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An extensive review of all the avoidance apparatus which have been used to date in chemical avoidance tests and also an analysis of the trends and needs for new improved designs led to the design of a new avoidance apparatus. The new apparatus is a viable tool for analyzing the behavioral responses of fish to potentially toxic effluents in three dimensions. The method is quick, sensitive and relatively easy to use if sufficient fish stock is available. / The variable parameters of the avoidance reaction were three: (a) the type of the pollutant, (b) the injection system, and (c) the concentration of the pollutant. / Statistical analysis for the experiments included preliminary calculations, time series analysis, analysis of variance and Duncan's tests. / Preliminary tests evaluated the general disposition of fish in the channel, the influence of a water acclimation jet and also the effects of noise, light and temperature. Experiments were performed to define the optimal number of fish per experiment, the influence of video analysis and also of variability, adaptability and continuity. / The types of pollutants used were D.S.S. (dodecyl sodium sulfate)--a well-known reference pollutant, polymers, monomers, a plating industrial effluent and its components (Cr, Ni, Fe). / For all of these, horizontal and vertical distributions were produced together with avoidance curves and avoidance reaction representations. / The role of the avoidance reaction in relation to lethal and sublethal levels has been defined; after the evaluation of synergism and antagonism phenomena, proposed adjustments for safe toxicant levels have been estimated.

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Previous studies have shown that rainbow trout exposed to dieldrin via diet for 9 to 12 weeks increased biliary excretion of a subsequent dose of [¹⁴C]dieldrin by 500%. This was not explained by induction of the cytochrome P-450 (CYP) system involved in oxidative metabolism of these compounds. We hypothesized that epoxide hydrolase activity increased in dieldrin fed-fish. Epoxide hydrolase is an enzyme that catalyzes the hydrolysis of epoxide compounds to their corresponding diols. For instance, dieldrin is metabolized to 6,7 trans-aldrindihydrodiol. This study investigated the activity of epoxide hydrolase in microsomes and cytosol of rainbow trout fed a diet that contained 0 or 15 ppm dieldrin. Fish were fed control or dieldrin diet (0.324 ug/g body weight/day) for 3, 6, or 9 weeks. There was a small increase in mortality and decrease in body weight among dieldrin-fed fish after 9 weeks. After week 9, dieldrin-fed fish were fed a control diet for an additional 3 weeks because of these signs of toxicity. At week 12, the difference of body weight between control and treated was not significant. Microsomal and cytosolic epoxide hydrolase activities were measured with a radiometric assay which determined differential partitioning of the parent compound (epoxide) in dodecane and the metabolite (diol) in the aqueous phase. Assays were run at optimal pH and temperature using [³H]trans-stilbene oxide (pH 7) as substrate for cytosol and [³H]cis-stilbene oxide (pH 8) as substrate for microsomes. In order to prevent competition for reaction with stilbene oxide, depletion of glutathione was efficiently achieved by dialysis at 4°C for 2 hours at room temperature in buffer [pH 7.5, potassium phosphate 10 mM, KCL 0.15 M, EDTA 1 mM, BHT 0.1 mM, 0.1 mM PMSF]. Protein quantification was determined by using BCA assay and concentrations were always between 5 and 25 ug/ml in the final assay volume. Epoxide hydrolase activities were not significantly different in cytosol or microsomes from control and dieldrin-fed fish. Dieldrin residues in liver were analyzed by gas chromatography with electron capture detection (GC/ECD). The concentration in the liver increased with time of exposure and declined markedly in fish fed dieldrin for 9 weeks and then fed control diet. No dieldrin was detected in livers from control fish. / Graduation date: 2003

This research focuses on the development, evaluation, and application of a mercury (Hg) mass balance model for predicting the accumulation of Hg in fish. This model requires accurate estimates of Hg elimination rate by fish and feeding rates to adequately predict Hg concentration in fish. An empirical model was developed to estimate Hg elimination by fish using data obtained from published experiments. This analysis showed that Hg elimination rate was overestimated in short-term experiments, positively correlated to water temperature, negatively correlated to body size, and that the elimination rate of inorganic Hg was faster than that of methylmercury. This empirical model was then incorporated in a Hg mass balance model to predict the concentration of Hg in fish. The Hg mass balance model accurately predicted Hg concentration in fish when it was combined with food consumption rates that were determined using a radioisotopic method. This analysis suggested that the parameters of the Hg mass balance model were adequate for predicting Hg concentration in fish. I also showed that Hg concentration tended to be underestimated by the Hg mass balance model when it was combined with feeding rates determined with a laboratory-derived bioenergetic model, probably because activity costs derived in the laboratory do not reflect activity costs of fish in the field. Beside predicting Hg concentration in fish, I showed that this mass balance model could also be used to estimate feeding rates of fish in the field by measuring the concentration of Hg in fish. This approach was validated using data obtained from a published experiment. It was also successfully tested using independent estimates of feeding rates obtained with a radioisotopic method. I applied this Hg mass balance model to compare the energy budget of sympatric populations of dwarf and normal whitefish (Coregonus clupeaformis). This analysis showed that dwarf whitefish consumed 40--50% more food than normal whitefi

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M.Sc. / Please refer to full text to view abstract

M.Sc. (Zoology) / Tilapia sparrmanii (Smith) obtained from the Provincial Fishery in Lydenburg was used for toxicological studies. The haematology of T. sparrmanii was investigated after exposure to sub-lethal doses of atrazine, zinc and iron for 72 hours in a continuous flow and static system, as well as prolonged (4 weeks) exposure in the continuous flow system. After exposure the blood of the experimental fish were sampled. Mean values and standard errors were obtained for several haematological parameters such as number of red and white blood cells, haematocrit, haemoglobin, mean corpuscular volume, pH, plasma glucose-, protein-, lipid- and lactate concentrations, as well as plasma Na+, K+, eland Ca+. Statistically significant differences were found between the values of haematological parameters of experimental and control fish. A tendency towards haemoconcentration was observed when fish were exposed to the low pollutant concentrations, while signs of haemodilution was found at higher concentrations. The results in this study demonstrate that, after prolonged exposure, T. sparrmanii develops a tolerance and partially recovers from the stress condition. The bioconcentration of atrazine in the blood of T. sparrmanii after exposure were determined by liquid chromatography and zinc and iron by atomic absorption spectrophotometry. During this study it was found that atrazine, zinc and iron concentrated in the blood of T. sparrmanii after short- and prolonged exposure to the pollutants. The routine oxygen consumption of "!'. sparrmanii exposed to atrazine, zinc and iron were determined in a continuous flow apparatus over 72 hours. The oxygen consumption of the experimental fish fluctuated for the first 8-12 hours during the acclimation period. The influence of atrazine on the routine oxygen consumption was not so drastic as that of zinc and iron. Zinc caused a decrease in the routine oxygen consumption of the experimental fish over 72 hours, while iron increased the routine oxygen consumption...

A research study was conducted to examine the relationship between speciation of heavy metals (Zn, Cd, Pb, Cu) and their biotoxicity. Therefore, static bioassay experiments were designed to determine LC-50 values for those metals on mosquitofish, Gambusia affinis. The standard bioassay scheme consisted of twelve 16-liter all-glass aquaria that provided duplicates of five different metal concentrations and a control for each run. Deionized tap water and filtered retention/detention water from Maitland Pond were used for bioassays. The effects of various physiochemical parameters such as pH, alkalinity, hardness and organic complexation on metal toxicity were determined. Also, synergistic and antagonistic effects resulting from metal mixtures were examined. It was concluded that metals in pond water were considerably less toxic than in deionized water for mosquitofish. Also, copper was consistently the most toxic metal tested for both deionized water and pond water.

The brain of fish, as in other vertebrates, is responsible for many functions basic to life and is also thought to be an anoxia-sensitive tissue. Therefore, during anoxia, the maintenance of energy within the brain is of paramount importance to the survival of the animal. Studies concerning energy levels and storage and the use of anaerobic metabolism in fish brains following exposure to anoxia are lacking. Rainbow trout (<u>Salmo gairdneri</u>) and brown bullhead catfish (<u>Ictalurus nebulosus</u>) occupy ecologically distinct habitats. Their tolerance of anoxia is different; trout survived l2 minutes while bullhead survived 62 minutes in anoxic water. Brains from control and anoxiaâ exposed trout and bullheads were analyzed using enzymatic assays and high pressure liquid chromatography (HPLC). Control bullhead brains had higher concentrations of glycogen, ATP, CrP, and glucose than control trout. With anoxia, bullheads showed a significant decrease in ATP, CrP, and glycogen with no change noted for glucose, ketone bodies (betaâ hydroxybutyrate and acetoacetate), or alternative anaerobic endproducts (succinate, alanine, propionate, isobutyrate, isovalerate, and ethanol). Lactic acid increased two-fold with anoxia. The bullhead was able to generate ATP by depleting its CrP stores and through classical anaerobic glycolysis. Death was most likely due to an inability to maintain ATP levels. Catfish may survive anoxia longer than trout in part due to greater fuel stores. Rainbow trout brain stored approximately one sixth the amount of glycogen as bullheads. With anoxia, these stores were depleted but there was no significant decrease in ATP, CrP, or glucose; the alternative endproducts also did not change. There was a l00% increase in lactic acid, suggesting that anaerobic glycolysis helped maintain ATP levels. Death may be due to factors other than ATP depletion such as lactic acid injury and increased intracellular free calcium. / Master of Science