Influence of season and pollution on the antioxidant defenses of the cichlid fish acará (Geophagus brasiliensis).

The livers of Geophagus brasiliensis collected from both a non-polluted site and a polluted site were analyzed for different antioxidant defenses, O2 consumption, thiobarbituric acid-reactive substance (TBARS) levels, and histological damage. Compared to controls (116.6 +/- 26.1 nmol g-1), TBARS levels were enhanced at the polluted site (284.2 +/- 25.6 nmol g-1), as also was oxygen consumption (86.6 +/- 11.3 and 128.5 +/- 9.8 micromol O2 min-1 g-1, respectively). With respect to enzymatic antioxidants, increased catalase activities (8.7 +/- 1.3 and 29.2 +/- 2.4 mmol min-1 g-1, respectively), unchanged superoxide dismutase activities (767.2 +/- 113.3 and 563.3 +/- 70.2 U g-1, respectively), and diminished glutathione S-transferase activities (29.0 +/- 3.2 and 14.9 +/- 3.2 micromol min-1 g-1, respectively) were detected. Reduced glutathione (1.91 +/- 0.17 and 1.37 +/- 0.25 mM, respectively), oxidized glutathione (1.50 +/- 0.20 and 0.73 +/- 0.17 mM, respectively), and total glutathione (3.40 +/- 0.26 and 2.07 +/- 0.27 mM, respectively) concentrations were also below control values at the polluted site. Nevertheless, the observed ethoxyresorufin-O-deethylase activities (1.34 +/- 0.11 and 16.7 +/- 0.21 pmol min-1 mg-1, respectively) showed enhanced values at the polluted site. The main histological damage observed in the hepatocytes from fish collected at the polluted site was characterized by heavy lipid infiltration. Fish collected at the end of spring showed higher O2 consumption, higher superoxide dismutase and glutathione S-transferase activities, and higher total and oxidized glutathione concentrations compared to the beginning of autumn. No seasonal changes were observed in catalase activities, glutathione or TBARS levels. Fish chronically exposed to relatively high pollution levels seem to be unable to set up adequate antioxidant defenses, probably due to severe injury to their hepatocytes. The higher antioxidant defenses found at the end of spring are probably related to the enhanced activities during high temperature periods in thermoconforming organisms.

[1]  S. De Flora,et al.  Enhanced liver metabolism of mutagens and carcinogens in fish living in polluted seawater. , 1991, Mutation research.

[2]  W B Jakoby,et al.  Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. , 1974, The Journal of biological chemistry.

[3]  D. Sheehan,et al.  Effects of seasonality on xenobiotic and antioxidant defence mechanisms of bivalve molluscs. , 1999, Comparative biochemistry and physiology. Part C, Pharmacology, toxicology & endocrinology.

[4]  K. Shepard,et al.  The use of bioindicators for assessing the effects of pollutant stress on fish , 1989 .

[5]  F. Tietze Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. , 1969, Analytical biochemistry.

[6]  D. Phillips,et al.  Biomarkers of marine pollution observed in species of mullet living in two eastern Mediterranean harbours , 1997 .

[7]  C. Pueyo,et al.  Biochemical Indicators of Oxidative Stress in Fish from Polluted Littoral Areas , 1993 .

[8]  T. Gabryelak,et al.  Seasonal variations in the activities of peroxide metabolism enzymes in erythrocytes of freshwater fish species. , 1983, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[9]  S. E. Bonga The stress response in fish , 1997 .

[10]  A. Bainy,et al.  Oxidative stress in gill, erythrocytes, liver and kidney of Nile tilapia (Oreochromis niloticus) from a polluted site , 1996 .

[11]  P. Thomas,et al.  Effect of xenobiotics on peroxidation of hepatic microsormal lipids from striped mullet (Mugil cephalus) and Atlantic croaker (Micropogonias undulatus) , 1988 .

[12]  R. Mayer,et al.  Direct fluorometric methods for measuring mixed function oxidase activity. , 1978, Methods in enzymology.

[13]  D. L. Cinti,et al.  Preparation of microsomes with calcium. , 1978, Methods in enzymology.

[14]  V. Palace,et al.  VARIATION OF HEPATIC ENZYMES IN THREE SPECIES OF FRESHWATER FISH FROM PRECAMBRIAN SHIELD LAKES AND THE EFFECT OF CADMIUM EXPOSURE , 1993 .

[15]  A. Goksøyr,et al.  The cytochrome P-450 system in fish, aquatic toxicology and environmental monitoring , 1992 .

[16]  L. Flohé,et al.  Superoxide dismutase assays. , 1984, Methods in enzymology.

[17]  J. Härdig,et al.  Seasonal and ontogenetic effects on methaemoglobin and reduced glutathione contents in the blood of reared baltic salmon , 1983 .

[18]  L. Dušek,et al.  Responses of carp hepatopancreatic 7‐ethoxyresorufin‐O‐deethylase and glutathione‐dependent enzymes to organic pollutants—a field study , 1997 .

[19]  R. Thurman,et al.  Biotransformation and Zonal Toxicity , 1986 .

[20]  D. W. Filho,et al.  Seasonal changes in antioxidant defenses of the digestive gland of the brown mussel (Perna perna) , 2001 .

[21]  A. Aksnes,et al.  Catalase, glutathione peroxidase and superoxide dismutase in different fish species , 1981 .

[22]  B. Halliwell,et al.  Free radicals in biology and medicine , 1985 .

[23]  D. Livingstone The fate of organic xenobiotics in aquatic ecosystems: quantitative and qualitative differences in biotransformation by invertebrates and fish. , 1998, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[24]  D. Larsson,et al.  Seasonal variations in the activities of selected hepatic biotransformation and antioxidant enzymes in eelpout (Zoarces viviparus). , 1999, Comparative biochemistry and physiology. Part C, Pharmacology, toxicology & endocrinology.

[25]  C. Giulivi,et al.  Antioxidant defences in marine fish—I. Teleosts , 1993 .

[26]  E. Beutler,et al.  Improved method for the determination of blood glutathione. , 1963, The Journal of laboratory and clinical medicine.

[27]  H. Aebi,et al.  Catalase in vitro. , 1984, Methods in enzymology.

[28]  大川 博,et al.  Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction , 1979 .

[29]  M. Machala,et al.  Glutathione‐dependent detoxifying enzymes in rainbow trout liver: Search for specific biochemical markers of chemical stress , 1997 .

[30]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[31]  D. W. Filho,et al.  Antioxidant defences in marine fish—II. Elasmobranchs , 1993 .

[32]  D. W. Filho Fish antioxidant defenses--a comparative approach. , 1996 .

[33]  R. Bird,et al.  Comparative studies on different methods of malonaldehyde determination. , 1984, Methods in enzymology.

[34]  J. Pedrajas,et al.  Antioxidant and detoxifying fish enzymes as biomarkers of river pollution. , 1997, Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals.

[35]  Mark S. Myers,et al.  Neoplastic and other diseases in fish in relation to toxic chemicals: an overview☆ , 1988 .

[36]  P. Thomas,et al.  Effects of metals and organic compounds on hepatic glutathione, cysteine, and acid-soluble thiol levels in mullet (Mugil cephalus L.). , 1984, Toxicology and applied pharmacology.

[37]  R. Wenning,et al.  Biochemical responses in aquatic animals: A review of determinants of oxidative stress , 1989 .