Toxicity of Herbicide Mixtures to Tropical Freshwater Microalgae Using a Multispecies Test.

Agriculture within the Great Barrier Reef catchment area has contributed to pesticide contamination of adjacent freshwater ecosystems that flow into the Great Barrier Reef World Heritage Area. A novel multispecies toxicity test was used to assess the toxicity of diuron and hexazinone, 2 herbicides commonly detected within the Great Barrier Reef catchment area, to a community of 3 tropical freshwater microalgae: Monoraphidium arcuatum, Nannochloropsis-like sp., and Pediastrum duplex. Diuron was the most toxic herbicide, with 10% inhibition concentration (IC10) values of 4.3, 7.1, and 29 µg/L for P. duplex, M. arcuatum, and Nannochloropsis-like sp., respectively, followed by hexazinone, with IC10 values of 15, 18, and 450 µg/L, respectively Toxicity testing on 2 commercial formulations (Barrage, 13.2% hexazinone and 48.6% diuron; Diurex, 90% diuron) showed that additives in the commercial formulations did not significantly increase the toxicity of diuron. Direct toxicity assessments were carried out on water samples from the herbicide-contaminated Sandy Creek, which discharges to the Great Barrier Reef lagoon, and a clean reference site, Tully Gorge in the Tully River. Toxicity was observed in several Sandy Creek samples. Artificial herbicide mixtures were assessed in synthetic soft water and natural freshwaters, with toxic responses being observed at environmentally relevant concentrations. The present study successfully applied a novel multispecies tropical microalgal toxicity test, indicating that it is an effective tool for the assessment of herbicide toxicity in both natural and synthetic freshwaters. Environ Toxicol Chem 2021;40:473-486. © 2020 SETAC.

Development and application of a multispecies toxicity test with tropical freshwater microalgae.

Microalgae are commonly used in ecotoxicity testing due to their ease of culturing and rapid cell division rates. These tests generally utilise a single species of algae; however, microalgae occur in the environment as complex communities of multiple species. To date, routine multispecies toxicity tests using tropical microalgae have not been available. This study investigated four tropical freshwater microalgal species for use in a chronic multispecies toxicity test based on the population growth (cell division) rate: Pediastrum duplex, Monoraphidium arcuatum, Nannochloropsis-like sp. and Chlorella sp. 12. Flow cytometric analysis identified the different fluorescence and light scattering properties of each algal species and quantified each species within multispecies mixtures. Following optimisation of test media nutrients and pH, a toxicity testing protocol was developed with P. duplex, M. arcuatum and Nannochloropsis-like sp. There were no significant differences in growth rates of each alga when tested over 72 h as single species or in multispecies mixtures. Atrazine and imazapic, two herbicides with different modes of action, were used to assess the sensitivity of the multispecies toxicity test. Atrazine was toxic to all species with 72-h IC10 values of 7.2, 63 and 280 µg/L for P. duplex, M. arcuatum and Nannochloropsis-like sp. respectively, while imazapic was not toxic to any species at concentrations up to 1100 µg/L. The toxicity of atrazine and imazapic to each microalgal species in the multispecies toxicity test was the same as that determined from single-species toxicity tests indicating that the presence of these microalgae in a mixture did not affect the toxicity of these two herbicides. This study is the first to develop a multispecies tropical microalgal toxicity test for application in freshwaters. This time- and cost-effective tool can be utilised to generate data to assist environmental decision making and to undertake risk assessments of contaminants in tropical freshwater environments.

Response of the hairy mussel Trichomya hirsuta to sediment-metal contamination in the presence of a bioturbator.

The accumulation of metals in tissue compartments of bivalve biomonitors is expected to reflect the phases in which metals are most bioavailable. In concurrent field and laboratory experiments we measured Zn, Cd and Pb concentrations in the gills and digestive glands of mussels exposed to sediments from Lake Macquarie in NSW, Australia. Mussels in the laboratory were also exposed to the bioturbating gastropod Batillaria australis. Zn, Cd and Pb concentrations in gills and digestive glands of mussels from both experiments were accumulated in proportion with levels of metal contamination in the sediments. An interaction in the field between site and tissue type was found for Cd and Pb suggesting variation in the phases in which metals are most bioavailable. No effect of bioturbation on metal accumulation in the bivalve was detected and we conclude that it is unlikely to be a significant factor in metal uptake when these species interact.

An assessment of five Australian polychaetes and bivalves for use in whole-sediment toxicity tests: toxicity and accumulation of copper and zinc from water and sediment.

The suitability of two polychaete worms, Australonereis ehlersi and Nephtys australiensis, and three bivalves, Mysella anomala, Tellina deltoidalis, and Soletellina alba, were assessed for their potential use in whole-sediment toxicity tests. All species except A. ehlersi, which could not be tested because of poor survival in water-only tests, survived in salinities ranging from 18 per thousand to 34 per thousand during the 96-hour exposure period. No mortality was observed in any of the species exposed to sediment compositions ranging from 100% silt to 100% sand for 10 days, thus demonstrating the high tolerance of the five species to a wide range of sediment types. All species showed decreased survival after exposure to highly sulfidic sediments in 10-day whole-sediment tests. In 96-hour water-only tests, survival decreased, and copper accumulation in body tissues increased with exposure to increasing copper concentration for all species except A. ehlersi, which again could not be tested because of its poor survival in the absence of sediment. S. alba and T. deltoidalis were the most sensitive species to aqueous copper (LC50s of 120 and 150 microg Cu/L, respectively). All species tested were relatively insensitive to dissolved zinc up to concentrations of approximately 1,000 microg/L. In addition and with the exception of N. australiensis, all species accumulated significant levels of zinc in their body tissues. Whole-sediment tests were conducted over a 10-day period with copper-spiked (1,300 microg/g) and zinc-spiked (4,000 microg/g) sediments equilibrated for sufficient time to ensure that pore water metal concentrations were well below concentrations shown to have any effect on organisms in water-only tests. Survival was decreased in the bivalves T. deltoidalis and S. alba after exposure to copper-spiked sediments, and all species-except T. deltoidalis, in which 100% mortality was observed-accumulated copper in their tissues. Exposure to zinc-spiked sediments significantly decreased the survival of only one species, T. deltoidalis. Both polychaetes appeared to regulate concentrations of zinc in their body tissues with no significant uptake of zinc occurring from the sediment phase. Of the five species assessed in this study, T. deltoidalis was found to be the most sensitive to copper- and zinc-contaminated sediments, and based on commonly used selection criteria (ASTM 2002a, ASTM 2002b, ASTM 2002c) is recommended for development as test species in whole-sediment toxicity tests.