CNU Journal of Higher Education,

Volume 9 (2015), p 15-27

Grain Protectant Efficacy of Makabuhay, Tinospora rumphii Extract

Against Maize Weevil, Sitophilus zeamais M.
Pedro M. Gutierrez, Jr.
Cebu Normal University

Date Submitted: September 30, 2015 Originality: 90%

Date Revised: December 22, 2015 Plagiarism Detection: Passed

ABSTRACT

The grain protectant efficacy of Makabuhay, Tinospora rumphii stem extract

was tested on adult maize weevil, Sitophilus zeamais M. Evaluation of phytochemicals
present in the extract was conducted to identify its bioactive compounds. The ethanolic
stem extract of T. rumphii contains the following phytochemicals: alkaloids,
flavonoids, steroids and tannins. These phytochemicals are known to possess
pesticidal activities against insects and other pests. The grain protectant efficacy of T.
rumphii against corn weevils was manifested by the percentage of mortality of
weevils, number of corn seed holes; weight loss and percentage corn germination.
Results revealed that the corn weevils treated with the highest concentration (18%) of
T. rumphii manifest the highest percentage of weevils’ mortality and significantly
higher (p =0.000) as compared to the negative control and the other treatment
concentrations. In addition, the mortality of corn weevils treated with the plant extract
is concentration-dependent. The average number of corn seed holes treated the T.
rumphii extract is statistically lower as compared to the negative control. Moreover,
the highest concentration (18%) of the plant extract showed the lowest average number
of holes. This implies that T. rumphii is responsible for the inhibition of the weevils
from infesting the corn seeds. Furthermore, result revealed significant decrease (p
=0.000) on the average weight loss of corn seeds previously treated with the plant
extract. The highest concentration of T. rumphii manifest the lowest percentage weight
loss. Result also showed a significant increase (p = 0.000) of percentage germination
of corn seeds randomly selected from the control and the treated groups. The highest
concentration of the plant extract reveals the highest percentage of germination. T.
rumphii stem extract therefore is an effective grain protectant against maize weevis, S.
zeamis due to the presence of phytochemicals flavonoids, alkaloids, steroids and
tannins which showed synergistic effects in terms of insecticidal action to maize
weevil.
Keywords: mortality, Sitophilus zeamais, grain protectant, phytochemicals,
Tinospora rumphii

INTRODUCTION (McFarlane, 1989; Garcia- Correia,

Corn, wheat and rice are among
the cereal products which constitute as
the main source of food for human
beings, including some animals. Reports
showed that about ninety percent (90%)
of such grains generated for their
consumption come from cereals that
characterized the sources of feeding
majority of the people in the world
2002). Taking into account the
continuous growth of human population
worldwide, the demand for food is also
increasing daily, therefore, there is a
critical need for the unremitting supply of
such cereal products to feed the
consumers on regular basis (Rupp et al.,
2011).
Unfortunately, various kinds of
pests infest into rice and corn stored
Gutierrez: Grainning Protectant Efficacy of Makabuhay, Tinospora rumphii Extract
products of which insects are among the
major agent contributors for the damage
after-crop period (Rupp et al., 2011).
Sori and Ayana (2012) stated that corn
weevil, Sitophilus zeamais is a major pest
responsible for damaging stored corn.
The said insect bores a hole in the grain
and lays its eggs which eventually
develop into adult weevils that feed on
the content resulting to total grain loss of
corn (Boxall et al., 2002).
Synthetic insecticides are found
effective inorder to control these pests
that reside and damage during the storage
of grains. However, Yankanchi and
Gadache (2010) and Dubey et al. (2007)
stated that synthetic/artificial insecticides
could lead to pest resistance, environmental
pollution, toxic effect to consumers, as
well as chronic and genetic problems.
This condition encourage researchers
to develop and discover alternative
methods in securing/ protecting corn grains
that are harmless/safer to consumers,
health and environment friendly. Huang
and Ho (1998) stated that there are
various kinds of natural/organic
pesticides that can be taken from higher
plants. In addition, Negahban et al.
(2006) stated that these products do not
generate harmful remains/residue to the
environment, instead they provide
medicinal value for humans. Moreover,
Pavela (2007) stated that the plant
derived products that have been applied
and used as potential seed protectant
often begins with the screening of plant
extracts.
Makabuhay, Tinospora rumphii,
locally known as panyawan (in Visayan
region) vine is used by some farmers as
natural plant pesticides. Various studies
16
showed that this kind of plant possesses
antimicrobial and parasitical activity. A
good example is the study conducted by
Salazar et al. (1988) on the affectivity of
the lotion from the ground stem of
Makabuhay in the treatment of scabies
which established the acaricidal property
of the said plant. Moreover, Fernandez
(1996) stated that T. rumphii is essential
in controlling intestinal worms of goats.
It provides an alternative to costly drugs
and potentially toxic chemical pesticides.
This study aims to investigate the
grain protectant efficacy of Tinospora
rumphii stem extract against corn weevil,
Sitophilus zeamais which may lead to the
revelation of new natural/organic
insecticides that help to control corn
weevil thus aiding in preserving cereal
products.
METHODOLOGY
Corn weevils were collected from
farmers’ stored corn grains in the
province of Cebu, Philippines. The
weevils were reared in a plastic box
containing corn seeds as their source of
food. Matured stems of Tinospora
rumphii were collected from Carcar,
Cebu, Philippines. The pulverized plant
samples were extracted by soaking with
concentrated (100%) ethanol (Analytical
Grade) within 48 hours. The filtered
ethanolic T. rumphii extracts was then
concentrated using a rotary evaporator.
Phytochemical analysis was done
following the procedure as described by
Harborne (1998).
The pesticidal activity of T.
rumphii extract against maize weevils, S.
zeamais was performed in 200 mL plastic
cups. The plastic cups were filled with 10
17
grams of mature corn seeds as their
source of food and the three (3) different
concentrations (6%, 12% and 18%) of
the plant extract. Range finding test was
conducted inorder to determine the
concentrations used in the definitive test.
Four replicates were carried out of each
concentration together with the negative
and positive (Malathion) control.
Mortality of weevils were monitored
every 24 hours for five days (Adedire et
al., 2011). After 45 days of exposure,
samples of corn seeds were randomly
drawn from each containers for the
evaluation of grain damage. Holes,
weight loss and germination rate.
The statistical tools that were
used in this study are the following: the
Arithmetic Mean to get the average
number of dead corn weevils; Analysis of
Variance (ANOVA) to determine the
significant difference on the mortality of
weevils between the control and the
experimental groups; and Post –Hoc
Analysis using the Tukey Test to
determine the degree of variability
between the control and different
concentrations of the plant extracts.
CNU Journal of Higher Education, Volume 9 (2015)
RESULTS AND DISCUSSION
Results revealed that Tinospora
rumphii stem extract contained the
following phytochemicals (Table 1):
alkaloids, flavonoids, steroids and
tannins. The plant’s phytochemicals/
secondary metabolites provide enormous
compartment that have strong biological
activity. Rupp et al. (2005) stated that
phytochemicals provide various biological
effects such as medicine, insecticide,
anti-microbial, repellent, larvae insecticide
and many others. In addition, Stangarlin
et al. (1999) stated that phytochemicals
performed plant’s protective responses;
their significant functions as anti-
herbivore and other insecticidal activity
are comparable to synthetic pesticides.
Phytochemicals such as alkaloids
and tannins manifest strong insecticidal
effect and therapeutic roles. Alkaloids
isolated from Piper longum plant
revealed very potential compounds
which are used to kill mosquito larvae
(Khanna and Kannabiran, 2007). Lee
(2000) also stated that tannins can be
used as insecticide against Culex
quinquefasciatus larvae. In addition,
Azmathullah et al. (2011) stated that
alkaloids and tannins are effective as
pesticides. On the otherhand, flavonoids
posses bioactive properties such as
antibacterial and insecticidal effects.
Hopkins and Huner (2009) stated that
steroids play a protective function by
disrupting the insect’s molting cycle
when ingested by insect herbivores.
Gutierrez: Grainning Protectant Efficacy of Makabuhay, Tinospora rumphii Extract 18

Table 1. Phytochemicals present in Tinospora rumphii stem extract.

Sample Alkaloids Cyanogenic Flavonoids Saponins Steroids Tannins
Glycosides
Tinospora +++ - +++ - +++ +++
rumphii
Legend: (-) absence; (+) - less abundant; (++) - average; (+++) - very abundant

The bioefficacy of T. rumphii treatment concentrations. In addition,

stem extract as grain protectant against results revealed that the mortality of corn
corn weevils was evaluated using weevils treated with the various
mortality test. Results revealed variations concentrations of T. rumphii is
of corn weevils’ average percentage concentration-dependent wherein as the
mortality which are treated with the concentration of the plant extract
various concentrations of the plant extract increases, the mortality is also increased.
including the positive control within 24, Kemabonta and Faludo (2013) stated that
48, 72, 96 and 120 hours of exposure most insects like maize weevil breathe
(Table 2). It is also noted that the using the trachea that continues to
negative control (untreated group) another opening of the spiracle. The plant
showed 0.00% mortality from 24 hours extract potentially damaged the said
to 72 hours of exposure and only 6.25% airways which resulted to suffocation.
mortality after 120 hours. On the other Tinzaaara et al. (2006) stated that four
hand, the weevils treated with the highest various plant extracts (Melia azedarach
concentration of T. rumphii showed the L., Tagates spp., Eichornia crassipes,
highest average percentage of mortality Martius and Ricinus communis L.) have
as compared to the other treatment been proven to display insecticidal
concentration. Meanwhile, the lowest activity and can kill pests by
concentration of the plant extract controlling/affecting their biological
manifest the least average percentage of machinery.
weevils’ mortality as compared to other

Table 2. Average Mortality in percent (%) of Maize Weevil, Sitophilus zeamais M.

Treated with Various Concentrations of Makabuhay, Tinospora rumphii Extract and
the Control Groups
Treatments Mean Percentage (%) Mortality
24 SD 48 Hours SD 72 Hours SD 96 SD 120 SD
Hours Hours Hours
Negative Control 0.00 0.00 0.00 0.00 0.00 0.00 3.75 4.79 6.25 4.79
Positive Control 27.50 15.55 52.50 26.30 68.75 29.00 73.75 25.00 77.50 23.30
(Malathion)
6% 8.75 8.54 26.25 8.54 43.75 9.46 53.75 9.46 66.25 8.54
12% 15.00 4.08 28.75 13.15 48.75 22.10 63.75 31.50 75.00 30.00
18% 23.75 13.77 41.25 11.09 62.50 15.00 76.25 21.40 91.25 11.81

The Two-Way ANOVA results rumphii extract and the control groups,
(Table 3) revealed a significant F 4, 75  39.43 , p  0.000 . That is, the
differences of percentage mortality of
maize weevil among treatments of various treatments significantly affect on
various concentrations of Tinospora the increase of the percentage mortality
19 CNU Journal of Higher Education, Volume 9 (2015)

of maize weevil. In addition, the revealed no significant interaction with

percentage mortality of maize weevil
varies significantly in different time of
 
each other, F 16, 75  1.48 , p  0.132 .

 
observations in hours, F 4, 75  26.23 ,
p  0.000 . This implies that the rising
percentage mortality of maize weevil, are
also dependent on the length of time of
observation. On the other hand, the
treatments and time of observation
This means that the treatments and time
of observations made during the
experiment do not affect simultaneously
on the increasing percentage mortality of
the weevil.

Table 3. Two – Way ANOVA Results for the Differences on the Percentage Mortality of Maize
Weevil, Sitophilus zeamais M. Among Treatments with Various Concentrations of Makabuhay,
Tinospora rumphii Extract and the Control Groups across Different Time of Observations

Result shows high significant activity against corn weevils from

differences, p = 0.000 (Table 5) on the infesting/ damaging the corn seeds.
number of holes in corn seeds exposed
to 6%, 12% and 18% concentrations of Table 4. Average Number of Holes of
the T. rumphii extract together with the Ten (10) seeds Randomly Selected
positive and negative control groups from Infected Corn Seeds Treated with
(Table 4). It is also noted that the Various Concentrations of Makabuhay,
negative control manifests the highest Tinospora rumphii Extract and the
number of holes (10.25 out of 10 seeds) Control Groups
infested with the corn weevils. In Treatments Average Standard
Number of Exit Deviations
addition, certain seeds in the negative Holes per 10
control are filled with more than one Seeds
Negative Control 10.25 5.68
holes. However, the highest Positive Control 0.50 0.58
concentration (18%) of the plant extract (Malathion)
6% 0.75 1.50
reveals the least number of holes (0.25
12% 0.75 0.96
out of 10 seeds). This implies that T. 18% 0.25 0.50
rumphii extract posseses grain protectant
Gutierrez: Grainning Protectant Efficacy of Makabuhay, Tinospora rumphii Extract 20

Table 5. One – Way ANOVA Results for the Differences on the Number of Holes of Ten (10)
seeds Randomly Selected from Infected Corn Seeds among Treatments of Various Concentrations
of Makabuhay, Tinospora rumphii Extract and the Control Groups

percentage weight loss

as compared to those
treated with the plant
extract and the positive
control. On the other
hand, the highest
concentration (18%) of
the treatment manifests
the lowest percentage
Table 6 presents the average weight loss. In addition,
percentage weight loss of corn seeds the decrease of corn seeds’ weight loss is
infested with corn weevils and treated affected by the increased of plant extract
with the different concentrations of the concentration. The significant decrease of
plant extract. Highly significant corn weight loss could be attributed to
variations (p =0.000) of percentage weight the pesticidal activity of T. rumphii
loss among the groups were noted. The extract causing the mortality of weevils.
negative control reveals the highest

Table 6. Average Weight loss in percent (%) of Corn Seeds Treated with Various
Concentrations of Makabuhay, Tinospora rumphii Extract and the Control Groups
Treatments Average Percentage (%) Standard
of Weight Loss Deviations
Negative Control 15.35 3.70
Positive Control 0.00 0.00
(Malathion)
6% 7.05 2.50
12% 6.52 0.99
18% 3.52 2.60
21 CNU Journal of Higher Education, Volume 9 (2015)

Table 7. One – Way ANOVA Result on the Differences of the Percentage (%) Weight loss of Corn
Seeds among Treatments of Various Concentrations of Makabuhay, Tinospora rumphii Extract
and the Control Groups

Results shows that the corn seeds
randomly selected from the control
groups (positive and negative) and from
the 6%, 12% and 18% concentrations of
the plant exract (Table 8) manifest high
significant variations on average percentage
germination (Table 9). The negative
control reveals the lowest percentage
(15%) of germination as compared to the
positive control and those which are
exposed to various plant extract
concentration. However, the highest
percentage concentrations of the T.
rumphii reveals the highest percentage of
germination (95.00%) as compared to 6%
and 12% with a percentage germination of
92.50%.

Table 8. Average Percentage (%) Germination of Infested Corn Seeds by Maize

Weevil After Treatment with Various Concentrations of Makabuhay, Tinospora
rumphii Extract and the Control Groups.
Treatments Average Percentage (%) of Standard
Germination Deviations
Negative Control 15.00 17.32
Positive Control 87.50 5.00
(Malathion)
6% 92.50 15.00
12% 92.50 9.57
18% 95.00 5.77

Table 9. One – Way ANOVA Results for the Differences of the Percentage (%) Germination of
infested Corn Seeds by Maize Weevil After Treatment among Various Concentrations of
Makabuhay, Tinospora rumphii Extract and the Control Groups
Gutierrez: Grainning Protectant Efficacy of Makabuhay, Tinospora rumphii Extract
Results of the study provide
evidence that the ethanolic stem extract of
Makabuhay, T. rhumpii is an effective
corn grain protectant against maize
weevil, S. zeamais. The presence of
secondary metabolites such alkaloids,
flavonoids, steroids and tannins can be the
strong causative factors of the plant extract
as a killing chemical means against maize
weevil.
A study conducted by Adeniyi et al
(2010) showed that Bryophyllum
pinnatum and Eucalyptus globules leaf
extracts showed insecticidal effect against
rice weevil, Sitophilus oryzae. Results on
phytochemical screening of these plants
reveal the presence of flavonoids,
saponins, alkaloids, steroids, tannins and
terpenoids. Umadevi and Sujatha (2013)
showed that different extracts of Curcuma
longa, Gymnema sylvestre, Azadira
chtaindica, Chrysanthemum trifurcatum are
effective as insecticide against red flour
beetle (Tribolium castaneum).
Phytochemical examinations showed that
these plants contain alkaloids, amino acids,
terpenoids, flavonoids, saponins, steroids,
and tannins. Another study conducted by
Gemechu et al. (2013) showed that oils of
Ethiopian mustard and cotton caused
mortality of maize weevil, Sitophilus
zeamais M. using various concentrations.
A separate study performed by Dejen
(2002) revealed that Chenopodium leaf
powder mixed with maize and sorghum
grains at 2 and 4% w/w concentrations
resulted to the complete reduction in
maize weevil F1 progeny. In addition,
Parugrug and Roxas (2008) evaluated the
pesticidal efficacy of the following plant
species: A. indica, C. citratus, L. camara,
O. basilicum and T. erecta against maize
weevil, Sitophilus zeamais. Results
revealed that all plant samples showed
22
repellent/repulsive properties against
maize weevil.
Various researches suggested that
Makabuhay, T. rumphii is valuable in
controlling some pests, like insects. A
notable example is the study conducted by
Re jesus et al. (1987) which showed that
the use of makabuhay aqueous extract for
seedling root soaking and spreading of
chopped vines are efficient in controlling
pests such as brown plant hopper, green
leaf hopper and rice stem borers. Gutierrez
et al. (2014) showed that Tinospora
rumphii together with other plant extracts
are also potent larvicides on Aedes aegypti
mosquito larvae. Moreover, T. rumphii
was found effective in controlling rice
stem borers, green leaf hoppers and black
bugs. The lotion derived from the
macerated stem of the said plant is also
effective in treating scabies (Salazar et al.,
1988).
Natural products/phytochemicals
are identified for their insecticidal/
pesticidal activity. Through the years,
several studies revealed strong evidences
that plants’ natural products perform
significant environmental roles, such as
protection against microbial and pests
attack, together with environmental
protection (Hopkins and Huner, 2009).
Various researches show that
flavonoids demonstrate a broad range of
biological warfare including antimicrobial,
23 CNU Journa
insecticidal and other pesticidal activities.
Alkaloids and tannins are also
known to possess medicinal and pesticidal
properties (Azmathullah, 2011). Alkaloids
isolated from Piper longum plant found to
have very potential compounds that are
used to kill mosquito larvae (Khanna and
Kannabiran, 2007). In addition, Nenaah
23 CNU Journal of Higher Education, Volume 9 (2015)

(2011) revealed the toxic effect of a certain on the average weight loss of corn seeds
kind of alkaloid known as glycoalkaloids previously treated with the plant extract.
from Solanum tuberosum and The highest concentration of T. rumphii
lycopersicon esculentum against manifest the lowest percentage weight
Tribolium castaneum beetles and rice loss. Result also showed a significant
weevils, S. oryzae. Lee (2000) also stated increase (p = 0.000) of percentage
that tannins can be used as insecticide germination of corn seeds randomly
against Culex quinquefasciatus larvae. selected from the control and the treated
Hopkins (2009) stated that steroids play a groups. The highest concentration of the
protective function by disrupting the plant extract reveals the highest
insect’s molting cycle when ingested by percentage of germination. T. rumphii
insect herbivores stem extract therefore is an effective grain
protectant against maize weevils, S.
CONCLUSION zeamis.

The ethanolic stem extract of T. rumphii ACKNOWLEDGMENT

contains the following phytochemicals: alkaloids,
flavonoids, steroids and tannins. These The author would like to
phytochemicals are known to possess pesticidal acknowledge the Center for Research and
activities against insects and other pests. Development of Cebu Normal University
for the financial support, Department of
The grain protectant efficacy of T. Environment and Natural Resources –
rumphii against corn weevils was Region 7 for granting the Gratuitous
manifested by the percentage of mortality Permit and City Environment and Natural
of weevils, number of corn seed holes; Resources – Argao, Cebu for
weight loss and percentage corn accompanying in the collection of
germination. Results revealed that the Makabuhay, T. rumphii used in the study.
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