P.

Umamaheswari and CVN Murthy

Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (43-47)

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Advance Research Journal of Biological Sciences and Molecular Techniques
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Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (43-47) (ISSN:2395-7689)

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ANTI MICROBIAL ACTIVITIES OF ANDROGRAPHICS PANICULATA

AND TURMERIC USED IN THE TREATMENT OF INFECTIOUS FISH
DISEASE.
UMAMAHESWARI, P AND C.V.NARASIMHA MURTHY*
*Department of Zoology, Sri Vivekananda Degree College, Vinjamur
Department of Botany, V.S.University P.G.Center, Kavali
Abstract
Plants are rich in a wide variety of secondary metabolites such as tannins, alkaloids and
flavonoids. Many of the spices and herbs used today have been valued for their antimicrobial
effects and medicinal powers. Aquaculture has been a growing activity for the last 20 years
worldwide and this impressive development has been attended by some practices potentially
damaging to human and animal health. Bacterial infections are considered the major cause of
mortality in aquaculture Treatments of bacterial diseases with various herbs have been safely
used widely in organic agriculture, veterinary and human medicine. Since ancient times,
medicinal plants have been used for the treatment of common infectious diseases and treatments
with plants having antibacterial activity are a potentially beneficial alternative in aquaculture. In
addition, plant-derived phytomedicines provide a cheaper source for treatment and greater
accuracy than chemotherapeutic agents in the field. Medicinal plant Rhizome curcuma
(Turmeric) and Andragraphics paniculata are medicinal plants of which combination are
traditionally used in the treatment of infectious fish disease. In this study turmeric and
Andragraphics paniculata extract with distil water and chloroform is used as solvent. These
plants assayed for antimicrobial activity against two bacteria that is pseudomonas and E. coli.
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P. Umamaheswari and CVN Murthy

Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (43-47)

The plant extract shows various levels of antimicrobial activity on different microorganism. The
most potent extract was formed from turmeric and Andragraphics paniculata with chloroform.
Key word: Antimicrobial activity, Medicinal plants, Fresh water fish, Microbial pathogens
Turmeric and Andragraphics paniculata.
--------------------------------------------------------------------------------------------------------------------Introduction
The bacterial infections are considered the major cause of mortality in aquaculture
(Grisez and Ollevier 1995). Among the common fish pathogens are pseudomonas and E. coli. .
Fish are susceptible to several bacterial infections, mainly when reared in high density
conditions. Disease outbreaks elevated the mortality rate and decrease the productivity
efficiency, causing high economic loss of the fish farmers. Due to the use of a wide variety of
antibiotics, aquaculture has been implicated as potential environment to the development and
selection of resistant bacteria and a source of these pathogens to other animals and humans
(Figueiredo et al 2005, Hatha et al 2005 ).Some bacterial fish pathogens are also associated to
diseases in humans, making the aquaculture products a potential risk to the customers due to
zoonotic or food borne diseases(Yanong et al 2006). S. agalactiae is a dangerous pathogen to
freshwater and marine fish. The infection is characterized by brain invasion, nervous signs and
septicemia(Russo et al 2006). The continuous use of antimicrobial agents in aquaculture has
resulted into resistant bacterial strains in the aquatic environment. Treatment of bacterial diseases
with different herbs has been safely used in organic agriculture, veterinary and human medicine.
Treatment with medicinal plants having antibacterial activity is a potentially beneficial
alternative in the aquaculture. These herbs mitigate many of the side effects which are associated
with synthetic antimicrobials. Additionally, the plant-derived phytomedicines provide a cheaper
source for treatment and greater accuracy than chemotherapeutic agents. Plants have been used
as traditional medicine since time immemorial to control bacterial, viral and fungal diseases. In
India, more medicinal plant species are used against pathogenic bacteria. Recently, research has
been initiated to evaluate the feasibility of herbal drugs in fish diseases. Because of the growing
bacterial resistance against commercial standard and reserve antibiotics, the search for new
active substances with antibacterial activity against pathogenic bacteria is of increasing
importance.

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P. Umamaheswari and CVN Murthy

Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (43-47)

Materials and methods

Sampling
Fresh leaves of Andragraphics paniculata and rhizome curcuma were collected from
different localities of Kavali .These medicinal plants were identified by a taxonomist at the
Department of Botany, V.S.University P.G.Centre, and Kavali SPSR Nellore district. A.P.
Samples of all medicinal plants were thoroughly rinsed with running tap water to remove sand
and other debris. Rinsed plant materials were air dried under shade for complete drying and
preserved at room temperature.
Extract preparation
Solvent extracts of medicinal plants were prepared through conventional solvent
extraction methods. Dried leaves of Andragraphics paniculata and rhizome of turmeric were
crushed, 30 and 20 mg of crushed leaves were soaked separately in 200 ml of organic solvents in
increasing order of polarity (Diethyl ether, chloroform, ethanol and methanol) for 15-25 days.
Each extract was filtered by Whatman No.1 filter paper, concentrated and stored at room
temperature for further processing (Nasim et al.,2012).
Test microorganisms
For current research work, the fish associated bacterial pathogens pseudomonas and E.
coli. were taken from Microbiology Department and Antibacterial activity was assayed by agar
disks diffusion method the antibacterial activity of all solvent extracts was tested against
bacteria. The microorganisms were activated by inoculating a loop full of strain in 25 ml of
nutrient broth medium (NBM; Oxoid CM001) and incubated at 37C on a rotary shaker for 24 h.
Next day, the old inoculated culture was mixed with freshly prepared nutrient agar medium
(NAM; Oxoid CM1) at 45C and poured the sterilized plates. All the plates were placed at room
temperature in laminar flow to solidify. The discs of 5 mm were prepared soaked with 200 l of
a particular extract or the corresponding solvent was applied on discs and then allowed to dry for
assay.
Presoaked discs were placed in Petri dishes at their labeled position. These prepared
plates were left for incubation at 37C for 48 h. Discs of diethyl ether, chloroform; ethanol and
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P. Umamaheswari and CVN Murthy

Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (43-47)

methanol were also used as negative control. The inhibition of tested bacteria was measured by
determining the diameter of the zone of inhibition after 24-48 h in millimeter (Seeley et al.,
2001). The results of the sensitivity tests were expressed as (0) for no sensitivity, + (below 4
mm) for low sensitivity, ++ (4-8 mm) for moderate sensitivity and +++ (9-19 mm) for high
sensitivity.
Sensitivity test
Sensitivity of antibiotics (positive control) such as ampicillin and trimethroprim against
test microbial strains was assessed through agar disc diffusion method (Bauer et al., 1966).
Sensitivity was predicted with degree of clear zone surrounding the disc.
Statistical analysis
Each experiment was repeated in triplicates and Mean Standard Deviation from
absolute data was calculated. The comparison of antibacterial activity of solvent extracts of
medicinal plants with standard antibiotics was done
Resultd and Discussions
Table I. - Effect of Andragraphics paniculata and Rhizome curcuma on the growth of various
bacterial pathogens. Medicinal plants Extracts Zone of inhibition in mm (M SD)

S.no. Extract

Medicinal plant

Zone of Inhibition in mm (M SD)

Pseudomonas

E.coli

1.

Diethyl ether Andragraphics paniculata 4.660.58**

1.660.58*

2.

Chloroform

Andragraphics paniculata 2.330.58*

12330.58*

Ethanol

Andragraphics paniculata 1.330.58*

1.730.58*

Methanol

Andragraphics paniculata 1.570.58*

1.110.58*

1.

Diethyl ether Turmeric

4.330.58**

3.440513**

2.

Chloroform

Turmeric

1.240.48*

1.210.58*

ethanol

Turmeric

1.180.35*

1.170.43

Methanol

Turmeric

1.660.16*

1.560.13*

The date is presented in Table -1.The antibacterial activity of naturally dried medicinal plants has
been elucidated against fish associated bacterial pathogens Pseudomonas and E. coli through

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P. Umamaheswari and CVN Murthy

Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (43-47)

agar disc diffusion method. Results found. The inhibition of tested bacteria was illustrated in
varying degrees by Andragraphics paniculata and turmeric.
Conclusions
It was concluded that the leaf of Andragraphics paniculata and turmeric rhizome extracts of
medicinal plants have high antibacterial effect as an individual agent .Future studies can be based
on this research to know the exact mechanism of inhibition and to develop new antimicrobial
agents.
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