Skip to main content
Log in

Genotoxicity monitoring of freshwater environments using caged carp (Cyprinus carpio)

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

The present study deals with genotoxicity assessment of freshwaters using caged carp (Cyprinus carpio). Carps were transplanted from a fish-farm to three differently polluted sites in eastern Croatia. Two polluted sites were situated in the river Drava, downstream from the cities of Belišće and Osijek, while the reference site was in the Nature Park Kopački rit, a preserved wetland area with limited anthropogenic influence. Exposure lasted for 3 weeks and was repeated for 3 years (2002–2004). DNA damage was assessed in erythrocytes of the exposed animals by the Comet assay and micronucleus test (MNT). In order to evaluate possible differences in stress responses to polluted water in situ and in aquaria a laboratory exposure was performed with water from the studied location in the second year of the study. Carp from the sites with high anthropogenic influence (Belišće and Osijek) had higher average DNA damage as expressed in both the MNT and Comet assay. Of the two, the Comet assay appeared to be more sensitive following both caging and aquaria exposures. The results from this study suggest that 3 weeks caging exposure of C. carpio may be a useful strategy to monitor for genotoxic agents in freshwater ecosystems.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Al-Sabti K (1986) The karyotypes of Cyprinus carpio and Leuciscus cephalus. Cytobios 47:19–25

    CAS  Google Scholar 

  • Al-Sabti K (1991) Handbook of genotoxic effects and fish chromosomes. Krištoff, Ljubljana

    Google Scholar 

  • Al-Sabti K, Metcalfe CD (1995) Fish micronuclei for assessing genotoxicity in water. Mutat Res 343:121–135

    Article  CAS  Google Scholar 

  • Avishai N, Rabinowitz C, Moiseeva E, Rinkevich B (2002) Genotoxicity of the Kishon River, Israel: the application of an in vitro cellular assay. Mutat Res 518:21–37

    CAS  Google Scholar 

  • Barbee GC, Barich J, Duncan B, Bickham JW, Matson CW, Hintze CJ, Autenrieth RL, Zhou G-D, McDonald TJ, Cizmas L, Norton D, Donnelly KC (2008) In situ biomonitoring of PAH-contaminated sediments using juvenile coho salmon (Oncorhynchus kisutch). Ecotoxicol Environ Saf 71:454–464

    Article  CAS  Google Scholar 

  • Belpaeme K, Cooreman K, Kirsch-Volders M (1998) Development and validation of the in vivo alkaline comet assay for detecting genomic damage in marine flatfish. Mutat Res 415:167–184

    CAS  Google Scholar 

  • Bickham JW, Smolen MJ (1994) Somatic and heritable effects of environmental genotoxins and the emergence of evolutionary toxicology. Environ Health Perspect 102:25–28

    Google Scholar 

  • Bombail V, Aw D, Gordon E, Batty J (2001) Application of the comet and micronucleus assays to butterfish (Pholis gunnellus) erythrocytes from the Firth of Forth, Scotland. Chemosphere 44:383–392

    Article  CAS  Google Scholar 

  • Bony S, Gillet C, Bouchez A, Margoum C, Devaux A (2008) Genotoxic pressure of vineyard pesticides in fish: field and mesocosm surveys. Aquat Toxicol 89:197–203

    Article  CAS  Google Scholar 

  • Brown JS, Steinert SA (2004) DNA damage and biliary PAH metabolites in flatfish from Southern California bays and harbors, and the Channel Islands. Ecol Indic 3:263–274

    Article  CAS  Google Scholar 

  • Buschini A, Martino A, Gustavino B, Monfrinotti M, Poli P, Rossi C, Santoro M, Dörr AJM, Rizzoni M (2004) Comet assay and micronucleus test in circulating erythrocytes of Cyprinus carpio specimens exposed in situ to lake waters treated with disinfectants for potabilization. Mutat Res 557:119–129

    CAS  Google Scholar 

  • Chu S, Xi Z, Xu X, Zhang Y, Xu Y (1996) Induction of micronuclei in peripheral erythrocytes of Misgurnus anguillicaudatus by polychlorinated biphenyls. Bull Environ Contam Toxicol 57:179–182

    Article  CAS  Google Scholar 

  • de Andrade VM, da Silva J, da Silva FR, Heuser VD, Dias JF, Yoneama ML, de Freitas TRO (2004) Fish as bioindicators to assess the effects of pollution in two southern Brazilian rivers using the Comet assay and micronucleus test. Environ Mol Mutagen 44:459–468

    Article  Google Scholar 

  • Devaux A, Pesonen M, Monod G (1997) Alkaline comet-assay in rainbow trout hepatocytes. Toxicol In Vitro 11:71–79

    Article  CAS  Google Scholar 

  • Devaux A, Flammarion P, Bernardon V, Garric J, Monod G (1998) Monitoring of the chemical pollution of the river Rhône through measurement of DNA damage and cytochrome P4501A induction in Chub (Leuciscus cephalus). Mar Environ Res 46:257–262

    Article  CAS  Google Scholar 

  • Flammarion P, Devaux A, Nehls S, Migeon B, Noury P, Garric J (2002) Multibiomarker responses in fish from the Moselle River (France). Ecotoxicol Environ Saf 51:145–153

    Article  CAS  Google Scholar 

  • Frenzilli G, Nigro M, Lyons BP (2009) The Comet assay for the evaluation of genotoxic impact in aquatic environments. Mutat Res 681:80–92

    Article  CAS  Google Scholar 

  • Grisolia CK, Starling FLRM (2001) Micronuclei monitoring of fishes from Lake Paranoá, under influence of sewage treatment plant discharges. Mutat Res 491:39–44

    CAS  Google Scholar 

  • Guilherme S, Válega M, Pereira ME, Santos MA, Pacheco M (2008) Erythrocytic nuclear abnormalities in wild and caged fish (Liza aurata) along an environmental mercury contamination gradient. Ecotoxicol Environ Saf 70:411–421

    Article  CAS  Google Scholar 

  • Gustavino B, Scornajenghi KA, Minissi S, Ciccotti E (2001) Micronuclei induced in erythrocytes of Cyprinus carpio (Teleostei, Pisces) by X-rays and colchicine. Mutat Res 494:151–159

    CAS  Google Scholar 

  • Hartmann A, Speit G (1997) The contribution of cytotoxicity to DNA-effects in the single cell gell test (comet assay). Toxicol Lett 90:183–188

    Article  CAS  Google Scholar 

  • Jarić D, Stepić S (2005) Differences in enzymes (biomarkers) activities in fish after cage exposure in Drava and Danube Rivers (Croatia). Verh Int Ver Limnol 29:873–876

    Google Scholar 

  • Kirsch-Volders M, Sofuni T, Aaderma M, Albertini S, Eastmond D, Fenech M, Ishidate M, Lorge E, Norppa H, Surallés J, Von der Hude W, Wakata A (2000) Report from the in vitro micronucleus assay working group. Environ Mol Mutagen 35:167–172

    Article  CAS  Google Scholar 

  • Kirsch-Volders M, Sofuni T, Aaderma M, Albertini S, Eastmond D, Fenech M, Ishidate M, Kirchner S, Lorge E, Morita T, Norppa H, Surallés J, Vanhauwaert A, Wakata A (2003) Report from the in vitro micronucleus assay working group. Mutat Res 540:153–163

    CAS  Google Scholar 

  • Klobučar GIV, Pavlica M, Erben R, Papeš D (2003) Application of the micronucleus and comet assays to mussel Dreissena polymorpha haemocytes for genotoxicity monitoring of freshwater environments. Aquat Toxicol 64:15–23

    Article  Google Scholar 

  • Lee RF, Steinert S (2003) Use of the single cell gel electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. Mutat Res 544:43–64

    Article  CAS  Google Scholar 

  • Llorente MT, Martos A, Castaño A (2002) Detection of cytogenetic alterations and blood cell changes in natural populations of carp. Ecotoxicology 11:27–34

    Article  CAS  Google Scholar 

  • Medina M, Correa JC, Barata C (2007) Micro-evolution due to pollution: possible consequences for ecosystem responses to toxic stress. Chemosphere 67:2105–2114

    Article  CAS  Google Scholar 

  • Murdoch MH, Herbert PDN (1994) Mitochondrial DNA diversity of brown bullhead from contaminated and relatively pristine sites in the Great Lakes. Environ Toxicol Chem 13:1281–1289

    Article  CAS  Google Scholar 

  • Nacci DE, Cayula S, Jackim E (1996) Detection of DNA damage in individual cells from marine organisms using the single cell gell assay. Aquat Toxicol 35:197–210

    Article  CAS  Google Scholar 

  • Oikari A (2006) Caging techniques for field exposures of fish to chemical contaminants. Aquat Toxicol 78:370–381

    Article  CAS  Google Scholar 

  • Pandrangi R, Petras M, Ralph S, Vrzoc M (1995) Alkaline single cell gel (comet) assay and genotoxicity monitoring using bullheads and carp. Environ Mol Mutagen 26:345–356

    Article  CAS  Google Scholar 

  • Peichel CL, Nereng KS, Ohgi KA, Cole BLE, Colosimo PF, Buerklet CA, Schluter D, Kingsley DM (2001) The genetic architecture of divergence between threespine stickleback species. Nature 414:901–905

    Article  CAS  Google Scholar 

  • Pellacani C, Buschini A, Furlini M, Poli P, Rossi C (2006) A battery of in vivo and in vitro tests useful for genotoxic pollutant detection in surface waters. Aquat Toxicol 77:1–10

    Article  CAS  Google Scholar 

  • Powers DA (1989) Fish as model systems. Science 246:352–358

    Article  CAS  Google Scholar 

  • Rank J, Jensen K, Jespersen PH (2005) Monitoring DNA damage in indigenous blue mussels (Mytilus edulis) sampled from coastal sites in Denmark. Mutat Res 585:33–42

    CAS  Google Scholar 

  • Russo C, Rocco L, Morescalchi MA, Stingo V (2004) Assessment of environmental stress by the micronucleus test and the Comet assay on the genome of teleost populations from two natural environments. Ecotoxicol Environ Saf 57:168–174

    Article  CAS  Google Scholar 

  • Sapone A, Gustavino B, Monfrinotti M, Canistro D, Broccoli M, Pozzetti L, Affatato A, Valgimigli L, Cantelli Forti G, Pedulli GF, Biagi GL, Abdel-Rahman SZ, Paolini M (2007) Perturbation of cytochrome P450, generation of oxidative stress and induction of DNA damage in Cyprinus carpio exposed in situ to potable surface water. Mutat Res 626:143–154

    CAS  Google Scholar 

  • Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  Google Scholar 

  • Štambuk A, Pavlica M, Vignjević G, Bolarić B, Klobučar GIV (2009) Assessment of genotoxicity in polluted freshwaters using caged painter’s mussel, Unio pictorum. Ecotoxicology 18:430–439

    Article  Google Scholar 

  • Sumathi M, Kalaiselvi K, Palanivel M, Rajaguru P (2001) Genotoxicity of textile dye effluent on fish (Cyprinus carpio) measured using the comet assay. Bull Environ Contam Toxicol 66:407–414

    Article  CAS  Google Scholar 

  • Theodorakis CW, Shugart LR (1997) Genetic ecotoxicology II: population genetic structure in mosquitofish exposed in situ to radionuclides. Ecotoxicology 6:335–354

    Article  CAS  Google Scholar 

  • Udroiu I (2006) The micronucleus test in piscine erythrocytes. Aquat Toxicol 79:201–204

    Article  CAS  Google Scholar 

  • Van der Oost R, Lopes SCC, Komen H, Satumalay K, van den Bos R, Heida H, Vermeulen NPE (1998) Assessment of environmental quality and inland water pollution using biomarker responses in caged carp (Cyprinus carpio): use of a bioactivation:detoxication ratio as a biotransformation index (BTI). Mar Environ Res 46:315–319

    Article  Google Scholar 

  • Whitehead A, Kuivila KM, Orlando JL, Kotelevtsev S, Anderson SL (2004) Genotoxicity in native fish associated with agricultural runoff events. Environ Toxicol Chem 23:2868–2877

    Article  Google Scholar 

  • Winter M, Day N, Hayes RA, Taylor EW, Butler PJ, Chipman JK (2004) DNA strand breaks and adducts determined in feral and caged chub (Leuciscus cephalus) exposed to rivers exhibiting variable water quality around Birmingham, UK. Mutat Res 552:163–175

    CAS  Google Scholar 

  • Wirzinger G, Weltje L, Gercken J, Sordyl H (2007) Genotoxic damage in field-collected three-spined sticklebacks (Gasterosteus aculeatus L.): a suitable biomonitoring tool? Mutat Res 628:19–30

    CAS  Google Scholar 

  • Žaja R, Klobučar GIV, Sauerborn Klobučar R, Hackenberger K, Smital T (2006) Haemolymph as compartment for efficient and non-destructive determination of P-glycoprotein (Pgp) mediated MXR activity in bivalves. Comp Biochem Physiol (part C) 143:103–111

    Google Scholar 

  • Zhu Y, Wang J, Bai Y, Zhang R (2004) Cadmium, chromium and copper induce polychromatocyte micronuclei in carp (Cyprinus carpio L.). Bull Environ Contam Toxicol 72:78–86

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We acknowledge support by the Research Council of Norway, for the project no. 150463 with Norwegian Institute for Water Research (NIVA) and by the Croatian Scientific Research Council for the project no. 119-0982934-3110.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Göran I. V. Klobučar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Klobučar, G.I.V., Štambuk, A., Pavlica, M. et al. Genotoxicity monitoring of freshwater environments using caged carp (Cyprinus carpio). Ecotoxicology 19, 77–84 (2010). https://doi.org/10.1007/s10646-009-0390-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-009-0390-6

Keywords

Navigation