The Murray-Darling basin is the largest river system in Australia with significant economic, social, recreational and cultural value. It supplies water for drinking and agriculture to a large inland area of the eastern and southern states of Australia. It is also the ultimate sink for many environmental contaminants that result from human activities within the catchment. Aquatic organisms live intimately with their environment and may be continuously exposed to these contaminants through the water column or the food chain. Some chemicals are bioaccumulated and biomagnified in tissue to reach high body burdens. Populations of native fish species within the Murray-Darling basin have been in decline since human settlement, yet little is known about the lethal and sublethal effects of environmental pollutants on native freshwater fish and many of the Australian water quality guidelines are based on data from exotic fish species. Researchers have correlated levels of pollution with immune dysfunction and an increased incidence of disease amongst wildlife populations. Many of the pollutants of the Murray-Darling basin have known immunotoxicity in both mammals and exotic fish species. The immune system is a sensitive target organ because, in order to maintain integrity, it requires constant renewal through the rapid proliferation and differentiation of cells. Efforts to increase numbers of native fish in the wild have led to an aquaculture industry that produces fingerlings for the restocking of waterways. In more recent years, this industry has matured and now produces table-size native freshwater fish for local and international markets. Although the industry has researched areas of reproduction, nutrition and stocking, there is little understanding of the immunology or immunotoxicology of Australian freshwater fish. This research project investigated the immunology of three large native fish species (i.e. 2 Murray cod, golden perch and silver perch), which are the basis of the native freshwater aquaculture industry. Additionally, a small fish species native to the basin (i.e. crimsonspotted rainbowfish) was studied as an alternative to the use of large fish. Of the four species, Murray cod possessed characteristics that made it an excellent candidate for ecoimmunotoxicity testing.

The Murray cod (Maccullochella peelii peelii Mitchell 1838) is Australia�s largest freshwater fish. Once highly abundant in the Murray-Darling river system, populations have drastically declined in recent decades. Many causes for this decline have been proposed, including over-fishing, habitat loss and altered river flow regimes. This study hypothesised that elevated salinities have led to selective mortality in some developmental stages, which have in turn depleted stock recruitment and adult populations. The objectives of this study were to determine the optimal, threshold, upper sublethal and lethal salinities for development of eggs, yolk-sac larvae, fry and fingerlings of M. peelii peelii. Investigation the impact of salinity on fertilisation utilised gametes of trout cod (M. macquariensis, Cuvier 1829) instead of M. peelii peelii. Studies were carried out in a controlled laboratory environment using test media prepared from commercial sea salt. The results showed that the eggs of the trout cod hatched only when fertilised and incubated in freshwater, and only larvae hatched in freshwater survived through the yolk absorption period of 12 days. Yolk utilisation efficiencies were not significantly different among the salinities of 0-0.30 g/L. There was no effect of pre- or post- fertilising processes on the salinity tolerances of yolk-sac larvae. No larvae survived at salinities higher than 0.30 g/L during the yolk utilisation period. Lethal salinity concentration in Trout cod and Murray cod larvae was exposure time dependent. The 1 day LC50 of the larvae was 1.97 and 2.33 g/L respectively, compared with the 12 day LC50 values of 0.50 and 0.35 g/L respectively. The threshold (no effect level) salinities of larvae of Trout cod and Murray cod were 0.46 and 0.34 g/L respectively at 12 days exposure. The salinity sensitivities of fry of Murray cod were moderated by increasing pH between pH 6.2 and 8.8, and stimulated by increasing temperatures from 15 to 30°C. The optimal salinity was only slightly affected by temperature. The threshold and upper sublethal salinities varied slightly depending on feeding regime. The salinity sensitivities of fingerlings of Murray cod were: LC50 = 13.7 g/L; optimal salinity from 4.6 to 5.0 g/L ; threshold salinity from 5.9 to 7.4 g/L, and upper sub-lethal salinity from 9.2 to 9.9 g/L � with the range in all cases affected by acclimation period salinity. The blood osmolality at LC50 of the fingerlings was 444 mOsmol/kgH2O or equivalent to 14.2 g/L, and the dehydration rate was 4.8%. The osmolality increased significantly in salinities higher than 9.0 and 6.0 g/L when fish were exposed for a period of 1 day and 41 days respectively. The oxygen consumption increased significantly in salinities higher than 8.0 g/L. Distortion of the notochord and corrosive skin syndrome were major symptoms describing sub-lethal effects found in the embryos, and fry and fingerlings of Murray cod respectively. Noting the risks of extrapolating directly from laboratory to field conditions, it is predicted that when salinity in natural habitats increases above 0.34 g/L a significant impact on Murray cod recruitment will result.