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Sequential expression of biomarkers in Bluegill Sunfish exposed to contaminated sediment

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Abstract

The temporal expression of various biological rsponses was determined in Bluegill SunfishLepomis macrochirus exposed under controlled laboratory conditions to sediment containing high concentrations of polynuclear aromatic hydrocarbons, polychlorinated biphenyls and heavy metals. Liver, gill, blood, kidney, brain, spleen and intestine were removed from Sunfish sampled at 1, 2, 4, 8, 16, and 40 weeks post-exposure. Biomarker data were recorded for specific proteins, enzymatic activities, DNA integrity, and histopathology. Biomarkers in the laboratory exposed fish were similar to those of indigenous Sunfish sampled from the site of origin of the contaminated sediment. Several patterns of development of biomarkers over time were also evident. For example, the responses of certain biomarkers are not time-dependent (i.e., intestine and gill ATPase activities) while that of others, such as brain ATPase activity, liver cytochrome P450 and NADPH content, stress proteins, chromatin proteins and DNA strand breaks, fluctuate over time. Still other biomarkers, such as EROD activity, zinc protoporphyrin content of the blood, and DNA adducts, showed marked increases over time. Such patterns need to be considered when comparing laboratory and field results and deciding which biomarkers to use for biomonitoring programs. Implications for natural selection and population/community level responses are also discussed.

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Abbreviations

Ache:

acetylcholinesterase

ALAD:

δ-aminolevulenic acid dehydratase

BaP:

benzo[a]pyrene

BaPDE:

benzo[a]pyrene diol-epoxide

DDW:

double distilled water

EFPC:

East Fork Poplar Creek

EROD:

ethoxyresorufin-O-deethylase

PAH:

polycyclic aromatic hydrocarbon

PCB:

polychlorinated biphenyl

PAS:

periodic acid Schiff

ZPP:

zinc protoporphyrin

References

  • Adams, S.M., Shepard, K.L., Greeley, M.S. Jr., Jimenez, B.D., Ryon, M.G. Shugart, L.R. and McCarthy, J.F. (1989) The use of bioindicators for assessing the effects of pollutant stress on fish.Mar. Environ. Res. 28, 459–64.

    Google Scholar 

  • Adams, S.M., Shugart, L.R. and Southworth, G.R. (1990) Application of bioindicators in assessing the health of fish populations experienced contaminant stress. In McCarthy, J.F. and Shugart, L.R. eds,Biomarkers of Environmental Contamination, pp. 333–54. Boca Raton: Lewis Publishers.

    Google Scholar 

  • Alberts, B., Bray, D., Lewis, J., Raff, M., Roberts, K. and Watson, J.D. (1983)Molecular Biology of the Cell. New York: Garland Publishing.

    Google Scholar 

  • Ames, B.N. and Dubin, D.T. (1960) The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid.J. Biol. Chem. 235, 769–73.

    Google Scholar 

  • Bancroft, J.D. (1977)Theory and Practice of Histological Techniques. New York: Churchill Livingstone.

    Google Scholar 

  • Bickham, J.W. (1990) Flow cytometry as a technique to monitor the effects of environmental genotoxins on wildlife populations. In Sandu, S.S. ed.,In Situ Evaluations of Biological Hazards of Environmental Pollutants. New York: Plenum.

    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 (Psuedemys 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 

  • Blaxhall, P.C. (1972) The hematological assessment of the health of freshwater fish: a review of selected literature.J. Fish Biol. 4, 539–604.

    Google Scholar 

  • Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of protein utilizing the principle of protein dye binding.Anal. Biochem. 72, 248–54.

    Google Scholar 

  • Bradley, B.P. (1992) Are the stress proteins indicators of exposure or effect?Mr. Environ. Res., in pres.

  • Bradley, B.P. and Ward, J.B. (1989) Detection of a major stress protein using a peptide antibody.Mar. Environ. Res. 28, 471–5.

    Google Scholar 

  • Burnette, W.H. (1981) Western blotting: electrophoretic transfer of proteins from SDS-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.Anal. Biochem. 112, 195–203.

    Google Scholar 

  • Cory-Slechta, D.A., Weis, B. and Cox, C. (1989) Tissue distribution of Pb in adult vs. old rats: a pilot study.Toxicology 59, 139–50.

    Google Scholar 

  • Cyriac, P.J., Antony, A. and Nambison, P.N.K. (1989) Hemoglobin and hematocrit values in the fishOreochromis mossambicus (Peters) after short term exposure to copper and mercury.Bull. Environ. Contam. Toxicol. 43, 315–20.

    Google Scholar 

  • Danga, A.D. and Masurekar, A.B. (1989) Effect of naphthalene exposure on activity of some enzymes in the cichlid fish Tilapia,Sanatherodon mossambicus (Peters).J. Anim. Morphol. Physiol. 31, 159–68.

    Google Scholar 

  • Dillon, T.M. and Burton, W.D.S. (1991) Acute toxicity of PCB congeners toDaphnia magna andPimephales promelas.Bull. Environ. Contam. Toxicol. 46, 208–15.

    Google Scholar 

  • Drummond, R.A. and Russom, C.L. (1990) Behavioral toxicity syndromes: A promising tool for asessing toxicity mechanisms in juvenile Fathead Minnows.Environ. Toxicol. Chem. 9, 37–46.

    Google Scholar 

  • Finnelli, V.N., Klauder, D.S., Karaffa, M.A. and Petering, H.G. (1975) Interaction of zinc and lead on δ-aminolevulinate dehydratase.Biochem. Biophys. Res. Commun. 65, 303–11.

    Google Scholar 

  • Fletcher, G.L., King, M.J., Kideniuk, J.W. and Addison, R.F. (1982) Liver hypertrophy in winter Flounder following exposure to experimentally oiled sediments.Comp. Biochem. Physiol. 73C, 457–62.

    Google Scholar 

  • Flora, S.J.S., Dhawan, M. and Tandon, S.K. (1991) Effects of combined exposure to aluminum and ethanol on aluminum body burden and some neuronal, hepatic and hematopoietic biochemical variables in the rat.Human Exp. Toxicol. 10, 45–8.

    Google Scholar 

  • Freeman, S.E. and Thompson, B.D. (1990) Quantitation of ultraviolet radiation-induced cyclobutylpyrimidine dimers in DNA by video and photographic densitometry.Anal. Biochem. 186, 222–8.

    Google Scholar 

  • Greenaway, P.J. and Murray, K. (1971) Heterogeneity and polymorphism in chicken erythrocyte fraction V.Nature 229, 233–8.

    Google Scholar 

  • Gromman, D.B. (1982)Histology of the Striped Bass. Bethesda, MD: American Fisheries Society.

    Google Scholar 

  • Halbrook, R.S., Guzman, C.E., Wilkinson, K.J., Shugart, L.R., Watson, A.P. and Munro, N.B. (1992) A rapid whole blood cholinesterase assay with potential use in biological monitoring during chemical weapons disposal.J. Assoc. Off. Anal. Chem 75, 549–53.

    Google Scholar 

  • Hewish, D.R. and Burgoyne, L.A. (1973) Chromatin sub-structure. The digestion of chromatin DNA at regularly spaced sites by a nuclear deoxyribonuclease.Biochem. Biophys. Res. Commun. 52, 504–10.

    Google Scholar 

  • Higgins, R.C. and Dahmus, M.E. (1979) Rapid visualization of protein bands in preparative SDS-polyacrylamide gels.Anal. Biochem. 93, 257–60.

    Google Scholar 

  • Hodson, P.V., Blunt, B.R. and Whittle, D.M. (1980) Biochemical monitoring of fish blood as an indicator of biologically available lead.Thalassia Jugosl. 16, 389–96.

    Google Scholar 

  • Hollander, M. and Wolfe, D.A. (1973)Nonparametric Statistical Methods. New York: Wiley.

    Google Scholar 

  • Jimenez, B.D., Burtis, L.S., Ezell, G.H., Egan, B.Z., Lee, N.E., Beauchamp, J.J. and McCarthy, J.F. (1988) The mixed function oxidase system of Bluegill Sunfish,Lepomis macrochirus: Correlation of activities in experimental and wild fish.Environ. Toxicol. Chem. 7, 623–34.

    Google Scholar 

  • Jimenez, B.D., Oikari, A., Adams, S.M., Hinton, D.E. and McCarthy, J.F. (1990) Hepatic enzymes as biomarkers: interpreting the effects of environmental physiological and toxicological variables. In McCarthy, J.F. and Shugart, L.R. eds,Biomarkers of Environmental Contamination. Boca Raton: Lewis Publishers.

    Google Scholar 

  • Johannsen, K.A.M. and DePierre, J.W. (1978) Measurement of cytochrome P450 in the presence of large amounts of contaminating hemoglobin and methemoglobin.Anal. Biochem. 86, 725–32.

    Google Scholar 

  • Kinubagaran, R. and Joy, K.P. (1988) Toxic effects of three mercurial compounds on survival and histology of the kidney of the catfishClarias batrachus (L.)Ecotox. Environ. Safety 15, 171–9.

    Google Scholar 

  • Klaassen, C.D., Amdur, M.O. and Doul, J. (1986)Toxicology: the basic science of poisons. New York: Macmillan.

    Google Scholar 

  • Kornegay, F.C. (1991)Oak Ridge Reservation Environmental Report for 1990. Volume 1:Narrative, Summary, and Conclusions. Oak Ridge: Oak Ridge National Laboratory.

    Google Scholar 

  • Krahn, M.M., Myers, M.S., Burrows, D.G. and Malins, D.C. (1984) Determination of metabolites of xenobiotics in bile of fish from polluted waterways.Xenobiotica 8, 633–46.

    Google Scholar 

  • Krahn, M.M., Moore, L.K. and MacLeod, W.D. (1986)Standard analytical procedures of the NOAA National Analytical Facility, 1986: Metabolites of Aromatic Compounds in Fish Bile. NOAA Technical Memorandum NMFS F/NWC-102, Seattle, WA.

  • Labbe, R.F. and Rettmer, R.L. (1989) Zinc protoporphyrin: a product of iron-deficient erythropoiesis.Seminars Hematol. 26, 40–6.

    Google Scholar 

  • Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227, 680–5.

    Google Scholar 

  • Lakshmi, R., Kundu, R., Thomas, E. and Mansur, A.P. (1991) Mercuric-chloride-induced inhibition of different ATPase in the intestine of MudskipperBoleophthalmus dentatus.Ecotox. Environ. Safety 21, 18–24.

    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 

  • La Roca, P.T. and Carlson, G.P. (1979) The effect of polychlorinated biphenyls on adenosine triphosphate activity.Toxicol. Appl. Pharmacol. 48, 185–91.

    Google Scholar 

  • Loar, J.M., Adams, S.M., Boston, H.L., Jimenez, B.D., McCarthy, J.F., Smith, J.G., Southworth, G.R. and Stewart, A.J. (1988)First Annual Report on the Y-12 Plant Biological Monitoring and Abatement Program. Oak Ridge: ORNL/TM, Oak Ridge National Laboratory.

    Google Scholar 

  • Lowe-Jinde, L. and Niimi, A.J. (1986) Hematological characteristics of Rainbow Trout,Salmo gairdneri (Richardson) in response to cadmium exposure.Bull Environ. Contam. Toxicol. 37, 375–81.

    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 

  • McCarthy, J.F. and Shugart, L.R. (1990) Biological markers of environmental contamination. In McCarthy, J.F. and Shugart, L.R. eds,Biomarkers of Environmental Contamination. Boca Raton: Lewis Publishers.

    Google Scholar 

  • McKee, M.J., Hendricks, A.C. and Ebel, R.E. (1983) Effects of naphthalene on benzo[a]pyrene hydroxylase and cytochrome P-450 inFundulus heteroclitus.Aquat. Toxicol. 3, 103–14.

    Google Scholar 

  • Maccubin, A.E., Black, P., Trzeciak, L. and Black, J.J. (1984) Evidence for polynuclear aromatic hydrocarbons in the diet of bottom-feeding fish.Bull. Environ. Contam. Toxicol. 34, 876–80.

    Google Scholar 

  • Mahajan, C.L. and Dheer, J.M.S. (1980) An autoradiographic and cytochemical study of erythropoiesis in freshwater fish.J. Fish Biol. 17, 641–8.

    Google Scholar 

  • Missenar, C., Hansen, G., Kutter, D. and Kremer, A. (1989) Vanadium induced impairment of haem synthesis.Br. J. Ind. Med. 46, 744–7.

    Google Scholar 

  • Mitchell, R.A., Drake, J.E., Wittlin, L.A. and Rejent, T.A. (1977) Erythrocyte porphobilinogen synthase (delta aminolevulinate dehydratase) activity: a reliable and quantitative indicator of lead exposure in humans.Clin. Chem. 23, 105–11.

    Google Scholar 

  • Nath, K. and Kumor, N. (1989) Nickel-induced histopathological changes in gill architecture of a tropical freshwater perchColisa fasciatus.Sci. Total Eniron. 80, 293–6.

    Google Scholar 

  • Palyga, J. (1990) Polymorphism of histone H1 in Goose erythrocytes.Biochem. Genet. 28, 359–65.

    Google Scholar 

  • Phillips, A.W. and Langdon, R.G. (1962) Hepatic triphosphopyridine nucleotide-cyt-c reductase isolation, characterization, and kinetic studies.J. Biol. Chem. 237, 2652–60.

    Google Scholar 

  • Shaw, B.D. and Panigrahi, B.R. (1990) Brain acetylcholine-esterase activity studies in some fish species collected from a mercury contaminated estuary.Water Air Soil Pollut. 53, 327–34.

    Google Scholar 

  • Shaw, G.R. and Connell, D.W. (1986) Factors controlling bioaccumulation of PCBs. In Waid, J.S. ed.,PCBs in the Environment, Vol. I. Boca Raton: CRC Press.

    Google Scholar 

  • Shugart, L.R. (1985) Quantitating exposure to chemical carcinogens:in vivo alkylation of hemoglobin by benzo[a]pyrene.Boxicology 34, 211–20.

    Google Scholar 

  • Shugart, L.R. (1986) Quantifying adductive modification of hemoglobin from mice exposed to benzo[a]pyrene.Anal. Biochem. 152, 365–9.

    Google Scholar 

  • Shugart, L.R. (1987) Analysis of adduct formation in the Bluegill Sunfish (Lepomis macrochirus) between benzo[a]pyrene and DNA of the liver and hemoglobin of the erythrocyte.Aquatic Toxicol,9, 319–25.

    Google Scholar 

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

    Google Scholar 

  • Shugart, L.R. (1990) DNA damage is an indicator of pollutant-induced genotoxicity. In Landis, W.G. and van der Schalie, W.H. eds,Aquatic Toxicology and Risk Assessment, 13th Vol., ASTM STP 1096. Philadelphia: American Society for Testing and Materials.

    Google Scholar 

  • Stahr, H.M., Hyde, H. and Anderson, A. (1977)Analytical Toxicology Methods Manual. Ames: Iowa State University Press.

    Google Scholar 

  • Steele, C.W. (1985) Latent behavioral toxicity of copper to Sea Catfish,Arius felis, and Sheepshead,Archosargus probatoce phalus.J. Fish Biol. 27, 643–54.

    Google Scholar 

  • US EPA (1983)Methods for Chemical Analysis of Water and Wastes. Washington, DC: US Environmental Protection Agency, Office of Research and Development.

    Google Scholar 

  • US EPA (1986)Test Methods for Evaluating Solid Waste: Physical/Chemical Methods. Washington DC: US Environmental Protection Agency, Office of Solid Waste Emergency Response.

    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 

  • Winston, S.E. (1989) Analysis of proteins.In Ausubel, E.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. eds,Current Protocols in Molecular Biology, Vol. II. New York: Wiley.

    Google Scholar 

  • Zaugg, W.S. (1982) A simplified preparation for adenosine triphosphatase determination in gill tissue.Can. J. Fish. Aquat. Sci. 39, 215–7.

    Google Scholar 

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Authors and Affiliations

  1. Graduate Program in Environmental Toxicology, University of Tennessee, 37916, Knoxville, TN, USA

    C. W. Theodorakis & W. L. Farkas

  2. Environmental Sciences Division, Oak Ridge National Laboratory, 37831, Oak Ridge, TN, USA

    S. J. D'surney, J. F. McCarthy & L. R. Shugart

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

    J. W. Bickham

  4. LGL Ecological Genetics Inc., 77801, Bryan, TX, USA

    J. W. Bickham

  5. Department of Soil and Crop Sciences, Texas A & M University, 77843, College Station, TX, USA

    T. B. Lyne

  6. Department of Biological Sciences, University of Maryland (Baltimore County Campus), 21228, Catonsville, MD, USA

    B. P. Bradley

  7. Gulf Coast Research Laboratory, 39564, Ocean Springs, MS, USA

    W. E. Hawkins

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  1. C. W. Theodorakis
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  2. S. J. D'surney
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  3. J. W. Bickham
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  4. T. B. Lyne
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  5. B. P. Bradley
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  7. W. L. Farkas
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  8. J. F. McCarthy
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  9. L. R. Shugart
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Theodorakis, C.W., D'surney, S.J., Bickham, J.W. et al. Sequential expression of biomarkers in Bluegill Sunfish exposed to contaminated sediment. Ecotoxicology 1, 45–73 (1992). https://doi.org/10.1007/BF00702655

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  • DOI: https://doi.org/10.1007/BF00702655

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