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Flow Cytometric Determination of Genotoxic Effects of Exposure to Petroleum in Mink and Sea Otters

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Abstract

Three experiments were conducted to investigate the genotoxic effects of crude oil on mink and sea otters. In the first experiment, the effects on mink of chronic exposure to weathered Prudhoe Bay crude oil were studied. Female mink were fed a diet that included weathered crude oil for a period of 3 weeks prior to mating, during pregnancy and until weaning. Kits were exposed through lactation and by diet after weaning until 4 months of age. Kidney and liver tissues of the kits were examined using flow cytometry (FCM) and it was found that the genome size was increased in kidney samples from the experimental group compared to the control group. This effect was probably due to some type of DNA amplification and it could have been inherited from the exposed mothers or have been a somatic response to oil exposure in the pups. No evidence of clastogenic effects, as measured by the coefficient of variation (CV) of the G1 peak, was found in kidney or liver tissue. In the second experiment, yearling female mink were exposed either by diet or externally to crude oil or bunker C fuel oil. Evidence for clastogenic damage was found in spleen tissue for the exposure groups, but not in kidney tissue. No evidence of increased genome size was observed. In the third experiment, blood was obtained from wild-caught sea otters in Prince William Sound. The sea otters represented two populations: one from western Prince William Sound that was potentially exposed to oil from the Exxon Valdez oil spill and a reference population from eastern Prince William Sound that did not receive oil from the spill. The spill had occurred 1.5 years prior to obtaining the blood samples. Although the mean CVs did not differ between the populations, the exposed population had a significantly higher variance of CV measurements and five out of 15 animals from the exposed population had CVs higher than the 95% confidence limits of the reference population. It is concluded that FCM is a sensitive indicator of the clastogenic effects of oil exposure and that haematopoietic tissues and blood are best for detecting clastogenic damage. Moreover, the observed differences in the genome size of the kidney cells were possibly heritable effects, but this needs further investigation. Lastly, sea otters exposed to spilled oil 1.5 years earlier showed evidence of clastogenic damage in one-third of the individuals sampled.

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References

  • Ballachey, B.E., Bodkin, J.L. and De Gange, A.R. (1994) An overview of sea otter studies. In T.R. Loughlin (ed.) Marine mammals and the Exxon Valdez, pp. 47–59. San Diego: Academic Press.

    Google Scholar 

  • Bickham, J.W. (1990) Flow cytometry as a technique to monitor the effects of environmental genotoxins on wildlife populations. In S.S. Sandhu, W.R. Lower, F.J. de Serres, W.A. Suk and R.R. Tice (eds) In situ evaluations of biological hazards of environmental pollutants, pp. 97–108. New York: Plenum.

    Google Scholar 

  • Bickham, J.W. (1994) Genotoxic responses in blood detected by cytogenetic and cytometric assays. In M.C. Fossi and C. Leonzio (eds) Nondestructive biomarkers in vertebrates, pp. 141–52. Boca Raton, FL: Lewis Publishers.

    Google Scholar 

  • Bickham, J.W. and Smolen, M.J. (1994) Somatic and heritable effects of environmental genotoxins and the emergence of evolutionary toxicology. Environ. Health Perspect. 102 (Suppl. 12), 25–8.

    Google Scholar 

  • Bickham, J.W., Hanks, B.G., Smolen, M.J., Lamb, T. and Gibbons, J.W. (1988) Flow cytometric analysis of the effects of low level radiation exposure on natural populations of slider turtles (Pseudemys scripta). Arch. Environ. Contam. Toxicol. 17, 837–41.

    Google Scholar 

  • Bickham, J.W., Sawin, V.L., Burton, D.W. and McBee, K. (1992) Flow-cytometric analysis of the effects of triethylenemelamine on somatic and testicular tissues of the rat. Cytometry 13, 368–73.

    Google Scholar 

  • Bickham, J.W., Sawin, V.L., Smolen, M.J. and Derr, J.N. (1994) Further flow cytometric studies of the effects of triethylenemelamine on somatic and testicular tissues of the rat. Cytometry 15, 222–9.

    Google Scholar 

  • Brown, E.D., Baker, T.T., Hose, J.E., Kocan, R.M., Marty, G.D., McGurk, M.D., Norcross, B.L. and Short, J. (1996) Injury to the early life stages of Pacific herring in Prince William Sound after the Exxon Valdez oil spill. In S.D. Rice, R.B. Spies, D.A. Wolfe and B.A. Wright (eds) Proceedings of the Exxon Valdez Oil Spill Symposium, pp. 448–62. Bethesda: American Fisheries Society Symposium 18.

    Google Scholar 

  • Bue, B.G., Sharr, S., Moffit, S.D. and Craig, A.K. (1996) Effects of the Exxon Valdez oil spill on pink salmon embryos and preemergent fry. In S.D. Rice, R.B. Spies, D.A. Wolfe and B.A. Wright (eds) Proceedings of the Exxon Valdez Oil Spill Symposium, pp. 619–27. Bethesda: American Fisheries Society Symposium 18.

    Google Scholar 

  • Burton, D.W. and Bickham, J.W. (1989) Heterochromatin variation and DNA content conservatism in Geomys attwateri and G. breviceps (Rodentia: Geomyidae). J. Mamm. 70, 580–91.

    Google Scholar 

  • Burton, D.W., Bickham, J.W. and Genoways, H.H. (1989) Flow cytometric analysis of the nuclear DNA content in four families of Neotropical bats. Evolution 43, 756–65.

    Google Scholar 

  • Colborn, T., Dumanoski, D. and Myers, J.P. (1996) Our Stolen Future. New York: Penguin Books.

    Google Scholar 

  • Cronin, M.A. and Bickham, J.W. (in press) A population genetic analysis of the potential for a crude oil spill to induce heritable mutations and impact natural populations. Ecotoxicology.

  • Custer, T.W., Bickham, J.W., Lyne, T.B., Lewis, T., Ruedas, L.A., Custer, C.M. and Melancon, M.J. (1994) Flow cytometry for monitoring contaminant exposure in black-crowned nightherons. Arch. Environ. Contam. Toxicol. 27, 176–9.

    Google Scholar 

  • Custer, T., Hines, R.K., Melancon, M.J., Hoffman, D.J., Wickliffe, J.K., Bickham, J.W., Martin, J.W. and Henshel, D.S. (1997) Contaminant concentrations and biomarker response in great blue heron eggs from 10 colonies on the Upper Mississippi River, USA. Environ. Toxicol. Chem. 16, 260–71.

    Google Scholar 

  • Davis, R.W., Williams, T.M., Thomas, J.A., Kastelein, R.A. and Cornell, L.H. (1988) The effects of oil contamination and cleaning on sea otters (Enhydra lutris): II. Metabolism, thermoregulation, and behavior. Can. J. Zool. 66, 2782–90.

    Google Scholar 

  • Deaven, L.L. (1982) Application of flow cytometry to cytogenetic testing of environmental mutagens. In T.C. Hsu (ed.) Cytogenetic assays of environmental mutagens, pp. 325–51. Totowa, NJ: Allanheld, Osmun and Company, Publishers.

    Google Scholar 

  • Djomo, J.E., Ferrier, V., Gauthier, L., Zoll-Moreux, C. and Marty, J. (1995) Amphibian micronucleus test in vivo: evaluation of the genotoxicity of some major polycyclic aromatic hydrocarbons found in crude oil. Mutagenesis 10, 223–6.

    Google Scholar 

  • Engelhardt, F.R. (1983) Petroleum effects on marine mammals. Aquatic Toxicol. 4, 199–217.

    Google Scholar 

  • Estes, J.A., Bacon, C.E., Jarman, W.M., Norstrom, R.J., Anthony, R.G. and Miles, A.K. (1997) Organochlorines in sea otters and bald eagles from the Aleutian archipelago. Mar. Pollut. Bull. 34, 486–90.

    Google Scholar 

  • Grafodatskii, A.S., Lushnikova, T.P., Romashchenko, A.G. and Radzhabli, S.I. (1985) Distribution of structural heterochromatin and receptive sequences of DNA on chromosomes of a number of mustelid species (Carnovora, Mustelidae). Soviet Genet. 21, 147–52.

    Google Scholar 

  • Guerin, M.R., Ho, C.-H., Rao, T.K., Clark, B.R. and Epler, J.L. (1980) Polycyclic aromatic primary amines as determinant chemical mutagens in petroleum substitutes. Environ. Res. 23, 42–53.

    Google Scholar 

  • Hsu, T.C. (1983) Genetic instability in the human population: a working hypothesis. Hereditas 98, 1–9.

    Google Scholar 

  • Hsu, T.C. (1987) A historical outline of the development of cancer cytogenetics. Cancer Genet. Cytogenet. 28, 5–26.

    Google Scholar 

  • IARC (1989) IARC Monographs on the Evaluation of Carcinogenic Risks to Humans, Occupational Exposures in Petroleum Refining; Crude Oil and Major Petroleum Fuels. Lyon: IARC.

    Google Scholar 

  • Khan, R.A. and Ryan, P. (1991) Long term effects of crude oil on common murres (Uria aalge) following rehabilitation. Bull. Environ. Contam. Toxicol. 46, 216–22.

    Google Scholar 

  • Khan, S., Irfan, M. and Rahimtula, A.D. (1987) The hepatotoxic potential of a Prudhoe Bay crude oil: effect on mouse liver weight and composition. Toxicology 46, 95–105.

    Google Scholar 

  • Lamb, T., Bickham, J.W., Gibbons, J.W., Smolen, M.J. and McDowell, S. (1991) Genetic damage in a population of slider turtles (Trachemys scripta) inhabiting a radioactive reservoir. Arch. Environ. Contam. Toxicol. 20, 138–42.

    Google Scholar 

  • Lamb, T., Bickham, J.W., Lyne, T.B. and Gibbons, J.W. (1995) The slider turtle as an environmental sentinel: multiple tissue assays using flow cytometric analysis. Ecotoxicology 4, 5–13.

    Google Scholar 

  • Lipscomb, T.P., Harris, R.K., Rebar, A.H., Ballachey, B.E. and Haebler, R.J. (1994) Pathology of sea otters. In T.R. Loughlin (ed.) Marine mammals and the Exxon Valdez, pp. 265–79. San Diego: Academic Press.

    Google Scholar 

  • Lyons, K.J. (1989) Individual variation in diet in the female California sea otter, Enhydra lutris. MS thesis, University of California, Santa Cruz.

    Google Scholar 

  • McBee, K. and Bickham, J.W. (1988) Petrochemical-related DNA damage in wild rodents detected by flow cytometry. Bull. Environ. Contam. Toxicol. 40, 343–9.

    Google Scholar 

  • McBee, K. and Bickham, J.W. (1990) Mammals as bioindicators of environmental toxicity. In H.H. Genoways (ed.) Current mammalogy, pp. 37–88. New York: Plenum.

    Google Scholar 

  • McBee, K., Bickham, J.W., Donnelly, K.C. and Brown, K.W. (1987) Chromosomal aberrations in native small mammals (Peromyscus leucopus) and Sigmodon hispidus) at a petrochemical waste disposal site. I. Standard karyology. Arch. Environ. Contam. Toxicol. 16, 681–8.

    Google Scholar 

  • McCarthy, J.F., Jimenez, B.D., Shugart, L.R., Sloop, F.V. and Oikari, A. (1990) Biological markers in animal sentinels: laboratory studies improve interpretation of field data. In S.S. Sandhu, W.R. Lower, F.J. de Serres, W.A. Suk and R.R. Tice (eds) In situ evaluations of biological hazards of environmental pollutants, pp. 163–75. New York: Plenum.

    Google Scholar 

  • Matsuoka, A., Shudo, K., Saito, Y., Sofuni, T. and Ishidate, M., Jr (1982) Clastogenic potential of heavy oil extracts and some azaarenes in Chinese hamster cells in culture. Mutat. Res. 102, 275–83.

    Google Scholar 

  • Mazet, J.A.K. (1996) Mink exposed to petroleum products as a model for sea otters. PhD dissertation, University of California, Davis.

    Google Scholar 

  • Otto, F.J. and Oldiges, H. (1980) Flow cytogenetic studies in chromosomes and whole cells for the detection of clastogenic effects. Cytometry 1, 13–17.

    Google Scholar 

  • Otto, F.J., Oldiges, H., Gohde, W. and Jain, V.K. (1981) Flow cytometric measurement of nuclear DNA content variations as a potential in vitro mutagenicity test. Cytometry 2, 189–91.

    Google Scholar 

  • Pathak, S. and Stock, A.D. (1974) The X chromosomes of mammals: karyological homology as revealed by banding techniques. Genetics 78, 703–14.

    Google Scholar 

  • Piatt, J.F. and Ford, R.G. (1996) How many seabirds were killed by the Exxon Valdez oil spill? Am. Fish. Soc. Symp. 18, 712–19.

    Google Scholar 

  • Shugart, L., Bickham, J.W., Jackim, G., McMahon, G., Stein, J., Steinert, S.A. and Ridley, W.P. (1992) DNA alterations. In R.J. Huggett, P.M. Mehrle, H.L. Bergman and R.A. Kimerle (eds) The existing and potential value of biomarkers in evaluating exposure and environmental effects of toxic chemicals, pp. 125–53. Boca Raton, FL: Lewis Publishers.

    Google Scholar 

  • Stegeman, J.J., Brouwer, M., Di Giulio, R.T., Forlin, L., Fowler, B.A., Sanders, B.M. and Van Veld, P.A. (1992) Molecular responses to environmental contamination: enzyme and protein systems as indicators of chemical exposure and effect. In R.J. Huggett, R.A. Kimerle, P.M. Mehrle and H.L. Bergman (eds) Biomarkers: biochemical, physiological, and histological markers of anthropogenic stress, pp. 235–335. Boca Raton, FL: Lewis Publishers.

    Google Scholar 

  • Stubblefield, W.A., Hancock, G.A., Ford, W.H. and Ringer, R.K. (1995a) Acute and subchronic toxicity of naturally weathered Exxon Valdez crude oil in mallards and ferrets. Environ. Toxicol. Chem. 14, 1941–50.

    Google Scholar 

  • Stubblefield, W.A., Hancock, G.A., Prince, H.A. and Ringer, R.K. (1995b) Effects of naturally weathered Exxon Valdez crude oil on mallard reproduction. Environ. Toxicol. Chem. 14, 1951–60.

    Google Scholar 

  • Tice, R.R., Ormiston, B.G., Boucher, R., Luke, C.A. and Paquette, D.E. (1987) Environmental biomonitoring with feral rodent species. In S.S. Sandhu, D.M. Demanine, M.J. Mass, M.M. Moore and J.L. Mumford (eds) Short-term bioassays in the analysis of complex environmental mixtures, Vol. V, pp. 175–9. New York: Plenum Press.

    Google Scholar 

  • Vindelov, L.L., Christensen, I.J. and Nissen, N.I. (1983) A detergent-trypsin method for the preparation of nuclei for flow cytometric DNA analysis. Cytometry 3, 323–7.

    Google Scholar 

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Authors and Affiliations

  1. Department of Wildlife and Fisheries Sciences, Texas A&M University, College Station, TX, 77843, USA

    John W. Bickham

  2. Wildlife Health Center, School of Veterinary Medicine, University of California, Davis, CA, 95616, USA

    Jonna A. Mazet

  3. Institute of Arctic Biology, University of Alaska, Fairbanks, AK, 99775, USA

    John Blake

  4. World Wildlife Fund, 1250 Twenty-fourth St NW, Washington, DC, 20037, USA

    Michael J. Smolen

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  1. John W. Bickham
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  5. Brenda E. Ballachey
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Bickham, J.W., Mazet, J.A., Blake, J. et al. Flow Cytometric Determination of Genotoxic Effects of Exposure to Petroleum in Mink and Sea Otters. Ecotoxicology 7, 191–199 (1998). https://doi.org/10.1023/A:1008930626834

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