Development of a Site-Specific Guideline Value for Copper and Aquatic Life in Tropical Freshwaters of Low Hardness.

Copper (Cu) is a contaminant of potential concern for a uranium mine whose receiving waters are in the World Heritage-listed Kakadu National Park in northern Australia. The physicochemical characteristics of the freshwaters in this region enhance metal bioavailability and toxicity. Seven tropical species were used to assess the chronic toxicity of Cu in extremely soft freshwater from a creek upstream of the mine. Sensitivity to Cu was as follows: Moinodaphnia macleayi > Chlorella sp. > Velesunio sp. > Hydra viridissima > Amerianna cumingi > Lemna aequinoctialis > Mogurnda mogurnda. The 10% effect concentrations (EC10s) ranged from 1.0 µg/L Cu for the cladoceran Moinodaphnia macleayi to 9.6 µg/L for the fish M. mogurnda. The EC50s ranged from 6.6 µg/L Cu for the mussel Velesunio sp. to 22.5 µg/L Cu for M. mogurnda. Geochemical modeling predicted Cu to be strongly bound to fulvic acid (80%-99%) and of low bioavailability (0.02%-11.5%) under these conditions. Protective concentrations (PCs) were derived from a species sensitivity distribution for the local biota. The 99% PC (PC99), PC95, PC90, and PC80 values were 0.5, 0.8, 1.0, and 1.5 µg/L Cu, respectively. These threshold values suggest that the current Australian and New Zealand default national 99% protection guideline value for Cu (1.0 µg/L) would not provide adequate protection in freshwaters of low hardness, particularly for this area of high conservation value. The continuous criterion concentration predicted by the Cu biotic ligand model for conditions of low pH (6.1), low dissolved organic carbon (2.5 mg/L), low hardness (3.3 mg/L), and 27 °C was 0.48 µg/L Cu, comparable with the PC99. Consideration of the natural water quality conditions of a site is paramount for protective water quality guidelines. Environ Toxicol Chem 2022;41:2808-2821. © 2022 Commonwealth of Australia. Environmental Toxicology and Chemistry © 2022 SETAC.

Resolving ecosystem complexity in ecological risk assessment for mine site rehabilitation.

CONTEXT: Ecological Risk Assessments (ERAs) are important tools for supporting evidence-based decision making. However, most ERA frameworks rarely consider complex ecological feedbacks, which limit their capacity to evaluate risks at community and ecosystem levels of organisation. METHOD: We used qualitative mathematical modelling to add additional perspectives to previously conducted ERAs for the rehabilitation of the Ranger uranium mine (Northern Territory, Australia) and support an assessment of the cumulative risks from the mine site. Using expert elicitation workshops, separate qualitative models and scenarios were developed for aquatic and terrestrial systems. The models developed in the workshops were used to construct Bayes Nets that predicted whole-of-ecosystem outcomes after components were perturbed. RESULTS: The terrestrial model considered the effect of fire and weeds on established native vegetation that will be important for the successful rehabilitation of Ranger. It predicted that a combined intervention that suppresses both weeds and fire intensity gave similar response predictions as for weed control alone, except for lower levels of certainty to tall grasses and fire intensity in models with immature trees or tall grasses. However, this had ambiguous predictions for short grasses and forbs, and tall grasses in models representing mature vegetation. The aquatic model considered the effects of magnesium (Mg), a key solute in current and predicted mine runoff and groundwater egress, which is known to adversely affect many aquatic species. The aquatic models provided support that attached algae and phytoplankton assemblages are the key trophic base for food webs. It predicted that shifts in phytoplankton abundance arising from increase in Mg to receiving waters, may result in cascading effects through the food-chain. CONCLUSION: The qualitative modelling approach was flexible and capable of modelling both gradual (i.e. decadal) processes in the mine-site restoration and the comparatively more rapid (seasonal) processes of the aquatic ecosystem. The modelling also provides a useful decision tool for identifying important ecosystem sub-systems for further research efforts.

Sediment spiking and equilibration procedures to achieve partitioning of uranium similar to contamination in tropical wetlands near a mine site.

The derivation of sediment quality guideline values (SQGVs) presents significant challenges. Arguably the most important challenge is to conduct toxicity tests using contaminated sediments with physico-chemistry that represents real-world scenarios. We used a novel metal spiking method for an experiment that ultimately aims to derive a uranium SQGV. Two pilot studies were conducted to inform the final spiking design, i.e. percolating a uranyl sulfate solution through natural wetland sediments. An initial pilot study that used extended mixing equilibration phases produced hardened sediments not representative of natural sediments. A subsequent percolation method produced sediment with similar texture to natural sediment and was used as the method for spiking the sediments. The range of total recoverable uranium (TR-U) concentrations achieved was 8-3200 mg/kg. This reflected the concentrations found in natural wetlands and water management ponds found on a uranium mine site and was above natural levels. Dilute-acid extractable uranium (AE-U) concentrations were >80% of total concentrations, indicating that much of the uranium in the spiked sediment was labile and potentially bioavailable. The portion of TR-U extractable as AE-U was similar at the start and end of the 4.5-month field-deployment. Porewater uranium (PW-U) analyses indicated that partition coefficients (Kd) were 2000-20,000 L/kg, and PW-U was greater in post- than pre-field-deployed samples when TR-U was ≤1500 mg/kg, indicating the binding became weaker during the field-deployment period. At higher spiked-U concentrations, the PW-U was lower post-field-deployment. Comparing the physico-chemical data of the spiked sediments with environmental monitoring data from sediments in the vicinity of a uranium mining operation indicated that they were representative of sediments contaminated by mining and that the U-spiked sediments had a clear U concentration gradient. This confirmed the suitability of the spiking procedure for preparing sediments that were suitable for deriving a SQGV for uranium.

Assessing the Toxicity of Mine-Water Mixtures and the Effectiveness of Water Quality Guideline Values in Protecting Local Aquatic Species.

Six tropical freshwater species were used to assess the toxicity of mine waters from a uranium mine adjacent to a World Heritage area in northern Australia. Key contaminants of potential concern for the mine were U, Mg, Mn, and total ammonia nitrogen (TAN). Direct toxicity assessments were carried out to assess whether the established site-specific guideline values for individual contaminants would be protective with the contaminants occurring as mixtures. Metal speciation was calculated for contaminants to determine which were the major contributors of toxicity, with 84 to 96% of Mg predicted in the free-ion form as Mg2+ , and 76 to 92% of Mn predicted as Mn2+ . Uranium, Al, and Cu were predicted to be strongly bound to fulvic acid. Uranium, Mg, Mn, and Cu were incorporated into concentration addition or independent action mixture toxicity models to compare the observed toxicity in each of the waters with predicted toxicity. For >90% of the data, mine-water toxicity was less than predicted by the concentration addition model. Instances where toxicity was greater than predicted were accompanied by exceedances of individual metal guideline values in all but one case (i.e., a Mg concentration within 10% of the guideline value). This indicates that existing individual water quality guideline values for U, Mg, Mn, and TAN would adequately protect ecosystems downstream of the mine. Environ Toxicol Chem 2021;40:2334-2346. © 2021 Commonwealth of Australia. Environmental Toxicology and Chemistry © 2021 SETAC.

Development of a Sublethal Chronic Toxicity Test for the Northern Trout Gudgeon, Mogurnda mogurnda, and Application to Uranium, Magnesium, and Manganese.

Many international guidance documents for deriving water quality guideline values recommend the use of chronic toxicity data. For the tropical fish northern trout gudgeon, Mogurnda mogurnda, 96-h acute and 28-d chronic toxicity tests have been developed, but both tests have drawbacks. The 96-h toxicity test is acute and has a lethal endpoint; hence it is not a preferred method for guideline value derivation. The 28-d method has a sublethal (growth) endpoint, but is highly resource intensive and is high risk in terms of not meeting quality control criteria. The present study aimed to determine the feasibility of a 7-d larval growth toxicity test as an alternative to the 96-h survival and 28-d growth tests. Once the method was successfully developed, derived toxicity estimates for uranium, magnesium, and manganese were compared with those for other endpoints and tests lengths within the literature. As a final validation of the 7-d method, the sensitivity of the 7-d growth endpoint was compared with those of 14-, 21-, and 28-d exposures. Fish growth rate, based on length, over 7 d was significantly more sensitive compared with existing acute toxicity endpoints for magnesium and manganese, and was similarly sensitive to existing chronic toxicity endpoints for uranium. For uranium, the sensitivity of the growth endpoint over the 4 exposure periods was similar, suggesting that 7 d as an exposure duration is sufficient to provide an indication of longer term chronic growth effects. The sensitivity of the 7-d method, across the 3 metals tested, highlights the benefit of utilizing the highly reliable short-term 7-d chronic toxicity test method in future toxicity testing using M. mogurnda. Environ Toxicol Chem 2021;40:1596-1605. © 2021 Commonwealth of Australia. Environmental Toxicology and Chemistry © 2021 SETAC.

Integrating laboratory and field studies to assess impacts of discharge from a uranium mine and validate a water quality guideline value for magnesium.

Magnesium (Mg) is a primary contaminant in mine water discharges from the Ranger Uranium Mine (north Australia). Site-specific water quality guideline values (WQGVs) for Mg have been derived from laboratory and field studies. Contaminated groundwater with elevated electrical conductivity and metals (Mg, Mn, U, SO4 , and Ca) was detected flowing from the mine site into adjacent surface waters. This provided an opportunity to investigate the protectiveness of the Mg WQGV by conducting an integrated laboratory and field study. A direct toxicity assessment (DTA) of the groundwater was conducted with local tropical freshwater species: duckweed (Lemna aequinoctialis), green hydra (Hydra viridissima), and the aquatic snail Amerianna cumingi. An in situ toxicity assessment was carried out in the creek receiving diluted groundwater by use of the same species of snail, to aid interpretation of laboratory-derived data. The toxicity of the contaminated groundwater was higher than Mg-only toxicity testing for H. viridissima, with other elevated metals and major ions contributing to toxicity. However, for duckweed and snail, the contaminated groundwater was less toxic than the Mg-only testing. In situ snail monitoring supported laboratory exposures, showing no effect on reproduction of A. cumingi exposed to an average of approximately 5 mg/L Mg; however, a very small effect was noted closer to the groundwater source, probably associated with other contaminants. The minimal toxicity observed for L. aequinoctialis and A. cumingi, despite the elevated Mg, can be explained by the high calcium (Ca) concentration of the water and the potential amelioration of metal toxicity. The extent of Ca amelioration of Mg toxicity was organism dependent. This study affirms the proposed environmental rehabilitation standard of 3 mg/L Mg for surface waters with a Ca concentration typical of water from this mine site. Integr Environ Assess Manag 2019;15:64-76. © 2018 SETAC.

Insights from the Genomes of Microbes Thriving in Uranium-Enriched Sediments.

Elevated uranium dose (4 g kg-1) causes a shift in billabong sediment communities that result in the enrichment of five bacterial species. These taxa include Geobacter, Geothrix and Dyella species, as well as a novel-potentially predatory-Bacteroidetes species, and a new member of class Anaerolineae (Chloroflexi). Additionally, a population of methanogenic Methanocella species was also identified. Genomic reconstruction and metabolic examination of these taxa reveal a host of divergent life strategies and putative niche partitioning. Resistance-nodulation-division heavy metal efflux (RND-HME) transporters are implicated as potential uranium tolerance strategies among the bacterial taxa. Potential interactions, uranium tolerance and ecologically relevant catabolism are presented in a conceptual model of life in this environment.

Effects of uranium concentration on microbial community structure and functional potential.

Located in the Northern Territory of Australia, Ranger uranium mine is directly adjacent to the UNESCO World Heritage listed Kakadu National Park, with rehabilitation targets needed to ensure the site can be incorporated into the park following the mine's closure in 2026. This study aimed to understand the impact of uranium concentration on microbial communities, in order to identify and describe potential breakpoints in microbial ecosystem services. This is the first study to report in situ deployment of uranium-spiked sediments along a concentration gradient (0-4000 mg U kg-1 ), with the study design maximising the advantages of both field surveys and laboratory manipulative studies. Changes to microbial communities were characterised through the use of amplicon and shotgun metagenomic next-generation sequencing. Significant changes to taxonomic and functional community assembly occurred at a concentration of 1500 mg U kg-1 sediment and above. At uranium concentrations of ≥ 1500 mg U kg-1 , genes associated with methanogenic consortia and processes increased in relative abundance, while numerous significant changes were also seen in the relative abundances of genes involved in nitrogen cycling. Such alterations in carbon and nitrogen cycling pathways suggest that taxonomic and functional changes to microbial communities may result in changes in ecosystem processes and resilience.

Development and implementation of a site-specific water quality limit for uranium in a high conservation value ecosystem.

Water quality guideline values (GVs) are a key tool for water quality assessments. Site-specific GVs, which incorporate data relevant to local conditions and organisms, provide a higher level of confidence that the GV will protect the aquatic ecosystem at a site compared to generic GVs. Site-specific GVs are, therefore, considered particularly suitable for sites of high sociopolitical or ecological importance. The present paper provides an example of the refinement of a site-specific GV for high ecological value aquatic ecosystems in Kakadu National Park, Northern Territory, Australia, to improve its site specificity and statistical robustness, thereby increasing confidence in its application. Uranium is a contaminant of concern for Ranger U mine, which releases water into Magela Creek and Gulungul Creek in Kakadu National Park. A site-specific GV for U has been applied, as a statutory limit, to Magela Creek since 2004 and to Gulungul Creek since 2015. The GV of 6 µg/L U was derived from toxicity data for 5 local species tested under local conditions. The acquisition of additional U data, including new information on the effect of DOC on U toxicity, enabled a revision of the site-specific U GV to 2.8 µg/L U and an ability to adjust the value on the basis of environmental concentrations of DOC. The revised GV has been adopted as the statutory limit, with the regulatory framework structured so the GV requires adjustment based on DOC concentration only when an exceedance occurs. Monitoring data for Magela Creek (2001-2013) and Gulungul Creek (2003-2013) downstream of the mine show that dissolved U has not exceeded 1 µg/L. Integr Environ Assess Manag 2017;13:765-777. © 2016 SETAC.

Increasing uranium exposure durations to the aquatic snail Amerianna cumingi does not result in lower toxicity estimates.

Reproductive inhibition (egg production) of the aquatic snail Amerianna cumingi over 4 d has been used to derive toxicity estimates for toxicants of concern in tropical Australia. Toxicity estimates from this test have been used as chronic data points in species sensitivity distributions (SSDs) for deriving site-specific guideline values. However, revised guidance for the Australian and New Zealand Water Quality Guidelines advises that test durations for adult macroinvertebrates should be ≥14 d to be considered chronic. Hence, to strengthen the data set underpinning the site-specific guideline value for uranium (U) in Magela Creek, which receives water from the Ranger Uranium Mine in northern Australia, the toxicity of U to A. cumingi was compared after 4 d, 9 d, and 14 d. Daily U concentrations were measured because of expected U loss during testing, providing extensive chemical analyses of the U exposure during the toxicity tests. Comparison of the U concentrations causing 50% reproductive inhibition (IC50) after 4 d, 9 d, and 14 d showed no difference in toxicity (4 d IC50 = 161 µg L-1 , confidence interval = 133-195; 9-d IC50 = 151 µg L-1 , confidence interval = 127-180; 14-d IC50 = 153 µg L-1 , confidence interval = 29-180). The present study provides evidence that test durations of <14 d are suitable for assessing chronic toxicity to U for this species and supports the use of the 4-d toxicity estimate in the SSD for U. Environ Toxicol Chem 2016;35:2851-2858. © 2016 Commonwealth of Australia.

Highly treated mine waters may require major ion addition before environmental release.

Mining operations often use passive and/or active water treatments to improve water quality prior to environmental release. Key considerations in choosing a treatment process include the extent to which the water quality is actually improved, and the potential residual environmental risks of the release of such water. However, there are few published studies concerning the environmental impacts of treated waste waters. This study used toxicity identification evaluation (TIE) methods to quantify and identify the "toxic" constituents of a highly-treated water (distillate) produced by brine concentration of a mining process water. Exposure of five freshwater species (Chlorella sp., Lemna aequinoctialis, Hydra viridissima, Moinodaphnia macleayi and Mogurnda mogurnda) to a concentration range of the distillate (0, 25, 50 and 100%) found that it was toxic to H. viridissima (50-100% effect when exposed to 100% distillate). TIE tests demonstrated that the effect wasn't due to residual ammonia (~1 mg L(-1)N) or trace organics, and unlikely to be due to manganese (Mn; 130-230 µg L(-1)). Conversely, addition of 0.2 and 0.5 mg L(-1) calcium improved the growth rate of H. viridissima by 61 and 66%, respectively, while addition of calcium, sodium and potassium (0.5, 1.0 and 0.4 mg L(-1), respectively) to levels comparable to that in the local aquatic environment resulted in 100% recovery. Further assessment on the likelihood of residual metal toxicity indicated that Mn concentrations in the distillate were at levels that could inhibit the growth of H. viridissima. Ultimately, the results demonstrated that ion deficiency should be considered as a potential stressor in risk/impact assessments of the discharge of treated wastewaters, and these may need to be supplemented with the deficient ions to reduce environmental impacts. The findings have highlighted the need for water managers to consider the possibility of unintended environmental risks from the discharge of highly-treated wastewaters.

Uranium exposure to the tropical duckweed Lemna aequinoctialis and pulmonate snail Amerianna cumingi: fate and toxicity.

The discharge of catchment-management water from the Ranger uranium (U) mine into Magela Creek upstream of the Ramsar-listed Magela Floodplain in Kakadu National Park is an important part of the mine's water-management system. Because U is one of the primary toxicants associated with this water, a receiving-water trigger value (TV), based on chronic toxicity data from five local native species, was derived for U. To strengthen the data set underpinning the derivation of the TV, the chronic toxicity of U to two additional tropical freshwater species, duckweed Lemna aequinoctialis (96-hour growth rate), and pulmonate gastropod, Amerianna cumingi (96-hour reproduction), was determined. The fate of U within the test systems was an important component of the study because analysis of U concentrations during the snail tests indicated that a substantial proportion of U (approximately 25%) was being lost from the test solutions when integrated during the entire test duration. Analysis of the snails and their food for U indicated that only a small proportion that was lost from solution was being taken up by the snails. Therefore, the majority of U that was lost was considered unavailable to the snails, and thus the exposure concentrations used to calculate the toxicity estimates were adjusted downward. Integrating the loss of U from the L. aequinoctialis test solutions over time showed that only a small proportion (6% to 13%) was lost during the test: Of that, almost half (2-5%) was taken up by the plants (constituting exposure). Uranium was only moderately toxic to L. aequinoctialis, with no observed-effect concentrations, lowest observed-effect concentrations, and inhibition concentrations causing 10% and 50% effects (IC10 and IC50) values of 226, 404, 207, and 1435 microg/l, respectively. A. cumingi was found to be more sensitive to U than L. aequinoctialis, with NOEC, LOEC, IC10, and IC50 values of 60, 61, 15, and 278 microg/l, respectively. The data for these two additional species will be used to revise the current TV for U in Magela Creek.