Cage Exposure of European Sea Bass (Dicentrarchus Labrax) for in Situ Assessment of Pollution-Related Genotoxicity

Šrut, Maja; Štambuk, Anamaria; Pavlica, Mirjana; Klobučar, Göran

doi:10.2478/10004-1254-61-2010-1988

Abstract

Genotoxic effects are often the earliest signs of pollution-related environmental disturbance. In this study, we used the comet assay and micronucleus test to assess DNA damage in the erythrocytes of the European sea bass (Dicentrarchus labrax) exposed to environmental pollution in situ. Fish were collected from a fish farm in the Trogir Bay and their cages placed at an unpolluted reference site Šolta (Nečujam Bay) and a polluted site Vranjic (Kaštela Bay) for four weeks. A group of fish which remained at the fish farm Trogir Bay were used as the second control group. Fish exposed at the Vranjic site showed a significantly higher erythrocyte DNA damage, measured by the comet assay, than either control group. Micronucleus induction showed a similar gradient of DNA damage, but did not reach statistical significance. Our results show that cage exposure of a marine fish D. labrax can be useful in environmental biomonitoring and confirm the comet assay as a suitable tool for detecting pollution-related genotoxicity.

References

Vlahogianni T, Dassenakis M, Scoullos MJ, Valavanidis A. Integrated use of biomarkers (superoxide dismutase, catalase and lipid peroxidation) in mussels Mytilus galloprovincialis for assessing heavy metals' pollution in coastal areas from the Saronikos Gulf of Greece. Mar Pollut Bull 2007;54:1136-71.10.1016/j.marpolbul.2007.05.018
Search in Google Scholar
Jakšić Ž, Batel R, Bihari N, Mičić M, Zahn RK. Adriatic coast as a microcosm for global genotoxic marine contamination - a long-term field study. Mar Pollut Bull 2005;50:1314-27.10.1016/j.marpolbul.2005.04.046
Search in Google Scholar
Knezić S, Margeta J. Integrated management of coastal sewerage systems: the case of Kaštela Bay, Croatia. Water Resour Manag 2002;16:279-305.10.1023/A:1021957308487
Search in Google Scholar
Milun V, Barić A, Zvonarić T. Temporal and spatial distribution of chlorinated hydrocarbons in mussels from the Kaštela Bay (Adriatic Sea). Fresenius Environ Bull 2004;13:1237-43.
Search in Google Scholar
Kljaković-Gašpić Z, Odžak N, Ujević I, Zvonarić T, Horvat M, Barić A. Biomonitoring of mercury in polluted coastal area using transplanted mussels. Sci Total Environ 2006;368:199-209.10.1016/j.scitotenv.2005.09.080
Search in Google Scholar
Russo C, Rocco L, Morescalchi MA, Stingo V. 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 2004;57:168-74.10.1016/S0147-6513(03)00027-7
Search in Google Scholar
Ahmad I, Maria VL, Oliveira M, Serafim A, Bebianno MJ, Pacheco M, Santos MA. DNA damage and lipid peroxidation vs. protection responses in the gill of Dicentrarchus labrax L. from a contaminated coastal lagoon (Ria de Aveiro, Portugal). Sci Total Environ 2008;406:298-397.10.1016/j.scitotenv.2008.06.027
Search in Google Scholar
Gustavino B, Scornajenghi KA, Minissi S, Ciccotti E. Micronuclei induced in erythrocytes of Cyprinus carpio (teleostei, pisces) by X-rays and colchicine. Mutat Res 2001;494:151-9.10.1016/S1383-5718(01)00191-7
Search in Google Scholar
de Lemos CT, Rödel PM, Terra NR, Oliveira NCD'A, Erdtmann B. River water genotoxicity evaluation using micronucleus assay in fish erythrocytes. Ecotoxicol Environ Saf 2007;66:391-401.10.1016/j.ecoenv.2006.01.004
Search in Google Scholar
Pandrangi R, Petras M, Ralph S, Vrzoc M. Alkaline single cell gell (Comet) assay and genotoxicity monitoring using bullheads and carp. Environ Mol Mutagen 1995;26:345-56.10.1002/em.2850260411
Search in Google Scholar
Devaux A, Flammarion P, Bernardon V, Garric J, Monod G. 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 1998;46:257-62.10.1016/S0141-1136(97)00105-0
Search in Google Scholar
Brown JS, Steinert SA. DNA damage and biliary PAH metabolites in flatfish from Southern California Bays and harbors, and the Channel Islands. Ecol Indic 2004;3:263-74.10.1016/j.ecolind.2003.11.004
Search in Google Scholar
de Andrade VM, da Silva J, da Silva FR, Heuser VD, Dias JF, Yoneama ML, de Freitas TRO. Fish as bioindicators to assess the effects of pollution in two southern Brazilian rivers using the Comet assay and micronucleus test. Environ Mol Mutagen 2004;44:459-68.10.1002/em.20070
Search in Google Scholar
Whitehead A, Kuivila KM, Orlando JL, Kotelevtsev S, Anderson SL. Genotoxicity in native fish associated with agricultural runoff events. Environ Toxicol Chem 2004;23:2868-77.10.1897/03-649.1
Search in Google Scholar
Winter M, Day N, Hayes RA, Taylor EW, Butler PJ, Chipman JK. 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 2004;552:163-75.10.1016/j.mrfmmm.2004.06.014
Search in Google Scholar
Guilherme S, Vélega M, Pereira ME, Santos MA, Pacheco M. Erythrocytic nuclear abnormalities in wild and caged fish (Liza aurata) along an environmental mercury contamination gradient. Ecotoxicol Environ Saf 2008;70:411-21.10.1016/j.ecoenv.2007.08.016
Search in Google Scholar
Oikari A. Caging techniques for field exposures of fish to chemical contaminants. Aquat Toxicol 2006;78:370-81.10.1016/j.aquatox.2006.03.010
Search in Google Scholar
Pellacani C, Buschini A, Furlini M, Poli P, Rossi C. A battery of in vivo and in vitro tests useful for genotoxic pollutant detection in surface waters. Aquat Toxicol 2006;77:1-10.10.1016/j.aquatox.2005.10.010
Search in Google Scholar
Frenzilli G, Nigro M, Lyons BP. The Comet assay for the evaluation of genotoxic impact in aquatic environments. Mutat Res Rev Mutat 2009;681:80-92.10.1016/j.mrrev.2008.03.001
Search in Google Scholar
Hartl MGJ, Kilemade M, Sheehan D, Mothersill C, O'Halloran J, O'Brien NM, van Pelt FNAM. Hepatic biomarkers of sediment-associated pollution in juvenile turbot, Scophthalmus maximus L. Mar Environ Res 2007;64:191-208.10.1016/j.marenvres.2007.01.002
Search in Google Scholar
Akcha F, Leday G, Pfohl-Leszkowicz A. Potential value of the Comet assay and DNA adduct measurement in dab (Limanda limanda) for assessment of in situ exposure to genotoxic compounds. Mutat Res 2003;534:21-32.10.1016/S1383-5718(02)00244-9
Search in Google Scholar
Akcha F, Vincent Hubert F, Pfohl-Leszkowicz A. Measurement of DNA adducts and strand breaks in dab (Limanda limanda) collected in the field: effects of biotic (age, sex) and abiotic (sampling site and period) factors on the extent of DNA damage. Mutat Res 2004;552:197-207.10.1016/j.mrfmmm.2004.06.020
Search in Google Scholar
Lyons BP, Stentiford GD, Bignell J, Goodsir F, Sivyer DB, Devlin MJ, Lowe D, Beesley A, Pascoe CK, Moore MN, Garnacho E. A biological effects monitoring survey of Cardigan Bay using flatfish histopathology, cellular biomarkers and sediment bioassays: Findings of the Prince Madog Prize 2003. Mar Environ Res 2006;62(Suppl 1): S342-6.10.1016/j.marenvres.2006.04.064
Search in 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. In situ biomonitoring of PAH-contaminated sediments using juvenile coho salmon (Oncorhynchus kisutch). Ecotoxicol Environ Saf 2008;71:454-64.10.1016/j.ecoenv.2008.01.001
Search in Google Scholar
Bony S, Gillet C, Bouchez A, Margoum C, Devaux A. Genotoxic pressure of vineyard pesticides in fish: Field and mesocosm surveys. Aquat Toxicol 2008;89:197-203.10.1016/j.aquatox.2008.06.017
Search in Google Scholar
Maria VL, Correia AC, Santos MA. Genotoxic and biochemical responses in caged eel (Anguilla anguilla L.) after short-term exposure to harbour waters. Environ Int 2004;29:923-9.10.1016/S0160-4120(03)00057-6
Search in Google Scholar
Maria VL, Gravato C, Correia AC, Santos MA. Biotransformation and genotoxicity responses to PAHs in two teleost species. Fresenius Environ Bull 2002;11:609-15.
Search in Google Scholar
Gravato C, Santos MA. β-naphthoflavone liver EROD and erythrocytic nuclear abnormality induction in juvenile Dicentrarchus labrax L. Ecotoxicol Environ Saf 2002;52:69-74.10.1006/eesa.2002.2151
Search in Google Scholar
Gravato C, Santos MA. Juvenile sea bass liver biotransformation induction and erythrocytic genotoxic responses to resin acids. Ecotoxicol Environ Saf 2002;52:238-47.10.1006/eesa.2002.2161
Search in Google Scholar
Gravato C, Santos MA. Genotoxicity biomarkers' association with B(a)P biotransformation in Dicentrarchus labrax L. Ecotoxicol Environ Saf 2003;55:352-8.10.1016/S0147-6513(02)00070-2
Search in Google Scholar
Teles M, Gravato C, Pacheco M, Santos MA. Juvenile sea bass biotransformation, genotoxic and endocrine responses to β-naphthoflavone, 4-nonylphenol and 17β-estradiol individual and combined exposures. Chemosphere 2004;57:147-58.10.1016/j.chemosphere.2004.02.02315294438
Search in Google Scholar
Teles M, Pacheco M, Santos MA. Biotransformation, stress and genotoxic effects of 17β-estradiol in juvenile sea bass (Dicentrarchus labrax L.). Environ Int 2006;32:470-7.10.1016/j.envint.2005.11.00616483653
Search in Google Scholar
Maria VL, Ahmad I, Santos MA. Juvenile sea bass (Dicentrarchus labrax L.) DNA strand breaks and lipid peroxidation response following 17β-estradiol two mode of exposures. Environ Int 2008;34:23-9.10.1016/j.envint.2007.06.008
Search in Google Scholar
Gravato C, Santos MA, Magalhães I. Juvenile Dicentrarchus labrax L. biochemical and genotoxic responses after short-term exposure to β-naphthoflavone and contaminated harbour waters. Fresenius Environ Bull 2000;9:269-74.
Search in Google Scholar
Gravato C, Santos MA. Juvenile sea bass liver biotransformation and erythrocytic genotoxic responses to pulp mill contaminants. Ecotoxicol Environ Saf 2002;53:104-12.10.1006/eesa.2002.2196
Search in Google Scholar
Gravato C, Santos MA. Dicentrarchus labrax biotransformation and genotoxicity responses after exposure to a secondary treated industrial/urban effluent. Ecotoxicol Environ Saf 2003;55:300-6.10.1016/S0147-6513(03)00024-1
Search in Google Scholar
Mohmood I, Maria VL, Oliveira M, Ahmad I, Pacheco M, Santos MA. Seasonal assessment of a contaminated coastal lagoon (Ria de Aveiro, Portugal) using Dicentrarchus labrax L. erythrocytic nuclear abnormalities. Fresenius Environ Bull 2008;17:1924-31.
Search in Google Scholar
Stien X, Percic P, Gnassia-Berelli M, Roméo M, Lafaurie M. Evaluation of biomarkers in caged fishes and mussels to assess the quality of waters in a Bay of the NW Mediterranean Sea. Environ Pollut 1998;99:339-45.10.1016/S0269-7491(98)00013-X
Search in Google Scholar
Klobučar GIV, Štambuk A, Hylland K, Pavlica M. Detection of DNA damage in haemocytes of Mytilus galloprovincialis in the coastal ecosystems of Kaštela and Trogir Bays, Croatia. Sci Total Environ 2008;405:330-7.10.1016/j.scitotenv.2008.05.015
Search in Google Scholar
Klobučar GIV, Pavlica M, Erben R, Papeš D. Application of the micronucleus and Comet assays to mussel Dreissena polymorpha haemocytes for genotoxicity monitoring of freshwater environments. Aquat Toxicol 2003;64:15-23.10.1016/S0166-445X(03)00009-2
Search in 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. Report from the in vitro micronucleus assay working group. Environ Mol Mutagen 2000;35:167-72.10.1002/(SICI)1098-2280(2000)35:3<;167::AID-EM3>3.0.CO;2-G
Search in Google Scholar
Cotelle S, Férard JF. Comet assay in genetic ecotoxicology: a review. Environ Mol Mutagen 1999;34:246-55.10.1002/(SICI)1098-2280(1999)34:4<;246::AID-EM4>3.0.CO;2-V
Search in Google Scholar
Odžak N, Zvonarić T, Kljaković-Gašpić Z, Horvat M, Barić A. Biomonitoring of mercury in the Kaštela Bay using transplated mussels. Sci Total Environ 2000;261:61-8.10.1016/S0048-9697(00)00595-7
Search in Google Scholar
Ujević I, Odžak N, Barić A. Trace metal accumulation in different grain size fractions of the sediments from semi enclosed bay heavily contaminated by urban and industrial wastewaters. Water Res 2000;34:3055-61.10.1016/S0043-1354(99)00376-0
Search in Google Scholar
Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988;175:184-91.10.1016/0014-4827(88)90265-0
Search in 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. Report from the in vitro micronucleus assay working group. Mutat Res 2003;540:153-63.10.1016/j.mrgentox.2003.07.005
Search in Google Scholar
Klobučar GIV, Štambuk A, Pavlica M, Sertić-Peroć M, Kutuzović-Hackenberger B, Hylland K. Genotoxicity monitoring of freshwater environments using caged carp (Cyprinus carpio). Ecotoxicology 2010;19:77-84.10.1007/s10646-009-0390-6
Search in Google Scholar
Wirzinger G, Weltje L, Gercken J, Sordyl H. Genotoxic damage in field-collected three-spined sticklebacks (Gasterosteus aculeatus L.): a suitable biomonitoring tool? Mutat Res 2007;628:19-30.
Search in Google Scholar
Udroiu I. The micronucleus test in piscine erythrocytes. Aquat Toxicol 2006;79:201-4.10.1016/j.aquatox.2006.06.013
Search in Google Scholar
Sola L, Bressanello S, Rossi AR, Laselli V, Crosetti D, Cataudella S. A karyotype analysis of the genus Dicentrarchus by different staining techniques. J Fish Biol 1993;43:329-37.10.1111/j.1095-8649.1993.tb00567.x
Search in Google Scholar
Chu S, Xi Z, Xu X, Zhang Y, Xu Y. Induction of micronuclei in peripheral erythrocytes of Misgurnus anguillicaudatus by polychlorinated biphenyls. Bull Environ Contam Toxicol 1996;57:179-82.10.1007/s001289900172
Search in Google Scholar
Peichel CL, Nereng KS, Ohgi KA, Cole BLE, Colosimo PF, Buerklet CA, Schluter D, Kingsley DM. The genetic architecture of divergence between threespine stickleback species. Nature 2001;414:901-5.10.1038/414901a
Search in Google Scholar
Lee RF, Steinert S. Use of the single cell gel electrophoresis/Comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. Mutat Res 2003;544:43-64.10.1016/S1383-5742(03)00017-6
Search in Google Scholar
Bombail V, Aw D, Gordon E, Batty J. Application of the Comet and micronucleus assays to butterfish (Pholis gunnellus) erythrocytes from the Firth of Forth, Scotland. Chemosphere 2001;44:383-92.10.1016/S0045-6535(00)00300-3
Search in Google Scholar

Cage Exposure of European Sea Bass (Dicentrarchus Labrax) for in Situ Assessment of Pollution-Related Genotoxicity

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