Methanolic Leaf Extracts of Certain Plants as Larvicides against Teak skeletonizer, Eutectona machaeralis

Abstract

Aim: Aim: The coveted Indian hardwood teak (Tectona grandis) grows well in particular tropical temperatures and soil types. With 8.9 million hectares, India's teak forests are essential to the luxury market. Indian teak trees are seriously threatened by the teak skeletonizer (Eutectona machaeralis Walker), which causes extensive defoliation. As a result, less timber is produced and trees grow slower. Materials and Methods Materials and Methods: In this work, methanolic leaf extracts of Wrightia tinctoria, Murraya koenigii and Prosopis juliflora were tested for their larvicidal efficacy against the fourth instar larvae of E. machaeralis. Results Results: All extracts had high levels of alkaloids, flavonoids and polyphenols, according to phytochemical analyses. The findings showed that mortality rates were concentration-dependent. M. koenigii had the maximum effectiveness, with mortality rates of 61.43±1.76% at 200 ppm and 92.86±2.40% at 400 ppm. Conclusion: Conclusion: These results highlight the potential of these plant extracts as efficient, environmentally beneficial teak pest management options.

Full text

Asian Journal of Biological and Life Sciences, Vol 13, Issue 1, Jan-Apr, 2024 61
Research Arcle
Correspondence:
Mahesh Lingakari
Department of Zoology,
Tara Government
Degree and PG College,
Sangareddy, Telangana,
INDIA.
Email: maheshlinga-
kari@gmail.com
Methanolic Leaf Extracts of Certain Plants as
Larvicides against Teak skeletonizer,
Eutectona machaeralis
Madhavi Maddala1, Mahesh Lingakari2,*, Srikanth Bandi1
1Department of Zoology, Entomology Laboratory, Osmania University, Hyderabad, Telangana, INDIA.
2Department of Zoology, Tara Government Degree and PG College, Sangareddy, Telangana, INDIA.
Submission Date: 11-01-2024; Revision Date: 14-02-2024; Accepted Date: 18-04-2024.
ABSTRACT
Aim:Aim: The coveted Indian hardwood teak (Tectona grandis) grows well in particular tropical
temperatures and soil types. With 8.9 million hectares, India's teak forests are essential to the
luxury market. Indian teak trees are seriously threatened by the teak skeletonizer (Eutectona
machaeralis Walker), which causes extensive defoliation. As a result, less timber is produced
and trees grow slower. Materials and MethodsMaterials and Methods: In this work, methanolic leaf extracts of Wrightia
tinctoria, Murraya koenigii and Prosopis juliora were tested for their larvicidal efcacy against the
fourth instar larvae of E. machaeralis. ResultsResults: All extracts had high levels of alkaloids, avonoids
and polyphenols, according to phytochemical analyses. The ndings showed that mortality rates
were concentration-dependent. M. koenigii had the maximum effectiveness, with mortality rates
of 61.43±1.76% at 200 ppm and 92.86±2.40% at 400 ppm. Conclusion:Conclusion: These results highlight
the potential of these plant extracts as efcient, environmentally benecial teak pest management
options.
Keywords:Keywords: Tectona grandis, Wrightia tinctoria, Murraya koenigii, Prosopis juliora,
Teak leaf skeletonizer, Eutectona machaeralis, Larvicidal activity.
INTRODUCTION
Teak, scientically known as Tectona grandis, is a tree
species that has a broad but fragmented distribution in
India. It grows well in humid, warm, tropical regions
and does well on deep uvial soil that has a pH of
roughly 6.5. It grows below 24ºN latitude, in southern
as well as certain eastern and western regions of India.
[1] The teak-bearing forests in India cover nearly 8.9
million hectares; falling within a precipitation range
of 800 to 2500 mm. Teak is a highly sought-after
hardwood species with a growing demand in luxury
markets. As a result, teak plantations have expanded
rapidly outside of their natural habitats to countries in
Asia, Latin America, Africa and Oceania. Currently, teak
is grown in approximately 70 tropical countries, with
Myanmar, India and Indonesia accounting for most
of the plantations.[2] Teak plantations have a signicant
presence in India, with the rst one established in 1846
at Nilambur, Kerala. According to,[3] India has teak
plantations on over 1.5 million hectares, with 50,000
hectares being added yearly.
In India, the teak skeletonizer (Eutectona machaeralis
Walker) is a serious pest of teak trees. It is regarded as
one of the most damaging teak pests, causing extensive
defoliation every year in nurseries, plantations and
wild forests throughout teak-growing regions.[4-6] The
larvae of this insect have a distinct feeding behavior,
consuming only the eshy leaf tissues while excluding
the veins. This feeding pattern has detrimental effects
on the overall growth and vitality of the teak tree.
Furthermore, it may cause several anomalies that
ultimately result in a decline in the amount and quality
of timber production. Over the years, numerous
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Maddala, et al.: Methanolic Leaf Extracts of Certain Plants as Larvicides against Teak skeletonizer, Eutectona machaeralis
62 Asian Journal of Biological and Life Sciences, Vol 13, Issue 1, Jan-Apr, 2024
researchers have conducted in-depth studies and
compiled signicant documentation on the effects of
the teak skeletonizer on teak trees.[7,8]
Since the start of the Green Revolution, synthetic
insecticides have become a prevalent tool for controlling
agricultural pests. However, these insecticides, while
aiding in increasing agricultural productivity, also
showed lethal consequences on non-target organisms.
[9] The surge in pest populations is a direct consequence
of these insects’ developing resistance to insecticides, a
situation aggravated by the overuse of these synthetic
pesticides. The existence of hazardous residues in
various environmental compartments is a result of
indiscriminate pesticide application and has a major
negative inuence on human well-being.[10] In light of
these concerns, there is a growing recognition of the
signicance of botanical pesticides, or biopesticides,
as viable alternatives to synthetic insecticides. These
organic plant extracts hold the advantage of being
biodegradable and pose fewer hazards to benecial
organisms in comparison to synthetic pesticides.
Several studies were conducted to test the efcacy of
botanicals against the larvae of E. machaeralis. In one
study[11] the extracts from thirteen different medicinal
plants were tested at concentrations of 5% and 10%.
Among these, Azadirachta indica, Nerium oleander, Strychnos
nux-vomica and Tylophora indica exhibited remarkable
effectiveness, resulting in signicant inhibition of
the pest. Conversely, the extract from Mirabilis jalapa
showed the least impact. In another study[12] 5%
concentration of crude extract from fresh leaves of 32
distinct medicinal plants was evaluated for antifeedant
and insecticidal properties against the third instar larvae
of E. machaeralis. Calotropis procera emerged as the most
effective, demonstrating potent biopesticidal properties.
Following closely were extracts from Datura metel and A.
indica. In this study, methanolic extracts were prepared
from the leaves of W. tinctoria, M. koenigii and P. juliora
and their larvicidal efcacy was evaluated against the
fourth instar larvae of E. machaeralis.
MATERIALS AND METHODS
Plant materials
W. tinctoria, M. koenigii and P. juliora leaves were obtained
from adjacent villages in Sangareddy town, Telangana,
India. They were then shade-dried and powdered using
an electric grinder before being stored separately in
airtight containers
Preparation of leaf extracts
In the Soxhlet apparatus, 100 grams of plant powders
were steeped in 200 mL of methanol for 6 hours at
60ºC. The methanol was later evaporated using a rotary
evaporator to get semisolid extracts of the test plants.
These semisolid extracts were then preserved in clean
bottles at 4ºC until use.
Phytochemical Analysis
The prepared plant extracts were subjected to qualitative
phytochemical analyses to nd out the presence
of various secondary metabolites. Mayor’s reagent
test (Alkaloids), Alkaline reagent test (Flavonoids),
Salkowski test (Terpenoids), Froth test (Saponins),
Keller-Killiani Test (Glycosides) and NaOH test
(Phenols) were employed to conrm the presence of
secondary metabolites
Preparation of test solutions
To make the 1000 ppm stock solutions of the test plant
extracts, 1 g of the extract was thoroughly mixed in 10
mL of methanol, later 990 mL of distilled water was
added to it. Using the serial dilution procedure, test
solutions of 50, 100, 200 and 400 ppm were produced
from the stock solutions. The control solution was made
with identical proportions of methanol and distilled
water as the test solution but without the extracts.
Collection of larvae
Early instar larvae of E. machaeralis were recovered from
the teak plants on the Osmania University campus. They
were raised by providing fresh teak leaves. The fourth
instar larvae from the reared population were employed
in the larvicidal bioassays.
Larvicidal activity
Topical application and leaf spray method were
followed in the larvicidal bioassay. 10 larvae were taken
in each test batch. They and the fresh teak leaves were
sprayed with test solutions, once the leaves were air-
dried, they were placed in separate Plastic jars. The
number of dead larvae was counted and noted. The
experiment continued until all the larvae were dead or
they metamorphosed into the next instar. The same
experiment was replicated ve times. Mortality results
were corrected by using the[13] formula.
Corrected mortality=(% test mortality-% control
mortality)/(100-control mortality)x100
RESULTS
The results of the phytochemical analyses of the test
plant extracts (Table 1) indicated that all three extracts are
rich in Alkaloids, Flavonoids and Polyphenols content.
Terpenoids were abundantly present in P. juliora and
W. tinctoria extracts, while they are moderately present in

Maddala, et al.: Methanolic Leaf Extracts of Certain Plants as Larvicides against Teak skeletonizer, Eutectona machaeralis
Asian Journal of Biological and Life Sciences, Vol 13, Issue 1, Jan-Apr, 2024 63
M. koenigii extracts. Saponins were moderately present
in W. tinctoria and M. koenigii extracts and absent in P.
juliora extracts. Glycosides were moderately observed
in M. koenigii extracts; however, they are absent in both,
P. juliora and W. tinctoria extracts.
Table 1: Identied secondary metabolites through
phytochemical analyses. Absence and presence are
denoted by ‘-’ and ‘+’, respectively. The number of
‘+’ is proportional to the relative abundance of the
secondary metabolites.
Extracts Alka
loids
Flavo
noids
Sapo
nins
Terpe
noids
Poly
phenols
Glyco
sides
M.
koenigii
+++ +++ ++ ++ +++ ++
P.
juliora
+++ +++ - +++ +++ -
W.
tinctoria
+++ +++ ++ +++ +++ -
The larvicidal bioassays results of the present study are
given in Table 2 and Figure 1. At 50 ppm, which is the
lowest concentration tested, M. koenigii extracts caused a
mortality rate of 21.43%±2.40, while P. juliora and W.
tinctoria recorded mortality rates of 17.86%±2.67 and
21.43%±2.40, respectively. Increasing the concentration
to 100 ppm resulted in higher mortality rates: M. koenigii
showed 39.29%±2.11, P. juliora recorded 32.14%±1.76
and W. tinctoria had a mortality rate of 32.14%±3.67.
At 200 ppm, M. koenigii demonstrated an even more
substantial effect, with a mortality rate of 61.43%±1.76.
P. juliora and W. tinctoria exhibited mortality rates of
46.43%±2.0 and 46.43%±2.83, respectively. The
highest concentration tested, 400 ppm, led to M. koenigii
achieving the highest mortality rate of 92.86%±2.40.
P. juliora and W. tinctoria still demonstrated substantial
efcacy, with mortality rates of 78.57%±2.83 and
71.43%±2.11, respectively.
Table 2: Larvicidal efcacy of different test extracts
against the 4th instar larva of E. machaeralis.
Extracts M. koenigii P. juliora W. tinctoria
Control 0±1.76 0±1.76 0±1.76
50 21.43±2.40 17.86±2.67 21.43±2.40
100 39.29±2.11 32.14±1.76 32.14±3.67
200 61.43±1.76 46.43±2.0 46.43±2.83
400 92.86±2.40 78.57±2.83 71.43±2.11
Regression
equation
y=0.2218x
+9.7335
y=0.1857x
+7.1435
y=0.1643
x+9.6429
R2 0.9545 0.9699 0.9396
The regression equations were derived through
regression analysis (Figures 2-4) to model the
relationship between the concentration of the plant
extract (x) and the corresponding mortality rate (y).
For M. koenigii, the equation is y=0.2218x+9.7335, for
P. juliora it is y=0.1857x+7.1435 and for W. tinctoria
it is y=0.1643x+9.6429. These equations allow for
the estimation of mortality rates at concentrations
not directly tested in the experiment. Furthermore,
the high R-squared (R2) values indicate a strong t of
the regression lines to the actual data points. For M.
koenigii, R2=0.9545, for P. juliora, R2=0.9699 and for
W. tinctoria, R2=0.9396. This underscores the reliability
of the regression equations in predicting mortality rates
based on the concentration of the plant extract
Figure 1: Larvicidal bioassay results of the methanolic leaf
extracts of M. koenigii, P. juliora and W. tinctoria against the
4th instar larvae of E. machaeralis.
Figure 2: Regression analysis of the M. koenigii’s methanolic
leaf extracts bioassays against the 4th instar larvae of
E. machaeralis.
Figure 3: Regression analysis of the P. julifora’s methanolic
leaf extracts bioassays against the 4th instar larvae of

Maddala, et al.: Methanolic Leaf Extracts of Certain Plants as Larvicides against Teak skeletonizer, Eutectona machaeralis
64 Asian Journal of Biological and Life Sciences, Vol 13, Issue 1, Jan-Apr, 2024
E.machaeralis.
Figure 4: Regression analysis of the W. tinctoria’s methanolic
leaf extracts bioassays against the 4th instar larvae of E.
machaeralis.
DISCUSSION
Secondary metabolites, formerly thought to be useless,
but now seen as vital plant components that provide
selection and adaptation benets. Plants have evolved
secondary metabolites to resist herbivorous animals
and insect pest attacks and adapt to their environment,
preventing them from feeding.[14,15] Countless studies
revealed the pesticidal properties of secondary
metabolites in various plant extracts. Secondary
metabolites like terpenes, phenolics, avonoids, saponins
and glycosides have various pesticidal properties and
can be used in plant protection.[16]
In a previous study[17] ethanolic leaf extracts of M. koenigii
were analyzed qualitatively revealing the presence of
saponins, tannins, alkaloids, glycosides, terpenoids and
phenols. In their study, Flavonoids were not detected.
However, we discovered the presence of avonoids
also including the other secondary metabolites. In our
study, we used methanol as a solvent in acquiring leaf
extracts of M. koenigii. This could be the reason for
avonoids being not identied in the analysis made
by.[17] Furthermore, the results of the present study are
supported by other studies where, M. koenigii leaf extracts
were reported to be rich in Alkaloids,[18] Terpenoids,[19,20]
Polyphenols[21] and Flavonoids.[22] In the study made
by,[23] methanolic leaf extracts of W. tinctoria identied
all secondary metabolites except glycosides are in line
with the results of the present study. Ethanolic leaf
extracts of P. juliora revealed the presence of all other
phytochemicals except, Saponins and Terpenoids.[24] In
our study, saponins and glycosides were not detected in
methanolic extracts of P. juliora leaves, suggesting the
role of solvent in acquiring the secondary metabolites.
Extracts of various plants were reported to possess good
pesticidal properties against the larvae of E. machaeralis.
In a study by,[25] ve-leaf extracts (Adhatoda vasica, Vitex
negundo, Azadiracta indica, Ricinus communis and Pongamia
glabra), A. indica seed kernel extract and A. indica seed
oil were investigated for their larvicidal properties
against E. machaeralis larvae to nd ve-leaf extracts
at 6% showed better larvicidal efcacy than neem oil
and neem seed kernel extract. However, the commercial
formulation Grub kill and Bacillus thuringiensis,
demonstrated marginally superior larvicidal activity.
In another study[26] Panchagavya with crude extracts
of Seaweed, Sargassum wightii, was tested against E.
machaeralis and reported the highest larvicidal efcacy
of 71% at 5000 ppm concentration after 72 hr of the
application. However, in the present study, all three
tested plant extracts showed better larvicidal efcacy at
much lower concentrations suggesting their potential in
the control of E. machaeralis. It is evident from the results
of the larvicidal bioassays that M. koenigii, P. juliora and
W. tinctoria extracts demonstrated varying degrees of
larvicidal activity. Notably, at higher concentrations,
the mortality rates substantially increased, suggesting
a concentration-dependent effect. The regression
equations and the high R-squared (R2) values indicate
a strong t of the regression lines to the actual data,
afrming the reliability of the results.
It is a well-established fact that Alkaloids, Polyphenols,
Flavonoids, Terpenoids, Saponins and Glycosides
possess pesticidal properties[27] and other biologically
important activities. The larvicidal bioassay results of the
present study, combined with the phytochemical analysis,
suggest that secondary metabolites in the tested plant
extracts contribute to the observed larvicidal activity
against the larvae of E. machaeralis. Comparatively, M.
koenigii extracts showed higher larvicidal efciency than
the remaining two plant extracts. The synergistic effect
of all six secondary metabolites-Alkaloids, Polyphenols,
Flavonoids, Terpenoids, Saponins and Glycosides in
the M. koenigii leaf extracts might be the reason for its
superior larvicidal efcacy over the other two extracts.
ACKNOWLEDGEMENT
The authors are thankful to the Principal, Tara
Government College, Sangareddy, Telangana, India
and The Head, Dept. of Zoology, Osmania University,
Hyderabad, Telangana, India for their support in
conducting this research.
AUTHORS CONTRIBUTIONS
Madhavi Maddala: Conceived the idea, conducted
experiments and edited the manuscript.

Maddala, et al.: Methanolic Leaf Extracts of Certain Plants as Larvicides against Teak skeletonizer, Eutectona machaeralis
Asian Journal of Biological and Life Sciences, Vol 13, Issue 1, Jan-Apr, 2024 65
Mahesh Lingakari: Conducted experiment and prepared
the rst draft.
Srikanth Bandi: Conducted experiment and analyzed
results.
CONFLICT OF INTEREST
The authors declare that there is no conict of interest.
ABBREVIATIONS
pH: Potential of Hydrogen; N: North; mm: Millimetre;
ºC: degree Celsius; NaOH: Sodium Hydroxide; ppm:
Parts Per Million; ml: Millilitre.
SUMMARY
The study evaluated the larvicidal efcacy of W. tinctoria,
P. juliora and M. koenigii methanolic extracts against
E. machaeralis, a teak tree pest. Alkaloids, avonoids
and polyphenols were discovered by phytochemical
examination of the extracts in this Telangana, India-
based study. Larvicidal bioassays showed effects that
varied with concentration; at 400 ppm, M. koenigii
extracts showed the maximum effectiveness, with
92.86% mortality. Signicant effectiveness was also
demonstrated by W. tinctoria and P. juliora, which
exhibited 71.43% and 78.57%, respectively, at the
greatest concentration. The results of the study
demonstrated the possibility of these extracts as
environmentally acceptable substitutes for teak leaf
skeletonizer management; the greater performance
of M. koenigii was related to its synergistic metabolite
actions. Further research is needed to determine the
secondary metabolites responsible for the observed
results in the present study.
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Cite this article: Maddala M, Lingakari M, Bandi S. Methanolic Leaf Extracts of Certain Plants as Larvicides against Teak
skeletonizer, Eutectona machaeralis. Asian J Biol Life Sci. 2024;13(1):61-6.