Behaviour, development and metal accumulation in striped marsh frog tadpoles (Limnodynastes peronii) exposed to coal mine wastewater

Fabbro, LD ORCiD: 0000-0001-8893-5115Coal mining generates large quantities of complex effluent, and this often contains high levels of dissolved solids, suspended solids, metals, hydrocarbons, salts and other compounds. Substantial volumes of mine wastewater are periodically discharged into the environment, through both planned and accidental releases, and this raises concerns about the potential for adverse impacts on aquatic wildlife. There have been few attempts to explore sub-lethal effects of coal mine wastewater on amphibians compared to other organisms, and this is particularly true for Australian species. To address existing knowledge gaps, we exposed striped marsh frog (Limnodynastes peronii) tadpoles to 25, 50 and 100% coal mine wastewater collected from two holding dams (CMW1 and CMW2) located at an open cut mine in Central Queensland, Australia. The exposure lasted for four weeks, after which survival, growth and development, swimming behaviour, and concentrations of metals and metalloids in tail and liver tissues were assessed. Physico-chemical parameters varied considerably between sites, with higher turbidity, nutrients, total and dissolved organic carbon, alkalinity and arsenic (As) concentrations at CMW1, and higher conductivity, salinity, dissolved solids, hardness and sulfate levels at CMW2. There was no mortality in controls and less than 5% mortality in CMW1 treatments, whereas survival was significantly decreased in tadpoles exposed to CMW2 with 40 and 55% mortality in the 50 and 100% treatments, respectively. Development was significantly delayed in 100% CMW1 wastewater, but tadpole size (growth) was not influenced by the exposure. Hepatosomatic indices were significantly increased in tadpoles exposed to 25 and 50% CMW1 but not the 100% treatment group. Exposed tadpoles (predominantly those exposed to CMW1) exhibited increased activity after very short-term exposure (24h), but this did not persist as animals approached metamorphic climax. At the end of the experiment, tadpoles exposed to both wastewaters had elevated levels of selenium (Se), cobalt (Co) and As in tail and liver tissue compared to controls. Manganese (Mn) levels were also elevated in livers and tails of CMW2 exposed tadpoles. Hepatic tissue accumulated 8-9 times higher concentrations of Co, Mn and Se compared to tail tissue, irrespective of treatments. Future research is warranted to explore possible relationships between metal bioaccumulation, morpho-physiological effects during development, and subsequent higher-level outcomes related to individual performance and population fitness