Skip to main content
Log in

Impact of Oxidative Stress Indicated by Thiobarbituric Acid Reactive Substances (TBARS) and Protein Carbonyl Levels (PC) on Ethoxyresorufin-O-deethylase (EROD) Induction in Common Carp (Cyprinus carpio)

  • Published:
Water, Air, & Soil Pollution Aims and scope Submit manuscript

Abstract

This study examined the impact of oxidative stress indicated by thiobarbituric acid reactive substances (TBARS) and protein carbonyl (PC), induced by intensive exercise and cadmium chloride (CdCl2) on ethoxyresorufin-O-deethylase (EROD) activity in juvenile carp (Cyprinus carpio). In the first experiment, fish were divided into three groups: (1) control, (2) carp exposed to intensive exercise, and (3) carp that was not exercised but previously, as well as carp in group 2, received single dose of 3-methylcholantrene (3-MC). The third and sixth day fish were sacrificed and the measurements were conducted. In the second experiment, fish were divided into (1) control, (2) carp in water containing CdCl2, and (3) carp in dechlorinated tap water (2 and 3 received single dose of 3-MC on the seventh day after exposure to CdCl2). The carp were killed 6 days later and livers were excised for biochemical analyses. In the first experiment, on the sixth day after treatment with 3-MC, results show statistically significant increase in EROD activity in non-exercised carp, while that increase in carp exposed to intensive exercise was significantly lower. Three days after exposure to 3-MC, statistically significant increase in TBARS was observed in both exercised and non-exercised carp. Six days after exposure to 3-MC, PC levels were significantly higher in exercised carp. Pretreatment with CdCl2, in the second experiment, caused oxidative stress and reduction of EROD activity. Results show linkage between expression of EROD activity and oxidative stress biomarkers and possible influence of oxidative stress on the cell membrane structures and consequently on EROD activity.

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
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ahmad, I., Pacheco, M., & Santos, M. A. (2004). Enzymatic and nonenzymatic antioxidants as an adaptation to phagocyte-induced damage in Anguilla anguilla L. following in situ harbor water exposure. Ecotoxicology and Environmental Safety, 57, 290–302.

    Article  CAS  Google Scholar 

  • Alessio, H. M., Goldfarb, A. H., & Culter, R. G. (1988). MDA content increases in fast- and slow-twitch skeletal muscle with intensity of exercise in a rat. The American Journal of Physiology, 6(1), 874–877.

    Google Scholar 

  • Almeida, J. A., Diniz, Y. S., Marques, S. F. G., Faine, L. A., Ribas, B. O., Burneiko, R. C., & Novelli, E. L. B. (2002). The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination. Environmental International, 27, 673–679.

    Google Scholar 

  • Almroth, B. C., Sturve, J., Berglund, A., & Förlin, L. (2005). Oxidative damage in eelpout (Zoarces viviparus), measured as protein carbonyls and TBARS, as biomarkers. Aquatic Toxicology, 73, 171–180.

    Article  CAS  Google Scholar 

  • Ando, S., & Yanagida, K. (1999). Susceptibility to oxidation of copper-induced plasma lipoproteins from Japanese eel: Protective effect of vitellogenin on the oxidation of very low density lipoprotein. Comparative Biochemistry and Physiology. C, 123, 1–7.

    Article  CAS  Google Scholar 

  • Arniç, E., Sen, A., & Bozcaarmutlu, A. (2000). Cytochrome P4501A and associated mixed-function oxidase induction in fish as a biomarker for toxic carcinogenic pollutants in the aquatic environment. Pure and Applied Chemistry, 72(6), 985–994.

    Article  Google Scholar 

  • Bagnyukova, T. V., Vasylkiv, O. Y., Storey, K. B., & Lushchak, V. I. (2005). Catalase inhibition by amino triazole induces oxidative stress in goldfish brain. Brain Research, 1052, 180–186.

    Article  CAS  Google Scholar 

  • Banjerdkij, P., Vattanaviboon, P., & Mongkolsuk, S. (2005). Exposure to cadmium elevates expression of genes in the OxyR and OhrR regulons and induces cross-resistance to peroxide killing treatment in Xanthomonas campestris. Applied and Environmental Microbiology, 71(4), 1843–1849.

    Article  CAS  Google Scholar 

  • Barker, C. W., Fagan, J. B., & Pasco, D. S. (1994). Down-regulation of P4501A1 and P4501A2 mRNA expression in isolated hepatocytes by oxidative stress. Journal of Biological Chemistry, 269, 3985–3990.

    CAS  Google Scholar 

  • Beamish, F. W. H. (1978). Swimming capacity. In W. S. Hoar & D. J. Randall (Eds.), Fish physiology. Vol. 7 (pp. 101–187). New York: Academic.

    Google Scholar 

  • Belviranli, M., & Gökbel, H. (2006). Acute exercise induced oxidative stress and antioxidant changes. European Journal of General Medicine, 3(3), 126–131.

    CAS  Google Scholar 

  • Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.

    Article  CAS  Google Scholar 

  • Burgeot, T., Bocquéné, G., Truquet, P., Le Dean, L., & Galgani, F. (1994). Induction of EROD activity in red mullet (Mullus barbatus) along the French Mediterranean coasts. Science of the Total Environment, 142, 213–220.

    Article  CAS  Google Scholar 

  • Burke, M. D., & Mayer, R. T. (1974). Ethoxyresorufin: Direct fluorometric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylcholanthrene. Drug Metabolism and Disposition, 2(6), 583–588.

    CAS  Google Scholar 

  • Chang, C.-K., Huang, H.-Y., Tseng, H.-F., Hsuuw, Y.-D., & Tso, T. K. (2007). Interaction of vitamin E and exercise training on oxidative stress and antioxidant enzyme activities in rat skeletal muscles. Journal of Nutritional Biochemistry, 18, 39–45.

    Article  CAS  Google Scholar 

  • Choi, J., & Oris, J. T. (2000). Evidence of oxidative stress in bluegill sunfish (Lepomis macrochirus) liver microsomes simultaneously exposed to solar ultraviolet radiation and anthracene. Environmental Toxicology & Chemistry, 19, 1795–1799.

    CAS  Google Scholar 

  • Delaporte, E., & Renton, K. W. (1997). Cytochrome P4501A1 and cytochrome P4501A2 are downregulated at both transcriptional and post-transcriptional levels by conditions resulting in interferon-alpha/beta induction. Life Sciences, 60, 787–796.

    Article  CAS  Google Scholar 

  • Fenet, H., Casellas, C., & Bontoux, J. (1998). Laboratory and field-caging studies on hepatic enzymatic activities in European eel and rainbow trout. Toxicology and Environmental Safety, 40, 137–143.

    Article  CAS  Google Scholar 

  • Flammarion, P., & Garric, J. (1997). Cyprinids EROD activities in low contaminated rivers: A relevant statistical approach to estimate reference levels for EROD biomarker? Chemosphere, 35(10), 2375–2388.

    Article  CAS  Google Scholar 

  • Floor, E., & Wetzel, M. G. (1998). Increased protein oxidation in human substantia nigra pars compacta in comparison with basal ganglia and prefrontal cortex measured with an improved dinitrophenylhydrazine assay. Journal of Neurochemistry, 70, 268–275.

    Article  CAS  Google Scholar 

  • Förlin, L., Haux, C., Karlsson-Norrgren, L., Runn, P., & Larsson, A. (1986). Biotransformation enzyme activities and histopathology in rainbow trout, Salmo gairdneri, treated with cadmium. Aquatic Toxicology, 8, 51–64.

    Google Scholar 

  • Fortuniak, A., Zadrinski, R., Bilinski, T., & Bartosz, G. (1996). Glutathione depletion in the yeast Saccharomyces cervisiae. Biochemistry and Molecular Biology International, 38, 901–910.

    CAS  Google Scholar 

  • Gabrzelak, T., & Klekot, J. (1985). The effect of paraquat on the peroxide metabolism enzymes in erythrocytes of freshwater fish species. Comparative Biochemistry and Physiology, 81C, 415–418.

    Google Scholar 

  • Grzebyk, D., & Galgani, F. (1991). Measurement of the effect of organic pollution on marine organisms: Rapid determination of EROD induction using plate readers. Aquatic Living Resources, 4, 53–59.

    Article  Google Scholar 

  • Halliwell, B., & Chirico, S. (1993). Lipid peroxidation: Its mechanism, measurement and significance. American Journal of Clinical Nutrition, 57, 715S–725S.

    CAS  Google Scholar 

  • Hassanain, M. A., Abdel-Rahman, E. H., Abo-Hegab, S., Tawfik, M. A. A., & Abbas, W. T. (2007). Induction of cytochrome P450 1A1 as a biomarker of benzo-a-pyrene pollution in Egyptian fresh water fish. Pakistan Journal of Biological Sciences, 10(8), 1161–1169.

    Article  CAS  Google Scholar 

  • Ji, L. L. (2000). Antioxidants and oxidative stress in exercise. Proceedings of the Society for Experimental Biology and Medicine, 222, 283–292.

    Article  Google Scholar 

  • Kelle, M., Diken, H., Şermet, A., Atmaca, M., & Koçyiğit, Y. (1998). Changes in blood antioxidant status and lipid peroxidation following distance running. Turkish Journal of Medical Sciences, 28, 643–647.

    Google Scholar 

  • Konstandi, M., Johnson, E., Lang, M. A., Camus-Radon, A. M., & Marselos, M. (2000). Stress modulates the enzymatic inducibility by benzo[α]pyrene in the rat liver. Pharmacological Research, 42, 3.

    Article  Google Scholar 

  • Lafontaine, Y., Gagne, F., Blaise, C., Costan, G., Gagnogn, P., & Chan, H. M. (2000). Biomarkers in zebra mussels (Dreissena polymorpha) for the assessment and monitoring of water quality of the St Lawrence river (Canada). Aquatic Toxicology, 50, 51–71.

    Article  Google Scholar 

  • Lemaire-Gony, S., & Lemaire, P. (1992). Interactive effects of cadmium and benzo-a-pyrene on cellular structure and biotransformation enzymes of the liver of the European eel Anguilla anguilla. Aquatic Toxicology, 22, 145–159.

    Article  CAS  Google Scholar 

  • Lenz, A.-G., Costabel, U., Shaltiel, S., & Levine, R. L. (1989). Determination of carbonyl groups in oxidatively modified proteins by reduction with tritiated sodium borohydride. Analytical Biochemistry, 177, 419–425.

    Article  CAS  Google Scholar 

  • Liu, J., Yeo, H. C., Övervik-Douki, E., Hagen, T., Doniger, S. J., Chu, D. W., Brooks, G. A., & Ames, B. N. (2000). Chronically and acutely exercised rats: Biomarkers of oxidative stress and endogenous antioxidants. Journal of Applied Physiology, 89, 21–28.

    CAS  Google Scholar 

  • Liu, L., Bridges, R. J., & Eyer, C. L. (2001). Effect of cytochrome P450 1A induction on oxidative damage in rat brain. Molecular and Cellular Biochemistry, 223, 89–94.

    Article  CAS  Google Scholar 

  • Livingstone, D. R. (2001). Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Marine Pollution Bulletin, 42, 656–666.

    Article  CAS  Google Scholar 

  • Lushchak, V. I., Bagnyukova, T. V., Lushchak, O. V., Storey, J. M., & Storey, K. B. (2005). Hypoxia and recovery perturb free radical processes and antioxidant potential in common carp (Cyprinus carpio) tissues. International Journal of Biochemistry & Cell Biology, 37, 1319–1330.

    Article  CAS  Google Scholar 

  • Manna, I., Jana, K., & Samanta, P. K. (2003). Effect of intensive exercise-induced testicular gametogenic and steroidogenic disorders in mature male Wistar strain rats: A correlative approach to oxidative stress. Acta Physiologica Scandinavica, 178, 33–40.

    Article  CAS  Google Scholar 

  • Oakes, K. D., & Van Der Kraak, G. J. (2003). Utility of the TBARS assay in detecting oxidative stress in white sucker (Catostomus commersoni) populations exposed to pulp mill effluent. Aquatic Toxicology, 63, 447–463.

    Article  CAS  Google Scholar 

  • Parvez, S., & Raisuddin, S. (2005). Protein carbonyls: Novel biomarkers of exposure to oxidative stress-inducing pesticides in freshwater fish Channa punctata (Bloch). Environmental Toxicology and Pharmacology, 20, 112–117.

    Article  CAS  Google Scholar 

  • Peters, L. D., Porte, C., Albaiges, J., & Livingstone, D. R. (1994). 7-Ethoxyresorufin O-deethylase (EROD) and antioxidant enzyme activities in larvae of sardine (Sardina pilchardus) from the North coast of Spain. Marine Pollution Bulletin, 28(5), 299–304.

    Article  CAS  Google Scholar 

  • Rice-Evans, C. A., Diplock, A. T., & Symons, M. C. R. (1991). Techniques in free radical research. In R. H. Brton & P. H. Knippenberg (Eds.), Laboratory techniques in biochemistry and molecular biology (pp. 147–149). Amsterdam: Elsevier.

    Google Scholar 

  • Roméo, M., Bennani, N., Gnassia-Barelli, M., Lafaurie, M., & Girard, J. P. (2000). Cadmium and copper display different responses towards oxidative stress in kidney of the sea bass Dicentrachus labrax. Aquatic Toxicology, 48, 185–194.

    Article  Google Scholar 

  • Romero, F., Bosch-Morell, F., Romero, M. J., Jareno, J., Romero, B., Marin, N., & Roma, J. (1998). Lipid peroxidation products and antioxidants in human disease. Environmental Health Perspectives Supplements, 106, 1229.

    CAS  Google Scholar 

  • Sen, C. K., Atalay, M., Agren, J., Laaksonen, D. E., Roy, S., & Hänninen, O. (2005). Fish oil and vitamin E supplementation in oxidative stress at rest and after physical exercise. Journal of Applied Physiology, 83, 189–195.

    Google Scholar 

  • Shacter, E. (2000). Quantification and significance of protein oxidation in biological samples. Drug Metabolism Reviews, 32(3–4), 307–326.

    Article  CAS  Google Scholar 

  • Shacter, E., Williams, J. A., Lim, M., & Levine, R. L. (1994). Differential susceptibility of plasma proteins to oxidative modification: Examination by western blot immunoassay. Free Radical Biology and Medicine, 17, 429–467.

    Article  CAS  Google Scholar 

  • Shi, H., Sui, Y., Wang, X., Luo, Y., & Ji, L. (2005). Hydroxyl radical production and oxidative damage induced by cadmium and naphthalene in liver of Carassius auratus. Comparative Biochemistry and Physiology. C, 140, 115–121.

    Google Scholar 

  • Široka, Z., & Drastichova, J. (2004). Biochemical markers of aquatic environment contamination—cytochrome P450 in fish. A review. Acta Veterinaria Brno, 73, 123–132.

    Google Scholar 

  • Stanic, B., Andric, N., Zoric, S., Grubor-Lajsic, G., & Kovacevic, R. (2006). Assessing pollution in the Danube River near Novi Sad (Serbia) using several biomarkers in starlet (Acipenser ruthenus L.). Ecotoxicology and Environmental Safety, 65, 395–402.

    Article  CAS  Google Scholar 

  • Stegeman, J. J. (1989). Cytochrome P450 forms in fish: Catalytic, immunological and sequence similarities. Xenobiotica, 19(10), 1093–1110.

    Article  CAS  Google Scholar 

  • Stein, J. E., Collier, T. K., Reichart, K., Caillas, L., Hom, E., & Varanasi, U. (1992). Bioindicators of contaminant exposure and sublethal effects: Studies with benthic fish in Pudget Sound, WA. Environmental Toxicology & Chemistry, 11, 701–714.

    Article  CAS  Google Scholar 

  • Stohs, S. J., & Bagchi, D. (1995). Oxidative mechanisms in the toxicity of chromium and cadmium ions. Journal of Environmental Pathology, Toxicology, and Oncology, 20, 77–88.

    Google Scholar 

  • Van der Oost, R., Beyer, J., & Vermeulen, N. P. E. (2003). Fish bioaccumulation and biomarkers in environmental risk assessment: A review. Environmental Toxicology and Pharmacology, 13, 57–149.

    Article  Google Scholar 

  • Wang, Y., Fang, J., Leonard, S. S., & Rao, K. M. (2004). Cadmium inhibits the electron transfer chain and induces reactive oxygen species. Free Radical Biology & Medicine, 36, 1434–1443.

    Article  CAS  Google Scholar 

  • Whyte, J. J., Jung, R. E., Schmitt, C. J., & Tillitt, D. E. (2000). Ethoxyresorufin-O-deethylase (EROD) activity in fish as a biomarker of chemical exposure. Critical Reviews in Toxicology, 30(4), 347–570.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Ministry for Science and Technology of the Republic of Croatia, Project No. 285-0000000-3484.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Branimir K. Hackenberger.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stepić, S., Hackenberger, B.K., Hackenberger, D.K. et al. Impact of Oxidative Stress Indicated by Thiobarbituric Acid Reactive Substances (TBARS) and Protein Carbonyl Levels (PC) on Ethoxyresorufin-O-deethylase (EROD) Induction in Common Carp (Cyprinus carpio). Water Air Soil Pollut 223, 4785–4793 (2012). https://doi.org/10.1007/s11270-012-1234-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11270-012-1234-1

Keywords

Navigation