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Effect of nanocapsules and extract of Metarhizium anisopliae in inhibiting acetylcholine esterase enzyme in Musca domestica larvae.


  • Aliaa Abdul Aziz Hammed Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq.
  • Hazim Idan Al Shammari Department of Agricultural Researches, Ministry of Sciences and Technology, Baghdad, Iraq.
  • Soolaf A Kathiar Department of Biology, College of Science for Women, University of Baghdad, Baghdad, Iraq.



Acetylcholine esterase enzyme inhibition, Housefly, M. anisopliae, Nanocapsules, Fungi secondary metabolites


M. domestica is the most important insect that transmit pathogens for diseases in the world. The use of nanotechnology is eco-friendly method in control pests. The study aims to investigate the feasibility of bio-manufacturing nanocapsules of fungal secondary metabolites in order to improve the efficiency of metabolite and assess their inhibitory effect on the acetylcholine esterase enzyme in housefly larvae. An equal mixture of organic solvents, ethyl acetate and dichloromethane, was used to extract the metabolic products of the fungus M. anisopliae, (PEG4000) and chitosan was used in the preparation of nanocapsules. The results of the DLS granular size assay showed that the size of the extract particles and the size of the chitosan and (PEG 4000) nanocapsules were 610, 217 and 188 nm, respectively. The SEM images showed that the diameter of the extract and the nanocapsules chitosan and polyethylene glycol 4000 reached a rate 547.5, 17.8 and 26.2 nm, respectively. The FTIR showed that the extract of the second products of the fungus contains functional groups like: alkynes and alkenes, amines, carboxyl and aromatic groups, while the presence of groups of phenols, alcohol, amines, alkenes, and alkyl halides was recorded for nanocapsules of chitosan and PEG. The results showed that the extract of fungal metabolic and nanocapsules has an inhibitory effect on acetylcholinesterase enzyme and reached the highest inhibition rate 53.2 ,36.3,18.2% when treated with nanocapsules PEG at a concentration 500 ppm, extract of fungal metabolites at a concentration 50,000 ppm, chitosan nanocapsules at a concentration 500 ppm respectively. It is clear that acetylcholinesterase inhibition is one of the mechanisms of fungi metabolic action and the nanocapsules prepared from them.


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Onyenwe E , Okore O O, Ubiaru P C , Abel C . Housefly-borne helminth parasites of Mouau and its public health implication for the university community. Anim. Res. Int. 2016 ; 13(1): 2352–2358.

Geden C J , Nayduch D , Scott J G , Burgess IV E R , Gerry A C , Kaufman P E . House Fly (Diptera: Muscidae): Biology, Pest Status, Current Management Prospects, and Research Needs. J Integr Pest Manag, 2021; 12(1):1–38.

Köhl J, Kolnaar R , Ravensberg WJ. Mode of action of microbial biological control agents against plant diseases: Relevance beyond efficacy. Frontiers in Plant Science. 2019; Jul;10:1–19.

Yaseen AT. The Effect of Alcoholic and Aqueous Extract of Piper nigrum on the Larvae of Culex pipiens molestus Forskal (Diptera: Culicid). Baghdad Sci J. 2020 ; 17(1): 28-33 .

Casida JE, Durkin K A . Neuroactive insecticides: Targets, selectivity, resistance, and secondary effects. Annu Rev Entomol. 2013; 58, 99–117.

Huang W, Wang S , Jacobs-Lorena M . Use of microbiota to fight mosquito-borne disease. Front Genet. 2020; 11: 196.

Gul HT, Saeed S, Khan FZA , Manzoor SA . Potential of nanotechnology in agriculture and crop protection: A review. Appl Sci Bus Econ. 2014; 1: 23–28.

Zhang Y, Maoyu L, Xiaomei G, Yongheng C, Ting L . Nanotechnology in cancer diagnosis: progress, challenges and opportunities . J Hematol Oncol. 2019 ; 12:137.

Parisi C , Vigani M , Rodriguez-Cerezo E . Agricultural nanotechnologies: What are the current possibilities?. Nano Today. 2015; 10: 124–127.

Al Hilfy AA A, Al Shammari H I , Kathiar SA. Toxicity of Nanoemulsion of Castor Oil on the Fourth larval stage of Culex quinquefsciatus under Laboratory Conditions . Baghdad Sci J. 2022; 19 (5): 99-107 .

Kouzegaran VJ , Farhadi K . Green synthesis of sulphur nanoparticles assisted by a herbal surfactant in aqueous solutions. Micro Nano Lett.. 2017; 12: 329-334.

Malakar Y, Lacey J, Bertsch P M . Towards responsible science and technology: How nanotechnology research and development is shaping risk governance practices in Australia. Humanit Soc Sci Commun 2022; 9(17): 1-14.

Kumar MY, Ravi A. Extraction and characterization of chitosan from shrimp waste for application in the feed industry. Int J Environ Sci Technol.. 2017; 6(4): 2548-2557.

Morin-Crini , N , Lichtfouse E, Torri G, Crini G . Fundamentals and Applications of Chitosan. In: Crini, G., Lichtfouse, E. (eds) Sustainable Agriculture Reviews 35. Sustain. Agric Res. 2019 ; 35. Springer Cham.

Rebecca A O, Angie B K , Brent S S. . Enlightening advances in polymer bioconjugate chemistry: light-based techniques for grafting to and from bio macromolecules. Chem Sci., 2020; 11: 5142 -5156.

Munawar AM , Jaweria TMS , Kishor MW , Ellen KW. An Overview of Chitosan Nanoparticles and Its Application in Non-Parenteral Drug Delivery. J Pharmaceutics. 2017; 9(4): 1-26.

Zafar J , Shoukat R F , Zhang Y , Freed S , Xu X , Jin F . Metarhizium anisopliae Challenges Immunity and Demography of Plutella xylostella. Insects. 2020; 11(10): 694. insects11100694.

Vivekanandhan P, Swathy K, Kalaimurugan D, Ramachandran M , Yuvaraj A , Kumar A N. Larvicidal toxicity of Metarhizium anisopliae metabolites against three mosquito species and nontargetingorganisms. PLoSONE. 2020; 15(5): 1-18. 0232172. 10.1371/journal.pone.0232172

Berestetskiy A , Hu Q . The Chemical Ecology Approach to Reveal Fungal Metabolites for Arthropod Pest Management. Microorganisms. 2021; 9: 1397.

Hoang N H , LeThanh T , Sangpueak R , Treekoon J , Saengchan C, Thepbandit W, et al . Chitosan Nanoparticles-Based Ionic Gelation Method: A Promising Candidate for Plant Disease Management. Polymers 2022; 14: 62. 10.3390/polym14040662 .

Wu J, Du C, Zhang J, Yang B, Cuthbertson A G S, Ali S. Synthesis of Metarhizium anisopliae–Chitosan Nanoparticles and Their Pathogenicity against Plutella xylostella (Linnaeus). Microorganisms 2022; 10(1): 1.

Paralikar P, Rai M . Bio inspired synthesis of sulphur nanoparticles using leaf extract of four medicinal plants with special reference to their antibacterial activity. IET Nanobiotechnol. 2017; 12: 25-31.

Wang X S , Xu J, Wang X M , Qiu B L , Cuthbertson A G S. ,Du C L et al . Isaria fumosorosea-based-zero-valent iron nanoparticles affect the growth and survival of sweet potato whitefly, Bemisia tabaci (Gennadius). Pest Manag Sci. 2019; 75: 2174–2181.

Khairan K, Zahraturriaz , Jalil Z . Green synthesis of sulphur nanoparticles using aqueous garlic extract (Allium sativum). Rasayan J Chem. 2019 ;12: 50-57.

Łopusiewicz Ł , Mazurkiewicz-Zapałowicz K , Tkaczuk C . Chemical changes in spores of the entomopathogenic fungus Metarhizium robertsii after exposure to heavy metals, studied through the use of FTIR spectroscopy. J Elem. 2020; 25(2): 487-499.

Huang Q, Liu M, Feng J, Liu Y. Effect of dietary benzoxadiazole on larval development, cuticle enzyme and antioxidant defense system in housefly (Musca domestica L.). Pest Biochem Physiol. 2008; 90: 119–125.

Attaullah, Zahoor M K, Zahoor M A, Mubarik M S, Rizvi H, Majeed H N et al. Insecticidal, biological and biochemical response of Musca domestica (Diptera: Muscidae) to some indigenous weed plant extracts. Saudi J Biol Sci. 2020; 27(1): 106-116.

Wu J, Du C, Zhang J, Yang B, Cuthbertson A G S, Ali S. Synthesis of Metarhizium anisopliae–Chitosan Nanocapsules and Their Pathogenicity against Plutella xylostella (Linnaeus). Microorganisms 2022; 10. 1.

Vey A, Hoagland R, Butt T M. Toxic metabolites of fungal biocontrol agents. Fungi as biocontrol agents progress, problems and potential. International.2001. Chap 12: 311-346. CABI Publishing, Wallingford, UK.

Namasivayam S K R, Bharani R S A, Karunamoorthy K. Insecticidal fungal metabolites fabricated chitosan nanocomposite (IM-CNC) preparation for the enhanced larvicidal activity—An effective strategy for green pesticide against economic important insect pests. Int. J Biol Macromol. 2018; 120: 921–944.

Han J, Zhou Z , Yin R , Yang D , Nie J. Alginate chitosan /hydroxyapatite polyelectrolyte complex porous scaffolds: Preparation and characterization. Int J Biol Macromol. 2010; 46: 199–205.

Vivekanandhan P., Swathy K, Murugan A C, Krutmuang P. Insecticidal Efficacy of Metarhizium anisopliae Derived Chemical Constituents against Disease-Vector Mosquitoes. J Fungi. 2022; 8: 300.

Bhavya M L, Obulaxmi S., Devi, S S. Efficacy of Ocimum tenuiflorum essential oil as grain protectant against coleopteran beetle, infesting stored pulses. J Food Sci Technol. 2021; 58: 1611–1616.

Vivekanandhan P, Swathy K, Thomas A, Kweka E J; Rahman A, Pittarate S. et al. Insecticidal Efficacy of Microbial-Mediated Synthesized Copper Nano-Pesticide against Insect Pests and Non-Target Organisms. Int J Environ Res Public Health 2021; 18: 10536.

Liu, L J, Alam M S, Hirata K, Matsuda K, Ozoe Y. Actions of quinolizidine alkaloids on Periplanta Americana nicotinic acetylcholine receptors. Pest Manag Sci. 2008; 64: 1222–1228.

Knutsson S. Towards Mosquitocides for Prevention of Vector-Borne Infectious Diseases Discovery and Development of Acetylcholinesterase 1 Inhibitors. Doctoral Thesis, Department of Chemistry Umeå University, 2016 .

Šagud I , Hrvat NM , Grgičević A , Tena Č , Josipa H , Milena D. et al . Design, synthesis and cholinesterase inhibitory properties of new oxazole benzylamine derivatives, Journal of Enzyme Inhibition and Medicinal Chemistry, 2020; 35(1): 460-467.

Yadav R N, Mahtab M R, Kumar R , Singh H B , Zaidi N W. Secondary Metabolites of Metarhizium spp. and Verticillium spp. and Their Agricultural Applications . In book: Secondary Metabolites of Plant Growth Promoting Rizomicroorganisms . 2019: 27-58.

Samuels R , Reynolds SE , Charnley AK . Calcium channel activation of insect muscle by destruxins, insecticidal compounds produced by the entomopathogenic fungus Metarhizium anisopliae (Metch.). Comp Bio chem Physiol. 1988; 90:403–412

Wu Sh , Zhou Y , Li Z . Bio catalytic selective functionalization of alkenes via single-step and one-pot multi-step reactions. Chem. Commun.2019; 55: 883-896. .

Shi L w , Zhang J , Zhao M , Tang Sh , Xu Ch , Zhang W , et al . Effects of polyethylene glycol on the surface of nanoparticles for targeted drug delivery. J Nanoscale. 2021; 13: 10748-10764.

Jia M, Cao G, Yibo L, Xiongbing T, Wang G , Xiangqun N , et al . Biochemical basis of synergism between pathogenic fungus Metarhizium anisopliae and insecticide chlorantraniliprole in Locusta migratoria (Meyen). Sci Rep 2016; 6: 28424.

Archita Sh, Kritika S, Jasreen K, Madhu K. Agrochemical loaded biocompatible chitosan nanoparticles for insect pest management. Biocatal Agric Biotechnol. 2019;18: 101079.