly for biochemical and organoleptic quality analyses.
Bivale Pickles
The edible part such as foot and adductor mussel is separate out and cut into
small pieces of 3cm x 3 size. The meat pieces are cooked in a pressure cooker at
151lbs/square Inch for 30 minutes. The cooked meat is used preparation of pickle.
Recipe for the preparation of meat pickle

Meat - 500g
Garlic - 250g
Ginger - 100g
Chili powder - 50g
Coriander powder - 50g
Mustard seeds - 10g
Tumeric powder - 5g
Asafetida - 10g
Salt - 40g
Vinegar - 100ml
Gingili oil - 300ml

Preparation of meat pickle
The cooked meat of the Bivales was added with 2g of turmeric powder 10g of
salt and 20g of chili powder and mixed well and kept for 30 min. The meat was fried
in gingili oil to golden brown color and kept aside. The other ingredients were fried in
the remaining quantity of gingili oil under low flame. The fried meat was added to
this mixture are mixed thoroughly for 10 min. The meat mixture was added with 4 %
acetic acid and mixed well by stirring for uniform distribution of all ingredients and
allowed to cool for some time.
Packaging of meat pickles
After sufficient cooling, the meat pickles were packed separately in sterilized
glass bottles with plastic screw caps. Before sealing the bottle a layer of heated gingili
oil was poured on the top of the pickle to cover the solids to prevent the exposure of
meat pickle to air and the bottles were stored at ambient temperature.
Bivale chutney powder
The cleaned Bivale meat was cooked for 30 minutes to remove the mucus in
separate containers. It was then cut into small slices and deodorized following the
method of Sen and Rao (1966) after that the meat was dried in a hot air oven at 50-
60ºC for 2 days. The dried meat was powdered in a mechanical pulverizer, sieved and
the meat powder was used for the preparation of chutney powder.
Ingredients used for the preparation of chutney powder:
Meat powder - 300g

Black gran (skinned) - 150g
Red chilies - 15nos
Curry leaves - 10g
Asafoetida - 5g
Cumin seeds - 2g
The ingredient such as black gram was fried under low flame until it became
golden brown. It was kept for some time and then powder using a warring blender and
sieved. Then the other ingredients like jeera seeds, red chillies and curry leaves were
fried under low flame for few minutes. Cooled powdered and mixed with the black
gram powder. Then the meat powder, Together with sufficient salt and asafetida
powder was mixed with the above mixer and the chutney powder is prepared. It was
consumed by mixing the powder with a little edible gingili or coconut oil.
The ready to serve chutney powder was packed in 500g capacity sterilized
glass bottles and closed tightly with plastic caps and stored at ambient temperature.
Bivale wafers
The edible meat was washed thoroughly to remove mucus and pigment in the
foot region. The cleaned meat was again washed thoroughly and cut into small pieces.
Deodorization of the processed meat
The processed meat was deodorized separately by following method of Sen

and Rao(1966).
Preparation of wafers
The deodorized meat was dried, pulverized and sieved. The ingredients and
their quantity used for wafers. The meat powder was mixed with corn flour, green
chili paste, salt and food color. Water was added to the meat mixture and thoroughly
mixed. Then it was uniformly spread on small plates in a thin layer of 2-3 mm
thickness and steamed for 10-15 minutes. The cooked and gelatinized material is cut
into desired shapes and dried under sunlight or in a mechanical dryer at 45 to 55c. The
dried product was used for the shelf life studies.
Ingredients:
Meat powder - 300g
Corn flour - 450g
Green chili paste - 112.5g

Food color a pinch
The wafers were packed in laminated pouches and sealed storage at ambient
temperature.
RESULT
Pickles, Chutney powder and ready to serve soup powder were prepared and
served to the staffs and students of our department.
DISCUSSION
In the present study, it was found that, the preparation of pickle, chutney
powder and ready to serve soup powder is very simple and cheapest.When compared
with vegetarian pickle, chutney powder and soup and also with due pickles and other
products of gastropods and fin fishers, the products developed from clams are rich in
protein (the biochemical study is needed). Regarding the taste and digestion, the food
products from clams are tasteful and easily digger table.
Similar findings were observed from different molluses is different places by
various researchers(Amerine, 1965, Ayyekkannu, 1994, Jacoben and Rand, 1982,
Conhell, 1975, Renitta and Patterens, 2004). Amerine, 1965 analyzed the sensory
evaluation of foods.Ayyakkannu, 1994 reported two history status of Babylonia
spirata at protonovo, South East Coast of India.He strongly recommended that the
modules have rich amount of protein than any other animals and also stated that, the
molluscan meat and their relative products were delicious than other fin fish products.

Table 1 Economically important marine molluscs
Mytilus viridis
Clams Bivalve
Donax

Because of the short period, there is no true for analyzing the biochemical
constituent’s v/v the qualification and quantitative studies of protein, carbohydrate,
fats and minerals. But it was found that the clams are neutralized bivalve and the
species are not familiar seafood and it is presently consumed only by a smaller section
of this fisher folk is the world. Recently due to over exploitation of shell and fin
fishers from ocean, there is a high demand occur for marine animals. Hence it is a
time for searching a new food organisms most of the shell fishers non-conventional
groups viz gastropods bivalves and cephalopods. These non-conventional food
resources must be transformed into conventional foods.
In the present study the pickles, chutney power and ready to sure group power
were developed from the Donax sp and was packed at ambient temperature. The
Donax sp are easily available. If proper collection and preparation could be made, it
will play and take major and unavoidable food resources to human beings. The
preparations of food from clams also provide good opportunity to the self-help
woman groups for increasing their economical status. The food products of Donax sp
will also play vital role in earning of foreign currency through export. Studies of
cultural aspects are also needed.At last but not least, we recommend that, all must use
due food products of calms, definitely it increase protein contest in your body and
give taste your tongued.
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Amerine, M.A.R.M, Pangborn and cyprinoids (Grey). Indian

Roessler, E.B. 1965 Principles J.Mar. Sci., 10:128-131.
of sensory evaluation of foods, APHA. 1992. Compendium of
Academic Press, New Methods for the
York,pp349 Microbiological Examination
Ansariz. A., ParulekarA.H., and. of foods, 3rd ed., (Vanderzant,
Matondka G.P. 1981 C. and Splittstoesser (Eds.),
Seasonalchangesinmeat weight American Public Health
and biochemical composition Association, Washington, DC.
of the black clam Villorita Ayyakkannu. 1994. Fishery status of
Babylonia spirata at Port Nova,

Southeast Coast of India. survey,U.K.,Fishing News
Phuket (Books)Ltd 126pp.
Mar.Biol.cent.spec.publ.no.13: Durve, V. S. and Bal, D.V.1961.
53-56. Studies on the chemical of the
Beatty, S.A and Gibbons, N.E. Oyster Crassostrea gryphoides
1937.The measurement of (Schlotheim) J.Zool. Soc.
spoilage in fish. J.Biol. Board India, 13:70-77.
Can.,3:77 Ekambaranath Ayyar. T.N.
Bligh, E.G. and Dyer, W.S.1959. A rap AnthaKrishnan, 1982 Manual
id method Of total lipid of Zoology Volume 1- part I I
extraction and purification Eskin, N.A., H.M. Henderson and R.J.
Can.J.Biochem.Physiol.,37:911 Townsend, 1971. Biochemistry
-917 of Foods, Academic Press,
Charie, HJ., East, E. and VanDerveen, New York.24pp.
W.J.1962. Proc. of First Fretheim, K.,P.E. Gra num and E.
International conference on Vold. 1980. Influence of
Food Science and Technology, generation temperature on the
London, Vol.6. chemical composition, and ant
Clucas,I.J. and Ward A,R. 1996. oxidative and antimicrobial
Chemical, Microbiological and effects of wood smoke.food
physical Methods of Quality sci.,45:999-1002.
Assessment and Measurement Gopal,T.K.S., Thankamma, R.,
of Spoilage. In: Post-harvest Shenoy, A.V., Rao, C.V.N. and
fisheries Development: A Govindan, T.K.1988.
Guide to Handling, Development of Flexible
Preservation , Presentation, packaging for fish soup
Processing and Quality powder. In: Proceedings of the
Chatham Maritime, Kent first Indian Fisheries Forum,
ME44TB, United (Joseph, M.M.Ed.), Asian
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Connell, J J. 1975. Control of fish Branch, Mangalore. Pp.369-
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1222.

BIO-EFFICACY OF EUPHORBIAHIRTA L (EUPHORBIACEAE)
AGAINST COTTON BOLLWORM, HELICOVERPA ARMIGERA
HUB. (LEPIDOPTERA: NOCTUIDAE) IN THE LABORATORY
AND GREENHOUSE
SaranyaT and LingathuraiS1,*
PG Department of Zoology, Pachaiyappa’s College for Men, Kanchipuram – 631 501,

Tamil Nadu, India
1
Department of Zoology, Aditanar College of Arts and Science, Tiruchendur – 628
216, Tamil Nadu, India
ABSTRACT
Hexane, Chloroform, ethyl acetate and water extracts were preparedfrom
Euphorbiahirtatested against Helicoverpa armigera. All the extracts showed
insecticidal activity with dose dependent manner with four different concentrations
(0.625, 1.25, 2.5 and 5%). An insecticidal activity was observed in 5 percent
concentrations of chloroform extract showed 73.33% (H. armigera) mortality. Lethal
concentrations (LC50) values were calculated all the extracts of the treatment.
Chloroform extract showed lowest value of lethal concentration values i.e., gave LC50
value 14.61 % for H. armigera. Effective and active extract (chloroform) further
tested growth inhibitory activity was tested against insect pest. Larval weight, pupal
weight and adult durations were inhibited with increasing concentrations of
treatments. Further glutathione S-transferase and monooxygenase enzymes were
inhibited when increasing concentrations of treatments. Field bio-efficacy was also
tested in two agricultural field and exhibited high reduction of pest. Significant
activities were observed all the treatment groups. GC-MS analysis was showed
phytochemical contain high amount of Stigmasterol and n-hexadecanoic acid. Our
results indicate that E.hirta had potential for development as botanical insecticides,
especially for control insect pests.
KEYWORDS: Euphorbiahirta, solvent extract,Helicoverpa armigera, insecticidal

activity, Growth inhibitory, detoxification enzyme, GC MS analysis

INTRODUCTION
India has a large and diverse agricultural sector, accounting, on average, for
about 16 percent of GDP and 10 percent of export earnings. India's arable land area of
159.7 million hectares (394.6 million acres) is the second largest in the world, after
the United States. Its gross irrigated crop area of 82.6 million hectares (215.6 million
acres) is the largest in the world. India has grown to become among the top three
global producers of a broad range of crops, including wheat, rice, pulses, cotton,
peanuts, fruits, and vegetables.
Agriculture is being affected by certain Biotic and abiotic factors, soil erosion,
irrigation problems, plant diseases caused by fungi, bacteria and virus and insect
infestation. Approximately one-third of the global food production is destroyed
annually by field and storage pests. Despite expensive and often environmentally
hazardous control measures, insects remain the chief pests of crops and stored
products. Synthetic pesticides are currently the most effective means of pest control,
but the appearance of insect resistance and other negative side effects has prompted a
search for new alternatives. In this respect, plants are able to synthesize a broad range
of different chemical compounds called secondary metabolites many of them
providing new sources of natural pesticides.
Euphorbia hirta (Euphorbiaceae) is a small annual herb. The plant is
commonly called pill bearing spurge and asthma herb and the stem is slender. The
leaves are oppositely arranged, lanceolate and are usually greenish or reddish
underneath measuring about 5cm long. The plant leaves are used to treat colic
troubles, dysentery, cough, asthma, worms and vomiting. The white latex is used as
eye drops to cure conjunctivitis. Paste of leaf is applied externally (twice daily) on the
place of scorpion bite. Therefore present study was planned to evaluate laboratory,
field bio-efficacy and GC-MS analysis of phytochemicals.

Fresh leaves of Euphorbiahirta leaf powder (500 gm) and soaked with hexane,
chloroform, ethyl acetate solvent and water extracts was prepared. Phytochemical
analysis of E. hirta extracts was done by Horborne, 1984. The insecticidal activity
and growth inhibtory activity (Isman et al., 2001) experiments were conducted using

third instar larvae of H. armigera. The effective extract was studied detoxifying
enzyme (Glutathione –S transferase and monooxygensase) activity (Oppenoorth et al.,
1979) Field bio-efficacy and GC-MS studied were conducted with effective extract.

Chloroform extract contain alkaloids, steroids, flavonoids and terpenoids in
our previous work. Among different solvent extracts, chloroform extract was found to
be more active and potential followed by ethyl acetate, hexane and water extracts. At
5% concentration of treatment chloroform extract exhibited maximum and significant
larvicidal activity towards Helicoverpa armigera. Significant and dose dependent
mortality were observed all the treatment groups. No mortality was observed at 0.625
% concentration of hexane and water extracts towards H. armigera and 1.25 and 2.5
% concentrations of water extracts showed no larval mortality. Mortality of larvae
was calculated with different concentrations as variable revealed significant
difference in larval mortality (P ≤ 0.05 level). Growth inhibitory activity were
assayed effective extracts i.e., chloroform extract. When increasing concentration of
treatment showed decreased level of larval and pupal weights. All the treatments
showed concentration dependent. At 5% concentration treatment was larval and pupal
weight reduced significantly H. armigera.
In the present study two detoxification enzyme levels (Glutathione S-
transferase and Monooxygenase) was estimated in the larvae of H. armigera.When
increasing concentration of these enzyme levels were reduced. All the treatments
were showed dose depended activity. Field concentrations were sprayed. Pre count of
H. armigera after sprayed the chloroform extract counted again post count. Insect
pests were reduced all the treatment and two field crops. In control (un-treated) pest
population were increasing levels showed.
Totally 28 phyto-compound groups were present in GC-MS analysis. The
extracts represent mainly a mixture of phyto-compounds. The extract obtained from
the leaves of E. hirta had the Stigmasterol as the major component (59.13%),
followed by the n-hexadecanoic acid, which constituted 16.07%. The chloroform
extract treated pupal and adult abnormalities were produced and observed both H.
armigera.

Viewing the result, it was considered of interest to tackle the isolation and
identification of the involved metabolites. According to the results obtained on the
bioassays and field performanceclearly showed the potent larvicidal activityof
extracts derived from E. hirtaplant, due to the presenceof drimanes. As such, this
plant has a high potential as a source of phytochemicals useful to protect our crops
from herbivorous insects, with interesting perspectives on ecological systems of
foodproduction.
Table 1. Insecticidal activity of Euphrbia hirta against Helicoverpa armigera

larvae
Concentration (ppm)
Extract
0.625 1.25 2.50 5
Hexane 0.0±0.00a 10.00±2.34b 16.67±3.53c 30.00±2.55c
Chloroform 16.67±1.15c 27.00±1.12d 46.67±3.77e 73.33±4.20e
Ethyl acetate 10.00±1.56b 20.00±3.59c 26.67±2.49d 43.33±3.77d
Water 0.0±0.00a 0.0±0.00a 3.33±0.32b 3.33±0.32b
Control 0.00±0.00a
within columns, means (± SD) followed by a same letter do not differ significantly
(Turkey’s test, P< 0.05 level)
Table 2. Lethal concentrations of Euphorbia hirta treated larval mortality

towards Helicoverpa armigera
LC50 95% Fiducial limit LC90 95% Fiducial limit Chi-

Extracts
(%) Lower Upper (%) Lower Upper square
Hexane 2424.59 - - 32123.51 - - 6.525
14.61
Chloroform 45.161 14520.00 7416.267 953.483 120638.37 6.523
Ethyl
299.38 9.723 31.929 177.866 64.106 1507.423 0.551*
acetate
Water 3525.48 799.68 72829.22 52957.00 - - 4.166*
*χ2 values are significant at P<0.05 level

Table 3. Growth inhibitory activity of chloroform extract of Euphorbia hirta
treated
Helicoverpa armigera
Concentrations (%) Larval weight (mg) Pupal weight (mg) Adult duration (days)
0.625 103.52±3.35b 73.55±1.75b 5.64±0.27b

1.25 94.28±2.13c 61.40±2.10c 4.80±0.26c
2.5 89.36±2.28cd 53.42±2.25d 4.60±0.07d
5 83.18±1.47d 42.76±1.43e 2.27±0.14e
Control 118.06±3.25a 87.42±2.13a 7.22±0.81a
Table 4. Detoxifying enzyme activities of chloroform extract of Euphorbia hirta

treated Helicoverpa armigera
Detoxifying enzyme activity

Concentrations (%)
Glutathione S-transferase Monooxygenase
(nM/min/mg of protein) mOD/min/mg proteins
0.625 2.057±0.23b 8.864±0.49b
1.25 0.687±0.04c 6.190±0.68c
2.5 0.329±0.02d 4.115±0.36d
5 0.211±0.03e 2.460±0.25e
a
Control 7.238±1.32 11.422±2.46a

Table 5.Field efficacy of chloroform extract of Euphorbia hirtaagainst H.
armigera populations in groundnut and bhendi field
Mean number of larva per plant (Mean ± SD)

First spray count Second spray count
Plot
Pre count Post count Pre count Post count
GroundnutField
T1 2.59± 0.62ab 2.02 ± 0.62a 1.65 ± 0.05a 1.50 ± 0.26a
T2 2.33 ± 0.27ab 2.25 ± 0.23a 2.03 ± 0.52b 1.96 ± 0.23ab
C 1.75 ± 0.89a 3.81 ± 0.64ab 6.42 ± 1.54c 8.93 ± 0.79b
Bhendi Field
T1 2.96 ± 1.06ab 2.53 ± 0.43a 1.96 ± 0.76a 1.62 ± 0.12a
T2 2.43 ± 0.74a 3.43 ± 0.62b 3.69 ± 0.33b 4.06 ± 0.96b
C 2.18 ± 1.00a 2.37 ± 0.37a 3.29 ± 0.42b 3.67 ± 0.27b
(T1- Chloroform extract; T2- Commercial neem pesticide; C- untreated control)
Within the columns, means ± SD followed by the same letter do not differ
significantly using Tukey’s test at P=0.05.
Plate 1. GC-MS Spectrum analysis of chloroform extract of Euphorbia hirta
ACKNOWLEDGEMENT
The authors wish to thank Tamil Nadu State Council for Science and
Technology (TNSCST), Chennai, Tamil Nadu for providing the funding sources
(Student Project Scheme - Ref No: AR – 13; Letter No. TNSCST/SPS/AR/2013-
2014).

Table 6. GC-MS analysis of chloroform extract of Euphorbia hirta
PEAK R.TIME AREA AREA% NAME

1 10.083 17968650 0.31 5-allyl-2-methanol
2 10.339 4432300 0.08 Copane
3 10.959 3193601 0.06 Caryophyllene
4 11.890 22829713 0.40 Octadecane,1-chloro
5 12.220 10577838 0.18 Beta-cadinene
6 12.851 38277059 0.66 Dodecanoic acid
7 13.498 9889350 0.17 Cis-isoapiole
8 14.258 5474900 0.10 Heptadecane
9 15.073 57984378 1.01 Tetradecanoic acid
10 16.148 57439022 1.00 Pentadecanoic acid
11 17.360 925962078 16.07 n-hexadecanoic acid
12 18.232 28036868 0.49 9,12-octadecadienoic acid(z,z)-,methyl ester
13 19.191 3406838253 59.13 Stigmasterol
14 20.613 55259128 0.96 9-octadecenal,(z)
15 21.731 17720255 0.31 Eicosane
16 22.497 29761371 0.52 Hexacosane
17 23.240 42277804 0.73 n-hexatriacontane
18 23.950 45957278 0.80 Tetratriacontane
19 24.069 21296623 0.37 All-trans-squalene
20 24.639 52092408 0.90 Hexatriacontane
21 25.305 40012646 0.69 n-hexatriacontane
22 25.955 35843432 0.62 Tetracontane
23 26.274 59294309 1.03 Alba-tocopherol-beta,-d-mannoside
24 27.134 104665009 1.82 Ergot-5-en-3-lo,(beta,24R)
25 27.358 169774080 2.95 E,e,z-1,3,12-nonadecatrine-5,14-diol
26 27.955 417403806 7.24 Gamma-sitosterol
27 29.043 37862669 0.66 Vitamine E
28 30.055 24851156 0.43 (2E)-3,7,11,15-tetramethyl-2-hexadecen-1-ol
REFERENCES
Horborne SB. 1984. A guide to KT. 1979. Pestic. Biochem.
modern techniques of plant Physiol. 7, 34.
analysis. Chapman and Hall, Ferry, N., Edwards, M., Gatehouse, J.,
London. 4-80.3. Gatehouse, M., 2004. Plant-
Isman MB Andrew JW and Passreiter, insect interactions: molecular
2001 .Fitoterapia.72: 65-68. approaches to insect resistance.
Oppenoorth FJ Smissaert HR Welling Curr. Opin. Biotechnol. 15,
W van den Pas LJT Hitman 155–161.

SYNTHESIS OF SILVER NANO PARTICLES FROM CRAB
SHELLS
A.Kavitha*, N. KamalaDevi*, B. Megala and G.Lakshmanan*

ABSTRACT
copper nanoparticle. But only dearth of knowledge is available for synthesis of silver
nanoparticles. So that in the present study we have also concentlingrated and done the
synthesis of silver nanoparticles from the crab shell. Green chemistry approach for
synthesis of silver nanoparticles has many advantages such as ease with which the
process can be scaled up, economic viability etc.
INTRODUCTION
Nanotechnology is the technology by which an atomic and molecular scale
level matter is material was skillfully managed as tool for various applications. It
deals with nanometer sized object. Nanoparticles are the fundamental building blocks
of nanotechnology.Nanoparticles synthesized from microparticles which are present
at nanoscale level in different parts of plants and animals. For example nanoparticles
present in root, stem, leaves, flowers and seeds of plants.Especially antennas, cuticles,
hair, feathers,and shells of animals.
Recently the animals are used to synthesis functional nanoparticles has been of
great interest. Synthesis of nanoparticles using biological entities has greatest
attention from the scientists throughout the world. It is due to their unique properties
such as size and shape depending optical, electrical and magnetic properties that have
antimicrobial application, biosensor materials, composite fibers, cryogenic
superconducting materials, cosmetic products and electronic components. For the
reasons of many application in the field of synthesizing nanoparticles such as drug

delivery system, gene delivery system, destruction of tumor etc. The present study
was taken in synthesize of nanoparticles from crab shell. Though the crab shells are
harmful to the environment they may be transformed with valuable products.
Crab shell used in this study is three spotted crab known as Protunussanguinolentus.
It is a marine decapod crustacean. The body of the crab is covered by shells that act as
a exoskeleton which provide protection to the crab and secreted by mantle.
The exoskeletons are made up of chitin. Chitin is a linear amino
polysaccharide composed of β1, 4- linked N-acetyl –D glucosamine (GIC NAC). It is
natural biopolymer. The chitins have chemical properties, biocompatibility and low
toxicity.The chemically modified chitin act as suitable vehicle for drug delivery
development. It has the ability to absorb both metal ions and hydrophobic organic
compounds. Chitin is used to for waste water treatment and other industrial
application.
copper nanoparticle. But only dearth of knowledge is available for synthesis of lead
nanoparticles. So that in the present study we have also concentrated and done the
synthesis of lead nanoparticles from the crab shell.
The synthesis of lead nanoparticles by the reduction and thermolysis of organo lead
compounds in organic solvents and polymer matrices, the electrolytic deposition from
aqueous solutions, photolysis, radiolysis and the dispersion of molten lead is organic
or metallic media were the methods used. Green chemistry approach towards the
synthesis of lead nanoparticle has many advantages than other methods.
Collection of crab shell:
Crab shells were collected from fish market, coastal areas from vast shore
fauna.Then cleaned thoroughly with distilled water in order to remove debris. It is
kept for few minutes. Then it powdered and sieved. The fine powder collected and
stored for further analysis.
Preparation of extract:
5g of crab shell powder was weighed and dissolved in 30ml of distilled water. It
is stirred vigoursly for few minutes then filtered thrice. The crude filtered extract of
crab shell was stored for further analysis.

Synthesis of silver nanoparticles:
For the synthesis of silver nanoparticles 100ml of 1mM silver nitrate aqueous
solution was prepared freshly. Then 10ml of silver nitrate and 1ml of crude extract
was mixed and stirred well. It is kept as experimental. For control 10ml of silver
nitrate and 1ml of distilled water was mixed and stirred well. Colour changes are
observed.
Characterization of silver nanoparticles:
Spectral analysis:
After two days the reddish brown colour is appeared. It was measured by UV-
Visible spectrophotometer at wavelength ranging from 300 to 700nm and the values
are plotted on the graph.
Morphological analysis:
Morphological analysis of Silver nanoparticles was done by using Scanning
Electron Microscope (SEM). It was carried out in the institute of Forest Genetics and
Tree Breeding, Coimbatore.
RESULTS
UV-Visible Spectrum analysis of silver nanoparticles synthesis:
The present study reports the formation of silver nanoparticles. The colour change
indicates the reduction of silver ion into silver particles. The synthesis of silver
nanoparticles from 1mM aqueous silver nitrate solution in the presence of crab shell
extract can be easily observed by UV-Visible spectroscopy this is an significant
reaction to preview the morphology and stability of nanoparticles. Silver
nanoparticles exhibit reddish brown colour in aqueous solution due to the surface
Plasmon resonance phenomenon. The time taken for the reduction of silver nitrate
into silver nanoparticle is 48hrs.The formation of silver nanoparticles was observed
with change of colour to reddish brown due to the coherent oscillation of electron gas
at the surface of nanoparticle resulting in surface Plasmon resonance for the control
(distilled water)the peak ranges of 476nm was observed. The crab shell extract
showing an optical absorption band peaked at 474 nm, broadening of peak indicated
that the particles are polydispersed which is kept as experimental analysis the results
from UV-Vis of the samples was predicated in the fig: 1

Silver nanoparticles
Control
1.0 474 nm Experiment
476nm
0.8
Absorbance
0.6
0.4
a
0.2
0.0
300 400 500 600 700 800 900 1000 1100 1200
wavelength nm
Fig:1. UV-Vis Absorbance spectra of Silver nanoparticles
Plate: 1 Silver Nanoparticles
Colour observation.
Morphological characterization:
SEM analysis of the synthesized samples were performed in order to investigate the
morphology and distribution of silver nanoparticles. The SEM images are observed at
different magnifications such as X 5000, X 10,000, X20,000,X 30,000 with 20kv all
of them shows irregular and spherical in shape. From the SEM images it is evident
that the morphology of silver nano particle is spherical which is good agreement with
the shape of SPR band in the UV-Vis spectra. The SEM images of silver nano
particles is shown in the figure 2-5.

DISCUSSION
In the present study, formation of silver, nanoparticles was confirmed by UV-
Vis spectra analysis and SEM analysis for the synthesis of various metal
nanoparticles, the colour changes followed by time measures for reductioin reaction
and absorbance ranges in nanometric scale predict the conformation of nanoparticles
in the sample.
By using biological method the conformation of silver nanoparticles from the
crab shell evidently proved by UV-vis spectra. In our present work the crab shell
extract takes 48hrs for changes in colour which shows the silver ions converted into
silver nanoparticles and the absorbance showed peak at 476 nm this maybe due to the
presence of chitin in the crab shell that makes the particles to synthesis when the
reducing agent silver nanoparticles is added.
This could be compare with the work of Umayaparvathi.S, et al (2013). They
have synthesized silver nanoparticles in the oyster tissue sample. The time taken for
the reduction of silver nitrate is 24 hrs and the brown colour is appeared. The UV –
Vis spectrum showed peak at 430 nm for oyster tissue. This may be due to the nature
of organ selected for nanoparticle synthesis.
Akl.M.Awwad. et al, (2013) analysed silver nanoparticles in Loquat leaf
extract. The changes of colour from yellow to reddish occur within 2 minutes. It takes
and the SPR ranges at 425 nm. This is due to presence of same phytochemical
compound in the Loquat leaf extract.
Similar work was done by Laura Christensen et al (2011). They have
investigated biological synthesis of silver nanoparticles using Murraya Koenigii leaf
extract the reduction of silver ions was monitored using UV-Visible

Spectrophotometry and showed formation of silver nanoparticles within 15 minutes.
The colour changes from yellow to dark brown. The peak occurs at 435 nm.
SEM analysis of Silver nanoparticles:
In the work, the surface morphological appearance of Silver Nanoparticles
was studied by Scanning Electron Microscope. Then Morphology of silver
nanoparticles is spherical. This may be due to silver nanoparticles obtained from the
proposed bioreduction.
SUMMARY
The waste crab shells are used for synthesis of nanoparticles such as silver,
copper and lead.
For silver nanoparticles, the time taken for reduction reaction is 48 hrs.
Reddish brown colour developed from the extract after the introducing reducing after
silver nitrate.
Peak value for silver, copper and lead observed in UV spectrophotometer.
SEM Analysis have been done from the SEM analysis, it was found that the
crab shells are one of the main sources for the presence of silver, copper and lead
nanoparticles.
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Aditi P. Kulkarni, AnkitaA.Srivastava, Akl M. Awwad , Nida M. Salem ,
PravinM.Harpale and Rajendra S. Amany O. Abdeen (2013)
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tapping the unexploited sources. extract and its antibacterial
J.Nat.Prod.Plant Resour.,1 (4): PP- activity.
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Russ. Chem.. Bull., VOL.61,No- pp-36-40.
2.pp 225-229. Aparna Y, EnkateswaraRao K.V and
SrinivasaSubbarao P (2012)

Synthesis and Characterization of Jinwen, Jie Li, Shijun Liu, Qi-yuan
Copper oxide Nano Particles by chem. (2011) Preparation of copper
Novel sol-Gel Method. nanoparticle in a water\ollic acid
International conference on mixed solvent via two step
Environment science reduction method.
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160. and Hudlikar M (2011). Novel
Ashok kumar. D (2013) Rapid And route for rapid biosynthesis of lead
Green Synthesis Of Silver nanoparticles using aqeous extract
Nanoparticles Using The Leaf of JatrophaCurcas L. latex. Mater
Extracts Of Lett.,5,3170 – 3172.
PartheniumHysterophorus, A novel Junkano,Tokushikiuka., (2009) Pure
biologicalapproach International lead nanoparticles with stable
research journal of Pharmacy,pp- metallic surface, on perovskite lead
169-173. strontium titanate particles
Jhon. L, CuyaHuaman., Kimitaka nanotechnology.volume 20
Sato., Satoshi Kurita., Takatoshi number29.
Matsumoto and Kavitha K.S., Syed Baker,Rakshith D.,
BalachandranJeyadevan (2011) Kavitha H.U., YashwanthaRao
Copper nanoparticles Synthesized H.C., Harini B.P. and Satish S.
by Hydroxyl ion Assited Alcohol (2013) Plants as Green towards
Reduction For Conducting ink. Synthesis of Nanoparticles.
J.Mater.Chem.,Vol.21, 7062 – International Research Journal of
7069. Biological Sciences, 2(6): 66-76.

DIVERSITY OF ZOOPLANKTON IN THAYAMANGALAM POND,
SIVAGANGAI DISTRICT, TAMILNADU, INDIA.
Umamaheswari K 1, Chandran G2,*, Rajeshkannan S3, Lingathurai S4 and

Nagarajan S5
1
Research scholar, Department of Zoology, Thiagarajar College Madurai, Tamil
Nadu,
India – 625 009
2
PG and Research Department of Zoology, Yadava College, Madurai, Tamil Nadu,
India - 625014
3
Department of Biotechnology, The Madura College, Madurai, Tamil Nadu, India
4
Department of Biochemistry, SLS.MAVMM, Ayira Vaisya College, Kallampatti,
Madurai, Tamil Nadu, India
5
* Corresponding Author: chandranphd1983@gmail.com
ABSTRACT
The present studies were carried out on the Zooplankton diversity in thayamangalam
pond system, Sivagangai District, Sivagangai. During a period of six months from
November 2013 to April 2014. Sequentially to right to use the various category of
Zooplankton diversity to relation to water quality parameters. A total of 15 species
were identified which belonging to under classes of Zooplankton was represented by
Cladocera, Copepoda, Rotifera and Protozoa. Among zooplanktons, Copepoda was
found dominantly.
KEY WORDS: Zooplankton diversity, water quality parameters, pond system
INTRODUCTION
In generally, ponds are small, shallow, restricted bodies of standing water,
habitats of great significance providing water for domestic, industrial and agricultural
uses. Acquaintance regarding the ecology of pond water is vital tool for their

systematic study. The planktonic study is a very valuable tool for the measurement of
water quality and also contributes to learning (Pawar et.al 2006). But In India, ponds
are essential part of human arrangement and almost all villages have at least one
pond. Ponds have many uses, e.g. bathing, washing of clothing andanimals and
discharge of numerous religious rituals.
Due to negligence, discount of religious outlook, lack of ecological education,
pollution increase and discharge of local wastes, these aquatic bodies are degrading
day by day. For the last few decades many investigators have emphasized on the
hydrobiological profiles of several water bodies (ponds, reservoirs, lakes) with the
intention to assess the water quality (Shastri and Pendse, 2001; Singh, 2000; Azizul
Islam et al., 2001; Thakur et al., 2003). The information available on the status of
lentic water bodies in the Indian subcontinent shows the deterioration of water quality
in general (Chandrashkhar and Jafer, 1998).In the present study an attempt has been
made to assess the diversity of zooplankton and their distribution in the pond system.

Study Area
Thayamangalam pond is located about 15kms away from sivagangai city.
Field survey was implemented for a period of six months from November 2013 to
April 2014. The sampling was carried out during morning hours between 9.00 and
10.00 m hours. For physico-chemical examination, water samples were collected in 1
liter of plastic bottles using plankton nylon ne and samples were preserved in 5%
solution of formaldehyde on the site. Identification and enumerated zooplankton were
done by means of binocular light microscope.
Physico -chemical properties
The physicochemical properties of Thayamangalam pond are summarized in
Table 1 and 2. Air temperature and water temperature ranges from 30.0 - 27.0
respectively. Mean water temperature is observed to be lower than air temperature
which is attributed to less heating of the ponds. The pH values (9.2-8.1) didn’t show a
definite seasonal course and high value was recorded during February in ponds and
low in November. This may be because of turbidity of water which in turn reduce
photosynthetic activity of algae leading to accumulation of CO2 and hence reduction

of pH (Adibisi, 1980). Dissolved Oxygen (DO) indicates physical, chemical and
biological activities in a water body. It is an important indicator of water quality. DO
affect the solubility and availability of many nutrients and therefore productivity of
aquatic ecosystems (Wetzel, 1983).
Table: Seasonal variation of Physico-chemical parameters of thayamangalam
pond
Premonsoon Post monsoon
Parameters
(February-April) (November-January)
Air Temp 30 27
Water Temp 27 25
pH 9.2 8.4
TDS 42.2 48.1
DO 9.3 8.5
BOD 2.8 4.2
CO2 3.9 2.7
Chloride 42.67 23.08
Calcium 12.56 9.06
T.Hardness 56.3 12.0
T.acedity 19.1 13.4
Note: all the parameters are in Mg/L except pH, temperature (
Table 2: Distribution and abundance of phytoplankton of Thayamangalam pond

system
Total number
Cladocera
individuals
Alona pulchella 12
Daphnia carinata 14
Diaphanosoma excisum 09
Moina carinata 11
Ceriodaphnia cornuta 08
Copepoda
Heliodiaptomus sp. 32
Naupliar larve 13
Neodiaptomus stregilipes 12
Rotifera
Brachionus falcatus 28
Brachionus quadridentatus 22
Lepadella ovalis 08
Protozoa
Arcella sp. 12
Diffusia sp. 04
Paramecium sp 07
Vorticella sp. 11
Total no. of individuals 203

In the present study, DO values were found to be more than 8.0 mg/l which
shows that in pond system is optimal for aquatic life. The low values of BOD indicate
the low levels of biodegradable materials and absence of non-biodegradable
substances. The chloride varied between 42.67-23.08mg/l which indicates that water
appears to be suitable for irrigation purposes.
A decrease trend in the chloride content in ponds during winter season may be
related to the absence of dilution effect of water.Variation in sulphate content in
ponds might be due to variable organic input.Total hardness (mg/l CaCO3) and total
alkalinity were found to be low and ranged from 52.10 to 12.0mg/l respectively and
such water bodies can be considered as soft.
Zooplankton Diversity
Diversity, relative abundance and dominance of zooplankton community of
the thayamangalam pond were studied during November 2013 to April 2014. A total
number of species 15 zooplankton taxa belonging to four groups (Copepoda,
Cladocera, Rotifera and Protozoa) were quantified in two seasons depicted in Table 2.
54, 57, 58 and 34 % respectively, in which highest number of zooplankton taxa was
found in pre monsoon and minimum was found in post monsoon.
The variation of population of different zooplankton groups at different
seasons of the year could be attributed to the availability of food material and
preference towards the food in order to avoid competition (Singh, 2000). Welch
(1952) has observed that smaller the water body, more quickly to react the changes in
atmospheric temperature. The pH of the water body showed alkaline in nature i.e. 7.2
to 8.0. This range is good for growth of aquatic organisms (Lendhe and Yeragi,
2004). Bell (1971) has stated the pH ranges between 6.5 to 9.0 provides an adequate
protection to the life of fresh water organisms.
CONCLUSION
An inverse relationship was observed zooplankton abundance. The managed
fish culture pond which was periodically limed, manure and fertilized showed greater
zooplankton being the dominant group. Whereas the unmanaged village pond showed
a less diverse and eutrophic condition, zooplankton being the dominant group. It

implies that a large amount of ecological niches are remaining void and unutilized in
village ponds. Whereas all the available ecological niches are being effectively
utilized by the stocked fishes and periodically replenished by fertilization in the
managed fish culture pond. Therefore selective stocking with appropriate species at
low densities and extensive fish culture practices in the village ponds has ample
scope. Adoption and transformation of such village ponds by scientific management
practices into semi-intensive fish culture ponds may prove to be an ecologically
efficient, financially feasible and socially viable venture.
REFERENCES
Adibisi AA. 1980. The Pawar S K Pulle J S and Shendge K M
Physicochemical Hydrology of 2006. The study on
Tropical Seasonal Upper Ogun phytoplankton of Pethwadaj
River. Hydrobiologia,79: 157- Dam, Taluka Kandhar, District
65. Nanded, Maharashtra, J. Aqua.
APHA 1995. Standard Methods for Biol. Vol.21, pp.1-6.
the Examination of Water and Shastri Y and Pendse D C 2001.
Wastewater. 19th Ed. American Hydrobiological study of
Public Health Association, Dehikhuta Reservoir .J.
New York. pp. 1143. Environ. Biol. 22: 67- 70.
Azizul Islam M., Choudry, A N and Singh D N 2000. Seasonal variation of
Zaman, M 2001. Limnology of zooplankton in a tropical lake.
Fish Ponds in Rajshahi Geobios 27: 9-100.
Bangladesh. Ecol. Envir. Thakur, T S Bhuyan I K and Burva R
Conserve. 7: 1-7. 2003. Indian J. Env. Ecoplan.
Chandrasekhar S V and Muhammed 7(1): 83 - 86.
Jafer P. 1998. Limnological Wetzel R G 1983. Limnology. 2 nd
studies of a temple pond in Edition. Saunders Coll. Publ. pp. 767
Kerala. Environ. Ecol. 16: 463-
367.

TOXICITY AND SYNERGISTIC STUDY
ONAZADIRACHTAINDICAA. JUSS. AND MELIAAZEDARACHL.
ON CALLOSOBRUCHUS MACULATUS FABRICIUS.
D Sarasa1, P C Sathya Narayanan2, M Pushpalatha3 R Jalajagandhi1, M Vidhya1

and S Lingathurai4,*
1
PG & Department of Zoology, Quaid-e-Millath Government College for Women,
Chennai, Tamil Nadu, India
2
PG & Department of Zoology, Pachaiyappa’s College, Chennai, Tamil Nadu, India
3
PG & Department of Zoology, Pachaiyappa’s College for Men, Kanchipuram, Tamil
Nadu, India
4
ABSTRACT
In the present study bio-pesticide was used to control the stored food product pest
Callosobruchusmaculates. The natural pesticide extracts were extracted from two
different species neem Azadirachtaindica and Meliaazedarach. The shade dried plant
powder was soaked sequentially in 1000 ml with increasing polarity of solvents
(hexane, chloroform and ethyl acetate) for 48 hours with constant shaking. These
concentrated three solvent crude extracts were analyzed for bio-efficacy and active
crude extract was further tested for growth adult emergence bioassay. Among the two
plants totally six solvent extracts tested A. indica ethyl acetate extract was identified
as the most toxic treatment against Callosobruchus maculatus.M. azedarach hexane
extract showed least level of adult morality. Ethyl acetate and chloroform extracts of
both plants were equally mixed (1:1 ratio) ethyl acetate mixture showed potential
larval mortality 93.33% at 4000 ppm concentration. Adult emergence was observed
dose dependent manner at 2000 ppm concentration showed 43% only emerged C.
maculatus. Chloroform extracts of A. indica +M. azedarach showed 36% emergence
at 4000 ppm concentration level.

KEY WORDS: Azadirachtaindica,Meliaazedarach,Callosobruchusmaculateus,
toxicity, adult emergence, synergistic study
INTRODUCTION
Civilization began with agriculture. It has played a key role in the development
of human civilization. When our nomadic ancestors began to settle and grow their
own food, human society was forever changed.
Agriculture may affected by certain biotic and abiotic factors, soil erosion,
irrigation struggle, plant diseases caused by fungi, bacteria and virus and insect
infestation. They developed and raised most of their crops for consumption, storage,
exported, etc., Food storage is both a traditional domestic skill and is important
industrially. Food is stored by almost every human society. Storing of food has
several main purposes, to enabling a better balanced diet throughout the year,
preparedness for catastrophes, emergences and periods of food scarcity and protection
from animals or theft.
Stored product insects may cause significant damage and loss to stored foods.
An oil or food extract (non-nutritive products are preferred) or synthesized scent that
will attract a select group of insects. In some cases the food attractant is impregnated
into an artificial medium. Insect infestations in grains and other stored food and fibre
products cause annual losses worth many millions of dollars worldwide.
To overcome this problem pesticides have to play a major role in pest

management in sustainable agriculture production. They are renewable, non-persistent
in the environment, and relatively safe to natural enemies, non-target organisms and
human beings.
Pesticides is a substance or mixture of substances intended for preventing,

destroying or controlling any pest, including vectors of human or animal disease,
unwanted species of plants or animals causing harm during or otherwise interfering
with the production, processing, storage, transport or marketing of food, agricultural
commodities, wood and wood products or animal feedstuffs, or substances which may
be administered to animals for the control of insects, arachnids or other pests in or on
their bodies.

Pesticides have two levels of benefits for pesticide use, primary and
secondary. Primary benefits are direct gains from the use of pesticides and secondary
benefits are effects that are more long-term. But they have environmental impact
often greater than what is intended by those who use them. The use of chemical
pesticides and fertilizers in Indian agriculture has seen a sharp increase in recent years
and in some areas has reached alarming levels with grave implications for human
health, the ecosystem and ground water. It is therefore increasingly urgent that
environmentally friendly methods of improving soil fertility and pests and disease
control are used.Over 98% of sprayed insecticides and 95% of herbicides reach a
destination other than their target species, including non-target species, air, water,
bottom sediments, and food (Miller, 2004). Pesticides can contaminate unintended
land and water when they are sprayed aerially or allowed to run off fields, or when
they escape from production sites and storage tanks or are inappropriately discarded
(Tashkent, 1998).
Neem (Azadirachta indica and Meliaazedarach) is a very effective and

extensively used pesticide. Pesticides based on azadirachtin may have direct adverse
effects on aquatic organisms and their toxicity depends on various factors. It has been
reported that neem extracts in aquatic environments are lethal to benthic populations
and drastically decrease the number of organisms in the food web and nutrient cycling
process (Goktepe et al., 2002 and El- Shazly et al., 2000). Even though
Azadirachtinis not likely to accumulate or cause long-term side effects, toxicity to fish
is only moderate and may not kill fish under normal use (Miller and Uetz, 1998).
Pesticides containing bioactive compounds from the neem plant, A. indica are
reported to be target specific and comparatively less toxic. Plants are virtually
inexhaustible sources of structurally diverse and biologically active substances
(Istvan, 2000). Some plants contain compounds of various classes that have
insecticidal, piscicidal and molluscicidal properties. Unlike synthetic chemical
pesticides, which leave harmful residues in the aquatic environment (Koesomadinata,
1980; Cagauan, 1990; Cagaun and Arce, 1992), botanical insecticides are believed to
be more environmentally friendlier because they are easily biodegraded and leave no
residues in the environment. Most of the pesticides, both plant based and chemical,
applied in the various agro ecosystems reach water bodies through runoff affecting

fish, the most abundant aquatic organism and a variety of other fauna. Persistent
chemical molecules with long half-life periods found in chemical pesticides pose a
threat to fish and also to the human population consuming the affected fish.
The present study is to establish whether the use of the less toxic pesticide
could be promoted among the agriculturists. It is possible to substitute chemical
pesticides with pesticides of plant origin. This needs an extensive study on the
properties of the two types of plant A. indica and M. azedarach, especially a study of
their synergistic effect on Callosobruchus maculatus
Plant Collection
Plants Melia azedarach and Azadirachta indica (Meliaceae) leaves were

collected from in and around Kanchipuram District, Tamil Nadu, India.
Extraction of Plant Materials
Each plant materials leaves were shade-dried, ground into powder by an

electronic blender and 300g of plant powder was soaked sequentially in 1000ml with
increasing polarity of solvents (Hexane, Chloroform and Ethyl acetate) for 48h with
constant shaking. The soaked powder material was filtered through filter paper. The
solvent in the filtrate was evaporated under reduced pressure by vacuum rotary
evaporator. These concentrated three solvent crude extracts were analyzed for bio-
efficacy and active crude extract was further tested for growth inhibition bioassay.
Two plants of crude solvent extracts were weighed and stored 4 oC until treatments.
Rearing Procedure
The initial culture of Callosobruchus maculatus was taken from the laboratory
of Department of Zoology, reared at 0±5˚C and 60±5% R.H. on Mung grains
(Vignaradiata) for the last 10 years, to minimize the biological variation factors like
age and size. The insects were kept in 1 lb glass jars. The mouth of jars was covered
with a piece of muslin cloth tied by means of rubber band. The wet cotton plugs were
placed over the mouth of jars for moisture. Grains of Vigna radiata were used as food
and egg laying media when the eggs were laid by the adults they soon died and the
new adults emerged in about 25-30 days. After emergence the new adults were
transferred to the jar containing fresh grains.

Insect Bioassays
Seeds were weighed and placed with 2–3 day-old fertilized weevil females
(three females per seed) in a glass vial containing 50 seeds for 24 h, in the dark at
28ºC, 60% relative humidity. Only three eggs remained on each seed after eggs laid in
excess were removed. After total emergence of adult insects, infested seeds were
opened and both seed mass and feces were weighed. Four experiments were
developed and the mean was calculated.
Insect Culture and Treatments
Each crude extracts were treated separately with leaf disc bioassay. Effective
crude extracts were mixed with respective solvents and finally growth inhibitory
studies were studied. Concentrations 0.25, 0.50, 1 and 2 % concentrations were used
for all treatments. Synergistic study were conducted and mixed each effective crude
extracts 1:1 (%) ratio.
The following treatment groups were followed
Individual treatment
1. Azadirachta indica (Hexane, Chloroform and Ethyl acetate extract)

2. Melia azedarach (Hexane, Chloroform and Ethyl acetate extract)
3. Synergistic treatment A. indica (Ethyl acetate extract) + M. azedarach (Ethyl
acetate extract)
Insecticidal activity lethal concentrations of Probit analysis were estimated

EPA 1.5 software. Further adult emergence was analyzed using one way ANOVA.
Significant differences between treatments were determined using Tukey’s multiple
range tests (P ≤ 0.05).
RESULTS
Hexane, chloroform and ethyl acetate extracts weighed and calculated the
percent content of plant source both A. indica and M. azedarach (Table 1). Maximum
quantity was elucidated chloroform extract of A. indica and minimum quantity was
ethyl acetate extract of A. indica. Further analysis were carried out from these extracts
on C. maculatus

Among the two plants totally six solvent extracts tested A. indica ethyl acetate
extract was identified as the most toxic treatment against C. maculatus at 4000ppm
concentration. Ethyl acetate extract recorded 75% insecticidal activity at 96 h (Table
2). Ethyl acetate extract of M. azederach showed good insecticidal activity against C.
maculatus larvae. Different concentrations showed larval mortality viz., 500 (15%),
1000 (35%), 2000 (50%) and 4000 (67.5%) percent. M. azederach hexane extract
showed least level of adult mortality.
Chloroform extract of both of plants also recorded high insecticidal activity.

The A. indica ethyl acetate extract showed LC50 value of 1646.78ppm and LC90 value
of 10042.40ppm (Table 3) and 1990.99ppm and 12716.25ppm for M. azederach. The
Chi-square values were significant at P≤ 0.05 level. The high Chi-square values in the
bioassays probably indicated the heterogeneity of the test population. Different
solvent crude extracts influenced insect mortality differently. Both of solvent and
water control did not showed mortality.
Ethyl acetate and chloroform extracts of both these plants crude equally mixed
(1:1 ratio) and tested against insect pest of C. maculatus. Ethyl acetate mixture
showed potential larval mortality 93.33 percent at 4000ppm concentration.
Chloroform extract showed good larval mortality at 83.33 percent concentration. This
treatment showed dose dependent manner activity. No larval mortality were recorded
both solvent and water control. Synergistic efficacy was also recorded as lethal
concentration 50 and 90 values. The Ethyl acetate extracts of A. indica + M.
azedarach 1052.10ppm (LC50) and 3563.61ppm (LC90) and Chloroform extracts of A.
indica + M. azedarach showed 1380.22ppm (LC50 ) and 5507.62ppm (LC90) (Table 5).
The Chi-square values were significant at P≤ 0.05 level. The high Chi-square values
in the bioassays probably indicated the heterogeneity of the test population.
Normal (water control) adult emergence was 98 percent (Table 6). Solvent
control showed as 96 percent. At 4000ppm concentration of mixed ethyl acetate
extract of A. indica treated showed only 19 percent emergence. Adult emergence was
observed dose dependent manner. At 2000ppm concentration showed 43 percent only
emerged C. maculatus. Chloroform extracts of A. indica + M. azedarach showed 36
percent emergence at 4000ppm concentration level.

DISCUSSION
Neem ingredients affect insects in various ways including mainly repellent,

antifeedant, toxic and growth regulatory effects. Fecundity of insects was also
affected by various neem products. However, neem-based products were proved as
medium- to broad-spectrum insecticides against various field and store pests. Seed
kernel represents the most important source of active ingredients found in neem tree,
such as azadirachtin and many others. Ogunwolu and Idowu (1994) studied the
particle size effects on insecticidal activity of Zanthoxylum zanthoxyloides root bark
and Azadirachta indica leaves solvent extracts against C. maculatus in labortaory.
From the observed results the extracts from solvents were more effective when
applied as finely ground particles than when applied as coarse particles (4000ppm).
Results in this study confirm the insecticidal potency of A. indica and M. azederach
products against stored product coleopterans as reported by Lale (1995).
Various synthetic chemicals have been used to keep stored grain products free
from pest attack, Synthetic pesticides are currently the method of choice to protect
stored grain from insect damage. But, continuous or heavy uses of synthetic pesticides
has created serious problems arising from factors such as direct toxicity to parasites,
predators, pollinators, fish and man. It also develops pesticides resistance (Mohamed,
2002), susceptibility of crop plant to insect pests (Pimentel, 1977) and increased
environmental and social cost (Pimentel et al., 1980). Therefore, environment needs
some other alternatives of chemical pesticides. One alternative to synthetic
insecticides is the botanical pesticides i.e. insecticidal plants or plant compound and
the use of natural compounds, such as essential oils that result from secondary
metabolism in plants. Essential oil and their constituents have been shown to be a
potent source of botanical pesticide.
Furthermore, the latter author concluded that neem products are suitable for
integrated pest management because of their low toxicity to non target organisms,
easy preparation and compatibility with other bio-products. In Sudan, several
products (i.e., powder, and water and organic extracts) of neem seeds showed
significant control of insect pests on post harvests as well as on some vegetable crops
under field conditions. The optimum dosage rates of water extracts were indicated and
recommended for important vegetables, so that they can be prepared and applied by

subsistence farmers wherever needed (Siddig, 1987; Mohamed, 2002, and Satti and
Nasr, 2006). Active neem ingredients are responsible for several biological activities
in insects, including repellent, antifeedant, toxic and hormonal effects (Schmutterer,
1995).
The toxicity of a large number of essential oils and their constituents has been
evaluated against a number of bruchid pests (Tripathi et al., 2002). In the present
study the neem were used as biopesticides. According to the latter author the toxic
effect of neem ingredients generally acts through stomach action rather than contact
action. This interpreted why neem products exerted delayed mortality effects on
insects which increased with time and concentration as explained in the present
results, and other previous works (Satti et al., 2003, Satti and Nasr, 2006). In the
study it is observed that Among the two plants, A. indica and M. azederach totally
six solvent extracts tested A. indica ethyl acetate extract was identified as the most
toxic treatment against Callosobruchus maculatus at 4000ppm concentration. Ethyl
acetate extract recorded 75% insecticidal activity at 96 h
Botanical materials are a rich source of bioactive organic chemicals. Over

2000 plant species around the world are known to possess pest control properties
(Ahmed et al., 1984). Previous research indicated that some plant powders and
extracts have strong effects on stored grain insects such as toxicity and the inhibition
of reproduction (Regnault - Roger and Hamraoui, 1993; Talukder and Howse, 1995).
Various plant by-products from Asia, Africa and America have been tested recently
with a good degree of success as protectants against a number of stored grain insect
pests (Rajapakse, 1990).
Some indigenous plants in the kingdom of Saudi Arabia are known to possess
some biological activity against insects (Al-Moajel and Al-Dosary, 2002 and Al-
Moajel and Al-Fuhaid, 2003). The adult mortality was directly related to the
concentration of the treatments. Chloroform extract of both of plants also recorded
high insecticidal activity. The A. indica ethyl acetate extract showed LC50 value of
1646.78ppm and LC90 value of 10042.40ppm (Table 3) and 1990.99ppm and
12716.25ppm for M. azederach. The Chi-square values were significant at P≤ 0.05
level. The high Chi-square values in the bioassays probably indicated the
heterogeneity of the test population.

However, neem treatments also reflected good results in conserving the
prevailing predators, and sustained populations as high as that of the control in most
cases (Satti et al., 2010). Likewise in the present study Synergistic efficacy was also
recorded as lethal concentration 50 and 90 values. The Ethyl acetate extracts of A.
indica + M. azedarach 1052.10ppm (LC50) and 3563.61ppm (LC90) and Chloroform
extracts of A. indica + M. azedarach showed 1380.22ppm (LC50) and 5507.62ppm
(LC90) (Table 5). The Chi-square values were significant at P≤ 0.05 level.
The three extracts of S. sesban seeds demonstrated significant toxic effect

against S. granarius adults, in spite of the fact that LC 50 value of the chloroform
extract was about half that of petroleum ether and acetone extracts. Also, the LC 90
value of chloroform extract was about half that of petroleum ether extract and one
fourth that of acetone extract. The toxicity of a number of plant extracts has been
evaluated against stored products insects (Lale and Yusuf, 2001). This means that
many plant materials can be used as insect toxicants but these investigators gave no
details about the active components. Results reported in this study show that S. sesban
has an insecticidal effect on S. granarius.Different plant extracts was also tested
against S. granarius (Al-Moajel, 2000 and Ahmed et al., 2002a), and the results
indicated that wheat grains were well protected by some plant extracts.B. rapaseed
extracts have been reported to be very effective in protection cowpea seeds against C.
maculates (Ahmed et al., 2001). Capparis spinosa seed extracts gave complete
protection to cowpea seeds against C. maculatus (Ahmed et al., 2002b).
Adult bruchid survivorship after 7 days in the untreated control reflects the
relatively short adult lifespan of the bruchids under bioassay conditions in Petri dishes
(Boeke et al., 2004). The relatively high contact toxicity of the botanical standard
(neem) to the two bruchid species in this study is comparable to the mortality reported
by Ivbijaro, 1990 for C. maculatus exposed to the seed oil of this plant. However, the
physico-chemical properties of the neem preparation used in this study may influence
the level of lethal effect, as neem powder lacked contact toxicity on this bruchid
species (Ogunwolu and Odunlami, 1996). Adult emergence was 98 percent (Table 6).
Solvent control showed as 96 percent. At 4000ppm concentration of mixed ethyl
acetate extract of A. indica treated showed only 19 percent emergence. Adult
emergence was observed dose dependent manner. At 2000ppm concentration showed

43 percent only emerged C. maculatus. Chloroform extracts of A. indica + M.
azedarach showed 36 percent emergence at 4000ppm concentration level.In the
current investigation the ethyl acetae extract from the A. indica and M. azedarach
exhibited high insect control efficiency on bruchid tested.
CONCLUSION
The results obtained in this study suggest good potential for the use of these
extracts as an insect mortality factor. Concentrations at LC50 and LC90 of hexane,
chloroform and ethyl acetate extracts (500, 1000, 2000 and 4000ppm) are active as a
toxicants against C. maculatus attacking grain and toxic as well. They can give good
protection to wheat grain for nearly 1 months, and reduce loss in grain weight.
Considering the above results, these extracts have great potentiality in the
management of an important stored grain pest such as C. maculatus synergistically.
Table 1. Quantity of extract yielded from respective solvent extracts of

Azadirachta indica and Melia azederach
No Solvent Quantity of yield from plant source (gm)
A. indica
1 Hexane 2.60
2 Chloroform 3.42
3 Ethyl acetate 0.68
M. azederach
4 Hexane 1.44
5 Chloroform 2.57
6 Ethyl acetate 1.26
Table 2. Effect of different solvent extracts from Azadirachta indica and Melia
azederach on Callosobruchus maculatus adult mortality
Concentration (ppm)
Name of Plant Solvent 500 1000 2000 4000
Hexane 15 30 50 65
Azadirachta indica Chloroform 7.5 22.5 40 52.5
Ethyl acetate 20 37.5 52.5 75
Hexane 0 5 20 30
Melia azederach Chloroform 5 20 32.5 45

Ethyl acetate 15 35 50 67.5
Solvent Control 0
Water control 0
Values are represented as percentage of 40 replicates
Table 3. Toxicity (LC50 and LC90) of different solvent extracts from Azadirachta
indica and Melia azedarach on Callosobruchus maculatus
Chi square
Name of Plant Solvent LC50 LL- UL LC90 LL- UL
value
1778.55- 8617.38-
Hexane 2159.09 13918.58 0.309*
2748.18 30565.79
2605.07- 11878.31-
Chloroform 3275.61 20772.03 1.617*
Azadirachta 4566-16 53479.05
indica Ethyl 1370.31- 6636.84 –
1646.78 10042.40 0.545*
acetate 2009.80 19452.96
4763.04 – 14868.37 –
Hexane 6420.74 27716.07 3.742*
10706.28 82443.11
3284.71- 15475.13 –
Chloroform 4351.27 30211.15 2.371*
Melia 6823.85 99719.32
azederach Ethyl 1646.81 – 8015.14 –
1990.99 12716.25 0.828*
acetate 2498.81 26974.95
Solvent Control 0
Water control 0
LL: Lower limit, UL ; Upper limit, LC50 and LC90 values are expressed as ppm
(n= 0). * χ2 values are significant at P <0.05 levels.

Table 4. Synergistic activity Azadirachta indica and Melia azederach on
Callosobruchus maculatus adult mortality
Concentration (ppm)
Treatment
500 500 500 500
Ethyl acetate extracts of A. indica + M. azederach 20 53.33 70 93.33
Chloroform extracts of A. indica + M. azederach 13.33 46.66 60 83.33
Solvent control 0
Water control 0
Table 5. Synergistic toxicity (LC50 and LC90) of different solvent extracts from
Azadirachta indica and Melia azederach on Callosobruchus maculatus
Chi square
Name of Plant Solvent LC50 LL- UL LC90 LL- UL
value
Ethyl acetate extracts of A. 788.32 – 2529.72 –

1052.10 3563.61 0.889*
indica + M. azederach 1341.70 6645.48
Chloroform extracts of A. 1035.84 – 3575.31 –
1380.22 5507.62 1.391*
indica + M. azederach 1830.08 12911.65
Solvent control 0
Water control 0
LL: Lower limit, UL ; Upper limit, LC50 and LC90 values are expressed as ppm
(n= 0). * χ2 values are significant at P <0.05 levels.
Table 6. Synergistic activity Azadirachta indica and Melia azederach on
Callosobruchus maculatus percent adult duration
Treatment Concentration (ppm)

500 1000 2000 4000
Ethyl acetate extracts of A. indica + M. azederach 73bc 55d 43de 19f
Chloroform extracts of A. indica + M. azederach 79b 67c 58d 36e
Solvent control 96a
Water control 98a

Values in each column followed by the same alphabets are not significantly different
by Tukey’s test at P < 0.05.
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BIOLOGICAL ACTIVITIES OF EXTRACTS FROM
AZADIRACHTA INDICA A. JUSSAND MELIA AZEDARACH
LINN. ONSPODOPTERA LITURA FAB. (LEPIDOPTERA:
NOCTUIDAE)
D Sarasa1, P C Sathya Narayanan2, M Pushpalatha3 R Jalajagandhi1, M

Vidhya1and S Lingathurai4,*
1
PG & Department of Zoology, Quaid-e-Millath Government College for Women,
Chennai, Tamil Nadu, India
2
PG & Department of Zoology, Pachaiyappa’s College, Chennai, Tamil Nadu, India
3
PG & Department of Zoology, Pachaiyappa’s College for Men, Kanchipuram, Tamil
Nadu, India
4
ABSTRACT
In the present investigation the impact of botanical pesticide on the cluster

caterpillar,Spodoptera litura was studied. The leaf extracts of Azadirachta indica and
Melia azedarach extracted from by using the solventsviz., hexane, chloroform and
ethyl acetate. Six solvent extracts of both plant extractswere tested against S. litura
larvae.Ethyl acetate extract of M.azedarachshowed good larvicidal activity against S.
litura larvae. Different concentrations showed larval mortality viz., 0.25 (10 %), 0.5
(26.66 %), 1.0 (46.66 %), 2.0 (73.33 %).Ethyl acetate and chloroform extracts of both
these plants crude equally mixed (1:1 ratio) and tested against insect pest of S. litura
larvae. Ethyl acetate mixture showed potential larval mortality 93.33 percent at 2 %
concentration. Chloroform extract showed good larval mortality at 83.33 percent at
2% concentration. At 2 % concentration of mixed ethyl acetate extract of A. indica
treated showed all insects not emerged from pupal stage. This concentration showed
pupal mortality observed dose dependent manner. It could not emerge adult stage
from pupal stage. At 1 % concentration showed 1.14 day only survived S. litura
larvae which are not able to produce their next generation.

KEY WORDS: Azadirachta indica, Melia azedarach, Spodoptera litura, insecticidal,
growth inhibition
INTRODUCTION
In the past few decades agriculture has been characterized by

enhanced productivity, by use of synthetic fertilizers and pesticides. Pesticides are
substances or mixture of substances intended for preventing, destroying, repelling or
mitigating any pest. It may be a chemical, biological agent (such as a virus or
bacterium), antimicrobial, disinfectant or device used against any pest. They
include insects, plant pathogens, weeds, molluscs, birds, mammals, fish, roundworms
and microbes that destroy property, spread disease or vectors for disease or cause
nuisance.
Although there are benefits to the use of pesticides, some also have
drawbacks, such as potential toxicity to humans and other animals. The overuse of
pesticides and synthetic fertilizers damages the long-term fertility of the
soil. The environmental impact of pesticides is often greater than what is intended by
those who use them. Though there can be benefits using pesticides, inappropriate use
can counterproductively increase pest resistance and kill the natural enemies of pests.
Many users are inadequately informed about potential short and long-term risks, and
the necessary precautions in the correct application of such toxic chemicals are not
always made. Pesticides can contaminate unintended land and water when they are
sprayed aerially or allowed to run off fields, or when they escape from production
sites and storage tanks or are inappropriately discarded.
Natural pesticides are active principles derived from plants for the
management of human and animal pest organisms or it can be said to be biologically
active ingredients, principally derived from plants, for the management of human and
animal pest organisms. With the growing global demand for environmentally sound
pest management strategies; there is a need to develop alternative pesticides with
minimal or non-ecological hazards. They bio-degradable and their use in crop
protection is a practically sustainable alternative. It maintains biological diversity of
predators and reduces environmental contamination and human health hazards.

Biopesticides is a naturally occurring substances that control pests
(biochemical pesticides), microorganisms that control pests (microbial pesticides),
and pesticidal substances produced by plants containing added genetic material (plant-
incorporated protectants) or PIPs."Biopesticides are biochemical pesticides that are
naturally occurring substances that control pests by nontoxic mechanisms.
Conventional pesticides, by contrast, are generally synthetic materials that directly
kill or inactivate the pests. Biopesticides are considered eco-friendly and easy to
use. In the USA, the EPA regulates the registration and use of earth friendly bio-
pesticides. Biopesticides are key components of integrated pest management (IPM)
programmes, and are receiving much practical attention as a means to reduce the load
of synthetic chemical products being used to control plant diseases.
Spodoptera litura isoriental leafworm moth which is considered as an

agricultural pest. It is also known as the cluster caterpillar, cotton leafworm, tobacco
cutworm, and tropical armyworm. It is found in the Indo-Australian tropics. It is also
established on most Polynesian islands where it occurs in a variety of island forms.
The larva feed on a wide range of plants and has been recorded from over 40 mostly
dicotyledonous plant families. It is a major pest of many crops. It is vital to control
this pest for the sustainable yields.
The neem tree (Azadirachta indica A. Juss and Melia azadirachta), from the
Meliaceae (mahogany) family, known as margosa or Indian lilac, has long been
recognized for its properties both against insects and in improving human health. The
seed consist of a shell and 1-3 kernels which contain azadirachtin and its homologous.
Both the bark and the leaves also contain biologically active molecules but not high
levels of azadirachtin which is found mainly in the seed kernels. The tree is now
grown in most tropical and sub-tropical areas of the world for shade, for reforestation
programmes and in plantations for the production of compound which have toxic,
antifeedant and repellent properties against insects.The active compounds, from
various parts of the plant with pesticidal, nematicidal, fungicidal, bactericidal, anti
inflammatory, anti-tumor and other properties.
In the present investigation biopesticides were used to control the Spodoptera

litura. Here, the extracts from the two neem plants are collected and screened against
the Lepidopteran pest. Study of lethal concentration, larval duration, mortality, pupal

dauration, adult duration, synergistic efficacy were carried on using the extracts of the
neem at different concentrations was evaluated.
Plants Azadirachta indica and Melia azedarach (Meliaceae) leaves were

collected from Pachaiyappa’s College campus, Chennai, Tamil Nadu.Each plant
materials leaves were shade-dried, ground into powder by an electronic blender and
300g of plant powder was soaked sequentially in 1000ml with increasing polarity of
solvents (Hexane, Chloroform and Ethyl acetate) for 48h with constant shaking. The
soaked powder material was filtered through filter paper. The solvent in the filtrate
was evaporated under reduced pressure by vacuum rotary evaporator. These
concentrated three solvent crude extracts were analyzed for bioefficacey and active
crude extract was further tested for growth inhibition bioassay. Two plants of crude
solvent extracts were weighed and stored 4oC until treatments.
Insect culture
S. litura larvae were collected from Aarani (Periyapalayam), Thiruvallur

District near Chennai. They were maintained on castor leaves in the laboratory at 28 ±
1ºC :11 ± 1 hr photoperiod and 65 – 70 % R.H. adults were released into oviposition
chambers for egg laying. Eggs were collected, kept separately and newly hatched
larvae were maintained on castor leaves. Freshly emerged 3 rd instar larvae were used
for the experiment.
Treatment
Each crude extracts were treated separately with leaf disc bioassay. Effective
crude extracts were mixed with respective solvents and finally growth inhibitory
studies were studied. Concentrations 0.25, 0.50, 1 and 2 % concentrations were used
for all treatments. Synergistic study were conducted and mixed each effective crude
extracts 50: 50 (%) ratio.
The following treatment groups were followed

1. Azadirachta indica (Hexane, Chloroform and Ethyl acetate extract)
2. Melia azedarach (Hexane, Chloroform and Ethyl acetate extract)

3. Synergistic treatmentA. indica (Ethyl acetate extract) + M. azedarach (Ethyl
acetate extract)
Growth Inhibition
Third instar larvae (average weight: 12.4 mg) were used for growth inhibition
bioassay. Leaf discs (4 cm diameter) were dipped in two extracts mixed respective
plants at different doses with acetone; 0.25, 0.50, 1 and 2 % and acetone alone were
used as solvent control. Thirty replicates were maintained for each treatment and
control. The following parameters were considered: larval toxicity, larval period
duration, pupal duration, pupal period duration and adult duration (Zhong et al.,
2001).
Insecticidal activity lethal concentrations- Probit analysis were estimated EPA

1.5 software. Further larval, pupal and adult durations were analyzed using one way
ANOVA. Significant differences between treatments were determined using Tukey’s
multiple range tests (P ≤ 0.05).
RESULTS
Among the two plants totally six solvent extracts tested A. indica ethyl acetate
extract was identified as the most toxic treatment against third instar larvae of S. litura
at 2 % concentration. Ethyl acetate extract recorded 80 % larvicidal activity at 96 h
(Table 1). Ethyl acetate extract of M. azedarachshowed good larvicidal activity
against S. litura larvae. Different concentrations showed larval mortality viz., 0.25 (10
%), 0.5 (26.66 %), 1 (46.66 %) and 2 (73.33 %) percent. Hexane extract of M.
azederach least level of larval mortality showed.
The larval mortality was directly related to the concentration of the treatments.
Chloroform extract of both of plants also recorded high larval mortality. The A. indica
ethyl acetate extract showed LC50 value of 0.861 % and LC90 value of 3.393 % (Table
2) and 1.03 % and 4.29 % for M. azedarach. The Chi-square values were significant
at P≤ 0.05 level. The high Chi-square values in the bioassays probably indicated the
heterogeneity of the test population. Both of solvent and water control did not showed

larval mortality. Ethyl acetate and chloroform extracts of both these plants crude
equally mixed (1:1 ratio) and tested against insect pest of S. litura larvae (Table 3).
Ethyl acetate mixture showed potential larval mortality 93.33 percent at 2 % and
chloroform extract 83.33 percent 2 % concentration. No larval mortality were
recorded both solvent and water control.
Normal (water control) larval period was 16.8 days (Table 4). Solvent control
showed as 16.4 days. At 2 percent concentration of mixed ethyl acetate extract treated
showed 9.6 days only. This is the high larval duration inhibition which treated mixed
extract. Pupal duration also inhibited when mixed ethyl acetate extracts from A.
indica and M. azederach (Table 5). Dose dependent activity was observed.
Normal (water control) larval period was 6.04 days (Table 6). Solvent control
showed as 5.43 days. At 2 percent concentration of mixed ethyl acetate extract of A.
indica treated showed all insects not emerged from puapl stage. This concentration
showed pupal mortality observed dose dependent manner. It could not emerge adult
stage from pupal stage. At 1% concentration showed 1.14 day only survived S. litura
larvae. It is not possible to produce their next generation. The observed duration of all
satges were affected and highly inhibited their treatment of mixed crude extract. Life
cycle duration inhibited and growth inhibited gradually.
DISCUSSION
Results on the toxicity, biology and mortality effects of different

concentrations of the A. indica and M. azedarach leaves extracts on the S. litura
reported in the present work, confirm their potential against economic important
cosmopolitan pest Asian army worm. This report is the first for studying the A. indica
and M. azedarach species effects for control of the armyworm insect pest. Generally
the botanical insecticides were reported to have less environmental impact than the
most commercial synthetic insecticides, for example neem insecticides (Hoelmer et
al., 1990). Consequently, neem leaves crude extracts (i.e., hexane, chloroform and
ethyl acetate) were screened for the identification of insecticidal potentiality of these
plants.

Leatemia and Isman (2004) reported that in leaf disc bioassays, insect death
was occurred due to a combination of starvation and contact toxicity of these extracts
and larval mortality observed in all crude extract treated insects. However, ethyl
acetate crude extract have the higher toxicity with the lowest LC50 and caused more
larval mortalities. Similarly in the present work may indicate the presence of active
anti-insect phytocompounds in the ethyl acetate crude extract have the higher toxicity
with the lowest LC50 of the both of M. azedarach and A.indica and their synergistic
effect. TheA. indica ethyl acetate extract showed LC50 value of 0.861 % and LC90
value of 3.393 % and 1.03 % and 4.29 % for M. azederach. The Chi-square values
were significant at P≤ 0.05 level.
The development of insects' growth regulators (IGR) has received

considerable attention for selective control of insect for medical and veterinary
importance and has produced mortality due to their high neurotoxic effects
(Wandscheer et al., 2004; Senthil Nathan et al., 2005a).
The growth regulatory effect is the most important physiological effect of M.

azedarachon insects. It is because of this property that family Meliaceae has emerged
as a potent source of insecticides. Exposure of A. stephensi larvae to sub-lethal doses
of neem leaves extract in the laboratory prolonged larval development, reduced pupal
weight and ovipostion (Su and Mulla, 1999). In the field, delayed phenology of
surviving larvae and reduced pupal weight are common occurrence after treatment
with neem (Zebitz, 1984). The direct and indirect contribution of such effects to
treatment efficacy through reduced larval feeding and fitness need to be properly
understood in order to improve the use of M. azedarachfor management of A.
stephensi. The results of this study indicate the plant-based compounds such as
Azadirachtin (compounds present in the Meliaceae plant family seed) may be an
effective alternative to conventional synthetic insecticides for the control of Culex
pipiens. Undoubtedly, plant derived toxicants are valuable sources of potential
insecticides. These and other naturally occurring insecticides may play a more
prominent role in mosquito control programs in the future (Mordue and Blackwell,
1993). Synergistic activity on S. litura larval mortality Ethyl acetate and chloroform
extracts of both these plants crude equally mixed (1:1 ratio) and tested against insect
pest of S. litura larvae. Ethyl acetate mixture showed potential larval mortality 93.33

percent at 2 % concentration. Chloroform extract showed good larval mortality at
83.33 percent 2 % concentration.
The results of this study will contribute to a great reduction in the application
of synthetic insecticides, which in turn will increase the opportunity for natural
control of various medicinally important pests by botanical pesticides. Since these are
often active against a limited number of species including specific target insects, less
expensive, easily biodegradable to non-toxic products, and potentially suitable for use
in mosquito control programme (Alkofahi et al., 1989), they could lead to
development of new classes of possible safer insect control agents. Plant
allelochemicals may be quite useful in increasing the efficacy of biological control
agents because plants produce a large variety of compounds that increase their
resistance to insect attack (Berenbaum, 1988; Murugan et al., 1996; Senthil Nathan et
al., 2005a).
Results from the study of Lucantoni et al., (2006) clearly indicated that the
neem treated female mosquito, A. stephensi, displayed a delay in oocyte development
in the vitellogenesis. As discussed by Weathersbee III and Tang (2002), the disruption
of reproductive capability could lead to substantial population decline over time.
Furthermore, Dhar et al., (1996) revealed that the exposure to neem extract
suppressed rather than inhibited oviposition in mosquitoes. The efficacy of
Azadirachtin on larvae, pupae, and adult of Culex pipiens. Correspondingly in the
present study the mode of action and synergism with the biocides under laboratory
condition on S. litura significantly reduced at maximum concentration 2 % ethyl
acetate extracts.
Previously and traditionally in many countries simple crude extracts have also
been used as insecticides (Crosby, 1971). While plant crude extracts often consist of
complex mixtures of active and inactive phytochemicals. Hummelbrunner and Isman
(2001) have reported that the exposure of several plant extracts to the insects causes
delayed larval development through decreased growth rates. Similar result was
obtained in the present study; fractions treated larval weight was reduced and duration
of the larva and pupa were increased. Reduced feeding activity (or increased
antifeedant activity) i.e., decreased consumption of castor leaf area was led to the

larval weight reduction of larval and pupal duration extension. Hence, fraction eighth
has the higher inhibitory potential with increasing concentration on growth, larval and
pupal developmental stages of S. litura. However, this effective extract shows higher
antifeedant and larvicidal activities at higher concentrations. Similarly Audrey
Leatemia and Isman (2004) reported that the high concentrations of extracts caused
high mortality of larvae even though small portions of the leaf discs were consumed.
Hummelbrunner and Isman (2001) reported that botanical extracts protect

crops by reducing the fitness of insect herbivores via disruption of larval
development, inhibition of larval growth and failure in pupal eclosion. Similarly in
the present study most effective eighth fraction deterrent effect evident by the poor
feeding, growth and development, led to the development of abnormal pupa and
adults. Telang et al. (2003) stated that malformed adult insects that were produced as
a result of plant toxin treatments were short-lived and infertile and these effects could
be considered important in the pest population reduction.
From the present study results it can be understood that these (A. indica and
M. azedarach) extract treatment is promising in reducing the feeding rate ofS. litura
and might be toxic to the larvae. Among the studied fractions isolated from ethyl
acetate crude extract of (A. indica and M. azedarach) leaves, extracts shows
promising antifeedant activity, larvicidal activity and insect development inhibitory
activities than the other extracts. The main advantages of using botanical pesticides
like neem are reduced human toxicity (Raizada et al., 2001). Accordingly A. indica
and M. azedarach extracts is also has the important toxic effect to the S. litura and
may less effective to the humans. These present study data suggests that the ethyl
acetate extract of the leaves of A. indica and M. azedarach should be further
investigated in order to establish their chemical composition and may use in insect
pest control programmes. Likewise in present study 2 % concentration of mixed ethyl
acetate extract of A. indica treated showed all insects not emerged from pupal stage.
This concentration showed pupal mortality observed dose dependent manner. It could
not emerge adult stage from pupal stage. At 1% concentration showed 1.14 day only
survived S. litura larvae.

Table 1. Effect of different solvent extracts from Azadirachta indica and Melia
azedarach on Spodoptera litura larval mortality
Concentration (%)
Name of Plant Solvent 0.25 0.50 1.0 2.0
Hexane 10 20 30 50
Azadirachta indica Chloroform 6.66 26.66 40 56.66
Ethyl acetate 13.33 30 53.33 80
Hexane 3.33 10 16.66 30
Melia azedarach Chloroform 3.33 3.33 20 43.33
Ethyl acetate 10 26.66 46.66 73.33
Solvent Control 0
Water control 0
Table 2. Toxicity (LC50 and LC90) of different solvent extracts from Azadirachta
indica and Melia azedarachon Spodopteralitura
Chi
Name of Solvent LC50 LL- UL LC90 LL- UL square
Plant value
Hexane 5.63- 0.124*
Azadirachta 22.11 1.30-7.96 17.65 877.27
indica Chloroform 1.027- 3.850- 0.867*
1.46 2.827 8.44 65.41
Ethyl 2.138- 0.132*
acetate 0.861 0.658-1.172 3.393 8.538
Hexane 8.60- 0.087*
Melia 4.817 2.25-156.15 41.13 108.99
azedarach Chloroform 2.47 1.68-6.07 9.70 4.51-81.53 1.157*
Ethyl 0.085*
acetate 1.03 0.78-1.48 4.29 2.55-12.80
Solvent Control 0
Water control 0
LL: Lower limit, UL ; Upper limit, LC50 and LC90 values are expressed as percentage
(n=24). * χ2 values are significant at P <0.05 levels

Table 3. Synergistic activity Azadirachta indica and Melia
azedarachonSpodopteralitura larval mortality
Concentration (%)
Treatment 0.25 0.50 1 2
Ethyl acetate extracts of A.
indica + M. azedarach 20 53.33 70 93.33
Chloroform extracts of A.
indica + M. azedarach 13.33 46.66 60 83.33
Solvent control 0
Water control 0
Table 4. Synergistic activity Azadirachta indica and Melia azedarachon

Spodoptera litura larval duration
Concentration (%)
Treatment 0.25 0.50 1 2
Ethyl acetae extracts of A.
indica + M. azedarach 13.5±2.5ab 12.2±1.60c 10.4±1.42cd 9.6±0.67d
indica + M. azedarach 14.9±1.70a 13.6±1.94b 11.41±0.64c 10.5±0.38cd
Solvent control 16.4±1.54a
Water control 16.8±2.21a

Spodoptera litura pupal duration
Concentration (%)
Treatment 0.25 0.50 1 2
indica + M. azedarach 8.46±0.72b 7.14±0.61bc 5.04±0.36d 4.51±0.19e
indica + M. azedarach 10.08±0.54a 8.43±0.19b 6.40±0.07c 5.21±0.02cd

Spodoptera litura adult duration
Concentration (%)
Treatment 0.25 0.50 1 2

indica + M. azedarach 3.19±1.08ab 2.48±0.46c 1.14±0.02d -
indica + M. azedarach 4.72±1.15b 4.31±0.18bc 3.16±0.51ab 2.54±0.04c
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proteinase inhibitors: potential larvae of Aedes aegypti.
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reproduction of Diaprepes 105. Beltsville, MD.

BIODEGRADATION OF SYNTHETIC PYRETHROID-
CYFLUTHRIN BY ENTEROBACTER ASBURIAE FROM SOIL
S. Lingathurai1, 2*, P. Sivadurgadevi1 AND P. Hemavathi1

1
2
Post Graduate Department of Zoology, Pachaiyappa’s College for Men,
Kanchipuram, Tamil Nadu, India
Corresponding Author: lings02@gmail.com
ABSTRACT
Different agricultural soil samples were collected from fields where Cyfluthrin was
applied. Bacterial colonies were isolated from soil and screened for their ability to
grow in the medium with Cyfluthrin. The selected bacterial isolate was subjected to
taxonomic identification and identified as Enterobacter asburiae. E. asburiae and the
reference strain Pseudomonas stutzeri were tested for their efficiency of Cyfluthrin
degradation. 500, 1000, 1500, 2000 and 2500 ppm concentrations of the Cyfluthrin
were selected for the study. The degradation of Cyfluthrin was studied by the
measurement of pH, estimation of carbon dioxide, esterase activity and turbidity. For
both the isolate and the reference strain the degradation efficiency was maximum in
500 ppm of Cyfluthrin. The UV-Visible spectrometry predicts that the chemical
nature of the initial has been changed and this may be due to the process of
degradation. HPLC analysis was done on the 10th day of treatment for the 500 ppm
concentration of Cyfluthrin with the isolate and the reference strain. The peak with a
different retention time shows the presence of an intermediate compound.
KEYWORDS: Enterobacter asburiae, Cyfluthrin, degradation, pH, esterase activity,

HPLC
INTRODUCTION
A current environmental concern is the defect of aquatic ecosystem due to

pesticide discharges from manufacturing plant, agricultural runoff, leaching,
accidental spills and other sources. Microbes play a significant role in the degradation

of synthetic pyrethroid exclusively in Cyfluthrin. Cyfluthrin is an insecticide in the
synthetic pyrethroid family and over ninety percent of the Cyfluthrin manufactured
worldwide is used to kill pests of cotton. It is also used on pests of lettuce, pecans, etc
and to kill cockroaches, fleas and termites in houses (Chaudhuri et al., 1999). Because
of the potential risks from these pesticides there is a serious need to develop
remediation process to eliminate or minimize contamination in the environment.
Biodegradation could be reliable and cost effective technique for pesticide abatement
(Sayler and Blackburn, 1989). Cyfluthrin degrades more slowly under anaerobic and
water-logged conditions (Yu, 2002).
Microorganisms are more important than purely physical and chemical
process of degradation (Collins and Lyne, 1985). Among the different genera of
bacteria degrading pesticides, the genus pseudomonas has a special status, as strain of
pseudomonas species are known to metabolize a broad range of organic compounds
and therefore an ideal choice as the bacteria to be used for degradative
biotechnologies.
To date, some physicochemical and biological remedial strategies have been
described by researchers which lead to degradation of synthetic pesticides into both
toxic and non-toxic metabolites (Sutherland, 2000). That is why it is necessary to
develop strategies primarily based on biological and biochemical means to detoxify
the pesticide residues accumulated in soil and water environments. In this study, we
isolated different strains of bacteria capable of degrading cyfluthrin from cyfluthrin -
polluted soil and water environments through repetitive enrichment culture and
successive subculture. Among these bacterial cultures, efficient bacterial strains,
namely Enterobacter asburiae was further tested for their potential to degrade
cyfluthrin.
Reagents and chemicals

Technical (95.5%) cyfluthrin obtained from Bayer Crop Science, R-PT-
Analytics, FFM Germany. Stock solutions of cyfluthrin and its metabolites were
prepared in acetone. HPLC grade acetone and acetonitrile were purchased from J. T.
Backers, Holland. All other chemicals fine chemicals procured Hi Media chemicals.

Soil sample collection
The soil samples were collected from the agricultural field where cyfluthrin
was applied. The field was near 12 km away from Tambaram.Soil used for the
isolation of cyfluthrin -degrading bacteria was obtained from a vegetable-cropping
farm near Padappai, Kanchipuram district, Tamil Nadu, India. Surface soil (0–10cm)
was removed using a spade and placed in plastic bags. The soil was transported to a
laboratory, sealed in bags and stored at 4◦C in a refrigerator until use.
Soil used for the isolation of degrading bacteria
The soil samples that were collected in sterile containers were brought to the
laboratory within six hours for bacteriological analysis. Morphological and
biochemical tests were done isolated microbes followed by (De Berjac and Bonnefois,
1968). The soil samples were serially diluted up to a dilution of about 10 -7 and the
diluted samples (10-5, 10-6, 10-7) were then inoculated by spread plate technique on
nutrient agar plates.
Isolation of cyfluthrin degrading organisms
Pretreated soil (5g) was added to sterile distilled water. Pour plate method was
used for the isolation of bacteria using nutrient agar medium (beef extract 3 g,
peptone 5 g, dextrose 5 g, NaCl 5 g, Agar 15 g, distilled water 1litre, pH 7) and
incubated at room temperature (26◦C). Four strains were isolated and sub-cultured.
Pure cultures were obtained by repeated sub culturing. The strains were maintained on
nutrient agar slants. The colony characters were identified based on the colony
morphology and staining characters. During the course of the investigation all the
strains were tested for their ability to degrade cyfluthrin using pure culture isolates.
One isolate had a stable cyfluthrin -degrading ability and was used in the experiment.
The plates were then incubated at 37oC for 24 hours. The isolated strains were then
tested for their resistance to the pesticides. The isolated bacteria were then inoculated
on minimal medium containing different concentrations of cyfluthrin (200, 500, 1000,
1500 and 2000 ppm).
Reference strain
Pseudomonas stutzeri was obscured from MTCC, IMTECH, Chandigarh and
used as the reference strain. The reference strain was also subjected to all the
experiments that were done for the isolate, Enterobacter asburiae..

Bacterial identification
Gram’s staining was done and observed under microscope. The bacterium was
inoculated on the selective media like MacConkey Agar, Pseudomonas Agar and
EMB Agar. Biochemical tests like catalase, oxidase, MR-VP, Indole production,
Citrate utilization and TSI Agar test were done for strain identification.
Identification and characterization of bacterial isolate
On the basis of morphological, cultural and biochemical characteristics, the
bacterial isolate was identified as a member of the genus Pseudomonas according to,
Bergey’s Manual of Determinative Bacteriology (1994). Characterization studies of
the isolate from these experiments, as well as of those by other researchers, indicate
that bacteria belonging to the genus pseudomonas are gram negative, rodshaped,
highly oxidative and metabolically versatile, able to degrade aromatic hydrocarbons,
oil, petroleum products and pesticides (Hashmi, 2000, Martin et al., 2000)
Estimation of esterase enzyme activity
The microbial inoculum 2ml added with 0.1 M NaPO4 buffer (pH 7.5)
containing 10% glycerol, 1 mM EDTA, and 0.1 mM dithiothreitol. The homogenates
was centrifuged at 10,000g for 15 min. The resulting supernatants were filtered
through glass wool and centrifuged at 100,000g for 60 min to pellets. The final
supernatant served as homogenate for esterase assays. The homogenate immediately
frozen in liquid nitrogen and stored at -80ºC until assayed for enzymatic activities.
Esterase assays were conducted using two model substrates, α-naphthyl
acetate (α-NA) and α- naphthyl butrate (α-NB), according to the method of van
Asperen, 1962. Reaction mixtures consisted of 10µl tissue microbial homogenate and
0.25mM substrate in a final volume of 1.0ml phosphate buffer (50mM, pH 7.4).
Substrates were added in acetone, giving a final acetone concentration of 1% in the
mixture. All esterase assays were carried out in duplicate. Reaction mixtures were
incubated for 10minutes in disposable cuvettes in a water bath at 27ºC. Reactions
were stopped by the addition of 166µl Fast Blue B dye reagent (0.3% in 3.5% SDS).
Colour was allowed to develop for 15min before absorbance was measured at λ = 600
nm against a blank cuvette containing all ingredients except substrate. The
concentration of α-NA and α-NB was determined from a standard curve of α-
naphthol.

Protein Determination
In esterase estimation, protein was determined by the method of Bradford,
1976 using bovine serum albumin as the standard.
Degradation efficiency
The isolate was inoculated on minimal broth containing different
concentrations of cyfluthrin like 500, 100, 1500, 2000 and 2500 ppm. The flasks were
then incubated at room temperature for a period of about 14 days and the samples
were then subjected for the estimation of pH, carbon dioxide and HPLC analysis.
Statistical analysis
The data were subjected to one way Analysis of Variance (ANOVA) to find
out the significance of the treatments at 5% level. The mean values were separated by
Least Significant Difference (LSD) at 5 percent level to find out the effective
degradation.
RESULTS
During the first days of incubation, biodegradation of cyfluthrin was ≤ 10%
of the spiked amount. At the end of the incubation (12 d), the removal of cyfluthrin by
bacterial strains ranged between 57 and 84%. The strains identified as E. asburiae
showed the highest potential to degrade cyfluthrin (Table 1) and they were selected
for further studies. Abiotic degradation was only recorded beyond the 7 th day of
incubation. The pH of the broth decreased in parallel with progressive degradation of
cyfluthrin i.e., higher degradation resulted in a lower broth pH (Figure 1). A highly
negative relationship was found between percent degradation of cyfluthrin and
resultant decrease in pH of all the cultures media. The largest decrease in pH from 7.9
to 4.8 was recorded in the case of E. asburiae which degrade cyfluthrin amount.
Biodegradation of cyfluthrin by efficient bacterial strains (E. asburiae and P.
stutzeri) was further investigated at different initial carbon dioxide of broth culture
(Fig. 2). Biodegradation of cyfluthrin by these bacterial cultures was more
pronounced at carbon dioxide values of the broth but it was produced when increased
period of days. Maximum biodegradation of cyfluthrin by bacterial strains was
observed at an initial carbon dioxide of 1.8 to 9.8. These two selected bacterial strains
behaved similarly for biodegradation of cyfluthrin in culture broths of different
carbon dioxide. During detoxification enzyme also increased when degradation of

cyfluthrin. Both carbon dioxide and esterase increased level in increased days of
incubation. Maximum 38.45µg/ml was showed at 12th day of incubation.
Interestingly, accumulation of cyfluthrin ether appeared in little amounts as detected
by HPLC analysis using authentic standards and matching the retention times of the
respective peaks. HPLC data showed reference and treated degradation peak
estimated.
DISCUSSION
This study reveals the enrichment and isolation of highly efficient bacterial
strains capable of degrading cyfluthrin in to less toxic compounds. Among the 20
bacterial strains tested, there were great variations in their potential to degrade
cyfluthrin (data not given). The highly efficient bacterial strain E. asburiae.P. stutzeri
isolate used for as positive control for estimation of degradation. These bacterial
strains degrade cyfluthrin more than 70% within 12 days of incubation. The
biodegradation rates observed in the case of these strains were much higher than of
previously documented bacterial strains utilizing cyfluthrin and endosulfan (Awasthi
et al., 1997 and 2003). This might be due to their prolific growth during the
incubation period as evident from the higher optical densities or the presence of an
efficient enzymatic system responsible for the degradation. There was a parallel
decrease in pH of the cultured medium as the biodegradation proceeded. This
dramatic reduction in pH of the bacterial cultures might be due to dehalogenation of
pesticides and subsequent formation of acidic substances. These results confirmed the
findings of previous studies(Awasthi, 2003, Kwon et al., 2002 and Sutherland et al.,
2002). They reported that decrease in pH might be due to the formation of HCl or
organic acids by microorganisms. By using authentic standards, HPLC analysis
demonstrated the disappearance of both isomers of cyfluthrin.
These results revealed that these bacterial species adopted the hydrolytic
pathway of cyfluthrin biodegradation(Kim et al., 2001) contrary to oxidative pathway
of cyfluthrin and endosulfan biodegradation in which cyfluthrin and endosulfan is
formed (Kwon et al., 2002 and Kullman and Matsumura, 1996). In general, neutral to
slightly alkaline conditions are considered more favourable for bacterial growth than
acidic conditions (Alexander, 1977 and Sylvia et al., 2005). It is very likely that the
initial alkaline pH 8.0 changed to neutral during the initial period of incubation as

evident from Table 3, favouring luxury growth of bacteria and degradation of
cyfluthrin. A pH greater than 8.0 might have resulted in a shock for the bacteria at the
start of incubation, which slowed down the proliferation of bacteria.
The assumption is also supported by Chaudhry (1994), who reported that
biodegradation during treatment is greatly affected because of low solubility of
compounds in an aqueoussystem, which was also observed in this study. The present
research findings described that this may be the first instance in which high
concentration of cyfluthrin degradation has been achieved in short retention time of
48 h. Although Maloney et al., (1988), reported the transformation of permethrin (50
mg/L) by pure culture of Pseudomonas fluorescence in the presence of tween 80
under aerobic conditions with a half-life of less than 5 days. Grant, et al., (2002),
reported that technical grade cyfluthrin can be reduced from 60 mg/L to 6 mg/L by
Pseudomonas sp. in 20 days. The overall findings suggest that these micorbs used as
degradation of cyfluthrin by this isolate culture may be feasible and a reasonable
treatment option for the removal of pesticide wastes from soil as biodegradation
observed only in the presence of acclimated microorganism as well as under aerobic
conditions.
In conclusion some soil indigenous bacteria carry the ability to degrade
cyfluthrin at accelerated rates. Such bacteria could be useful in framing a
bioremediation strategy for pesticide contaminated soil and water environments.
Fig1. Structure of cyfluthrin cyano(4-flouro-3-phenoxyphenyl)methyl-3-(2,2-

dichloroethenyl)-2,2-dimethyl-cycloprpanaecarboxylate

Figure 1. Change in pH of the broth as a result of cyfluthrin degradation by two
efficient strains of bacteria over a period of time (Mean ±SD)
Figure 2. Degradation of cyfluthrin on two strains in broth total carbon dioxide
30 Pseudomonas stutzeri Enterobacter asburiae Control

25
Carbon dioxide
20
15
10
5
0
0 2 4 Da6ys 8 10 12
Figure. 3. Total esterase activity of 500 ppm cyfluthrin treated two bacterial
strains
120
Estrase activity (µg/ml)
Pseudomonas stutzeri Enterobacter asburiae Control

100
80
60
40
20
0
0 2 4 Da6ys 8 10 12

Fig 4. HPLC finger prints a) Control, b) Isolated sample and c) References strain
REFERENCES
Alexander M (1977) Introduction to Compounds. Mutat Res

soil microbiology. John Wiley 439:63–67
and Sons, New York Chaudhuri K, Selvaraj S, Pal AK
Awasthi N, Manickam N, Kumar A (1999) Mutat. Res. 439:63–67
(1997) Bull. Environ. Contam. Collins CH, Lyne PM (1985)
Toxicol 59:928–934 Microbiological Methods. 5th.
Awasthi N, Singh AK, Jain RK, Edition. Butterworth and Co
Khangarot BS, Kumar A (Publishers) Ltd.
(2003) Appl. Microbiol. De Berjac H, Bonnefois A (1968) J.
Biotechnol. 62:279–283 Invert. Pathol 11.335-347.
Bergey’s Manual of Determinative Grant RJ, Daninell TJ, Betts WB
Bacteriology (1994) Edited by (2002) J. Appl. Microbiol., 92
Hensyl W. R.), 9th. Edn. (3), 534-540.
Williams & Wilkins, Hashmi I (2000) Microbiological
Baltimore, Mass. transformation of hazardous
Chaudhuri K, Selvaraj S, Pal AK waste during biological
(1999) Biodegradation of treatment. Ph.D. Thesis.
Halogenated Organic Institute of Environmental

Studies, University of Karachi. Ed.CRC Press, Boca Raton,
Pakistan. FL. 53-71.
Kim YK, Kim SH, Choi SC (2001) Sutherland TD, Horne I, Harcourt RL,
Biotechnol. Lett. 23:163–166 Russell RJ, Oakeshott JG
Kullman SW, Matsumura F (1996) (2002) J. Appl. Microbiol.
Appl. Environ. Microbiol. 93:380–389.
62:593–600 Sutherland TD, Horne I, Lacey MJ,
Kwon GS, Kim JE, Kim TK, Sohn Harcourt RL, Russel RJ,
HY, Koh SC, Shin KS, Kim Oakeshott JG (2000) Appl.
DG (2002) FEMS Microbiol. Environ. Microbiol. 66:2822–
Lett. 215:255–259 2828
Kwon GS, Kim JE, Kim TK, Sohn Sylvia DM, Fuhrmann JJ, Hartel PG,
HY, Koh SC, Shin KS, Kim Zuberer DA (2005) Principles
DG (2002) FEMS Microbiol. and applications of soil
Lett 215:255–259 microbiology. Prentice Hall,
Maloney SE, Maule A, Smith AR, W New Jersey
(1988) Appl. Envir. Microbiol. van Asperen AK (1962) Study of
2874-2876. house fly esterases by means of
Martin M, Mengs G, Plaza E, Garbi C, a sensitive colourimetric
Sanchez M, Gibello A, method, J. Insect Physiol. 8,
Gutierrez F, Ferrer E (2000) 401.
Appl. Environ. Microbiol. 66 Yu JJ (2002) Removal of
(3), 1190-1194. organophosphate pesticides
Sayler GS, Blackburn JW (1989) from wastewater by
Modern biological methods: supercritical carbon dioxide
The role of biotechnology. In: extraction.Water Res 36 (4),
Biotreatment of agricultural 1095-1101
wastewater, ME Huntley,

DEVELOPMENT OF VALUE ADDED PRODUCT FROM TUNA
(MASI)
*Mohamed Ramlath Sabura, S., *Delighta Mano Joyce, M.I., **Hasna Naziya, I.
*Assistant Professors, Department of Zoology, Sadakathullah Appa College,
Tirunelveli.
**Scholar, Department of Physics, Sadakathullah Appa College, Tirunelveli.
ABSTRACT
Tuna is a type of saltwater fish that belongs to the same family as mackerel fish and
bonito fish. They are members of the Thunnini tribe, which includes 15 different tuna
species.This fish is a powerhouse of essentialnutrients such asomega-3 fatty acids,
potassium, magnesium, iron, vitamin A, B6 and B12, and as such is beneficial for
health. The meat of the tuna fish is often sold frozen, fresh or canned and is widely
consumed around the world as a popular ingredient for sandwiches, salads, casseroles
and sushi rolls.The fish is boiled with water, a handful of salt, two teaspoons of
turmeric powder for 20-30 minutes. After half an hour the cooked fish was taken out
and sundried for about a week.This is the main product called “Masi”. To make value
added products,the dried masi is ground with dried red chilli and salt which is called
as Masi podi.Masi sambal,Tuna Salad, Cheeni sambal,are some of the well known
dishes made from masi powder.
KEY WORDS:Tuna, nutrients,mackerel fish,masi, casseroles
INTRODUCTION
A tuna is a type of saltwater fish that belongs to the same family as mackerel
fish and bonito fish. They are members of the Thunnini tribe, which includes 15
different tuna species. Tuna fish are of varyingspeciesand are found all over the world
oceans such as blue fin, yellow fin, bullet tuna and to name a few. Life of tuna fish
varies from three to five years though some are known to have lived for more than
two decades. They are comparatively easy to catch and go in the making of delicious
dishes like steaks,salads, burgers and the like. They are nomadic, which means that
they keep migrating.

The fish is a powerhouse of essentialnutrients such asomega-3 fatty acids,
potassium, magnesium, iron, vitamin A, B6 and B12, and as such is beneficial for
health. The meat of the tuna fish is often sold frozen, fresh or canned and is widely
consumed around the world as a popular ingredient for sandwiches, salads, casseroles
and sushi rolls.
Tuna meat is literally loaded with protein 42 grams in each can. Combined
with just a few other high-protein foods, including even one serving of tuna fish in
your diet can help provide your body with the protein that you need. Though it is high
in protein but low in calories, making it an excellent addition to a weight loss diet.
One study published in the American Journal of Clinical Nutrition showed
that eating a high-protein meal led to greater reductions in levels of ghrelin, the
hormone responsible for stimulating hunger, than a high-carb meal. It also slowed
gastric emptying, resulting in increased satiety. (1) Another study from the University
of Washington School of Medicine found that upping protein intake by just 15 percent
decreased daily caloric intake by 441 calories and caused a significant reduction in
body weight and fat mass. (2)
Tuna nutrition is high in selenium, a mineral that plays a central role in the
health of your thyroid gland. Multiple studies have found that selenium
supplementation may be beneficial for conditions like autoimmune thyroiditis,
Graves’ disease and hypothyroidism. (3)
Tuna fish is high in several nutrients that have been directly linked to brain
health. Niacin, in particular, has been linked to a lower risk of Alzheimer’s disease
and age-related decline. (4) Tuna fish is also high in omega-3 fatty acids, which can
boost cognitive performance and may be protective against mental health problems
like depression. (5,6)
Tuna is packed with antioxidant-rich selenium, providing 190 percent of the
daily recommended value for selenium in each can. Selenium has antioxidant
properties and can help reduce oxidative stress and prevent damage to cells. (7)
According to a study published in the World Journal of Biological Chemistry, tuna is
also high in a selenium-containing compound called selenoneine, which fights off free
radicals and could even protect against chronic disease and aging. (8)
Although many people think of tuna as a last resort dinner option when the
fridge is starting to get empty, it can be so much more than that. We can easily use

tuna canned to make a delicious tuna fish sandwich or tuna fish salad. Other than the
canned we can prepare different dishes using dry masi powder.
METHODOLOGY
To get 1 kg of dry masi, we need 3kgs of fresh Tuna fish. Fresh Tuna fish is
deheaded and degutted and cleaned thoroughly .Then a big container is filled with
water, a handful of salt, two teaspoons of turmeric powder are added and boiled for
20-30 minutes. After half an hour the cooked fish was taken out and the vertebral
column and the spines are removed to some extent. Then cut into pieces.Then the
pieces are sundried for about a week. Then the outer skin is scrubbed out and washed
thoroughly then again dried for two days. Now the final product is obtained.
This is the main product called “Masi”. To make value added products,the
dried masi is ground with dried red chilly and salt which is called as Masi podi.
Tuna is also a popular ingredient in sushi rolls, pasta dishes, soups, poke
bowls and of course, tuna fish casserole. Tuna steaks can also be seasoned and grilled,
seared or baked and served alongside your favorite side dishes.
Here are some simple tuna fish recipes that we can try out of dry masi powder.
1. Masi sambal
Preparation:
Mix masi powder with grated coconut,small onions and green chillies, squeeze
well.This a very good side dish for rotti and rice.
2. Tuna Salad
Preparation:
Mix masi powder with your own choice of vegetables and make salads.
3. Cheeni sambal:
Preparation
Saute onions, tomatoes green chillies, red chilli powder, salt and add two
spoons of masi powder .This a very good side dish for chappathi and rice.For fish
lovers this is a very good ingredient for peculiar fishy flavor.
4. Masi vadai
Vaada (Kayalpattinam famous) are alsoof the dishes made out of masi
powder.

Tuna Nutrition Facts
The nutrition profile of tuna can vary slightly depending on the type of tuna as
well as how it is prepared. In general, however, tuna is low in calories but bursting
with protein, selenium, niacin and vitamin B12. About 165 grams of light tuna
contains approximately:191 calories
 42.1 grams protein
 1.4 grams fat
 133 micrograms selenium
 21.9 milligrams niacin
 4.9 micrograms vitamin B12
 0.6 milligram vitamin B6
 269 milligrams phosphorus
 558 milligrams sodium
 2.5 milligrams iron
 44.6 milligrams magnesium
 391 milligrams potassium
 1.3 milligrams zinc
 0.1 milligram riboflavin
In addition to the nutrients listed above, tuna also contains some thiamine, vitamin
E, pantothenic acid and copper.
SUMMARY
Tuna is one of the important saltwater fish preferred throughout the
country.Their contribution towards foreign exchange is more.We all know that
consuming fish is healthy and provides multiple benefits, from improving the eyesight
and hair quality to keep the heart healthy. Tuna also preferred for its taste and flavor
and it is a powerhouse of essentialnutrientssuch asomega-3 fatty acids, potassium,
magnesium, iron, vitamin A, B6 and B12, and as such is beneficial for health.The
demand for value added products are keep on increasing due to the modern feeding
habits of people and also this is reliable health foods.

REFERENCES
Hickman, Martin 2009,"How Tuna Roe, Sam; Hawthorne, Michael (2005-
Conquered the 12-13)."How safe is
World".www.independent.co.u tuna?".Chicago Tribune.
k. The Independent. Retrieved Archived fromthe originalon
30 January 2019. 2009-11-26.
Shapiro, Amanda 2017, "If You're Not Balshaws, S.; Edwards, J.W.; Ross,
Buying the Best Canned Tuna, K.E.; Daughtry, B.J. 2008,
Are You Really "Mercury distribution in the
Living?".www.bonappetit.co. muscular tissue of farmed
Bon Appetite. Retrieved 30 southern bluefin tuna (Thunnus
January 2019. maccoyii) is inversely related to
Ellis, Richard, 2009,Tuna, A Love the lipid content of
Story New York: Random tissues".Food Chemistry.
House, p. 119.ISBN0-307- 111(3): 616–621.
38710-0 Rachael Link, MS, RD, 2018, Tuna
The tuna processing industry"U.S. Fish: Brain-Boosting, Protein
Department of Labor. Powerhouse or Toxin-Filled
Retrieved11 August 2013. Danger Food?
CFR– Code of Federal Regulations
Title 21". Accessdata.fda.gov.
Retrieved 2010-09-22.

TOXIC, FEEDING DETERRENCE, NUTRITIONAL AND
DEVELOPMENTAL PHYSIOLOGY OF PIPER BETLE L.(PIPERACEAE)
TREATED SPODOPTERA LITURA FAB.(LEPIDOPTERA: NOCTUIDAE).
Lingathurai S*
*Department of Zoology and Research Centre, Aditanar College of Arts and Science,
Corresponding Author: lings02@gmail.com
ABSTRACT
The biological activity of Piperbetle extracts (Hexane, chloroform, ethyl acetate and
methanol) was used for treatment with Spodopteralitura. The experiments were
carried out with concentrations of 0.625, 1.25, 2.5 and 5 percent in a leaf disc no
choice method and compared with control S. litura. Ethyl acetate extract was high
feeding deterrent and larvicidal activity for third instar larvae of S. litura (2.5 and 5
percent concentrations, 63.74 and 72.8 percent respectively). The ethyl acetate extract
on third instar larvae of S. litura showed LC50 value was 2.41%. Food consumption,
digestion, relative consumption rate, efficiency of conversion of ingested food,
efficiency of conversion of digested food, and relative growth rate values declined
significantly but approximate digestibility of treated larvae was significantly higher as
a result of treatment. Larval survival, pupal survival, larval period duration, pupal
period duration and pupal weight also inhibited. Qualitative analysis of P. betle
ethylacetate extract revealed that contains phytochemical such as, steroids and
quinines.The high biological activity of these quinines from P. betle ethyl acetate
extract could be used as an active principle during the groundwork of botanical
insecticides for lepidopteran pests. Based on their growth inhibitory and feeding
deterrent properties, some of this plant extract have higher for use as alternative crop
protectants against a number of pest species.

KEY WORDS:Spodoptera litura, Piper betle, quinine, feeding deterrence, toxicity,
nutritional variation, development physiology
INTRODUCTION
Insect pests destroy about twenty five percent of the world’s annual crop
production (Oerke, 1994). Most of the lepidopteran insects cause their damages
caused vegetable crops and cereals and pulses. Therefore, in recent years, various
researchers have been concentrating their efforts on the search for natural products
derived from plants and plant sources as an alternative to conventional chemical
insecticides for insect control. Plant based secondary molecules have been the subject
of thorough exploration for the past 30 years in an effort to find out new sources of
botanical insecticides and antifeedants.
Among the plant families studied, the Meliaceae, Rutaceae, Asteraceae,

Labiateae, Piperaceae and Annonaceae are possibly the most promising
(Schoonhoven, 1982; Jacobson, 1989; Isman, 1995). Azadirachtin, a triterpenoid
compound, limonoid group from neem tree seeds (Azadirachta indica, Meliaceae),
possesses most potent antifeedant and growth inhibitory effects against various insect
pests (Isman, 1997). Screening for biological activity using simple and fast bioassays
has now been added to give a better indication of the usefulness of the plants.
Various species of Piper are used in traditional medicine (Pio-Corrêa, 1984),

and as food flavouring and pest control agents (Estrela et al., 2006). Phytochemical
Investigations of different Piper species and plant parts have led to the isolation of
numerous active components including alkaloids, amides, pyrones, dihydrochalcones,
flavonoids, phenylpropanoids and lignans (Parmar et al., 1997).
Piper betle Linn. (Piperaceae) is a perennial dioecious creeper, probably

native of Malaysia but cultivated in India for its leaves, used for chewing (CSIR,
1969). The leaf is carminative, aphrodisiac, tonic, laxative and improves appetite
(Kirtikar and Basu, 1998). This plant is found widely growing in the tropical humid
climate of South East Asia, and its leaves, with a strong pungent and aromatic flavour,
are widely consumed as a mouth freshener. Leaves contained caryophyllene,

cadinene, γ-lactone, allyl catechol,p-cymene and eugenol methyl ether in varying
amounts (Sarkar et al., 2000).
The Indian traditional system of medicine has identified the P. betel leaves
with digestive and pancreatic lipase stimulant activities (Chatterjee and Pakrashi,
1995; Prabhu et al., 1995).The alcoholic extract of the leaf-stalk showed significant
antifertility effects in both male and female rats (Adhikary et al., 1989; Adhikary et
al., 1998). Autran et al., 2009, leaves of Pipermarginatum Jacq essential oil showed
potential larval toxicity against Aedes aegypti.Some scientists also reported
gastrocytoprotective, antimicrobial prperties and healing properties of the leaf extract
on experimentally induced gastric lesions (Majumdar et al. 2003; Nalina and Rahim,
2007; Bhattacharya et al.,2007).
The cluster caterpillar, S. litura (Fabricious) (Lepidoptera; Noctuidae)

important polyphagous pest of cultivated crops primarily in tropical, subtropical
regions (Brown and Dewhurt, 1975) andWestern and Southeast Asia (Murata and
Tojo, 2002). It has a wide range of host, feeding on 112 species worldwide, of which
40 species are known from India (Singh et al., 1998 and Paulraj, 2001). Many
vegetables and other crops are damaged by S. litura crops like to be seriously
damaged various taros, cabbage and its relatives and tomatoes (Schreiner, 2000). It is
a polyphagous and has about 150 host species (Rao et al., 1993). S. litura South
Indian strains exhibited high resistant levels 61- to – 148 fold to organic pesticides
(Karanth et al., 2002).
Awareness on the deleterious effects of chemical insecticides has led to

emphasis on biological agents for insect pest management that are eco-friendly, safe,
economically viable and socially acceptable. There is growing interest in the use of
biochemical / botanical pesticides (Rao et al., 1990; Koul et al.., 2004; Teway et al.,
2005). Botanicals are active and such pest control tools that have been eliciting
interest in recent times. They possess a complex of bioactive compounds that cause
different behavioral and physiological responses in insects.
Plants in the Piperaceae are members of traditional pharmacopeia in many

Asian and African tradedtional healers and have been used for pest control.
Piperguineese (Ivbijaro and Bolaji, 1990) for insecticidal and molluskicides, Piper

longumL, Piper betle L and Piper cubeba (Miyakado et al., 1989) for insecticidal
activity against mosquitoes and flies. There is no report for lepidopteran pests. The
search for plant-derived chemicals that have potential use as crop protectants
(insecticides, antifeedants, and growth inhibitors) often begins with the screening of
plant extracts. Initially, the test insects are fed the extracts and effects on insect
behavior and development are monitored. We undertook this study to determine the S.
litura to establish the phytocompound for pest control management. We discover to
evaluate their antifeedant, development, growth regulation and nutritional indices
against cluster caterpillar S. litura.
Insects
Spodoptera litura egg mass and larvae were collected from Valajabad
agriculture field, Kancheepuram district, Tamil Nadu, India. Collected egg mass and
larvae were maintained on castor leaves (Ricinus communis) in the laboratory at 26 ±
10C: 11 ± 1hr photoperiod and 65 – 70% R.H. Adults were released into oviposition
chambers for egg laying. Eggs were collected, kept separately and newly hatched
larvae were maintained on castor leaves. Freshly emerged 3 rd instar larvae were used
for the experiment.
Preparation of hexane, chloroform ethyl acetate and methanol extracts
Fresh leaves of Piper betle was collected from Samayanallur, Madurai

District, Tamil Nadu, India and were washed twice with tap water and once with
distilled water and then shade-dried for two weeks. The dried leaves material ground
into powder by an electronic blender and 300g of plant powder was soaked
sequentially in 1000ml with increasing polarity of solvents (Hexane, Chloroform,
Ethyl acetate and Methanol) for 72h with constant shaking. The soaked powder
material was filtered through filter paper. The solvent in the filtrate was evaporated
under reduced pressure by vacuum rotary evaporator (Evator, Buchi type, The
Science House Instruments, Chennai, India) to yield crude hexane, chloroform and
ethyl acetate extract. These concentrated three solvent crude extracts were analyzed

for antifeedant bioassay and active crude extract was further tested for growth
inhibition bioassay.
Antifeedant activity for Piper betle leaf extracts.

Antifeedant activity of crude extracts was studied using leaf disc no choice
method (Isman, et al, 1990). The stock concentration of crude extracts (5%) was
prepared by dissolving in acetone and mixing with dechlorinated water. From the
stock, required concentrations were prepared and tested against S. litura. Fresh castor
leaf discs of 4-cm diameter were punched using cork borer and dipped in 0.625, 1.25,
2.5 and 5. % concentrations of hexane, chloroform and ethyl acetate and methanol
extracts individually and air dried for 5 minutes. After air-drying, treated leaf discs
were kept in petridishes (1.5 cm X 9 cm) separately and single 2hr pre-starved 3rd
instar larva of S. litura was introduced on each treated leaf discs. Leaf discs treated
with acetone were considered as positive control. Ten replications were maintained
for each treatment and control. Progressive consumption of leaf area by the larva in
24 hr period was recorded in control and treatments. Leaf area consumed in plant
extract treatment was corrected from the control. The percentage of antifeedant index
was calculated using the formula of Bentley et al. (1984):
Antifeedant activity = [(C-T)/C] x 100
Bioassay
The leaf disc method of bioassay was discussed with Binod et al., 2007. In
contrast, fresh castor leaf discs were dipped in the different concentrations of plant
extracts (0.625, 1.25, 2.5 and 5 %) of three solvent extracts. separately for 1min.
Control leaves were treated with water and air-dried. The leaves were allowed to dry
at room temperature for 1min and were then placed in 90cm diameter petri dishes.
The experiments were carried out with newly moulted 4hr starved third instars (12
larvae per concentration 3 replication). The larvae were allowed to feed treated leaves
as well as solvent control leaves. After 24 h, the larvae were transferred to fresh
untreated castor leaves and maintained until they developed or died. The larvae were
observed for mortality. The percent mortality data after correction (Abott, 1925) were
estimated for a period of 4 days continuously. Moribund larvae were also considered
as dead larvae.

Preliminary Phytochemical analysis
Ethyl acetate of P. belte extracts are examined preliminary phytochemical

analysis. This method followed by Mukergy (2002) and Harborne (1983).
Nutritive food utilization of ethyl acetate treated S. litura larvae
Growth inhibitory activity and food consumption of effective extracts were

studied for four days with the treatment and after treatment. The various food
utilization efficiency measures were adopted from the progression of Waldbauer
(1968). S. litura larval weight gain, food consumption, and faeces were measured
every 24h. All weights were measured using a Monopan balance (Mettler Toledo -
Switzerland) accurate to 0.2mg. The fresh castor leaves (Ricinus communis) sprayed
with 0.625, 1.25, 2.5 and 5.0 percent concentrations of ethyl acetate extract of P.
betle. Control leaves were treated with acetone and air-dried. The newly moulted third
instar S. litura were used and starved for 4 h. After measuring the initial weight of the
larvae, they were individually introduced into detach containers. The larvae (12 larvae
per concentration) were allowed to feed period of 24h on castor leaves of weighed
quantities of extracts treated and untreated.
The uneaten leaves were weighed and removed after 24h and replaced with
fresh untreated leaves. Larvae were again weighed and the difference in weight of the
larvae was used as fresh weight gained during the period of study. Sample larvae were
weighed fresh to found a percentage of the experimental larvae. The leaves remaining
at the end of each day were weighed to establish a percentage conversion value to
allow for the assessment of diet weight. The quantity of food ingested was estimated
by subtracting the diet (dry weight) residual at the end of each experiment from the
total dry weight of the diet provided. Faeces were collected and weighed, and then
oven dried, and re-weighed to estimate the dry weight of excreta. The experiment was
continued for 4 days and observations were recorded every 24 h. Consumption and
post-ingestive food utilization efficiencies (dry weight) were calculated. Relative
growth rate (RGR), consumption index (CI), approximate digestibility (AD),
efficiency of conversion of ingested food (ECI) and efficiency of conversion of
digested food (ECD) were estimated until pupation of treated and control insects.
Consumption index (CI) = E/ TA, Relative Growth rate (RGR) = P/ TA, Efficiency of

conversion of ingested food (ECI)= P/E X 100, Efficiency of conversion of digested
food (ECD) = P/ (E-F) X 100, Where, A is mean weight of animal during period, E is
the weight of food eaten, F is the weight of feces produced, p is the weight gain of
insect, T is the duration of experimental period.
Effect of ethyl acetate extracts of P. betle on S. litura larval development
Third instar larvae were used for S.litura larval development bioassay. Leaf
discs (4cm diameter) were dipped in ethyl acetate extractof P. betle at different doses.
Four concentrations, (0.625, 1.25, 2.5 and 5%) were dissolved in acetone and applied
individual leaf discs were used (3 groups of 10 insects each). Controls were treated
with acetone alone. The duration oflarval, pupal and adult stages were noted after
treatments with different concentrations of the ethyl acetate extracts were evaluated.
Every 24h the castor leaf were replaced and for each individual the weight and the
stage were recorded until it died.
Statistics
The lethal concentrations (both LC50 and LC90) were calculated using probit
regression analysis and values were expressed as means ± standard deviation (SD).
Data from nutritional indices, antifeedant activity and larval development were
subjected to analysis of variance (ANOVA). Probit analysis was done to calculate
median lethal concentration (LC50) and LC90 using SPSS 11.5 version software
package.
RESULTS
Impact of four different solvent extracts (Hexane, chloroform, ethyl acetate

and methanol) of leaves of P. betle extracts screened feeding deterrence tested against
third instar larvae of S.litura. Among the different extracts, ethyl acetate extract
exhibited promising result antifeedant activity followed by chloroform, hexane and
methanol. Five percent concentration of ethyl acetate extracts shows 72.8%, hexane
51.63% chloroform 38.36%, and methanol35.3%. Feeding deterrence of S. litura third
instar larvae on castor leaves measured percent leaf damage was significantly greater
on P. betle ethyl acetate extract-treated leaves than on solvent treated control leaves in
both experiments 22.85, 33.16, 65.74 and 72.8% feeding deterrent between control

and treatment with 0.625 – 5% were noted 24h feeding. The observed higher feeding
activity of larvae on control leaves compared to extract-treated leaves increased. The
ethyl acetate extract were tested phytochemical analysis followed by Harborne (1983)
and Mukergy (2002) contain steroids and quinines (Table 1).
So, we test toxicity, nutritional parameter and biology of S. litura assessed

ethyl acetate extracts. The LC50 and LC95 values, confidence limit (95%) and
regression slope at 96h exposure to P. betle ethyl acetate extract shown in Table 1.
The LC50 and LC95 for third instar larvae is 2.41 and 18.0 % concentration (table 2).
Consumption index (CI), relative growth rate (RGR) and nutritional efficiency
measured (ECI and ECD) of treated individuals were reduced in comparison to those
control. Relative growth rate did not show significant changes in treated larvae
compared to control. But approximate digestibility (AD) increased with increased
concentration of treated insects. Both ECI and ECD were significantly reduced at all
treated concentrations. Table 3 shows no significant difference in relative growth rate
between larvae in control (11.45mg), and larvae treated with 0.625 percent
concentration of extract (10.42mg). the difference was significant between control
and treatment with 1.25 % concentration (9.14 mg), 2.5% (7.00mg) and highly
significant with 5% extract (6.25mg). Consumption Index also same as at 5percent
concentration level showed 1.18 and control insects 5.82. The approximate
digestibility (AD) only increased by the increasing concentration of treatment. At 5%
level (69.35 percent) and lower concentration 0.625% (52.49 percent) with
comparisons of control 51.2 percent. The efficiency of ingested food was also
affected significantly at the different concentration (0.625 – 5%) compared with
control (28.3 percent). Differences were also found in the efficiency of digested food
between control and different P. betle ethyl acetate extract concentrations.
All treatments reduced RGR, CI, ECI and ECD of from third instar to
pupation. The treatment of P. betle into the castor leaves significantly reduced larval
growth of armyworm compared to controls (Table 3). There was a concentration-
dependent reduction in growth from 0.625 to 5%. Efficiency of conversion of
ingested and digested food (ECI and ECD) into biomass of S. litura larvae was
reduced except the control. The reduction in these parameters was irrespective of any
significant change in relative consumption rates and the only significant reduction in

consumption relative to controls was observed at the highest treatment dose of 5%
(Table 3). Approximate digestibility (AD) of 5% extract treated larvae was
significantly higher than the control in the 0.625 and 1.25% treatments during the
experimental periods. Ethyl acetate extracts on the total larval duration, pupal
duration, pupal weight, adult emergence and malformed adults are given Table 4.
The elongation 5.54 days was observed in larval period (19.75 d) at 5% ethyl
acetate extracts of P. betle. Lower concentrations of P. betle extracts showed
concentration dependent increased larval period (14.25, 17.10 and 18.50 days for
0.625, 1.25 and 2.5% concentration respectively), of which 0.625% concentration was
insignificant when compared to the control (p ≥ 5%). Experimental treated pupal
stages also increased, the highest elongation of is pupal period was observed in P.
betle (11.39 days) followed by (6.85 – 9.42 days) at 0.625 – 2.5% concentration of
treatment. Since the treatment showed concentration dependent positive response of
pual weight also decreased accordingly. Among the experimental insects minimum
pupal weight was recorded 126.67mg at 5% concentration of treatment followed by
146.29, 167.20 and 180.48 in the concentration of 2.5, 1.25 and 0.625%.
Adult duration was observed treated insect life span significantly decreased.
Lower concentration (0.625%) showed 6.35 days, higher concentration 5% treatment
2.62 days only. Deformed adult S. litura was also noted in their respective
concentration 5% level 38.66 abnormal adults were showed. Larval duration of
control insects showed 1 days. The larval duration increased insect (p ≤ 5% level)
except for comparison of control. (Table 4). Pupal life duration did show significant
difference in all treated groups as compared with control except at 0.625%
concentration of ethyl acetate extract of P. betle. Pupal weight also decreased by
increasing concentration of treatment. Adult life span sharply decreased at higher
concentrations did show pronounced differences as compared with the control insect.
Growth regulatory effect such as a deformed adults (deformed wings) occurred only
at higher concentrations. The deformed insect exhibited major growth retardation of
further development.

DISCUSSION
Plant secondary metabolites synthesized by plants an important role in

protecting plants against insect pests. These compounds affect insects by being toxic
causing a delay in larval growth and can act as antifeedant Isman (2006). S. litura
larvae consumed less foods and gained lesser weight after the P. betle treatment when
compared control. Reduced consumption of leaves in treated is likely to be the
consequence of toxicity rather than cause of growth inhibition.
The present study indicates that ethyl acetate extract of P. betle is reduced feeding
rate of S. litura. The rate of feeding varied significantly depending on the
concentration of the plant extract. Ethyl acetate extract of this plant caused
malformation of pupal and adult stages. Similar intermediates (larval –pupal and
pupal - adult) were obtained when treated larvae of S. litura, S.
mauritia,Ephestiakuehniella.and M. sexta (Gujar and Mehrotra, 1983; Jegannadh and
Nair, 1992; Schluter et al., 1985; Barby and Klocke, 1990; Kumar et al., 2001).
This information well supported by the data from nutritional experiments

where P. betle resulted in lower RGR and concomitant reductions in ECI and ECD.
Interestingly, the RGR was significantly reduced by ethyl acetate extract of P. betle
treatment which indicates that feeding depression was caused by behavioral effects
(Jeyabalan and murugan, 1995). Reduction of ECI and ECD confirms the deleterious
effects of post-ingestive toxicity.
This study clearly revealed that P. betle highly reduces the food consumption
index, growth rate, efficiency of conversion of ingested food and efficiency of
conversion of digested food. Hence P. betle leaf ethyl acetate extract can be explored
in S. litura management. The extended larval and pupal duration and reduced
longevity suggest that extract may disturb the endocrine function either to the
blockage of haemolymph ecdysteroid peak, or extracts interfere with other
biochemical / physiological processes through binding to critical macromolecules is
highly probable(Koul and Isman 1991; Mordue et al., 1986).
The excellent antifeedant activity of the ethyl acetate extract of P. betle

demonstrates their potential use as natural insecticides. Additionally, this extract

exhibited larval mortality against of S. litura. This antifeedant and growth-inhibiting
activity can therefore be incorporated into other insect control techniques in the
strategy of integrated pest management (IPM). It would be interesting to investigate
whether Piper betle contains substances similar to the antifeedant and growth
inhibiting compounds present in the fruits of Azadirachta indica (Rembold, 1984,
Schmutterrer, 1995) and Melia volkensii (Mwangi, R.W. and H. Rembold, 1998 and
Kabaru,1996)
In conclusion, our results indicate that P. betle extract has toxic, as well as
growth regulatory; feeding deterrence caused pupal and adult malformation in S.
litura. The use of this plant extract may play a more prominent role in integrated pest
management programs in the future.
Figure 1. Percent feeding deterrence of hexane, chloroform, ethyl acetate and

methanol solvent of P. betle leaf extracts treated against 3rd instar larvae of S.
litura
Table1. Preliminary phytochemical analysis P. betle extract
Extract Quantity of extract (gm) Phytochemicals
Hexane 6.83 Steroids
Chloroform 14.42 Steroids,

Ethyl acetate 8.71 Steroids and quinone

Methanol 12.58 Saponin
Table 2. Toxicity of P. betle ethyl acetate leaf extract against third instar larvae
of S. litura
Insect LC50a LC95a Slope ± SE Chi square (X2)
S. litura 2.41 (1.23 – 8.48) 18.0 (6.24 – 16225.9) 1.88 ± 0.72 0.170
Units LC50 and LC95 = % / w, applied for 96h. a95% lower and upper fiducial limits
are
shown in parenthesis.
Table 3. Nutritional indices of P. betle ethyl acetate extract treated with third
instar
larvae of S. litura
Treatment
RGR (mg) CI (mg) AD (%) ECI (%) ECD (%)
(%)
Control 11.45 ± 2.97a 5.82 ± 0.48a 51.2 ± 2.30a 28.34 ± 1.83a 48.4 ± 4.4
0.625 10.42 ± 2.75a 5.42 ± 0.99a 52.49 ± 3.82a 15.54 ± 2.46b 32.94 ± 5.6
1.25 9.14 ± 0.89b 4.59 ± 0.58b 59.04 ± 4.65ab 17.08 ± 1.35a 27.69 ± 1.2
2.50 7.00 ± 1.11b 2.60 ± 0.81bc 65.24 ± 3.49b 11.21 ± 0.58b 20.54 ± 2.0
5.00 6.25 ± 1.93bc 1.18 ± 0.02c 69.35 ± 5.56c 8.34 ± 1.68c 21.04 ± 3.5
(Mean ± SD)Values carrying same alphabets in a column are statistically not significant by
LSD at 5% level. RGR, relative growth rate: CI, consumption index: AD, approximate
digestibility; ECI, efficiency of conversion of ingested food; ECD, efficiency of conversion
of digested food.
Table 4. Biological characteristics of S. litura on ethyl acetate extract of P. betle

treated castorleaves.
Treatments Larval Pupal Pupal Adult Percent

(%) duration Duration weight duration deformed
(days) (days) (mg) (days)* adults
Control 13.21 ± 6.50 ± 219.24± 7.75 ± 0a
1.78a 2.11a 3.30a 0.91a
0.625 14.25 ± 6.85 ± 180.48 ± 6.35 ± 2.33 ± 0.24a
1.45a 1.62a 2.67a 1.47a

1.25 17.10 ± 8.43 ± 167.20 ± 6.22 ± 8.66 ± 0.52b
1.53b 1.88ab 2.88bc 1.04b
2.5 18.50 ± 9.42 ± 146.29 ± 5.83 ± 16.34 ± 1.86
1.86bc 2.38b 6.10c 1.30bc
5 19.75 ± 11.39 ± 126.67 ± 2.62 ± 0.64 38.66 ± 3.76
2.52 1.43 4.21
Values carrying same alphabets in a column are statistically not significant by LSD at
5% level
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Biology of Lacanobia oleracea

BIO-EFFICACY AND DETOXIFYING ENZYME ACTIVITIES OF
EUPHORBIAHIRTA L. (EUPHORBIACEAE) TREATED ARMYWORM,
SPODOPTERA LITURAFAB. (LEPIDOPTERA: NOCTUIDAE)
SaranyaTand LingathuraiS1,*
PG Department of Zoology, Pachaiyappa’s College for Men, Kanchipuram – 631 501,

Tamil Nadu
1
Department of Zoology, Aditanar College of Arts and Science, Tiruchendur – 628
216, Tamil Nadu
ABSTRACT
Toxic effect of crude seed extracts of Euphorbiahirta from Kanchipuram, Tamil
Nadu, India, was evaluated against the armyworm, Spodoptera litura (Lepidoptera:
Noctuidae) using different solvent extracts. All the four different viz., Hexane,
Chloroform, ethyl acetate and water extracts were subjected to preliminary
phytochemical assay. All the extracts showed insecticidal activity with dose
dependent manner with four different concentrations (0.625, 1.25, 2.5 and 5%). High
insecticidal activities were observed in 5 percent concentrations of chloroform extract
showed 65.33% mortality of S.litura. Chloroform extract showed lowest value of
lethal concentration values i.e., gave LC50 value 358.59 % S. litura. Effective and
active extract (chloroform) further tested growth inhibitory activity was tested against
S. litura. Larval weight, pupal weight and adult durations were inhibited with
increasing concentrations of treatments. Further glutathione S-transferase and
monooxygenase enzymes were inhibited when increasing concentrations of
treatments. Our results indicate that E.hirta had potential for development as botanical
insecticides, especially for local use.
KEYWORDS: Euphorbiahirta, solvent extract, Spodoptera litura, insecticidal

activity, Growth inhibitory, detoxification enzyme
INTRODUCTION
In recent technology, pesticide manufacture have focused research in develop
plant compound related pesticide Despite all the efforts exerted in protecting crops

from noxious pests all over the world, losses due to these causes can annually reach
10–20% (Ferry et al., 2004), still remaining a challenge to be resolved. Currently,
synthetic insecticides are the most used mechanisms to control pests. However,
concerns over the development of resistance, toxicity and environmental pollution
associated with conventional synthetic insecticides compel us to look for new
compounds. Attention is being directed towards plants that can be an alternative to
synthetic insecticides because they have evolved together with herbivorous insects,
developing mechanisms to interact and defend themselves. In this respect, plants are
able to synthesize a broad range of different chemical compounds called secondary
metabolites many of them providing new sources of natural pesticides.
Euphorbia hirta (Euphorbiaceae) is a small annual herb. The plant is
commonly called pill bearing spurge and asthma herb and the stem is slender. The
leaves are oppositely arranged, lanceolate and are usually greenish or reddish
underneath measuring about 5cm long. The plant leaves are used to treat colic
troubles, dysentery, cough, asthma, worms and vomiting. The white latex is used as
eye drops to cure conjunctivitis. Paste of leaf is applied externally (twice daily) on the
place of scorpion bite. Therefore present study was planned to evaluate laboratory
expriments of leaf extract of E. hirta Linn.

Fresh leaves of Euphorbiahirta leaf powder (500 gm) and soaked with hexane,
chloroform, ethyl acetate solvent and water extracts was prepared (Plate 1).
Phytochemical analysis of E. hirta extracts was done by Horborne, 1984. The test
insect of Spodoptera litura cultured and tested against E. hirta. The insecticidal
activity and growth inhibtory activity (Isman et al., 2001) experiments were
conducted using the third instar larvae of S. litura. The effective extract was studied
detoxifying enzyme (Glutathione –S transferase and monooxygensase) activity
(Oppenoorth et al., 1979) Field bio-efficacy and GC-MS studied were conducted with
effective extract.

The extract showed a greater yield for chloroform (3. 68gm). And then
hexane, water and ethyl acetate extracts were yielded 2.54, 1.82 and 1.25 gm (Table

1). Hexane extract contain tannins and terpenoids. Chloroform showed alkaloids,
steroids, flavonoids and terpenoids. Ethyl acetate and water extracts resulted
alkaloids, anthraquinones, phenolics and saponins, flavonoids and phenolics (Table
2). Among different solvent extracts, chloroform extract was found to be more active
and potential followed by ethyl acetate, hexane and water extracts.
At 5% concentration of treatment chloroform extract exhibited maximum and
significant larvicidal activity towards Spodoptera litura. Significant and dose
dependent mortality was observed all the treatment groups. Mortality of larvae was
calculated with different concentrations as variable revealed significant difference in
larval mortality (P ≤ 0.05 level) (Table and 4).
Growth inhibitory activity were assayed effective extracts i.e., chloroform
extract (Table 5). When increasing concentration of treatment showed decreased
level of larval and pupal weights. All the treatments showed concentration
dependent. At 5% concentration levels of treatment were larval and pupal weight
reduced significantly towards S. litura. And adult durations also decreased
significantly when increased concentration of treatment.
In the present study two detoxification enzyme levels (Glutathione S-

transferase and Monooxygenase) was estimated in the larvae of S. litura in
chloroform treated (Table 6). When increasing concentration of these enzyme levels
were reduced. All the treatments were showed dose depended activity. Malformed
insect development was formed (Plate 2). As such, this plant has a high potential as a
source of phytochemicals useful to protect our crops from herbivorous insects, with
interesting perspectives on ecological systems of foodproduction.
Plate 1. Euphorbiahirta plant

Table 1. Yield of Euphorbiahirta crude extracts
Solvent Quantity of Yield (gm)

Hexane 2.54
Chloroform 3.68
Ethyl acetate 1.25
Water 1.82
Table 2. Preliminary phytochemical analysis of Euphorbia hirta extracts
Extracts Phytochemicals
Alk Ste Sap Tan Fla Ter Ant Qui Phe Cou
Hexane - - - + - + - - - -
Chloroform + + - - + + - - - -
Ethyl + - - - - - + - + -
acetate
Water - - + - + - - - + -
(Alk: Alkaloids, Ste: Steroids, Sap: Saponins, Tan: Tannins, Fla: Flavonoids,
Ter: Terpenoids, Ant: Anthraquinones, Qui: Quinones, Phe: Phenolics and
Cou: Coumarines) (+ present and - absent)
Table 3. Insecticidal activity of Euphorbia hirta against Spodoptera litura larvae
Concentration (ppm)
Extract
0.625 1.25 2.50 5
Hexane 0.0±0.00a 3.33±0.72b 12.00±1.77b 26.67±2.36c
Chloroform 0.0±0.00a 16.67±3.59c 30.00±3.20c 65.33±4.00e
Ethyl acetate 0.0±0.00a 20.00±2.19d 36.67±2.07d 48.67±2.18d
Water 0.0±0.00a 0.0±0.00a 0.0±0.00a 10.00±1.53b
Control 0.00±0.00a
Table 4. Lethal concentrations of Euphorbia hirta treated larval mortality

towards Spodoptera litura
Extracts LC50 95% Fiducial limit LC90 95% Fiducial limit Chi-
(%) Lower Upper (%) Lower Upper square
Hexane 929.18 669.461 1705.553 3599.41 1896.22 12687.34 0.855*
Chloroform 358.59 215.906 1687.638 1079.93 513.43 15144.79 6.037

Ethyl 578.18 414.560 1030.516 5693.13 2450.37 29199.05 1.800*
acetate
Water - - - - - - -
*χ2 values are significant at P<0.05 level
Table 5. Growth inhibitory activity of chloroform extract of Euphorbia hirta

treated Spodoptera litura
Concentrations Larval weight Pupal weight Adult duration

(%) (mg) (mg) (days)
0.625 127.38±2.96b 94.17±2.80b 5.65±1.02a
1.25 107.22±2.46c 78.14±2.27c 4.62±0.36ab
2.5 86.20±2.15d 59.80±1.22de 3.92±0.33b
5 62.49±1.58e 57.44±1.44de 3.64±0.18bc
Control 150.70±4.66a 103.24±3.28a 6.20±0.84a
within columns, means (± SD) followed by a same letter do not differ
significantly (Turkey’s test, P< 0.05 level)
Table 6. Detoxifying enzyme activities of chloroform extract of Euphorbia hirta

treated Spodoptera litura
Concentrations (%) Detoxifying enzyme activity

Glutathione S-transferase Monooxygenase
(nM/min/mg of protein) mOD/min/mg proteins
0.625 2.176±0.52b 8.715±1.30b
1.25 1. 894±0.33c 2.937±1.12c
2.5 0.299±0.37d 0.145±0.83d
5 0.232±0.14e 0.128±0.42e
Control 4.380±0.42a 11.232± 1.61a

Plate 2. Abnormal pupae and adult after the treatment
ACKNOWLEDGEMENT
The authors wish to thank Tamil Nadu State Council for Science and
Technology (TNSCST), Chennai, Tamil Nadu for providing the funding sources
(Student Project Scheme - Ref No: AR – 13; Letter No. TNSCST/SPS/AR/2013-
2014).
REFERENCES
Horborne SB. 1984. A guide to KT. 1979. Pestic. Biochem.
modern techniques of plant Physiol. 7, 34.
analysis. Chapman and Hall, Ferry, N., Edwards, M., Gatehouse, J.,
London. 4-80.3. Gatehouse, M., 2004. Plant-insect
Isman MB Andrew JW and Passreiter, interactions: molecular approaches to
2001. Fitoterapia.72: 65-68. insect resistance. Curr. Opin.
Oppenoorth FJ Smissaert HR Welling Biotechnol. 15, 155–161.
W van den Pas LJT Hitman .

PRODUCTION AND MARKETING OF PALM LEAF ARTICLES – A STUDY
WITH SPECIAL REFERENCE TO UDANKUDI
S. Siril Arun* and M. Ruban Jesu Adaikalam
Dept of Commerce, Aditanar College of Arts & Science. Tiruchendur – 628 216.
ABSTRACT
Women occupy 50 percent of the employees in the field. Particularly in the
production of palm leaf articles, women play the major sole. In the Thoothukudi
District, Udankudi is considered as an important village, particularly for women of
palm sector and poor people. It is mainly marketed through “The Udankudi women
workers plamleaf industrial co-operative society” at Udankudi and local merchants.
Both the artisans and middlemen are confronted with many problems in marketing
their palm leaf products.
INTRODUCTION
Tamil Nadu is the pioneer state in palmgur industry next to agriculture and
handloom weaving in the state. Palmyrah tree is the “state tree” of Tamil Nadu. The
performance of production of the industry is well identified by the steady growth of
the production of palm based products. Palmyrah fibre and palm leaf articles are
exported from only Tamil Nadu and Andhra Pradesh. Palm industry in the largest
employment generating industry in the region. Women occupy 50 percent of the
employees in the field. Particularly in the production of palm leaf articles, women
play the major sole. In the Thoothukudi District, Udankudi is considered as an
important village, particularly for women of palm sector and poor people. It is mainly
marketed through “The Udankudi women workers plamleaf industrial co-operative
society” at Udankudi and local merchants. Both the artisans and middlemen are
confronted with many problems in marketing their palm leaf products. Hence the
research makes an attempt to study the “Production and marketing of palm leaf
articles at Udankudi”.
Marketing fancy and utility articles from palmyrah leaf is a traditional activity
in Tamil Nadu. But modern techniques in the production are very much sophisticated
and the economic importance attached to this industry is great. Two aspects of this

industry are worth mentioning here. One is the value of the articles as a foreign
exchange earner and the other that it has generated employment opportunities to
women. It has been stressed by eminent scientists and social workers that in order to
uplift the status of women, there is a need to generate opportunities for their
employment to make them economically independent. In our country a number of
development programme have been implemented for accomplishing effective growth
for enhancing women’s productive role. It is a well known fact that the palm leaf
articles have the greater risk of colour changing and fungus attack in the rainy season.
Therefore they are to be stored in air tight rooms fitted with heaters. As such the
artisans have the problem of immediate disposal of articles.
This study is based on both primary and secondary data. The primary data
were collected with help of the interview schedule. The secondary data were collected
from the books journal and websites. In order to primary data were collected directly
from the 82 artisans and middlemen were surveyed. The sample was selected on the
basis of simple random sampling and the sample respondents were selected by lottery
method. The data were collected, coded, tabulated and presented in a master table.
From the master table, sub-tables were prepared. The statistical techniques used in
this study are tables and percentage.
Tools / Equipments used
Attempts have been made successfully to evolve appropriate tools for the
sizing of palmyrah leaves. There are screw press, sewing machine, cutting knife, ears
removing needle, leaf sizing knife, stove, dyeing vessel, hammer, pliers, fork, blow
lamp, scissors, balance, sharpening, store and hand drilling machine.
Types of fancy articles produced
They are mango tree open, mango trey closed. Spoon tray, oval and round try
open, round plate, sweet try, oval and round tray open, octangular tray, mango plate,
oval plate square tray, round dish, Triangular tray, diamond share tray big, glass
holder, toy elephant, flower vase hanger, deluxe round dish, pithi round mate, deluxe
round bowl small, pilapetti small, flower vase hanger set etc.,
The articles produced by them may be prescribed or un-prescribed. Table: 1
shows the type of articles produced by the artisans.
Table 1 Types of articles produced by the artisans

Types Number of respondents Percentage
Prescribed 70 85
Un-prescribed 12 15
Total 82 100
Table 1 reveals that 85 percent of the respondent produce only prescribed articles and
the remaining 15 percent produce un-prescribed articles.
Sources of supply of leaves
Palm leaf articles are produced with palm leaves. Palm leaves are received
from various sources. The sources of supply of leaves are presented in table 2
Table 2 Sources of supply leaves
Source Number of Percentage
respondents
Family member being a tapper 20 24
Purchasing from any other tapper 15 18
From lease owners 40 49
Palmyrah cutters 7 9
Total 82 100
Table 2 reveals that about 24 percent of the respondents purchase their leaves
from family members, 18 percent from any other tapper 49 percent from lease owner
and 9 percent from palmyrah cutters.
Training
Though every female member of the artisan family is conversant with making
the common variety, training helps them in knowing a number of new varieties.
Almost all the artisans have undergone the training offered by IRDP,
Reasons for being in the occupation
Table 3 reveals the reasons for being in the occupation.
Table 3 Reasons for being in the business
Reasons Number of respondents Percentage
Hereditary 30 37
Low capital needed 33 40
Unable to do other jobs 6 7
Assistance from family 13 16
members
Total 82 100

From table 3 it is very clear that the major reason for doing this business by
the artisans are tow namely “Low capital needed” 40 percent and “Hereditary” 7
percent. The other reasons revealed by the respondents are “Assistance from family
members” 16 percent and “Unable to do other jobs” 17 percent.
Production of palm leaf articles
The production of palm leaf articles per year by the sample respondents is
given in the table 4.
Table 4 Production of palm leaf articles by sample respondents
Production (Rs.) Number of respondents Percentage
Below 2000 22 27
Rs. 2000 – 4000 15 18
Rs. 4000 – 6000 32 39
Rs. 6000 – 8000 5 6
Rs. 8000 – 10000 4 5
Above Rs. 10000 4 5
Total 82 100
It is inferred from the table 4 that 27 percent of the artisans produce the
articles worth less than Rs. 2000 and 18 percent of them produce the articles for Rs.
2000 – Rs. 4000. These two groups belong to the in active member of the field. The
artisans whose production exceed Rs. 4000 are of active members of the field. Most
the artisans 39 percent fall under the group of Rs. 4000 – Rs. 6000 in respect of their
income, 6 percent of the artisans fall under the group of Rs. 6000 – Rs. 8000. These
two groups are of active members. 5 percent fall under the group of Rs. 8000 – 10000
and 5 percent in the group of above Rs. 10000. These two groups get assistance from
their family members.
Income from output
Table 5 shows the income of the artisan from the output alone.
Table 5 Income from output per year
Income (Rs.) Number of respondents Percentage
Below 2000 37 45
Rs. 2000 – 4000 32 39
Rs. 4000 – 6000 5 6
Rs. 6000 – 8000 4 5
Rs. 8000 – 10000 4 5
Above Rs. 10000 - -
Total 82 100

From table 5 it is inferred that most of the respondents fall under the income
group of below Rs. 2000 45 percent. The artisan who fall under the category of Rs.
2000 – Rs. 4000 39 percent mainly constitute the active member of the field, and the
artisans who fall under the category of Rs. 8000 – Rs. 10000 are those who get
assistance from family members and are active.
Marketing of palm leaf articles by the artisans
The palm leaf articles are sold by the artisans through two ways.
 Through “The Undakudi Women Worker palm leaf Industrial cooperative
society.
 Through other middlemen.
Marketing Through the co-operative society
Table 6 reveals the number of respondents who are the members of the
society.
Table 6 Members of the co- operative society
Category Number of respondents Percentage
Yes 75 91
No 7 9
Total 82 100
From the table 7, it is very clear that 91 percent of the respondents are the
members of the society and the remaining 9 percent are the non members.
Marketing through other middlemen
The reasons are
 Immediate cash payment
 Initial stage artisans are benefitted as quality is not insisted
 For quick disposal of articles when there is heavy rain
 High price is paid for the articles.
Problems of this industry
The problems of this industry as per the artisans are grouped into four
categories namely production problems, marketing problems, financial problems and
training problems. They were included in the interview schedule. The opinions of the
respondents namely artisans who are the members of the society and artisans who are
not the members of the society were gathered they are presented in this part. Further
anlaysis of group, intergroup as well as over all perception about the problems of the
industry have also been taken into account.

Perception of the actors about the production problems
 Seasonal availability of raw materials.
 Quality of leaf decline due to moisture in the atmosphere.
 Time limit imposed by the society.
Perception of the actors about the financial problems
 Lack of finance.
 Lower prices for the article.
 Increase in the price of raw material.
 Loan under IRDP given to limited number of members.
Recommendations
 If the society, with adequate storage facilities can supply the leaves as and
when they are needed.
 Due to various constraints, only limited number of artisans are able to get
IRDP loan thorough the society.
 There is only one IRDP sponsored training centre by the society.
 Strict control is being enforced by the society. But the middlemen do not seem
to be bothered must about the quality of the products.
 The palm leaf articles normally produced at a lesser price form the artisan.
 The articles are transported only through the buses crossing the village. The
quantum transported thus is not so large.
CONCLUSION
The palm leaf products are till to date produced only in the old fashioned,
traditional style of the products are tailor-made to suit the popular demands of the
average customers. The individual creativity of the artisans is totally missing because
of lack of proper encouragement. Therefore the whole production process has to be
modernized embracing beautification coupled with creativity. For this the artisans are
to be trained through intensive training programme in the training centres.
Encouragement must also be there in the form of state and national awards to
recognize talents.
Most of the artisans are women folk who do not have adequate financial
facilities at their disposal. So Government sponsored agencies should come forward
with liberal financial aid at low rates of interest and allowing reasonable subsidies

when the industry is hit by natural constraints. So when this industry is allowed a
natural growth as the palm trees are allowed so, a real renaissance will soon be in the
offing.
REFERENCE
1. Kotler Philop, “Marketing Management”, Prentice Hall of India, New Delhi.
2. I. M. Pandey, “Financial Management”, Vikas publications, New Delhi.
3. Van Horne James C, “Financial Management Policy” Prentic Hall Inc, New
Jensey
4. Indian Journal of markeing vol. XXIV, No 2 to 3
5. Khadi Gramochog Vol. XXIV - XXXXI

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Department of Zoology and Research Centre

Website: www.aditanarcollege.in Phone: 04639 242232

In the present scenario, a spectacular and overwhelming utilization of Bio-resources

Editorial Board Members

Dr. C. SUNDARAVADIVEL Dr. D. VASUMATHI

Dr. P. AROCKIA MARY FERNADEZ MISS. P. JENIFER

This publication is in copyright. Subject to statuary exception and to the

First published 2019

Proceedings of International Conference on Bio-Commerce - 2019

Published by Dr.G.Lakshmanan for ‘Proceedings of International

Typeset & Printed by Robert Printers, Thoothukudi – 628 002. Ph:

USING THE WORLD’S DEEPEST CORAL NURSERY TO EXAMINE THE

EVALUATION OF ANTI-INFLAMMATORY AND ANTIBACTERIAL 02

A MOSQUITOCIDAL ACTIVITY OF MUSIZIN ISOLATED FROM 14

INVESTIGATION ON ETHNO PHARMACOLOGICAL AND 75

T.Jebarani Rajathy and T.Mohanraj

EFFECT OF PROBIOTICS ON CICHLID FISH, MAYLANDIA 101

BIO-EFFICACY OF EUPHORBIAHIRTA L (EUPHORBIACEAE) AGAINST 384

S. Siril Arun and M. Ruban Jesu Adaikalam

Daniel G. Merselis1, Thinesh Thangadurai 1, Anthony bellantuono 1, Katherine

1 Department of Biological Sciences, Florida International University, Miami, FL,

2 NASA Johnson Space Center, Houston, TX, USA.

Worldwide coral transplantation efforts are being undertaken to restore reefs

International Conference on Bio-Commerce 2019 Page 1

International Conference on Bio-Commerce 2019 Page 2

Muzafar Riyaz1 and S. Ignacimuthu1*

KEYWORDS: Pests, Climate, Global warming, Agriculture.

International Conference on Bio-Commerce 2019 Page 3

T.P Ajeesh Krishna1 and V. Duraipandiyan1*

KEYWORDS: Phosphorus, Climate changes, Elevated temperature, CO 2,

International Conference on Bio-Commerce 2019 Page 4

G. Victor Roch and V.Duraipandiyan*

International Conference on Bio-Commerce 2019 Page 5

T. Maharajan1 and V. Duraipandiyan1#

KEYWORDS: Finger millet, Shoot apex, In-vitro, Regeneration, ISSR.

International Conference on Bio-Commerce 2019 Page 6

F. Esther Isabella Eucharista*and A.Shanthi

*Department of PG Zoology, Aditanar College of Arts & Science, Tiruchendur.

KEYWORDS: Physico-chemical parameters, Phytoplankton.

International Conference on Bio-Commerce 2019 Page 7

J. Rajaselvam, I.Vasudhevan and T.Subi

KEYWORDS:Antimicrobial activity, Rhizophora mucronata, Disc diffusion method.

International Conference on Bio-Commerce 2019 Page 8

Pathakaraimuthu Balakumar1, Subramaniam Balakumar2, Perumal

International Conference on Bio-Commerce 2019 Page 9

K. Jesima* and S. Jesily

International Conference on Bio-Commerce 2019 Page 10

Devi Priya .S* and L. JeyaPraba

KEY WORDS: Physico-chemical parameters, pH,salinity,Manapadu fish diversity.

International Conference on Bio-Commerce 2019 Page 11

P. Rama Devi * ˡ K. Nandha Devi1, Arokiya Mary Fernandez1 and C. Babu ²

KEYWORDS: Lectin, Heamagglutination, Chromatography, SDS

International Conference on Bio-Commerce 2019 Page 12

International Conference on Bio-Commerce 2019 Page 13

William Raja Tharsius Raja1, Ganesan Pathalam1, Duraipandiyan Veeramuthu1*

International Conference on Bio-Commerce 2019 Page 14

S. Subburaman¹*, Nileshjoshi², D. Nagarajan¹, J.Nagarajan¹ and Ramakotti¹

International Conference on Bio-Commerce 2019 Page 15

R. Prema1 and BeenaSomanat2

International Conference on Bio-Commerce 2019 Page 16

KEY WORDS: Supplementation, Silk worm, Haemolymph, Bombyx mori

International Conference on Bio-Commerce 2019 Page 17

P. Subavathy, R. D. Thilaga and B. Jeba Nesam*