Port Curtis is one of Australia's leading ports for which substantial industrial expansion is proposed over the next decade. However, there has been little attempt to date to assess the extent of contamination in waters, sediments and biota or to characterize the potential impacts of contaminants on aquatic biota. Contaminants of potential concern to biota and human health were investigated in the Port Curtis estuary using a screening-level risk assessment approach. Dissolved metal concentrations in waters were below [ANZECC/ARMCANZ, 2000. Australian and New Zealand Guidelines for Fresh and Marine Water Quality, Vol. 1. The Guidelines, Australian and New Zealand Environment and Conservation Council and Agriculture and Resource Management Council of Australia and New Zealand] trigger values, suggesting low risk of these contaminants. In sediments, arsenic, nickel and chromium concentrations exceeded interim sediment quality guidelines-low (ISQG-low), but were also high in the reference zone suggesting a natural origin. Historical data on naphthalene in Harbour sediments showed that it was also of potential concern. Bioaccumulation of contaminants in a range of biota was also used as an indicator of contaminant exposure. Biota were generally enriched in metals and tributyltin, which was also elevated in water and sediments. Although not unique to Port Curtis, mercury in barramundi was identified as a potential risk to human health.