ISSN: 2391-3991, Volume 2 Issue 2, page 19 - 24Borneol is an essential oil with bicyclic organic compound that belong to essential oil, and has a natural insect repellent effect. It

International Journal of Agriculture & Agribusiness ISSN: 2391-3991, Volume 2 Issue 2, page 19 - 24

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Zambrut.com. Publication date, March 4, 2019.

Mar, K. M. 2019. Efficacy of Annona Squamosa L. Leaf Extracts ............ 19

Efficacy of Annona Squamosa L.

Leaf Extracts Against the Larvae

of Tobacco Caterpillar,

Spodoptera Litura

(Fabricius, 1775)

Khin Myint Mar

Ass. Prof. Dr. Khin Myint Mar

Associate Professor, Department of Zoology

University of Magway

Myanmar

Abstract: The experiment was carried out the efficacy of crude extracts of Annona squamosa leaves

for the control of the larvae of common major pest, Tobacco caterpillar Spodoptera litura in the

Department of Zoology, University of Magway, Myanmar during October 2014 to March 2015. The

leaves of fresh Annona squamosa were crushed and strained by filter paper to obtain crude extracts

and then diluted with the distilled water to treatment percent. The adult specimens of Spodoptera

litura were collected from groundnut and tomato fields in Magway environs cultured in the

laboratory with food plants to be reared for larval stages. The 4th instar larvae of S.litura were tested

with spraying method of A.squamosa leaves extract with only distilled water (control), 60%, 75% and

90% respectively. Three replications of ten larvae were tested for each treatment. The results showed

that the mortality rate of Spodoptera litura was the highest (33.33% mortality) at 90% A.squamosa

leaves crude extracts while the lowest at 60% (0% mortality) and the moderate (20% mortality) at

75% at room temperature of about 26.2˚C. The results from the experiment showed that the

insecticidal activities of A.squamosa leaves on the larvae tested was the most effective at the

concentration of 90% crude extract. Leave extract of A.squamosa is a promising candidate for

development as simple botanical insecticide for local use in rural Myanmar.

Keywords: Annona squamosa, Crude extract, 4th

instar Larvae, mortality, Spodoptera litura.

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1. INTRODUCTION

The indiscriminate use of chemical pesticides has given to many serious problems such as

intoxication of people and animals, environmental pollution, residues on food, high persistence in the

environment and impact on beneficial insects, among other effects, including genetic resistance of pest

species, toxic residues, increasing costs of application, environmental pollution and hazards. With a greater

awareness of the hazards associated with the use of synthetic organic pesticides, there has been an urgent

need to explore suitable alternative products for pest control. This is a world-wide interest in the

development of alternative strategies including the search for new types of insecticides and used for

traditional botanical pest control agents. Plant crude extracts often consist of complex mixtures of

compounds which may act synergistically (Berenbaum, 1985) and are highly effective, safe and

ecologically acceptable (Senthil Nathan and Kalaivani, 2005) (cited by Baskar et al., 2011).

Plants are rich sources of natural substances that can be utilized in the development of

environmentally safe methods for insect control. Numerous plant species have been identified as

possessing pesticidal properties and have shown potential as alternative to chemical pesticides (Singh,

2000). More than 2000 plants species have been known to produce chemical factor and metabolites of

value in pests control programme (Sreeletha and Geetha, 2012). Plant derived pesticides are eco-friendly,

non-toxic to non-target organisms, non-persistent in nature, besides they do promote drug resistance

(Kandagal and Khetagoudar, 2013). In fact, botanical pesticides from many plant origins including Annona

squamosa and neem are in use in Indian agriculture for over a century to minimize losses caused by pests

and diseases (Prakash, 2008). The phytochemical constituents in the leaves of Annona squamosa contain

alkaloids, protein, amino acid, carbohydrate, glysides, phytosteroles, tanins and phenolic compounds

(Kumar et al., 2010). Annona squamosa leaf content was possibility effective as insecticides or repellent is

borneol. Borneol is an essential oil with bicyclic organic compound that belong to essential oil, and has a

natural insect repellent effect. It has not been yet known how the about the mechanism of mechanisms

work borneol as insect repellent (Kesetyaningsih, 2012).

Among the polyphagous pests, the tobacco caterpillar Spodoptera litura Fabricius (1775) is a

serious pest of various economically important crops such as cotton, groundnut, chilli, tobacoo, caster, oil

seeds, vegetables, bendy and pulses etc. The fully grown caterpillars are most voracious feeders and can

cause extensive damage by defoliation. It has the ability to develop resistance to many conventional

insecticides used for its control (Deshmukhe et al., 2010).

Myanmar is an agricultural country. The agricultural sector accounts for 34% of its gross domestic

products (GDP) and 15.4 % of total export earnings. It also employs 61.2% of the total labor force. The

importance of agriculture in Myanmar is underscored by the stated objective of having agricultural as the

base of the country economy and the engine for the overall development of other sectors (Kan Zaw et al.,

1998). In Myanmar, most of the researchers studied the tobacco and neem based pesticides used in the

experiments and the research conducted with Annona squamosa as insecticides was very limited. The

present study involves the application of A. squamosa (sugar apple) leaves crude extracts to study its

effects on the mortality of Tobacco caterpillar at certain stage of development. In the present study, the

larvae of Tobacco caterpillar Spodoptera litura were tested with the application of Annona squamosa

(sugar apple) leaves crude extract as botanical pesticides. Therefore, the objective of the present study was

to evaluate the insecticidal efficacy of the crude extract of Annona squamosa leaves on Spodoptera litura.

2. REVIEW OF LITERATURE

Karunaratne and Arukwatta (2009) have published efficacy of three plants species on the mortality

and food consumption of Epilachna vigntioctopunctata. They reported the comparison of Annona

squamosa leaf extract with the other two plants extract, it was very strong antifeedant effect on the larvae

at all concentrations tested. Among the three plants tested, Annona squamosa was the most effective

against the Epilachna larvae causing significantly very high mortality even after 24 hours.

Deshmukhe et al. (2010) recorded bioefficacy of cold ethly alchol extract of Annona squamosa

against Spodoptea litura Fabricius. They observed the use of plant extracts to manage Spodoptea litura as


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an eco-friendly management strategy in organic farming, the need of the day. The result showed that the

larvae which were administered the plant extract through their food showed total mortality of 76.66 ± 4.41

at 25% concentration, of which 61.66 ± 1.66% was at the larval stage itself.

Kumar et al. (2010) described insecticidal activity of ethanolic extract of leaves of Annona squamosa

in which it showed potent activity against Sitophilus oryzae pest.

Sreeletha and Geetha (2012) published the pesticidal effects of Annona squamosa L. on male

Oryctes rhinoceros Linn. in relation to reproduction. In which the pesticidal effect of Annona squamosa L.

leaves on the fat body and testis of Oryctes rhinoceros Linn. was studied by incorporating the leaf powder

along with the culture medium, cow dung. 50% larval mortality, 20% pupal mortality and 20% abnormal

adult formation of O.rhinoceros were in 20% w/w proportion of leaf powder cow dung mixture and 30%,

20% and 20% and 10% w/w of it respectively.

Leatemia and Isman (2014) described toxicity and antifeedant activity of crude seed extract of

Annona squamosa (Annonaceae) against Lepidopteran pests and natural enemies. They recorded that crude

aqueous seed extracts of Annona squamosa had moderate to high toxicity to O. insidosus. In which the use

of A. squamosa seed extract was used in conjunction with biocontrol agent for pest management.

Online information Services For Non-chemical Pest Management in the Tropics (Oisat, 2015)

inform pesticidal effect of Annona squamosa leaves crude extract on Spodoptera litura, Spodoptera

exigua, Phenacoccus solenopsis and detailed procedure for preparation of crude aqueous extract.

3. MATERIALS AND METHODS

The experiment was conducted in the Department of Zoology, University of Magway, Myanmar.

The adult specimens of Spodoptera litura were collected from groundnut and tomato fields in Magway

environs. Then, they were cultured in the laboratory with food plant to be reared for larval stages. 4th

instar

larvae were used to test. Experiment was carried out from October to December 2014.

3.1 Preparation of Annona squamosa leaves crude extract

The fresh leaves of the plant Annona squamosa (sugar apple) were collected from the home

gardens near the campus of Magway University. The 100g of fresh leaves were washed with running tap

water to clean dust and other particles. Then, the fresh leaves of A.squamosa were crushed by using a

pestle and filtered the pulp by using filter paper for liquid of crude extracts. Different concentrations of

A.squamosa leaves extract (60%, 75%, and 90%) with their respective dosage were applied to be evaluated

their effects against the 4th

instar larvae of Tobacco caterpillar under laboratory condition. In the extract

method of A.squamosa, any chemical was not used.

3.2 Experimental Design

A contact bioassay was used to test the insecticidal activity of Annona squamosa leaves extract.

The 4th

instar larvae of Tobacco caterpillar Spodoptera litura were treated with the concentration of

Annona squamosa leaves extract. Three replicates for each concentration (60%, 75%, and 90%) were made

for five day period and control treated only with distilled water. Then, ten larvae of tested insects were

introduced into the glass petri-dish 12 cm in diameter. Then, the food leaves were washed with tap water

and added into the dish. The larvae of tested pests sprayed with the A.squamosa leave crude extracts were

introduced into the dish prepared. Mortality was tested every 24 h after the treatment and recorded at room

temperature. The mortality was determined under the dissecting microscope when they did not respond to

mechanical stimulation.


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3.3 Data analysis

Data calculation was conducted by using calculation of Abbott’s formula (1925). Analysis of data

was carried out with hand calculation method.

Abbott’s formula

Where, x is the % survivorship of the control group;

y is the % survivorship in the experimental group.

4. RESULTS

Effectiveness of A.squamosa leaves on some common pests

In the experiment, the results of the effectiveness of A.squamosa leaves crude extract as botanical

pesticides against the 4th

instar larvae of Tobacco caterpillar Spodoptera litura were recorded during five

days of exposure.

The results of tables 1 & 2 showed that the mortality rate of Spodoptera litura was the highest

mortality (ten individuals, 33.33%) in the group tested with 90% Annona squamosa leaves crude extract

while the lowest in the group tested with 60% (no mortality) and the moderate (six individuals, 20%

mortality) in the group tested with 75% at room temperature of about 26.2˚C. The crude extract 60% of

Annona squamosa seems ineffective to kill Spodoptera litura after five days of exposure. This appears to

be shown by 0% mortality in the control group. On the treatment groups, it seems that the higher the

concentration of the crude extract of A.squamosa leaves was the higher death rate of individuals (Fig. 1).

Table 1 Number of individual mortality of 4th

instar larvae of Spodoptera litura

Sr.

No. Day

Control 60% treatment 75% treatment 90% treatment Temperature°C

A B C A B C A B C A B C

1 1st

0 0 0 0 0 0 0 0 0 0 0 0 27

2 2nd

0 0 0 0 0 0 2 0 0 2 2 0 27

3 3rd

0 0 0 0 0 0 2 0 0 0 0 4 27

4 4th

0 0 0 0 0 0 0 2 0 2 0 0 27

5 5th

0 0 0 0 0 0 0 0 0 0 0 0 23

Table 2 Percent morality of S.litura larva during five days of exposure

Species Dose Total number of

individual mortality

Percent of average

mortality

Average

Temperature

S. litura

Control 0 0%

26.2˚C 60% 0 0%

75% 6 20%

90% 10 33.33%

% mortality= 𝑋−𝑌

𝑋 ×100


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0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

1 2 3 4

Dose Percent of average mortality

Fig. 1 Percent mortality of Annona squamosa leaves on Spodoptera litura larvae with respect to dose

5. DISCUSSION

The Family Annonaceae has drawn a lot of attention since the 1980s, due to the presence of

acetogenins, a class of natural products with a broad range of biological activities, among which

insecticidal activity stands out. The acetogenins are found in the leaves and branches and predominantly in

the seeds of annonaceous plants. Acetogenins for insect control will probably continue to be based on

crude or partially refined extracts obtained from plant sources. They can be useful against stem borers,

sucking pests, and scale insects (Isman and Seffrin, 2014).

In the present study, the effectiveness of A.squamosa leaves crude extract as botanical pesticides

against the larvae of Tobacco caterpillar Spodoptera litura were tested. In Spodoptera litura, the 60%

concentration of A.squamosa leaves extract led to 0% mortality, 75% concentration to 20% mortality and

90% concentration to 33.33% mortality.

In 2010, Deshmukhe et al. reported that the highest mortality at the larval stage was 61.66 ± 1.66%

for 20% concentration of A.squamosa leave (ethyl extract). Rajguru and Sarma, (2012) recorded that the

leaf extracts of Acacia arabica and Annona squamosa were found most promising causing 76.66 % and

83.33 % larval mortality respectively at 25 % concentration of within 3 days of treatment. In the present

study the leaves crude extract of A.squamosa 90% concentration led to 33.33% of larval mortality

Spodoptera litura that was lower effectiveness than the findings of Deshmukhe et al. (2010) and Rajguru

and Sharma (2012). It was suggested that the solvent used in the extract method was different.

The results obtained in this experiment can be utilized as an alternative to synthetic insecticides for

the management of pests. Synthetic insecticide application is more effective and immediate control than

botanical insecticide, however, this effort is not effective environmental friendly. The botanical

insecticides have been proven more environmental friendly. Public awareness and resulting environmental

protection agencies (EPA) rulings have led to the removal of some chlorinated, organophosphorus and

carbamate insecticides from the market. It is because of growing concern for clear environmental and

insect populations that are resistant to conventional chemicals (Singh and Saratchandra, 2005).


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6. CONCLUSION

Although A.squamosa leave extracts was little effectiveness on Spodoptera litura, it gives baseline

information to researchers and informs new botanical pesticides to local farmers. An application of the

crude extract is an inexpensive, readily available, attractive and effective technique and its easy approach

of this technology by local farmers. Crude extract of A.squamosa is a promising candidate for development

as simple botanical insecticide for local use in rural Myanmar.

7. RECOMMENDATION

a. A.squamosa (Sugar apple) leaves should be used for the control of some pests in the field.

b. Farmers should work hard to improve the quality of crude extract of sugar apple leaves.

c. Farmers should be recognized to use the botanical insecticide instead of chemical pesticides to value

our environment.

8. REFERENCES [1] Abbott, W. S., 1925. A method for computing the effectiveness of an insecticides. J. Ecological Entomol. 18: 265-267.

[2] Baskar, K., Maheshwaran, R., Kingsley, S., Ignacimuthu, S., 2011. Bioefficacy of plant extracts against Asian army

worm Spodoptera litura Fab. (Lepidoptera: Noctuidae). Journal of Agricultural Technology, 7(1): 123-131.

[3] Deshmukhe, P. V., Hooli, A. A., Holihosur, S. N., 2010. Bioefficacy of cold ethyl alchol extract of Annona squamosa

against Spodoptera litura Fabricius. Journal of biopesticides, (1): 271-274.

[4] Isman, M. B., Seffrin, R., 2014. Natural Insecticides from the Annonaceae: A Unique Example for Developing

Biopesticides. Advances in Plant Biopesticides, DOI 10.1007/978-81-322-2006-0_2, © Springer India.

[5] Kandagal, A. S., Khetagoudar, M. C., 2013. Study on larvicidal activity of weed extracts against Spodoptera litura.

Journal of environmental Biology, 39: 253-257.

[6] Kan zaw, Nu Nu Lwin, Khin Thida Nyine, Mya Thandar, 1998. Agricultural transformation, institutional changes and

rural development in Ayeyarwady Delta, Myanmar. Yangon Institue of Economics.

[7] Kesetyaningsih, T. W., 2012. Efficacy of Annona squamosa Leaf Extract as An Insecticide against Cockroach

(Periplaneta americana). International Conference: Research and Application on Traditional Complementary and

Alternative Medicine in Heath Care (TCAM), June, 22-23. Surakarta Indonesia.

[8] Kumar, J. A., Rhkha, T., Devi, S. S., Kannan, M., Jaswanth, A., Gopal, V., 2010. Insecticidal activity of ethanolic

exract of leaves of Annona squamosa. J.Chem. Pharm, Res., 2(5): 177-180.

[9] Leatemia, J. A., Isman, M. B., 2014. Efficacy of crude seed extracts of Annona squamosal against diamondback moth,

Plutella xylostella L. in the green house. International journal of pest management, 50(2): 129-133.

[10] Online information Service for Non-chemical Pest Management in the Tropic (internet). Pesticide Action Network

(PAN) Germany; (cited 2015 March). Available from: http:// www. Pan germany. Org. www. Oisat. Org.

[11] Prakash, A., Rao, J., Nandasgopal, V., 2008. Future of botanical pesticides in rice, wheat, pulses and vegetables pest

management. Journal of biopesticides, 1 (2): 154-169.

[12] Rajguru, M., Sharma, A. N., 2012. Comparative efficacy of plant extracts alone and in combination with Bacillus

thuringiensis sub sp. kurstaki against Spodoptera litura Fab. larvae. JBiopest., 5(1): 81-86.

[13] Singh, R. N., Saratchandra B., 2005. The Development of Botanical Products with Special Reference to Seri-

Ecosystem. Caspian J. Env. Sci., 3(1): 1-8.

[14] Sreeletha, C., Geetha, P. R., 2012. Pesticidal effects of Annona squamosa L. on male Oryctes rhinocerous Linn.

(Coleptera: Scarabaeidae) in relation to reproduction. Current Biotica, 6 (1): 8-21.

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ISSN: 2391-3991, Volume 2 Issue 2, page 19 - 24Borneol is an essential oil with bicyclic organic compound that belong to essential oil, and has a natural insect repellent effect. It