Sains Malaysiana 53(5)(2024): 1081-1091
http://doi.org/10.17576/jsm-2024-5305-09
Molluscicidal Activity and Inhibition of
Acetylcholinesterase Activity of Azadirachta indica Extract on Pomacea canaliculate
(Aktiviti Moluskisid dan Perencatan Aktiviti Asetilkolinesterase Ekstrak Azadirachta indica pada Pomacea canaliculate)
NIK AMALIA NASRATIENA MAT ZAIB1,
SAIYIDAH NAFISAH HASHIM2, WAN ROZIANOOR MOHD HASSAN1,3,* & CHIA CHAY TAY1
1Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam,
Selangor, Malaysia
2Faculty of Applied Science, Universiti Teknologi MARA Cawangan Perak Kampus Tapah, 35400 Tapah, Perak, Malaysia
3Human Genetic and Biochemistry (HuGeB),
Research Nexus UiTM (ReNeU), Universiti Teknologi MARA,
40450 Shah Alam, Selangor, Malaysia
Diserahkan: 12 Disember2023/Diterima: 5 April 2024
Abstract
The
Golden Apple Snail, Pomacea canaliculata, is a notorious pest in Malaysia, leading
to substantial agricultural and economic losses, while, the neem tree, Azadirachta indicahas
been identified as alternative source for bio-pesticides. There is currently
limited information on A. indica as natural molluscicide against P. canaliculata. This study aims to evaluate the effects of A. indica on P. canaliculata at various life stages
(eggs, juvenile and adult). The bioactive compound, azadirachtin in A. indica extracts was quantified using
high-performance liquid chromatography (HPLC). Egg hatchability was determined
using spot spraying on egg clusters while the molluscicidal activity of A. indica on juvenile and adult
specimens was evaluated using the immersion bioassay approach. The snail's
acetylcholinesterase (AChE) inhibition was assessed
using Ellman assay. The azadirachtin content in the ethanolic extract of A. indica leaves at a concentration of 1 µg/mL was found
to be 45.42%. Increasing the concentration of A. indica extracts resulted in lower egg hatchability and increased mortality of juvenile
and adult snails. Azadirachta indicaextracts showed the highest effectiveness
against adult P. canaliculata with an LC50 value of 0.52 mg/mL, compared to 0.92 mg/mL for juveniles and 0.74 mg/mL for
eggs. Additionally, A. indica extract exhibited greater inhibition of AChE in adult specimens compared to juvenile specimens and
eggs. The IC50 for adult is 0.60 μg/mL,
which is the lowest while the IC50 for juvenile is 0.68 μg/mL, the highest. Taken together, A. indica shows immerse potential as natural pesticide for
managing the population of P. canaliculata.
Keywords: Azadirachta indica; egg
hatchability; molluscicides and inhibition of AChE; Pomacea canaliculata
Abstrak
Siput Gondang Emas (Pomacea canaliculata) adalah perosak utama yang menyebabkan kerugian dalam sektor pertanian dan ekonomi Malaysia, manakala pokok Mambu, Azadirachta indica telah ditemui sebagai sumber alternatif untuk racun serangga berasaskan bahan semula jadi dalam beberapa kajian, termasuklah sebagai racun moluska semula jadi terhadap P. canaliculata. Oleh itu, kajian ini bertujuan untuk menilai kesan ekstrak A. indica terhadap perosak P. canaliculata pada pelbagai peringkat kehidupan (telur, juvana dan dewasa). Sebatian bioaktif, azadiraktin daripada ekstrak A. indica ditentukan menggunakan kromatografi cecair prestasi tinggi (HPLC). Kebolehtetasan telur dinilai dengan semburan tompok pada kelompok telur manakala teknik rendaman digunakan untuk menilai aktiviti moluskisida A. indica pada peringkat juvana dan dewasa. Kemudian, perencatan asetilkolinesterase (AChE) pada siput ditentukan menggunakan ujian Ellman. Di sini, kami melaporkan bahawa terdapat 45.42% azadiraktin dijumpai di dalam 1 µg/mL ekstrak etanol daun A. indica. Keputusan kajian menunjukkan bahawa apabila kepekatan ekstrak A. indica meningkat, kebolehtetasan telur berkurangan manakala, kematian siput juvana dan dewasa didapati meningkat. Ekstrak A. indica menunjukkan nilai LC50 terendah untuk siput dewasa (0.52 mg/mL) berbanding dengan juvana (0.92 mg/mL) dan telur (0.74 mg/mL). Ini menunjukkan potensi ekstrak yang tinggi terhadap P. canaliculata peringkat dewasa. Tambahan pula, ekstrak A. indica juga menunjukkan perencatan AChE yang lebih tinggi pada siput dewasa berbanding juvana dan telur.
IC50 untuk dewasa adalah yang paling rendah (0.60 μg/mL) manakala IC50 untuk juvana adalah yang tertinggi (0.68 μg/mL). Secara keseluruhannya, A. indica didapati sangat berpotensi sebagai racun perosak semula jadi dalam mengawal populasi P. canaliculata.
Kata kunci: Azadirachta indica; kebolehtetasan telur; moluskisida dan perencatan AChE; Pomacea canaliculata
RUJUKAN
Abubakar, A., Bala, A.Y. & Singh, K. 2017. Plant molluscicides and their modes of action: A review. International
Journal of Scientific Research in Technology, Applied Sciences & Health
Studies | IJSRTASHS 2(1): 37-49.
Al-hashemi, Z.S.S. & Hossain, M.A. 2016.
Biological activities of different neem leaf crude extracts used locally in Ayurvedic medicine. Pacific
Science Review A: Natural Science and Engineering 18(2): 128-131.
Araújo, M.C., Assis, C.R.D., Silva, K.C.C., Souza, K.S., Azevedo, R.S., Alves, M.H.M.E., Silva, L.C., Silva, V.L.,
Adam, M.L., Carvalho Junior, L.B., Souza Bezerra, R. & Oliveira, M.B.M. 2018. Characterization of brain
acetylcholinesterase of bentonic fish Hoplosternum littorale:
Perspectives of application in pesticides and metal ions biomonitoring. Aquatic Toxicology 205: 213-226.
Aromin, E.J.F., Akol, G.B.D.,
Soriano, Z.M.L. & Lucero, J.A. 2020. The molluscicidal potential of selected plants against Pomacea canaliculata. IAR Journal of Agriculture Research and Life Sciences 1(1): 26-34.
Bigatti, G.,
Maximiliano, G.B., Israel, A.V., Pablo E.P. & Alfredo, C.V. 2010. The
calcareous egg capsule of the patagonian neogastropod Odontocymbiola Magellanica: Morphology, secretion and mineralogy. Journal of Molluscan Studies 76(3):
279-288.
Chen, Y. 2012. Organophosphate-induced brain damage: Mechanisms,
neuropsychiatric and neurological consequences, and potential therapeutic
strategies. NeuroToxicology 33(3):
391-400.
Decuypere, E., Onagbesan, O., De Smit, L., Tona,
K., Everaert, N., Witters,
A., Debonne, M., Verhoelst,
E., Buyse, J., Hassanzadeh,
M., de Baerdemaeker, J., Arckens,
L. & Bruggeman, V. 2006. Hypoxia and hypercapnia
during incubation of chicken eggs: Effects on development and subsequent
performance. World’s Poultry Science
Journal XII Europe (June):
486-487.
Ellman, G.L., Diane
Courtney, K., Valentino, A. & Robert, M.F. 1961. A new and rapid
colorimetric determination of acetylcholinesterase activity. Biochemical Pharmacology 7(2): 88-95.
Fernandes, S.R., Barreiros, L., Oliveira, R.F., Cruz, A., Prudêncio, C., Oliveira, A.I., Pinho,
C., Santos, N. & Morgado, J. 2019. Chemistry,
bioactivities, extraction and analysis of azadirachtin:
State-of-the-art. Fitoterapia 134: 141-150.
Ismail, H.N. & Musa N.N. 2021. Molluscicidal activity of Zingiber officinale and Carica papaya on the egg hatchability of the
golden apple snail, Pomacea canaliculata (Gastropoda: Ampuliriidae). Malaysian Journal of Science 40(2):
40-50.
Ji, M., Cheng, P., Huai, E.H., Suganiya, R.R. & Yoon, Y.Y. 2018. Invasive apple snails
in wetlands of Selangor, Malaysia: Species, distribution, and ecological
associations. Journal of Tropical Biology
and Conservation 15(1): 43-60.
Joshi, R.C. 2007. Problems with the management of the golden apple snail Pomacea canaliculata:
An important exotic pest of rice in Asia. In Area-Wide Control of Insect Pests: From Research to Field
Implementation, edited by Vreysen, M.J.B., Robinson, A.S. & Hendrichs,
J. Dordrecht: Springer. pp. 257-264.
Kaushik, N. 2002. Determination of azadirachtin and fatty acid methyl esters of Azadirachta indica seeds by HPLC and GLC. Analytical and Bioanalytical Chemistry 374(7-8): 1199-1204.
Kilani-Morakchi, S., Morakchi-Goudjil, H. & Sifi, K. 2021. Azadirachtin-based
insecticide: Overview, risk assessments, and future directions. Frontiers in Agronomy 3: 676208.
Kristoff, G., Luis, C.C., Noemí, R.V.G. &
Adriana, C.C. 2011. Effects of the organophosphate insecticide azinphos-methyl on the reproduction and cholinesterase
activity of Biomphalaria glabrata. Chemosphere 84(5): 585-591.
Kumar, G.H. & Priyadarsini, V.R. 2010. The
neem limonoids azadirachtin and nimbolide inhibit cell proliferation and induce
apoptosis in an animal model of oral oncogenesis. Investigational
New Drugs. 28: 392-401.
Kumar, P., Singh, V.K. & Singh, D.K. 2012. Feeding of bait to snail Lymnaea acuminataand
their effect on certain enzyme in the nervous tissue. ISRN Biochemistry 2012: 343047.
Liu, C., Zhang, Y., Ren, Y., Wang, H., Li, S., Jiang, F., Yin, L., Qiao, X., Zhang, G., Qian, W., Liu, B. & Fan, W. 2018.
The genome of the golden apple snail Pomacea canaliculata provides insight into stress tolerance and
invasive adaptation. Gigascience 7(9): giy101.
Lushchak, V.I., Matviishyn, T.M., Husak,
V.V., Storey, J.M. & Storey,
K.B. 2018. Pesticide toxicity: A mechanistic approach. EXCLI
J. 17: 1101-1136.
Manikanta, P. & Dokuparthi, S.K. 2014. A review on role of Azadirachta indicaA. Juss as a biopesticide. International Journal of Universal Pharmacy
and Bio Sciences 3(2): 10.
Massaguni, R. & Md Latip, S.N.H. 2015. Assesssment the molluscicidal properties of azadirachtin against golden apple
snail, Pomacea canaliculata. Malaysian Journal of Analytical Sciences 19(4): 781-789.
Massaguni, R. & Md Latip, S.N.H. 2012. Neem crude
extract as potential biopesticide for controlling
golden apple snail, Pomacea canaliculata. Pesticides - Advances in Chemical and
Botanical Pesticides. InTech. doi:10.5772/48626.
Md. Latip, S.N.H., Keni,
M.F. & Rosli, R. 2017. Antifeedant activities of neem seed extracts for controlling golden appple snail, Pomacea canaliculata. The Social Sciences 12(2): 294-298.
Musman, M., Kamaruzzaman,
S., Karina, S., Rizqi, R. & Arisca,
F. 2013. A preliminary study on the anti hatching of
freshwater golden apple snail Pomacea canaliculata (Gastropoda: Ampullariidae) eggs from Barringtonia racemosa (Magnoliopsida: Lecythidaceae) seeds extract. AACL Bioflux 6(4): 394-398.
Narayanan, S.E., Abdul Rehuman, N., Harilal, S., Vincent, A., Rajamma, R.G., Behl, T., Uddin, M.S., Md Ashraf,
G. & Mathew, B. 2020. Molecular mechanism of zinc neurotoxicity in
Alzheimer’s disease. Environmental
Science and Pollution Research 27(35): 43542-43552.
Ramsay, R.R. & Tipton, K.F. 2017. Assessment of enzyme inhibition: A
review with examples from the development of monoamine oxidase and
cholinesterase inhibitory drugs. Molecules 22(7): 1192.
Rawi, S.M., Al-Hazmi, M. & Al Nassr, S.F.
2011. Comparative study of the molluscicidal activity
of some plant extracts on the snail vector of Schistosoma mansoni, Biomphalaria alexandrina. International Journal of Zoological Research 7(2): 169-189.
Salleh, N.H.M., Dachyar, A., Daud, M.Z.M., Pilus,
N. & Nawi, R. 2012. Distribution and management
of Pomacea canaliculata in the Northern Region of Malaysia: Mini review. APCBEE Procedia 2: 129-134.
Sami, A.J., Sehrish, B., Khalid, M., Shakoori, F.R., Rehman, F. & Shakoori, A.R. 2016. Effect of crude neem (Azadirachta indica)
powder and azadirachtin on the growth and
acetylcholinesterase activity of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). Pakistan Journal of Zoology 48(3):
881-886.
Senthil Nathan, S.,
Choi, M.Y., Seo, H.Y., Paik, C.H., Kalaivani, K. & Kim, J.K. 2008. Effect of azadirachtin on acetylcholinesterase (AChE)
activity and histology of the brown planthopper Nilaparvata lugens (Stål). Ecotoxicology and Environmental Safety 70(2):
244-250.
Shafeek, A., Jaya Prasanthi, R.P., Hariprasad Reddy, G., Chetty, C.S. & Rajarami Reddy, G. 2004. Alterations in acetylcholinesterase and electrical activity in
the nervous system of cockroach exposed to the neem derivative, azadirachtin. Ecotoxicology
and Environmental Safety 59(2): 205-208.
Sidhu, G.K., Singh, S., Kumar, V., Dhanjal,
D.S., Datta, S. & Singh, J. 2019. Technology
toxicity, monitoring and biodegradation of organophosphate pesticides: A
review. Critical Reviews in Environmental
Science and Technology 49(13): 1135-1187.
Silva, J.C.T., Gulab, N.J., Rosângela,
D.L.O. & Leslie, B. 2007. Purification of the seven tetranortriterpenoids in neem (Azadirachta indica) seed
by counter-current chromatography sequentially followed by isocratic
preparative reversed-phase high-performance liquid chromatography. Journal of Chromatography A 1151(1-2): 203-210.
Singh, A. & Singh, D.K. 2001. Molluscicidal activity of the custard apple (Annona squamosa L.) alone and in combination with other plant
derived molluscicides. Journal of Herbs, Spices and Medicinal Plants 8(1): 23-29.
Singh, K. & Singh, D.K. 2000. Toxicity to the snail Limnaea acuminata of
plant-derived molluscicides in combination with
synergists. Pest Management Science 56(10): 889-898.
Sinha, S., Murthy, P.S.N., Rao, C.V.N., Ramaprasad, G., Sitaramaiah, S., Kumar, D.G. & Suresh, K.S. 1999.
Simple method for enrichment of azadirachtin from
neem seeds. Journal of Scientific and
Industrial Research 58(12): 990-994.
Sisa, M.H., Aspani,
F., Massaguni, R., Awang Damit, H. & Joseph, H. 2016. Inhibition of egg hatching
of the golden apple snail by synthetic molluscicides.
In Regional Conference on Science, Technology and Social Sciences (RCSTSS
2014), edited by Yacob, N., Mohamed, M. & Megat Hanafiah, M. Singapore:
Springer.
Soni, H., Mishra, K.,
Sharma, S. & Singhai, A.K. 2012. Characterization
of azadirachtin from ethanolic extract of leaves of Azadirachta indica. Journal of Pharmacy Research 5(1):
199-201.
Sparling, D.W. & Fellers, G. 2007. Comparative toxicity of chlorpyrifos, diazinon, malathion
and their oxon derivatives to larval Rana boylii. Environmental Pollution 147(3): 535-539.
Teixeira, T., José, S.R., Nuno, R., José, B.
& Armindo, R. 2012. Assessment of molluscicidal activity of essential oils from five Azorean
plants against Radix peregra (Müller 1774). Chemosphere 87(1): 1-6.
Thanomsit, C., Maprajuab, A., Saowakoon, S., Prasatkaew, W., Ocharoen, Y., Wattakornsiri, A., Nanuam, J.
& Nanthanawat, P. 2018. Acetylcholinesterase (AChE): Potential biomarker for evaluating pesticide
exposure on egg and tissue of golden apple snail (Pomacea canaliculata) from Huai−Saneng Reservoir, Surin Province, Thailand. Thai J. Agric. Sci. 51(3): 104-117.
Vinotha Alex, A. &
Mukherjee, A. 2021. Review of recent developments (2018–2020) on
acetylcholinesterase inhibition based biosensors for organophosphorus
pesticides detection. Microchemical Journal 161: 105779.
Yahaya, H., Badrulhadza, A., Sivapragasam,
A., Nordin, M., Muhamad Hisham,
M.N. & Misrudin, H. 2017. Invasive apple snails
in Malaysia. In Biology and Management of Invasive Apple Snails, 2nd
ed., edited by Joshi, R.C., Cowie, R.H. & Sebastian, L.S. Philippine:
Philippine Rice Research Institute. pp. 187-613.
Zheng, Y., Wu, J., Wang, Y.,
Peng, X. & Zhang, Y. 2018. Seed yield and azadirachtin content of Azadirachta indica in
four ecosystems of Southwest China. Industrial
Crops and Products 122: 23-27.
*Pengarang untuk surat-menyurat;
email: rozianoor@uitm.edu.my
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