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Investigation of the genotoxicity of endosulfan to freshwater Cyprinid fish Crucian carp (Carassius carassius L.) using the micronucleus and chromosomal aberration as biomarkers

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Abstract

This study was aimed to evaluate the in vivo genotoxic effects of endosulfan in freshwater fish crucian carp, Carassius carassius L. The LC50–96h value of endosulfan (with 95 % confidence limits), determined by probit analysis, was 0.070 (0.046–0.093) ppm L−1 and the estimated safe levels of endosulfan varied from 00.70 × 10−2 to 00.70 × 10−6 ppm L−1. Based on the LC50–96h value, three test concentrations (viz. sub-lethal I, II, and III) were determined to be 0.052, 0.035 and 0.017 ppm L−1, respectively, and the fish specimens were exposed to these concentrations for 35 days. The mean concentration of endosulfan in water samples (aquaria) was always within 5 % of the intended concentration when analyzed by dispersive liquid-liquid micro extraction (DLLME) followed by gas chromatography–mass spectrometry (GC-MS). Autopsy was done on days 1, 2, 3, 4, 7, 14, 21, 28 and 35 of endosulfan exposure for assessment of micronucleus (MN) formation and chromosomal aberrations (CA). The MN formation in the peripheral erythrocytes, authenticated by scanning electron microscopy (SEM), was found to be significantly higher (p < 0.05) in all the treated groups, including positive control cyclophosphamide (4 ppm L−1) compared to negative control. Similar significant effects (p < 0.05) were also observed on CA in the head kidney cells. In general, both MN and CA exhibited a concentration and time-dependent response. The current study revealed the potential genotoxicity of endosulfan in fish; and that the micronucleus and chromosomal assays are useful tools in determining genotoxicity of water pollutants and might be appropriate as a part of environmental monitoring program.

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References

  1. Ali D, Nagpure NS, Kumar S, Kumar R, Kushwaha B, Lakra WS. Assessment of genotoxic and mutagenic effects of chlorpyrifos in freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single cell gel electrophoresis. Food Chem Toxicol. 2009;47:650–6.

    Article  CAS  PubMed  Google Scholar 

  2. Al-Sabti K, Fijan N, Kurelec B. A simple and fast technique for chromosome preparation in fish. Veterinarski Arch. 1983;53:283–90.

    Google Scholar 

  3. Al-Sabti K, Metcalfe CD. Fish micronuclei for assessing genotoxicity in water. Mutat Res. 1995;2:121–35.

    Article  Google Scholar 

  4. Ameur WB, Trabelsi S, El Megdiche Y, Hassine SB, Barhoumi B, Hammami B, et al. Concentration of polychlorinated biphenyls and organochlorine pesticides in mullet (Mugil cephalus) and sea bass (Dicentrarchus labrax) from Bizerte Lagoon (Northern Tunisia). Chemosphere. 2013;90:2372–80.

    Article  PubMed  Google Scholar 

  5. Ansari RA, Rahman S, Kaur M, Anjum S, Raisuddin S. In vivo cytogenetic and oxidative stress-inducing effects of cypermethrin in freshwater fish, Channa punctata Bloch. Ecotoxicol Environ Saf. 2011;74:150–6.

    Article  CAS  PubMed  Google Scholar 

  6. APHA, AWWA, WPCF. Standard methods for the examination of water and wastewater. 21st ed. Washington: American Publication of Health Association; 2005.

    Google Scholar 

  7. Assocham. Debunking theory of India’s pesticide consumption being highest. A Report on “Pesticide residues in Indian food and agricultural products” submitted to Govt. India 2007. Available from http://www.freshplaza.com/news_detail.asp?id=1265.

  8. Buikema JR, Naider-Lehner AL, Cairns JR. Biological monitoring: part IV. Toxicity testing Environ Mol Mutagen. 1982;33:287–92.

    Google Scholar 

  9. Bull S, Fletcher K, Boobis A, Batterrshill J. Evidence for genotoxicity of pesticides in pesticide applicators. Mutagenesis. 2006;21:93–103.

    Article  CAS  PubMed  Google Scholar 

  10. Burkepile DE, Moore MT, Holland MM. Susceptibility of five non-target organisms to aqueous diazinon exposure. Bull Environ Contam Toxicol. 2000;64:114–21.

    Article  CAS  PubMed  Google Scholar 

  11. Cabaleiro T, Caride A, Romero A, Lafuente A. Effects of in utero and lactational exposure to endosulfan in prefrontal cortex of male rats. Toxicol Lett. 2008;176:58–67.

    Article  CAS  PubMed  Google Scholar 

  12. Chaudhary R, Pandey S, Kushwaha B, Gaur KK, Nagpure NS. Fish micronucleus assay: a sensitive tool for ecogenotoxicity studies. J Ecophysiol Occup Health. 2006;6:143–7.

    CAS  Google Scholar 

  13. Chaurasia OP, Kumari C. Genotoxic effect of ground water salts rich in fluoride. Cytologia. 2007;72:141–4.

    Article  CAS  Google Scholar 

  14. Chorvatovicová D, Sandula J. Effect of carboxymethyl-chitinglucan on cyclophosphamide induced mutagenicity. Mutat Res. 1995;346:43–8.

    Article  PubMed  Google Scholar 

  15. Committee on Water Quality Criteria (CWQC), A Report of the Committee on Water Quality Criteria. Ecol Res Series, EPA-R3-73-003;1972.

  16. Cucchi C, Baruffaldi A. A new method for karyological studies in teleost fish. J Fish Biol. 1990;37:71–5.

    Article  CAS  Google Scholar 

  17. El Bakouri H, Palacios-Santander JM, Cubillana-Aguilera L, Ouassini A, Naranjo-Rodríguez I. Hidalgo-Hidalgo de Cisneros JL. Electrochemical analysis of endosulfan using a C18-modified carbon-paste electrode. Chemosphere. 2005;60:1565–71.

    Article  PubMed  Google Scholar 

  18. Farah MA, Ateeq B, Ahmad W. Antimutagenic effect of neem leaves extract in freshwater fish, Channa punctatus evaluated by cytogenetic tests. Sci Total Environ. 2006;364:200–14.

    Article  CAS  PubMed  Google Scholar 

  19. Fenech M, Chang WP, Kirsch-Volders M, Holland N, Bonassi S, Zeiger E. HUMN project: detailed description of the scoring criteria for the cytokinesis block micronucleus assay using isolated human lymphocyte cultures. Mutat Res. 2003;534:65–75.

    Article  CAS  PubMed  Google Scholar 

  20. Finney DJ. Probit analysis. Cambridge: Cambridge University Press; 1971. p. 333.

    Google Scholar 

  21. Ganeshwade RM, Dama LB, Deshmukh DR, Ghanbahadur AG, Sonawane SR. Toxicity of endosulfan on freshwater fish Channa striatus. Trends in Fish Res. 2012;1:29–31.

    Google Scholar 

  22. Garcia-Reyero NDenslow ND. Applications of genomic technologies to the study of organochlorine pesticide induced reproductive toxicity in fish. J Pestic Sci. 2006;31:252–62.

    Article  Google Scholar 

  23. Hussein SY, El-Nasser MA, Ahmed SM. Comparative studies on the effects of herbicide atrazine on fresh water fish Oreochromis niloticus and Chrysichthyes auratus at Assiut, Egypt. Bull Environ Contam Toxicol. 1996;57:503–10.

    Article  CAS  PubMed  Google Scholar 

  24. IJC. New and Revised Great Lakes Water Quality Objectives. Windsor, Ontario. 1977; p. 1.

  25. Johnson WW, Finley MT. Handbook of acute toxicity of chemicals to fish and aquatic invertebrates. In: Resource publication 137. U.S. Department of interior. Washington: Fish and Wildlife Service; 1980. p. 6–56.

    Google Scholar 

  26. Klobucar GIV, Stambuk A, Pavlica M, SerticPeric M, Kutuzovic-Hackenberger B, Hylland K. Genotoxicity monitoring of freshwater environments using caged carp (Cyprinus carpio). Ecotoxicology. 2010;19:77–84.

    Article  CAS  PubMed  Google Scholar 

  27. Knytl M, Kalous L, Petr R. Karyotype and chromosome banding of endangered crucian carp, Carassius carassius (Linnaeus, 1758) (Teleostei, Cyprinidae). Comp Cytogen. 2013;7:205–13.

    Article  Google Scholar 

  28. Kumar R, Nagpure NS, Kushwaha B, Srivastava SK, Lakra WS. Investigation of the genotoxicity of malathion to freshwater teleost fish Channa punctatus (Bloch) using the micronucleus test and comet assay. Arch Environ Contam Toxicol. 2009;58:123–30.

    Article  PubMed  Google Scholar 

  29. Marigoudar SR, Nazeer AR, David M. Impact of cypermethrin on behavioural responses in the freshwater teleost, Labeo rohita (Hamilton). World J Zool. 2009;4:19–23.

    Google Scholar 

  30. Marques de Miranda Cabral Gontijo Á, Barreto RE, Speit G, Valenzuela Reyes VA, Luiz Volpato G, Favero Salvadori DM. Anesthesia of fish with benzocaine does not interfere with comet assay results. Mutat Res. 2003;534:165–72.

    Article  Google Scholar 

  31. Minissi S, Ciccotti E, Rizzoni M. Micronucleus test in erythrocytes of Barbus plebejus (Teleostei, Pisces) from two natural environments: a bioassay for the in situ detection of mutagens in freshwater. Mutat Res. 1996;367:245–51.

    Article  CAS  PubMed  Google Scholar 

  32. NAS/NAE. Water Quality Criteria, US Government Printing Office, Washington, DC. EPA-R3-033; 1973.

  33. Nwani CD, Lakra WS, Nagpure NS, Kumar R, Kushwaha B, Srivastava SK. Toxicity of the herbicide atrazine: effects on lipid peroxidation and activities of antioxidant enzymes in the freshwater fish Channa Punctatus (Bloch). Int J Environ Res Public Health. 2010;7:3298–312.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  34. Ondarza PM, Gonzalez M, Fillmann G, Miglioranza KSB. PBDEs, PCBs and organochlorine pesticides distribution in edible fish from Negro River basin. Argentinean Patagonia Chemosphere. 2014;94:135–42.

    CAS  Google Scholar 

  35. Osman AGM, Wuertz S, Mekkawy IA, Exner HJ, Kirschbaum F. Lead induced malformations in embryos of the African catfish Clarias gariepinus (Burchell, 1822). Environ Toxicol. 2007;22:375–89.

    Article  CAS  PubMed  Google Scholar 

  36. Özkan F, Gündüz SG, Berköz M, Hunt AÖ. Induction of micronuclei and other nuclear abnormalities in peripheral erythrocytes of Nile tilapia, Oreochromis niloticus, following exposure to sublethal cadmium doses. Turk J Zool. 2011;35:585–92.

    Google Scholar 

  37. Pandey S, Kumar R, Sharma S, Nagpure NS, Srivastava SK, Verma MS. Acute toxicity bioassays of mercuric chloride and malathion on air breathing fish Channa punctatus (Bloch). Ecotoxicol Environ Saf. 2005;61:114–20.

    Article  CAS  PubMed  Google Scholar 

  38. Pandey S, Nagpure NS, Kumar R, Sharma S, Srivastava SK, Verma MS. Genotoxicity evaluation of acute doses of endosulfan to freshwater teleost Channa punctatus (Bloch) by alkaline single-cell gel electrophoresis. Ecotoxicol Environ Saf. 2006;65:56–61.

    Article  CAS  PubMed  Google Scholar 

  39. Pathak R, Suke SG, Ahmed T, Ahmed RS, Tripathi AK, Guleria K, et al. Organochlorine pesticide residue and oxidative stress in pattern delivery cases. Hum Exp Toxicol. 2010;29:351–8.

    Article  CAS  PubMed  Google Scholar 

  40. Raisuddin S, Jha AN. Relative sensitivity of fish and mammalian cells to sodium arsenate and arsenite as determined by alkaline single-cell gel electrophoresis and cytokinesis-block micronucleus assay. Environ Mol Mutagen. 2004;44:83–9.

    Article  CAS  PubMed  Google Scholar 

  41. Reimer L. Scanning electron microscopy: physics of image formation and of microanalysis. Berlin: Springer; 1985.

    Book  Google Scholar 

  42. Rishi KK, Grewal S. Chromosome aberration test for the insecticide, dichlorvos, on fish chromosomes. Mutat Res. 1995;344:1–4.

    Article  CAS  PubMed  Google Scholar 

  43. Rita AJJ, Milton JMC. Karyomorphological analysis of the fresh water cichlid Oreochromis mossambicus (Peter) exposed to carbamate pesticide methomyl (Lannate). J Adv Zool. 2008;29:57–61.

    CAS  Google Scholar 

  44. Saglio P, Trijasse S. Behavioural responses to atrazine and diuron in goldfish. Arch Environ Contam Toxicol. 1998;35:484–91.

    Article  CAS  PubMed  Google Scholar 

  45. Sharma S, Nagpure NS, Kumar R, Pandey S, Singh PJ, Srivastava SK, et al. Studies on the genotoxicity of endosulfan in different tissue of fresh water fish Mystus vittatus using the Comet assay. Arch Environ Contam Toxicol. 2007;53:617–23.

    Article  CAS  PubMed  Google Scholar 

  46. Shegefti S, Sereshti H, Samadi S. Determination of endosulfan in water samples using dispersive liquid-liquid micro-extraction and experimental design for optimization. Int J Environ Res. 2010;4:237–46.

    CAS  Google Scholar 

  47. Sprague JB. Measurement of pollutant toxicity to fish: III. Sublethal effects and “SAFE” concentrations. Water Res. 1971;5:245.

    Article  CAS  Google Scholar 

  48. Taylor D, Green N, Stout G. Biological science. 3rd ed. Australia: Cambridge University Press; 1997.

    Google Scholar 

  49. Velisek J, Svobodova Z, Piackova V, Sudova E. Effects of acute exposure of metribuzin on some haematological, biochemical and histopathological parameters of common carp (Cyprinus carpio L.). Bull Environ Contam Toxicol. 2009;82:492–5.

    Article  CAS  PubMed  Google Scholar 

  50. Vernier P, Maron S, Canova S. Detection of micronuclei in gill cells and haemocytes of mussels exposed to Benzo [a] pyrene. Mutat Res. 1997;390:33–44.

    Article  Google Scholar 

  51. Yadav KK, Trivedi SP. Chromosomal aberrations in a fish, Channa punctata after in vivo exposure to three heavy metals. Mutat Res Genet Toxicol Environ Mutagen. 2009;678:7–12.

    Article  CAS  Google Scholar 

  52. Zeljezic D, Garaj-Vrhovac V. Sister chromatid exchange and proliferative rate index in the longitudinal risk assessment of occupational exposure to pesticides. Chemosphere. 2002;46:295–303.

    Article  CAS  PubMed  Google Scholar 

  53. Zoller JF, Hohmann U, Herrmann RG, Wanner G. Ultrastructural analysis in meiosis I and II of rye (Secale cereale L.). Cytogenetic and. Genome Res. 2004;105:145–56.

    Article  CAS  Google Scholar 

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Acknowledgments

The authors are grateful to Director of the Centre of Research for Development (CORD), University of Kashmir, Srinagar, India for providing laboratory facilities in the course of this research and University Grants Commission (UGC), New Delhi, India for providing financial assistance to Mr. Sabzar in the form of Junior Research Fellowship (JRF). The authors also thank the technicians of Advanced Instrumentation Research Facility (AIRF), Jawaharlal Nehru University, New Delhi and University Sophisticated Instruments Facility (USIF), Aligarh Muslim University, U.P. for their cordial support in carrying out the GC-MS and SEM analysis, respectively.

Conflict of interest

The authors report no conflicts of interest.

Author contributions statement

First author carried out the experimental work and writing, author second and third designed and calibrated the experimental setup, fourth author carried out the statistical analysis and fifth author carried out the writing and critical evaluation, along with the first author, of the manuscript.

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  1. Limnology and Fisheries Laboratory, Centre of Research for Development (CORD), University of Kashmir, Srinagar, 190006, J & K, India

    Sabzar Ahmad Dar, Abdul Rehman Yousuf & Farooq Ahmad Ganai

  2. Division of Fisheries, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir (SKUAST-K), Srinagar, J & K, India

    Masood-ul-Hassan Balkhi & Farooz Ahmad Bhat

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  1. Sabzar Ahmad Dar
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Correspondence to Sabzar Ahmad Dar.

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Dar, S.A., Yousuf, A.R., Balkhi, MuH. et al. Investigation of the genotoxicity of endosulfan to freshwater Cyprinid fish Crucian carp (Carassius carassius L.) using the micronucleus and chromosomal aberration as biomarkers. Nucleus 57, 87–98 (2014). https://doi.org/10.1007/s13237-014-0110-3

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