The prospect of utilizing the cichlid teleost tilapia (Oreochromis niloticus) as an alternative experimental model to mammals for immunotoxicity risk assessment is currently being proposed. As such, the National Toxicology Program's (NTP) standard battery of rodent immunotoxicity assays is being developed for use in this fish species. Included in the testing series are the hemolytic plaque forming cell (PFC) and the cytotoxic T-lymphocyte (CTL) assays, quantitative indicators of antibody production and cell-mediated activity, respectively. The assays were modified in consideration of specific tilapian immune parameters, then tested using fourteen environmental contaminants or drugs, ten of which are classified by the NTP as immunotoxic in rodents. Reduced antibody production via a decrease in plaque number was observed in response to exposure of tilapia to eight of the nine humoral immunotoxicants, and five of the five non-immunotoxicants. Under specific immunization circumstances, immunostimulation (also a response to immunotoxicity) was noted via an increase in plaque number in benzo[a]pyrene (B[a]P) exposed fish using the PFC assay, a result noted in rodents as well. Reduced T-cell recognition and lysis of allogeneic tilapian lymphocytes via a decrease in the percentage of specific 51Chromium (51Cr) release was observed in response to exposure of tilapia to the nine of the ten cell-mediated immunotoxicants, and four of the four non-immunotoxicants. Although the normal teleost immune responsiveness was slightly weaker than seen with mice under comparable conditions (presumably due to differences in antibody structure and decreased cells counts), tilapia were found to exhibit well-defined humoral and cell-mediated immune responses, and responses to immunotoxic and non-immunotoxic chemicals comparable to the rodent model. / Master of Science

Sea Turtles are long-lived marine reptiles that have existed for more than 150,000 years. All seven turtle species extant today are characterised as endangered by the World Conservation Union Red List of Threatened Species. This PhD thesis investigates aspects of endocrinology and toxicology in green (Chelonia mydas) (Peninsular Malaysia) and flatback (Natator depressus) (Curtis Island, Queensland, Australia) turtles. This thesis aimed to identify if toxicants have an affect on reproduction and development in marine turtles. A high affinity steroid binding protein showing an affinity for testosterone and oestradiol was identified and described in nesting C. mydas and the thermal profile of sex steroid binding proteins in C. mydas was studied. This is the first study that has investigated sex steroid binding proteins in marine turtles. A sex steroid binding protein was also observed in male and female hatchling C. mydas and the effect of temperature on sex steroid binding proteins was investigated. This new, minor-invasive method of sex identification shows that different sex steroid binding proteins could reflect different hormonal patterns in female and male hatchling C. mydas. The steroid and metabolites profiles in nesting C. mydas (Peninsular Malaysia) and N. depressus (Curtis Island, Queensland) were established. Progesterone levels were the highest followed by testosterone, oestrone and dihydrotestosterone levels. The significant role of oestrone in nesting sea turtles was verified. The elevated concentrations of corticosterone, thyroxine, glucose and triglyceride indicated high metabolic demands placed upon C. mydas and N. depressus during nesting activities. The metabolite profiles and hormone concentrations were investigated to determine whether there was any relationship between hormonal changes during nesting activities in C. mydas and N. depressus. A methodology to measure heavy metal concentrations in the blood and eggs of N. depressus was developed. Moreover, this was the first time that tin compound derivatives have been identified in marine turtles in Australia. A complex heavy metal profile was identified in eggs. It is suggested that eggs may be an important, practical and reliable indicator of environmental contamination in nesting marine turtles. Pesticides were measured in the blood and eggs of N. depressus. Organochlorine and polychlorinated biphenyl concentrations were measured in the blood in N. depressus. Traces of OCs and PCBs were detected in eggs of N. depressus. Furthermore, the effects of heavy metals in developing C. mydas of both sexes are presented. During development, it was investigated whether heavy metal concentrations were affected by age, gender or the different clutch in C. mydas. The effect of a variety of toxicants (i.e. pesticides and heavy metals) on the sex steroid binding protein properties was investigated in nesting C. mydas. This important study identifies mechanisms by which toxicants may exert a physiological effect upon sex steroids (i.e. testosterone and oestradiol) in nesting turtles. The toxicants were studied at a wide range of concentrations. Some toxicants were shown to weaken or abolish the association of the testosterone and/or oestradiol binding proteins in nesting C. mydas. Correlations were observed between heavy metal concentrations found in blood and eggs and steroid and metabolite levels in nesting N. depressus. Zinc which was previously measured in the blood was positively correlated with testosterone concentrations in nesting N. depressus. Chromium which was found in eggs was negatively correlated corticosterone concentrations. Manganese was the only heavy metal (measured in eggs) which was negatively correlated with progesterone and positively correlated with triglyceride concentrations in nesting N. depressus. Finally, a quantified histological method was developed to determine the sex in C. mydas hatchlings by investigating specific histological criteria. A number of “undetermined sex” hatchlings were verified and it is hypothesised that abnormal reproductive development is related to the presence of environmental contaminants previously found in the studied C. mydas population of Peninsular Malaysia.