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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