Freshwater mussels play key roles in aquatic ecosystems, but are experiencing a global decline. Although studies have reported high acute sensitivity of mussels to some contaminants, chronic toxicity data are lacking for deriving high-reliability water quality guideline values. Ammonia is a contaminant of potential concern in some catchments of tropical northern Australia, where freshwater mussels are important ecological and cultural components. The extremely soft waters (hardness < 5 mg/L) of these environments can result in increased toxicity of many contaminants including ammonia, and regionally relevant tropical guideline values are needed to adequately protect these unique ecosystems. An optimized 14-d toxicity test protocol was used to assess the chronic toxicity of ammonia for 2 species, the lotic Velesunio sp. and the lentic Velesunio angasi. Ammonia exposures were conducted at pH 6.0 and 27 ± 0.5 °C to represent local environmental conditions, using shell length growth rate as the endpoint. Chronic toxicity estimates indicated high sensitivity to ammonia, with mean median effect concentrations (in total ammonia nitrogen) being 7.0 mg/L for V. angasi from the semi-urbanized Lake Bennett, 9.2 mg/L for V. angasi from Sandy Billabong, and 11.3 mg/L for Velesunio sp. from Gulungul Creek. When the 10% effect concentration values were compared with other chronic ammonia data (normalized to pH 7.0 and 20 °C), Velesunio spp. were found to be more sensitive than 8 of 16 other temperate and 7 of 9 tropical invertebrate and fish species. These chronic toxicity estimates will be used to further inform regionally relevant and site-specific guideline values. Environ Toxicol Chem 2019;38:841-851. © 2019 Commonwealth of Australia. Published by Wiley Periodicals Inc. on behalf of SETAC.

In aquatic toxicity testing of engineered nanoparticles (ENPs) the process of agglomeration is very important as it may alter bioavailability and toxicity. In the present study, we aimed to identify test conditions that are favorable for maintaining stable ENP suspensions. We evaluated the influence of key environmental parameters: pH (2-12) and ionic strength using M7, Soft EPA (S EPA) medium, and Very Soft EPA (VS EPA) medium; and observed the influence of these parameters on zeta potential, zeta average, and acute immobilization of Daphnia magna for three different ENPs. Despite being sterically stabilized, test suspensions of silver (Ag) ENPs formed large agglomerates in both VS EPA and M7 media; and toxicity was found to be higher in VS EPA medium due to increased dissolution. Low-agglomerate suspensions for zinc oxide (ZnO) could be obtained at pH 7 in VS EPA medium, but the increase in dissolution caused higher toxicity than in M7 medium. Titanium dioxide (TiO2) ENPs had a point of zero charge in the range of pH 7-8. At pH 7 in VS EPA, agglomerates with smaller hydrodynamic diameters (~200nm) were present compared to the high ionic strength M7 medium where hydrodynamic diameters reached micrometer range. The stable suspensions of TiO2 ENPs caused immobilization of D. magna, 48-h EC50 value of 13.7mgL(-1) (95% CI, 2.4mg-79.1mgL(-1)); whereas no toxicity was seen in the unstable, highly agglomerated M7 medium suspensions, 48-h EC50 >100mgL(-1). The current study provides a preliminary approach for methodology in testing and assessing stability and toxicity of ENPs in aquatic toxicity tests of regulatory relevance.

The present study investigated changes in suspension stability and ecotoxicity of engineered nanoparticles (ENPs) by addition of Suwannee River natural organic matter and aging of stock and test suspensions prior to testing. Acute toxicity tests of silver (Ag), zinc oxide (ZnO), and titanium dioxide (TiO2 ) ENPs with Daphnia magna were carried out following Organisation for Economic Co-operation and Development test guidelines. Daphnia magna was found to be very sensitive to Ag ENPs (48-h 50% effective concentration 33 μg L(-1) ), and aging of the test suspensions in M7 medium (up to 48 h) did not decrease toxicity significantly. Conversely, the presence of Suwannee River natural organic matter (NOM; 20 mg L(-1) ) completely alleviated Ag ENP toxicity in all testing scenarios and did not aid in stabilizing suspensions. In contrast, addition of Suwannee River NOM stabilized ZnO ENP suspensions and did not decrease toxicity. Aging for 48 h generated monotonous concentration-response curves in the presence and absence of Suwannee River NOM. At concentrations up to 100 mg L(-1) TiO2 ENPs did not cause immobilization of D. magna under any of the tested conditions. Presence of Suwannee River NOM caused agglomeration in stock suspensions. The authors\' results suggest that aging and presence of Suwannee River NOM are important parameters in standard toxicity testing of ENPs, which in some cases may aid in gaining better control over the exposure conditions but in other cases might contribute to agglomeration or elimination of ENP toxicity. Therefore, modifications to the current guidelines for testing ENPs should be evaluated on a case-by-case basis. Environ Toxicol Chem 2015;34:497-506. © 2014 SETAC.