, Volume 18, Issue 3, pp 325–333 | Cite as

Acetylcholinesterase: a potential biochemical indicator for biomonitoring of fertilizer industry effluent toxicity in freshwater teleost, Channa striatus

  • Archana Yadav
  • Anita Gopesh
  • Ravi S. Pandey
  • Devendra K. Rai
  • Bechan Sharma


Monitoring of acetylcholinesterase (EC:, AChE) activity has been widely used in aquatic and terrestrial systems as an indicator of pollutant exposure. The reports regarding impact of fertilizer industry effluent on the level of AChE activity are very scanty. In this paper, an attempt has been made to investigate the in vitro impact of fertilizer industry effluent upon the levels of AChE activity and protein content in different tissues of non-target aquatic fish, Channa striatus (Bloch). The fish when exposed to three sublethal concentrations (3.5, 4.7, and 7.0%; v/v) of fertilizer industry effluent for short (96 h) and long (15 days) durations registered sharp reduction in the levels of AChE activity (15–75%) and protein (10–71%) in different fish organs. The highest effluent concentration treatment for short or long duration, the fish brain and gills registered significant (P < 0.001) inhibition (64–75%) in the activity of AChE whereas other organs such as muscles, liver, and heart exhibited slightly lower inhibition (40–59%) in enzyme activity. However, kidney of C. striatus was the only organ where very less effect (14–18%) of the effluent was observed on the activity of AChE when the fish were exposed to all the three concentrations of the effluent for both treatment durations. This effluent also induced alterations in the level of protein in different fish organs; in kidney the effect was pronounced only at higher concentrations at both treatment durations. The most affected organs were muscle and gills where in 60–71% reduction in the protein content was recorded due to highest effluent concentration treatment at short or long durations. The results of present study indicated that the fertilizer industry effluents might significantly influence the neurotransmission system and protein turnover in the non-target organisms after exposure even at very low concentrations. Further, the data suggested that the fish AChE could be used as a potential biochemical marker for fertilizer industry effluent pollution in aquatic systems.


Fertilizer industry effluent Acetylcholinesterase Protein Inhibition Biomarker 



The authors express their gratitude to Professor Pratima Gaur, Head, Department of Zoology, University of Allahabad, Allahabad, for providing basic facilities for carrying out the present research work. The central research facility developed at the Department of Zoology with the help of financial assistance obtained from UGC-SAP and DST-FIST was utilized in this study


  1. Abarnou A, Burgeot T, Chevreuil M, Leboulenger F, Loizeau V, Madoulet-Jaouen A, Minier C (2000) Les contaminants organiques: quels risques pour le monde vivant? Programme Scientifique Seine Aval 13. Ifremer, ParisGoogle Scholar
  2. Adham KG (2002) Sublethal effect of aquatic pollution in Lake Maryut on the African sharptooth catfish, Clarias garipinus (Burchell, 1822). J Appl Ichthyol 18:87–94. doi: 10.1046/j.1439-0426.2002.00337.x CrossRefGoogle Scholar
  3. APHA/AWWA/WPCF (1975) Standard methods for the examination of water and wastewater, 14th edn. American Public Health Association/American water work association/water pollution control federation, Washington, DCGoogle Scholar
  4. Ausley LW (2000) Reflections on whole effluent toxicity: the Pellstone workshops. Environ Toxicol Chem 19:1–2. doi:10.1897/1551-5028(2000)019<0001:ROWETT>2.3.CO;2CrossRefGoogle Scholar
  5. Bobmanuel NOK, Gabriel UU, Ekweozor IKE (2006) Direct toxic assessment of treated fertilizer effluents to Orechromis niloticus, Clarias gariepinus and catfish hybrid (Heterobranchus bidorsalis(male) × Clarias gariepinus(female)). Afr J Biotechnol 5(8):635–642Google Scholar
  6. Bocquené G, Bellanger C, Cadiou Y (1995) Joint action of combinations of pollutants on the acetylcholinesterase activity of several marine species. Ecotoxicology 4:266–279. doi: 10.1007/BF00116345 CrossRefGoogle Scholar
  7. Bocquene G, Galgani F, Truquet P (1990) Characterization and assay conditions for use of AChE activity from several marine species in pollution monitoring. Marine species in pollution monitoring. Mar Environ Res 30:75–89. doi: 10.1016/0141-1136(90)90012-D CrossRefGoogle Scholar
  8. Chapman PM (2000) Whole effluent toxicity testing—usefulness, level of protection and risk assessment. Environ Toxicol Chem 19:3–19. doi:10.1897/1551-5028(2000)019<0003:WETTUL>2.3.CO;2CrossRefGoogle Scholar
  9. Chen C-J, Liao S-L (2003) Zinc toxicity on neonatal cortical neuron: involvement of glutathione chelate. J Neurochem 85:443–453. doi: 10.1046/j.1471-4159.2003.01691.x CrossRefGoogle Scholar
  10. Chiffoleau JF, Gonzalez JL, Miramand P, Thouvenin B (1999) Le cadmium, comportement d’un contaminant métallique en estuaire. Programme Scientifique Seine Aval, 10. Ifremer, ParisGoogle Scholar
  11. Chukwu LO (1993) Evaluation of pollutant load of WEMABOD treated industrial effluent, Lagos. Nig J Biotechnol 11:142–147Google Scholar
  12. De la Torre FL, Ferrari R, Salibian A (2002) Freshwater pollution biomarker: response of brain acetylcholinesterase activity in two fish species. Comp Biochem Physiol Toxicol Pharmacol 131(3):271–280. doi: 10.1016/S1532-0456(02)00014-5 CrossRefGoogle Scholar
  13. Effiok BJ (1993) Basic calculations for chemical and biological analyses. AOAC, Baltimore, p 150Google Scholar
  14. Ekweozor IKE, Bobmanuel NOK, Gabriel UU (2001) Sublethal effect of ammonical fertilizer effluents on three commercial fish species from Niger Delta Area, Nigeria. J Appl Sci Environ Mang 5(1):63–68Google Scholar
  15. Ellman GL, Courtney D, Andres V Jr, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–96. doi: 10.1016/0006-2952(61)90145-9 CrossRefGoogle Scholar
  16. Engel DW, Sundu WG, Fowler BA (1981) Factor effecting trace metal uptake and toxicity to estuarine organisms. Academic Press, LondonGoogle Scholar
  17. Escartin E, Porte C (1997) The use of cholinesterase and carboxylesterase activities from Mytilus galloprovincialis in pollution monitoring. Environ Toxicol Chem 16:2090–2095. doi:10.1897/1551-5028(1997)016<2090:TUOCAC>2.3.CO;2CrossRefGoogle Scholar
  18. Federal Environmental Protection Agency (FEPA) Regulations (1991) Abuja: S.1.8 effluent and S.1.9 pollution abatement in industries and facilities generating wastesGoogle Scholar
  19. Forget J, Pavillon JF, Beliaeff B, Bocquene G (1999) Joint action of pollutant combinations (pesticides and metals) on survival (LC50 values) and acetylcholinesterase activity of Tigriopus brevicornis (Copepoda, harpacticoida). Environ Toxicol Chem 18:912–918. doi:10.1897/1551-5028(1999)018<0912:JAOPCP>2.3.CO;2CrossRefGoogle Scholar
  20. Fulton MH, Key PB (2001) Acetylcholinesterase inhibition in estuarine fish and invertebrates as an indicator of organophosphorus insecticide exposure and effects. Environ Toxicol Chem 20:37–45. doi:10.1897/1551-5028(2001)020<0037:AIIEFA>2.0.CO;2CrossRefGoogle Scholar
  21. Gaitonde D, Sarkar A, Kaisary S, Silva CD, Dias C, Rao DP, Ray D, Nagarjan R, De sousa SN, Sarker S, Patill D (2006) Acetylcholinesterase activities in marine snail (Cronia contracta) as a biomarker of neurotoxic contaminants along the Goa coast, West coast of India. Ecotoxicology 15(4):353–358. doi: 10.1007/s10646-006-0075-3 CrossRefGoogle Scholar
  22. Kirby MF, Morris S, Hurst M, Kirby SJ, Neall P, Tylor T, Fagg A (2000) The use of cholinesterase activity in flounder (Platichthys flesus) muscle tissue as a biomarker of neurotoxic contamination in UK estuaries. Mar Pollut Bull 40:780–791. doi: 10.1016/S0025-326X(00)00069-2 CrossRefGoogle Scholar
  23. Kopecka J, Lehtonen KK, Barsiene J, Broeg K, Vuorinen PJ, Gercken J, Pempkowiak J (2006) Measurement of biomarker levels in flounder (Platichthys flesus) and blue mussel (Mytilus trossulus) from the Gulf of Gdansk (southern Baltic). Mar Pollut Bull 53:406–421. doi: 10.1016/j.marpolbul.2006.03.008 CrossRefGoogle Scholar
  24. Kupechella CE, Hyland MC (1989) Environmental science. Allyn and Baron, LondonGoogle Scholar
  25. Lionetto MG, Caricato R, Giordano ME, Pascariello MF, Marinosci L, Schettino T (2003) Integrated use of biomarkers (acetylcholinesterase and antioxidant enzymes activities) in Mytilus galloprovincialis and Mullus barbatus in an Italian coastal marine area. Mar Pollut Bull 46(3):324–330. doi: 10.1016/S0025-326X(02)00403-4 CrossRefGoogle Scholar
  26. Lloyd R (1992) Pollution and freshwater fish. Fishing-news books. Division of Blackwell Scientific Publications, Ltd, UK, p 176Google Scholar
  27. Lorz HW, McPherson BP (1976) Effects of copper or zinc in fresh water on the adaptation to seawater and ATPase activity and the effects of copper on migratory disposition of coho salmon (Oncorhynchus kisutch). J Fish Res Board Can 33:2023–2030Google Scholar
  28. Lowry OH, Rosebrough NJ, Farr AL, Randel RJ (1951) Protein measurement with folin-phenol reagent. J Biol Chem 192:265–275Google Scholar
  29. Matthiessen P, Thain J, Law RJ, Fileman TW (1993) Attempts to assess the environmental hazard posed by complex mixtures of organic chemicals in UK estuaries. Mar Pollut Bull 26:90–95. doi: 10.1016/0025-326X(93)90097-4 CrossRefGoogle Scholar
  30. Najimi S, Bouhaimi A, Daubèze M, Zekhnini A, Pellerin J, Narbonne JF, Moukrim A (1997) Use of acetylcholinesterase in Perna perna and Mytilus galloprovincialis as a biomarker of pollution in Agadir marine bay (South of Morocco). Bull Environ Contam Toxicol 58(6):901–908. doi: 10.1007/s001289900419 CrossRefGoogle Scholar
  31. Nussey G (1998) Metal ecotoxicology of the upper Olifants river at selected localities and the effect of copper and zinc on fish blood physiology. Ph.D. thesis, RAUGoogle Scholar
  32. Obodo GA (2002) The bioaccumulation of heavy metals in fish from the lower reaches of river Niger. J Chem Soc Niger 27(2):173–176Google Scholar
  33. Ojuola FA, Onuoha GC (1987) The effect of liquid petroleum refinery effluent on fingerlings of Sarotherodon melanotheron (Ruppel 1852) and Oreochromis niloticus (Linn. 1757). Aquaculture working paper ARAC/87/WP, 8Google Scholar
  34. Parvez S, Raisuddin S (2005) Protein carbonyls: novel biomarkers of exposure to oxidative stress inducing pesticides in freshwater fish Channa punctata Bloch. Environ Toxicol Pharmacol 20:112–117. doi: 10.1016/j.etap.2004.11.002 CrossRefGoogle Scholar
  35. Payne JFA, Mathieu WM, Fancey LL (1996) Acetylcholinesterase, an old biomarker with a new future. Field trilas in association with two urban rivers and a paper mill in Newfoundland. Mar Pollut Bull 32:225–231CrossRefGoogle Scholar
  36. Ramakritinan CM, Kumaraguru AK, Balasubramanian MP (2005) Impact of distillery effluent on carbohydrate metabolism of freshwater fish, Cyprinus carpio. Ecotoxicology 14:693–707. doi: 10.1007/s10646-005-0019-3 CrossRefGoogle Scholar
  37. Rand GM, Petrocelli SM (1984) Fundamentals of aquatic toxicology methods and applications. McGraw-Hill International Book Company, New York, p 666Google Scholar
  38. Roger TD (1980) Mechanism and regulation of protein degradation in animal cells. In: Clemens HJ, Ashwell M (eds) Biochemistry of cellular regulation, vol 2. CRS Press, Florida, pp 101–122Google Scholar
  39. Sarkar A, Ray D, Shrivastava AN, Sarker S (2006) Molecular biomarkers: their significance and application in marine pollution monitoring. Ecotoxicology 15(4):333–340CrossRefGoogle Scholar
  40. Scholz NL, Truelove NK, Labenia JS, Baldwin DH, Collier TK (2006) Dose-additive inhibition of chinook salmon acetylcholinesterase activity by mixtures of organophosphate and carbamate insecticides. Environ Toxicol Chem 25(5):1200–1207. doi: 10.1897/05-030R1.1 CrossRefGoogle Scholar
  41. Sharma B (1999) Effect of carbaryl on some biochemical constituents of the blood and liver of Clarias batrachus, a fresh water fish. J Toxicol Sci 24:157–164Google Scholar
  42. Singh RK, Sharma B (1998) Carbofuran induced biochemical changes in Clarias batrachus. Pestic Sci 53:285–290. doi:10.1002/(SICI)1096-9063(199808)53:4<285::AID-PS771>3.0.CO;2-0CrossRefGoogle Scholar
  43. Singh RK, Sharma B (2004) In vivo alteration in protein metabolism by subacute carbofuran intoxication in the freshwater teleost, Clarias batrachus. Bull Environ Contam Toxicol 73(5):919–926. doi: 10.1007/s00128-004-0514-x CrossRefGoogle Scholar
  44. Singh RK, Sharma B (2005) Sub-acute toxicity of carbofuran on acetylcholinesterase activity in the freshwater catfish, Clarias batrachus. J Environ Occup Med 22(5):403–407Google Scholar
  45. Singh RK, Singh RL, Sharma B (2003) Acute toxicity of carbofuran to a freshwater teleost, Clarias batrachus. Bull Environ Contam Toxicol 70(6):1259–1263. doi: 10.1007/s00128-003-0118-x CrossRefGoogle Scholar
  46. Tamburlini G, Ehrenstein OV, Bertollini R (2002) Children’s health and environment: a review of evidence. In: Environmental issue report no. 129, WHO/European Environment Agency, WHO Geneva, 223Google Scholar
  47. Thurston RV, Russo TL, Smith CE (1978) Acute toxicity of ammonia and nitrate to cutthroat fry. Trans Am Fish Soc 107:361–368. doi:10.1577/1548-8659(1978)107<361:ATOAAN>2.0.CO;2CrossRefGoogle Scholar
  48. Tilak KS, Vereraiah K, Rao DK (2005) The effect of chloropyrifos, an organophosphate in acetylcolinestrase activity in fresh water fishes. J Environ Biol 26(1):73–78Google Scholar
  49. Tripathi G, Verma P (2004) Endosulfan-mediated biochemical changes in the fresh water Clarias batrachus. Biomed Environ Sci 17(1):47–56Google Scholar
  50. Valarmathi S, Azariah J (2003) Effect of copper chloride on the enzyme activities of the Crab, Sesarma quadratum (Fabricius). Tark J Biol 27:253–256Google Scholar
  51. Wepener V, Van Vuren JHJ, Du Preez HH (2001) Uptake and distribution of a copper, iron and zinc mixturein gill, liver and plasma of a freshwater teleost, Tilapia sparmanii. Water SA 27(1):99–108Google Scholar
  52. WHO (2002) Water pollutants. Biological agents, dissolved chemicals, non-dissolved chemicals, sediments, heat. WHO CEHA, AmmanGoogle Scholar
  53. Williams AK, Sova RC (1966) Acetylcholinesterase levels in brains of fishes from polluted waters. Bull Environ Contam Toxicol 1:198–204. doi: 10.1007/BF01684097 CrossRefGoogle Scholar
  54. Yadav A, Gopesh A, Pandey RS, Rai DK, Sharma B (2007) Fertilizer industry effluent induced biochemical changes in freshwater teleost Channa striatus (Bloch). Bull Environ Contam Toxicol 79:588–595. doi: 10.1007/s00128-007-9294-4 CrossRefGoogle Scholar
  55. Yadav A, Neraliya S, Singh R (2005) Effect of fertilizer industrial effluent on the behavior and morphology of fresh water catfish, Heteropeneustes fossilis (Bloch). Proc Nat Acad Sci India 75(B)(111):191–195. Google Scholar
  56. Yadav A, Singh RK, Sharma B (1998) Interaction of carbofuran with acetylcholinesterase from the brain of the teleost, Clarias batrachus. Toxicol Environ Chem 65:245–254. doi: 10.1080/02772249809358574 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Archana Yadav
    • 1
  • Anita Gopesh
    • 1
  • Ravi S. Pandey
    • 1
  • Devendra K. Rai
    • 2
  • Bechan Sharma
    • 2
  1. 1.Department of ZoologyUniversity of AllahabadAllahabadIndia
  2. 2.Department of BiochemistryUniversity of AllahabadAllahabadIndia

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