• Shrinivas S. Jadhav Environmental and Molecular Toxicology Laboratory, Department of PG Studies in Zoology, Karnatak University, Dharwad, Karnataka, India 580003
  • David M Department of higher studies in Zoology, Karnatak University, Dharwad


Objective: Flubendiamide is extensively used in agriculture practices as foliar application pesticide. Due to long persistency in the soil, flubendiamide pose serious health concern in non-target organisms. Our main objective was to examine flubendiamide impact on burrowing animal Eudrilus eugeniae with special emphasis on avoidance behaviour and neurotoxicity.

Methods: Acute toxicity study of flubendiamide (Diamide pesticide) was conducted on earthworm, Eudrilus eugeniae through direct paper contact method and artificial soil method. Median lethal concentration (LC50) of flubendiamide was calculated by following probit analysis. The neurotoxic potential of flubendiamide was studied with marker enzyme Acetyl cholinesterase (AChE) levels in both In vivo and In vitro experiments.

Results: LC50 in earthworms was found to be 94.4 µg cm-2 at 48h paper contact test and 332.21 mg kg-1and 238.31 mg kg-1 respectively at 7 and 14 d artificial soil exposure. Morphological and physiological alterations in earthworms attribute to inhibition of AChE levels. The kinetic study of AChE activity in presence and absence of inhibitor suggests the enzyme reaction is competitive in nature.

Conclusion: Present study establishes concentration-dependent flubendiamide toxicity in earthworm E. eugeniae. No clear conclusive remarks were made on earthworm avoidance behaviour as the worms were located both in toxic and control soil after 48h of exposure. Further studies may be needed in this aspect to establish clear understanding on avoidance nature of E. eugeniae in different soil types. 

Keywords: Earthworm, Acetylcholinesterase, Flubendiamide, Enzyme kinetics, Avoidance behaviour, Eudrilus eugeniae


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1. Jadhav SS, David M. Biodegradation of flubendiamide by a newly isolated Chryseobacterium sp. strain SSJ1. 3 Biotech 2016;6:31.
2. Chauhan SS, Agrawal S, Srivastava A. Effect of Imidacloprid insecticide residue on biochemical parameters in potatoes and its estimation by HPLC. Asian J Pharm Clin Res 2013;6:114-7.
3. Venkateswara Rao J, Surya Pavan Y, Mahadevendra SS. Toxic effects of chlorpyrifos on morphology and acetylcholinesterase activity in the earthworm Eisenia foetida. Ecotoxicol Eniviron Saf 2003;54:296-301.
4. Eguchi S, Hatano R, Sakuma T. Toshio effect of earthworms on the decomposition of soil organic matter. Nippon Dojo-Hiryogaku Zasshi 1995;66:165–7.
5. Dash MC, Senapathi BK. National Seminar on Organic Waste Utilize, Vermi Comp. Part-13, Proceedings; 1986. p. 157–77.
6. Vishwanathan R. Physiological basis of assessment of ecotoxicology of pesticides to soil organisms. Chemosphere 1997;35:323–34.
7. Helling B, Reinecke SA, Reinecke AJ. Effects of the fungicide copper oxychloride on the growth and reproduction of Eisenia foetida (Oligochaeta). Ecotoxicol Environ Saf 2000;46:108–16.
8. Yeardley RB, Lazorchak JM, Gast LC. The potential of an earthworm avoidance test for evaluation of hazardous waste sites. Environ Toxicol Chem 1996;15:1532-7.
9. ISO. International Organization for Standardization. Draft ISO-17512: Soil Quality e Avoidance test for evaluating the quality of soils and the toxicity of chemicals. Test with earthworms (Eisenia fetida/andrei). Geneve, Switzerland; 2006.
10. Organisation for Economic Co-operation and Development (OECD). OECD Guidelines for Testing of Chemicals, Earthworm, Acute Toxicity Tests (Filter paper test and Artificial soil test); 1984. p. 1–9.
11. Venkateswara Rao J, Kavitha P. Toxicity of azodrin on the morphology and acetylcholinesterase activity of the earthworm Eisenia foetida. Environ Res 2004;96:323-7.
12. Finney DJ. Probit analysis 2nd Edition. Cambridge University Press, Cambridge, UK; 1953.
13. Garcia M, Rombke J, Torres de Brito M, Scheffczyk A. Effects of three pesticides on the avoidance behaviour of earthworms in laboratory tests performed under temperate and tropical conditions. Environ Poll 2008;153:450-6.
14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with folin phenol reagent. J Biol Chem 1951;193:265–75.
15. Ellman GL, Courtney KD, Andres Jr VV, Featherstone RM. A new and rapid colorimetric determination of acetyl cholinesterase activity. Biochem Pharmacol 1961;7:88–95.
16. Dixon M, Webb CE. ENZYMES. 3rd Edition. Longman Group Limited, New York; 1965.
17. Calow P, Sibly R. A physiological basis of population processes: Ecotoxicological implications. Funct Ecol 1990;4:283-8.
18. Parvez S, Raisuddin S. Protein carbonyls: novel biomarkers of exposure to oxidative stress-inducing pesticides in freshwater fish Channa punctate (Bloch). Environ Toxicol Pharmacol 2005;20:112-7.
19. Ramaswami V, Subbram V. Effect of selected textile dye on the survival, morphology, and burrowing behavior of the earthworm Polypheretima elongata. Bull Environ Contam Toxicol 1992;48:253-8.
20. Reinecke AJ, Maboeta MS, Vermeulen LA, Reinecke SA. Assessment of lead nitrate and mancozeb toxicity in earthworms using the avoidance response. Bull Environ Contam Toxicol 2002;68:779-86.
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How to Cite
Jadhav, S. S., and D. M. “EFFECT OF FLUBENDIAMIDE ON MORPHOLOGY, AVOIDANCE BEHAVIOUR AND ACETYLCHOLINESTERASE ACTIVITY IN EARTHWORM EUDRILUS EUGENIAE”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 9, no. 9, Sept. 2017, pp. 233-8, doi:10.22159/ijpps.2017v9i9.20684.
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