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DNA strand breaks, cytochrome P-450-dependent monooxygenase system activity and levels of chlorinated biphenyl congeners in the pyloric caeca of the seastar (Asterias rubens) from the North Sea

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Abstract

Seastars (Asterias rubens L.) were collecte d at sampling locations in different areas along transects radiating into the southern North Sea, representing areas impacted by contaminants to different degrees. Strand breakage in DNA isolated from tissue of the pyloric caeca was measured by the alkaline unwinding assay, modified to allow for the isolation of highly intact DNA. The interpretation of the results is based on the incidence of DNA strand breaks (expressed as the double:total DNA ratio or F value, indicating the degree of double-strandedness). The cytochrome P-450 concentration and the benzo [a] pyrene (BaP) hydroxylase activity were measured in microsomal fractions of the pyloric caeca. The chlorinated biphenyl (CB) congeners in tissue samples from the digestive gland were determined by temperature-programmed gas-chromatography, with a CPSil8 capillary column as the stationary phase, hydrogen as the carrier gas and 63Ni-electron capture detection. Areas where seastars showed different DNA integrity could be described. The highest integrity (0.75 < F < 0.85) was found in the off-shore reference sites of the Dogger Bank and Southern Bight. The mean F value in seastars from most sampling locations varied between 55 and 75%. The lowest DNA integrity (0.35 < F < 0.55) was found in specimens obtained from sampling locations near the river Rhine delta, along the Dutch coast and at two expected uncontaminated offshore areas. The BaP hydroxylase activity was relatively high near the mouth of the rivers Rhine and Scheldt, but also at a supposedly clean site near Dogger Bank. The concentration of CB congeners in the pyloric caeca of seastars decreased along transects radiating into the southern North Sea from the coastal areas of the Netherlands; the highest concentrations were in the nearby coastal areas and the lowest were in the open sea sampling locations. The data indicate that there might be a relationship between pollution from the Rivers Rhine, Meuse and Scheldt and the incidence of DNA strand breaks and/or BaP hydroxylase activity

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References

  • Anonymous (1984) PCB Regulation. The Hague: Dutch Staatscourant.

  • Anonymous (1988) Verordnung über Höchstmengen an Schadstoffen in Lebensmitteln. Bundesgesetzblatt 1, 422–4.

    Google Scholar 

  • Anonymous (1987) Biological markers in environmental health research. Environ. Health Perspect. 74, 3–9.

    Google Scholar 

  • Ballschmiter, K. and Zell, M. (1980) Analysis of polychlorinated biphenyls (PCB) by glass capillary gas chromatography. Composition of technical Arochlor and Clophen–PCB mixtures. Fresenius' Z. Anal. Chem. 302, 20–31.

    Google Scholar 

  • Boon, J.P., Van Zantvoort, M.B., Govaert, M.J.M.A. and Duinker, J.C. (1985) Organochlorines in benthic polychaetes (Nephtys spp.) and sediments from the Southern North sea. Identification of individual PCB components. Neth. J. Sea Res. 19, 93–109.

    Article  Google Scholar 

  • Boon, J.P., Van Arnhem, E., Jansen, S., Kannan, N., Petrick, G., Schultz, D.E., Duinker, J.C., Reijnders, P.J.H. and Goksoyr, A. (1992a) The toxicokinetics of PCBs in marine mammals with special reference to possible interactions of individual congeners with the cytochrome P450-dependent monooxygenase system-an overview. In C.H. Walker and D.R. Livingstone (eds) Persistant pollutants in marine ecosystems, pp 119–54. Oxford, New York, Seoul, Tokyo: Pergamon Press.

    Google Scholar 

  • Boon, J.P., Everaarts, J.M., Hillebrand, M.T.J., Eggens, M.L., Pijnenburg, J. and Goksoyr, A. (1992b) Changes in levels of hepatic biotransformation enzymes and haemoglobin levels in female plaice (Pleuronectes platessa) after oral administration of a technical PCB mixture (Clophen A40). Sci. Total Environ. 114, 113–33.

    Article  PubMed  Google Scholar 

  • De Boer, J. and Dao, Q.T. (1991) Analysis of seven chlorobiphenyl congeners by multidimensional gas chromatography. J. High Resolut. Chromatogr. 14, 593–6.

    Google Scholar 

  • Dehnen, W., Tomongas, R. and Roos, J. (1973) A modified method for the assay of benzo[a]pyrene hydroxylase. Anal. Biochem. 53, 373–83.

    PubMed  Google Scholar 

  • Den Besten, P.J., Herwig, H.J., Van Donselaar, E.G. and Livingstone, D.R. (1990) Cytochrome P-450 monoxygenase system and benzo[a]pyrene metabolism in echinoderms. Mar. Biol. 107, 171–7.

    Google Scholar 

  • Den Besten, P.J., Elenbaas, J.M.L., Maas, J.R., Dieleman, S.J., Herwig, H.J. and Voogt, P.A. (1991) Effects of cadmium and polychlorinated biphenyls (Clophen A50) on steroid metabolism and cytochrome P-450 monooxygenase system in the sea star Asterias rubens L. Aquatic Toxicol. 20, 95–110.

    Article  Google Scholar 

  • Den Besten, P.J., Lemaire, P., Livingstone, D.R., Woodin, B., Stegeman, J.J., Herwig, H.J. and Seinen, W. (1993) Time-course and dose-response of the apparent induction of the cytochrome P450 monooxygenase system of pyloric caeca microsomes of the female sea star Asterias rubens L. by benzo[a]pyrene and polychlorinated biphenyls. Aquatic Toxicol. 26, 23–40.

    Article  Google Scholar 

  • Den Besten, P.J., Lemaire, P. and Livingstone, D.R. (1994) NADPH-, NADH-and cumene hydroperoxide-dependent metabolism of benzo[a]pyrene by pyloric caeca microsomes of the seastar Asterias rubens L. (Echinodermata: Asteroidea). Xenobiotica 24, 989–1001.

    PubMed  Google Scholar 

  • Duinker, J.C. and Hillebrand, M.Th.J. (1978) Minimizing blank values in chlorinated hydrocarbon analysis. J. Chromatogr. 150, 195–9.

    Article  Google Scholar 

  • Duinker, J.C. and Hillebrand, M.Th.J. (1983) Characterization of PCB components in Clophen formulations by capillary GC-MS and GC-ECD techniques. Environ. Sci. Technol. 17, 449–56.

    Google Scholar 

  • Everaarts, J.M. (1995) DNA integrity as a biomarker of marine pollution: strand breaks in seastar (Asterias rubens) and dab (Limanda limanda). Mar. Pollut. Bull. 31, 431–8.

    Article  Google Scholar 

  • Everaarts, J.M. and Fischer, C.V. (1989) Microcontaminants in surface sediment and macrobenthic invertebrates of the North Sea. NIOZ-Rapport 1989–6, 1–44.

    Google Scholar 

  • Everaarts, J.M., Otter, E. and Fischer, C.V. (1990) Cadmium and polychlorinated biphenyl: different distribution pattern in North Sea benthic biota. Neth. J. Sea Res. 26, 75–82.

    Article  Google Scholar 

  • Everaarts, J.M. Shugart, L.R., Gustin, M.K., Hawkins, W.E. and Walker, W.W. (1993) Biological markers in fish: DNA integrity, hematological parameters and liver somatic index. Mar. Environ. Res. 35, 101–7.

    Article  Google Scholar 

  • Everaarts, J.M., Sleiderink, H.M., Den Besten, P.J., Halbrook, R.S. and Shugart, L.R. (1994) Molecular responses as indicators of marine pollution: DNA damage and enzyme induction in Limanda limanda and Asterias rubens. Environ. Health Perspect. 102(Suppl. 12):37–43.

    Google Scholar 

  • Everaarts, J.M., Sarkar, A., Rigterink, H., Appelman, J., Van Weerlee, E. and Den Besten, P.J. (1995) DNA strand breaks, MO activity and contaminant levels in pyloric caeca and gonads of the seastar (Asterias rubens) from the North Sea. In J.M. Everaarts (ed.) EC Indo-Dutch joint research project 1994–1997, First annual progress report, pp. 9–16. Netherlands Institute for Sea Research: Texel, The Netherlands.

    Google Scholar 

  • Grahl-Nielsen, O., Staveland, J.T. and Wilhelmsen, S. (1978) Aromatic hydrocarbons in benthic organisms from coastal areas polluted by Iranian crude oil. J. Fish. Res. Bd Can. 35, 615–23.

    Google Scholar 

  • Holden, A.V. and Marsden, K. (1969) Single-stage clean-up of animal tissue extracts for organochlorine residue analysis. J. Chromatogr. 44, 481–92.

    Article  PubMed  Google Scholar 

  • Interdepartmental Co-ordinating Committee for North Sea Affairs (1992) North Sea Atlas for Netherlands Policy and Management. Amsterdam: Stadsuitgeverij.

    Google Scholar 

  • Kanter, P.M. and Schwartz, H.S. (1982) A fluorescence enhancement assay for cellular DNA damage. Mol. Pharmacol. 22, 145–51.

    PubMed  Google Scholar 

  • Klungsøyr, J. and Wilhelmsen, S. (1993) Concentrations of PAHs and PCBs in total sediment. NIOZ-Rapport 1993–9, 59–62.

    Google Scholar 

  • Knickmeyer, R., Landgraff, O. and Steinhart, H. (1992) Cyclic organochlorines in the sea star Asterias rubens from the German Bight, December 1988–May 1989. Mar. Environ. Res. 33, 127–43.

    Article  Google Scholar 

  • Knutzen, J. and Sortland, B. (1982) Polycyclic aromatic hydrocarbons (PAH) in some algae and invertebrates from moderately polluted parts of the coast of Norway. Water Res. 16, 421–8.

    Article  Google Scholar 

  • Kohn, H.W. (1983) The significance of DNA-damage assays in toxicity and carcinogenicity assessment. Ann. NY Acad. Sci. 407, 106–18.

    PubMed  Google Scholar 

  • Kurelec, B., Garg, A., Krca, S. and Gupta, R.C. (1989) DNA adducts as biomarkers in the genotoxic risk assessment in the aquatic environment. Mar. Environ. Res. 28, 317–21.

    Article  Google Scholar 

  • Livingstone, D.R. (1991) Organic xenobiotic metabolism in marine invertebrates. Adv. Comp. Environ. Physiol. 7, 45–185.

    Google Scholar 

  • McCarthy, J.F., Jacobson, D.N., Shugart, L.R. and Jimenez, B.D. (1989) Pre-exposure to 3-methylcholanthrene increases benzo[a]pyrene adducts on DNA of bluegill sunfish. Mar. Environ. Res. 28, 323–8.

    Article  Google Scholar 

  • Martineau, D., Lagacé, A., Béland, P., Higgins, R., Armstrong, D. and Shugart, L.R. (1988) Pathology of stranded Beluga whales (Delphinapterus leucas) from the St. Lawrence estuary, Québec, Canada. J. Comp. Pathol. 98, 287–311.

    Article  PubMed  Google Scholar 

  • Nacci, D. and Jackim, E. (1989) Using the DNA alkaline unwinding assay to detect DNA damage in laboratory and environmentally exposed cells and tissues. Mar. Environ. Res. 28, 333–7.

    Article  Google Scholar 

  • Shugart, L.R. (1988a) An alkaline unwinding assay for the detection of DNA damage in aquatic organisms. Mar. Environ. Res. 24, 321–5.

    Article  Google Scholar 

  • Shugart, L.R. (1988b) Quantitation of chemically induced damage to DNA of aquatic organisms by alkaline unwinding assay. Aquatic Toxicol. 13, 43–52.

    Article  Google Scholar 

  • Shugart, L.R. (1990) DNA damage as an indicator of pollutant-induced genotoxicity. In W.G. Landis and W.H. van der Schalie (eds) Aquatic toxicology and risk assessment, Vol. 13, pp. 348–55. Philadelphia: American Society for Testing and Materials.

    Google Scholar 

  • Shugart, L.R., Bickham, J., Jackim, G., McMahon, G., Ridley, W., Stein, J. and Steinert, S. (1992) DNA alterations. In R.J. Huggett, R.A. Kimerle, P.M. Mehrle Jr. and H.L. Bergman (eds) Biomarkers: biochemical, physiological and histological markers of anthropogenic stress, pp. 125–53. Chelsea, MI: Lewis Publishers.

    Google Scholar 

  • Shugart, L.R., Peakall, D.B., Walker, C.H. and Everaarts, J.M. (1993) Overview. In D.B. Peakall and L.R. Shugart (eds) Biomarkers. Research and application in the assessment of environmental health, NATO ASI series. Series H: Cell Biology, 68: XIII–XIX.

  • Sikka, H.C., Rutkowski, J.P. and Kandaswami, C. (1990a) Comparative metabolism of benzo[a]pyrene by liver microsomes from brown bullhead and carp. Aquatic Toxicol. 16, 101–12.

    Article  Google Scholar 

  • Sikka, H.C., Rutkowski, J.P., Kandaswami, C., Kumar, S., Early, K. and Gupta, R.C. (1990b) Formation and persistence of DNA adducts in the liver of brown bullheads exposed to benzo[a]-pyrene. Cancer Lett. 49, 81–7.

    Article  PubMed  Google Scholar 

  • Theodorakis, C.W., D'Surney, S.J., Bickham, J.W., Lyne, T.B., Bradley, B.P., Hawkins, W.E., Farkas, W.L., McCarthy, J.F. and Shugart, L.R. (1992) Sequential expression of biomarkers in bluegill sunfish exposed to contaminated sediment. Ecotoxicology 1, 45–73.

    Google Scholar 

  • Varanashi, U., Stein, J.E., Nishimoto, M., Reichert, W.L. and Collier, T.K. (1987) Chemical carcinogenesis in feral finfish; uptake, activation and detoxication of organic xenobiotics. Environ. Health Perspect. 71, 155–70.

    PubMed  Google Scholar 

  • Wells, D.E., De Boer, J., Tuinstra, L.G.M.T., Reutergårdh, L. and Griepink, B. (1988) Improvements in the analysis of chlorobiphenyls prior to the certification of seven CBs in two fish oils. Fresenius' Z. Anal. Chem. 332, 591–7.

    Google Scholar 

  • Witbaard, R. (1989) The distribution of a number of macrobenthos species in the North Sea. NIOZ-Rapport 1989–1, 1–99 (in Dutch).

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Authors and Affiliations

  1. Netherlands Institute for Sea Research, PO Box 59, 1790 AB, Den Burg Texel, The Netherlands

    J.M. Everaarts & M.Th.J. Hillebrand

  2. Research Institute of Toxicology, PO Box 80058, 3508 TB, Utrecht, The Netherlands

    P.J. Den Besten

  3. Oak Ridge National Laboratory, Environmental Sciences Division, PO Box 2008, Oak Ridge, USA

    R.S. Halbrook & L.R. Shugart

Authors
  1. J.M. Everaarts
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  2. P.J. Den Besten
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  3. M.Th.J. Hillebrand
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  4. R.S. Halbrook
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  5. L.R. Shugart
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Everaarts, J., Den Besten, P., Hillebrand, M. et al. DNA strand breaks, cytochrome P-450-dependent monooxygenase system activity and levels of chlorinated biphenyl congeners in the pyloric caeca of the seastar (Asterias rubens) from the North Sea. Ecotoxicology 7, 69–79 (1998). https://doi.org/10.1023/A:1008811802432

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