Mercury is a highly toxic trace metal that readily biomagnifies in food webs where it is inaccessible to current bioremediation methods. Animals could potentially be engineered to detoxify mercury within their food webs to clean up impacted ecosystems. We demonstrate that invertebrate (Drosophila melanogaster) and vertebrate (Danio rerio) animal models can express organomercurial lyase (MerB) and mercuric reductase (MerA) from Escherichia coli to demethylate methylmercury and remove it from their biomass as volatile elemental mercury. The engineered animals accumulated less than half as much mercury relative to their wild-type counterparts, and a higher proportion of mercury in their tissue was in the form of less bioavailable inorganic mercury. Furthermore, the engineered animals could tolerate higher exposures to methylmercury compared to controls. These findings demonstrate the potential of using engineered animals for bioremediation and may be applied to reduce the burden of methylmercury in impacted ecosystems by disrupting its biomagnification or to treat contaminated organic waste streams.

Sulfur oxidizing bacteria (SOB) play a key role in sulfur cycling in mine tailings impoundment (TI) waters, where sulfur concentrations are typically high. However, our understanding of SOB sulfur cycling via potential S oxidation pathways (sox, rdsr, and S4I) in these globally ubiquitous contexts, remains limited. Here, we identified TI water column SOB community composition, metagenomics derived metabolic repertoires, physicochemistry, and aqueous sulfur concentration and speciation in four Canadian base metal mine, circumneutral-alkaline TIs over four years (2016 – 2019). Identification and examination of genomes from nine SOB genera occurring in these TI waters revealed two pH partitioned, metabolically distinct groups, which differentially influenced acid generation and sulfur speciation. Complete sox (csox) dominant SOB (e.g., Halothiobacillus spp., Thiomonas spp.) drove acidity generation and S2O32- consumption via the csox pathway at lower pH (pH ~5 to ~6.5). At circumneutral pH conditions (pH ~6.5 to ~8.5), the presence of non-csox dominant SOB (hosting the incomplete sox, rdsr, and/or other S oxidation reactions; e.g. Thiobacillus spp., Sulfuriferula spp.) were associated with higher [S2O32-] and limited acidity generation. The S4I pathway part 1 (tsdA; S2O32- to S4O62-), was not constrained by pH, while S4I pathway part 2 (S4O62- disproportionation via tetH) was limited to Thiobacillus spp. and thus circumneutral pH values. Comparative analysis of low, natural (e.g., hydrothermal vents and sulfur hot springs) and high (e.g., Zn, Cu, Pb/Zn, and Ni tailings) sulfur systems literature data with these TI results, reveals a distinct TI SOB mining microbiome, characterized by elevated abundances of csox dominant SOB, likely sustained by continuous replenishment of sulfur species through tailings or mining impacted water additions. Our results indicate that under the primarily oxic conditions in these systems, S2O32- availability plays a key role in determining the dominant sulfur oxidation pathways and associated geochemical and physicochemical outcomes, highlighting the potential for biological management of mining impacted waters via pH and [S2O32-] manipulation.

In mine wastewaters, three microbial sulfur oxidation pathways have the potential to cause different water quality outcomes. These outcomes can differ from abiotic models of sulfate and acidity predictions currently used to monitor potential sulfur risks. However, studies integrating microbiology and geochemistry in active mine tailings impoundments are very limited. Here, we developed a novel diagnostic approach to detect microbially driven sulfur pathways. Within this 28-day study, eight on-site, 500 L mesocosms were filled with water extracted directly from the water cap of an active Ni/Cu mine tailings impoundment. Diverse combinations of tailings, sulfur compounds, and nitrate amendments were added to the mesocosms simulating common operational variations experienced by active tailings impoundments. Mesocosm results linked complete SOx, S4I, and incomplete SOx + rDSR pathway occurrence (metagenomes, inferred from the identity, i.e. 16S rRNA) and activity (mRNA) to physiochemistry and sulfur geochemistry. By integrating the three lines of evidence, the diagnostic approach was able to identify which sulfur pathways were active under varying physiochemical conditions and how geochemical outcomes were affected. A relationship emerged between acid generation and soxCD expression (soxCD expression indicates the complete SOx pathway activity). However, observed proton yields and sulfate concentrations were less than those predicted by complete SOx pathway activity alone. This indicates other sulfur pathways, e.g. the partial S4I pathway (within Thiomonas and Halothiobacillus), and/or activity of the incomplete SOx pathway (within Thiobacillus and Desulfurivibrio) when either not coupled to rDSR, or paired with use of nitrate, influenced overall sulfur outcomes along with the complete SOx pathway.

Environmental context Metal concentrations can build up to potentially harmful levels in marine mammals as they are at the top of the food chain. This review summarises the information available on metal concentrations in marine mammals, birds and turtles from around Australia. Despite large data gaps, the available data suggest that metal concentrations are similar to those encountered in other regions of the world. Abstract A comprehensive compilation of the published data for trace element concentrations (metals and metalloids) in Australian marine mammals, birds and turtles is presented. The majority of studies have relied on the utilisation of opportunistically collected samples, animal strandings and bycatch. This has resulted in large gaps in geographical, temporal and species coverage data. For instance, little or no data are available for cetaceans in New South Wales or the Northern Territory, and out of 14 endemic species of dolphins, data only exist for seven species. The aforementioned data gaps make it hard to identify statistically significant trends, a problem compounded by data being reported in the form of ranges without raw data. Trace element concentrations measured in various marine species and their tissue types are extremely variable, with ranges typically spanning several orders of magnitude, but are generally comparable with international data. Trends in contaminant concentrations with tissue type follow generally accepted patterns of behaviour for higher organisms, with the highest mercury concentrations in liver and cadmium in kidney tissues. Herbivores have lower contaminant loadings than carnivores, reflecting the importance of diet, and there are identifiable age-related trends for elements such as mercury. The lack of supporting pathology on dead and stranded animals and data on specimens from uncontaminated locations restrict conclusions on organism health impacts. There have been some attempts to use non-invasive sampling of indicator tissues such as fur, bristle and feathers. However, it is currently difficult to extrapolate these data to estimate contaminant concentrations in major organs. Recommendations for future investigations are made.

The acidification of water in mining areas is a global environmental issue primarily catalyzed by sulfur-oxidizing bacteria (SOB). Little is known about microbial sulfur cycling in circumneutral pH mine tailing impoundment waters. Here we investigate biological sulfur oxidation over four years in a mine tailings impoundment water cap, integrating aqueous sulfur geochemistry, genome-resolved metagenomics and metatranscriptomics. The microbial community is consistently dominated by neutrophilic, chemolithoautotrophic SOB (relative abundances of ~76% in 2015, ~55% in 2016/2017 and ~60% in 2018). Results reveal two SOB strategies alternately dominate across the four years, influencing acid generation and sulfur speciation. Under oxic conditions, novel Halothiobacillus drive lower pH conditions (as low as 4.3) and lower [S 2 O 3 2− ] via the complete Sox pathway coupled to O 2 . Under anoxic conditions, Thiobacillus spp. dominate in activity, via the incomplete Sox and rDSR pathways coupled to NO 3 − , resulting in higher [S 2 O 3 2− ] and no net significant acidity generation. This study provides genomic evidence explaining acidity generation and thiosulfate accumulation patterns in a circumneutral mine tailing impoundment and has significant environmental applications in preventing the discharge of sulfur compounds that can impact downstream environments. These insights illuminate opportunities for in situ biotreatment of reduced sulfur compounds and prediction of acidification events using gene-based monitoring and in situ RNA detection.

Environmental context This special issue is a tribute to Graeme Batley’s career and his many contributions to the field of environmental chemistry.

Cyanobacterial blooms have major impacts on the ecological integrity and anthropogenic value of freshwater systems. Chrysosporum ovalisporum, a potentially toxic cyanobacteria has been rare in Australian waters until recently when is has bloomed in a number of lake and river systems. The aim of this study was to determine drivers of its growth and growing dominance. We performed regular monitoring of Mannus Lake, a small freshwater reservoir in South-Eastern Australia that has recently undergone extremely dense bloom events. Blooms of the diazotrophic Chrysosporum ovalisporum occurred in both summers of the 19 month study during periods of persistent thermal stratification. Following the C. ovalisporum blooms, non-diazotrophic taxa (Microcystis aeruginosa and Woronichinia sp.) dominated the phytoplankton community under less stratified conditions. Thermal stratification and nitrogen availability appeared to be the primary drivers of changes in cyanobacterial community structure. We propose that the observed transition from C. ovalisporum to M. aeruginosa and/or Woronichinia sp. may be a result of nitrogen limitation in early summer, which combined with persistent thermal stratification led to an ecological advantage for the nitrogen-fixing C. ovalisporum. Mixing events caused the senescence of the C. ovalisporum bloom, likely supplementing the nutrient budget of the lake with atmospherically derived N and alleviating N limitation to non-diazotrophic taxa. Non-diazotrophic cyanobacterial growth then increased, albeit at much lower biovolumes compared to the initial bloom. Overall, the results demonstrate the role of thermal stratification and nutrient cycling in structuring the cyanobacterial community and provide insights into the environmental factors driving the proliferation of the relatively new, potentially toxic cyanobacterium C. ovalisporum in Australian waters.

Rationale. There are very few high-quality chronic inorganic arsenate (As-V) toxicity data to assess the risks to marine ecosystems. We aimed to determine the range in chronic toxicity of As-V to marine biota and derive reliable water quality guideline values (GVs) for the long-term protection of marine ecosystems. Methodology We generated chronic toxicity data based on measured dissolved (<0.45 mu m filtered) As-V concentrations for 13 marine species representing seven taxonomic groups from temperate and tropical environments. Effect concentrations at the 10% level (EC10) were used in a species sensitivity distribution (SSD) to derive water quality GVs. Results The range of concentrations causing chronic 10, 20 and 50% adverse effects were 13-26 000, 18-34 000 and 32-330 000 mu g As-V L-1, respectively. Increased phosphate and nitrate concentrations were found to reduce the toxicity of As-V to certain microalgal, sea urchin and bivalve species. The range in effect concentrations for tropical versus temperate species overlapped at all effect levels. The GVs for the long-term protection of 80, 90, 95 and 99% of marine biota were: 48, 22, 12 and 4.8 mu g As-V L-1, respectively. Discussion Recommendations on performing toxicity tests with arsenic to prevent artefacts associated with arsenic speciation were made to improve future research on arsenic toxicity. The new data will improve the reliability status of the Australian and New Zealand As-V GVs for marine water quality and fill a data gap for global risk assessments of As-V for marine biota.

Environmental context High-quality ecotoxicology data are required to derive reliable water quality guideline values that ensure long-term protection of marine biota from arsenate. Tropical and temperate marine biota have sensitivity to arsenate covering three to four orders of magnitude due to the range of arsenate detoxification mechanisms used to reduce toxicity. The water quality guideline values derived in this study will contribute to robust risk assessments of arsenate in marine environments. Rationale There are very few high-quality chronic inorganic arsenate (AsV) toxicity data to assess the risks to marine ecosystems. We aimed to determine the range in chronic toxicity of AsV to marine biota and derive reliable water quality guideline values (GVs) for the long-term protection of marine ecosystems. Methodology We generated chronic toxicity data based on measured dissolved (<0.45 µm filtered) AsV concentrations for 13 marine species representing seven taxonomic groups from temperate and tropical environments. Effect concentrations at the 10% level (EC10) were used in a species sensitivity distribution (SSD) to derive water quality GVs. Results The range of concentrations causing chronic 10, 20 and 50% adverse effects were 13–26 000, 18–34 000 and 32–330 000 µg AsV L–1, respectively. Increased phosphate and nitrate concentrations were found to reduce the toxicity of AsV to certain microalgal, sea urchin and bivalve species. The range in effect concentrations for tropical versus temperate species overlapped at all effect levels. The GVs for the long-term protection of 80, 90, 95 and 99% of marine biota were: 48, 22, 12 and 4.8 µg AsV L–1, respectively. Discussion Recommendations on performing toxicity tests with arsenic to prevent artefacts associated with arsenic speciation were made to improve future research on arsenic toxicity. The new data will improve the reliability status of the Australian and New Zealand AsV GVs for marine water quality and fill a data gap for global risk assessments of AsV for marine biota.

Spills of hydraulic fracturing (HF) fluids and of produced water during unconventional gas extraction operations may cause soil contamination. We studied the degradation and microbial toxicity of selected HF chemical components including two biocides (methylisothiozolinone- MIT, chloromethylisothiozolinone- CMIT), a gel-breaker aid (triethanolamine -TEA), and three geogenic chemicals (phenol, m-cresol and p-cresol) in ultrapure water, HF fluid and produced water in five different soil types (surface and subsurface soils). The degradation of the two biocides (in soils treated with HF fluid or ultrapure water) and of the three geogenic chemicals (in soils treated with produced water) was rapid (in all cases DT50 values < 2 days in surface soils). In contrast, the loss of TEA was much slower in soils, especially in those treated with HF fluid (DT50 > 30 days). Sorption coefficients (Koc in L/Kg) in these soils ranged from 71 to 733 for TEA, 64-408 for MIT and 11-72 for CMIT. In terms of soil microbial toxicity, exposure to HF fluid and produced water reduced microbial respiration, albeit temporarily. The overall microbial activities in surface soils contaminated with produced water had fully recovered in most soils. In contrast, the HF fluid addition to soils completely inhibited the nitrification in all soils, with little recovery over the 60 day experimental period. In the case of produced water exposure, three out of five surface soils showed complete recovery in nitrification during the study period. The functional genes for nitrogen fixation (nifH) and carbon cycling (GA1) and microbial community composition (16 S rRNA) were significantly affected by HF fluid in some soils. Overall, the study shows that the HF fluid can have significant detrimental impact on soil microbial functions, especially on nitrogen cycling. More work is needed to identify the exact cause of microbial toxicity in soils contaminated with HF fluid.

Environmental context The oil and gas industry has a significant liability in decommissioning offshore infrastructure. Following decommissioning, subsea pipelines could be left on the seabed to provide artificial reefs. Mercury is a contaminant of concern which could remain within pipelines. There are gaps in our knowledge on how mercury moves through the marine environment. We review the current science and identify future research needs to understand potential impacts from mercury in subsea pipelines which will better inform decommissioning activities globally. Abstract In the coming years, the oil and gas industry will have a significant liability in decommissioning offshore infrastructure such as subsea pipelines. The policies around decommissioning vary depending on regional policies and laws. In Australia, the ‘base case’ for decommissioning is removal of all property and the plugging and abandonment of wells in line with the Offshore Petroleum and Greenhouse Gas Storage (OPGGS) Act 2006. Options other than complete removal may be considered where the titleholder can demonstrate that the alternative decommissioning activity delivers equal or better environmental outcomes compared to complete removal and meets all requirements under the OPGGS Act and regulations. Recent research has demonstrated that decommissioning in situ can have significant environmental benefits by forming artificial reefs, increasing marine biodiversity, and providing a potential fishery location. An issue, which has been given less attention, is around contaminants remaining within decommissioned infrastructure and their potential risks to the marine environment. Mercury is a contaminant of concern known to be present in some oil and gas pipelines, but the potential long-term impacts on marine ecosystems are poorly understood. We present a synthesis of information on mercury cycling in the marine environment including key drivers of methylation in sediments and ocean waters, existing models to predict methylmercury concentrations in sediments, and toxicological effects to marine biota. We discuss the applicability of existing water and sediment quality guidelines, and the associated risk assessment frameworks to decommissioning offshore infrastructure contaminated with mercury. Globally, research is needed to provide a comprehensive risk assessment framework for offshore infrastructure decommissioning. We recommend future areas of research to improve our understanding of the potential risks associated with mercury in subsea oil and gas pipelines.

Flowback water from shale gas operations contains formation-derived compounds, including trace metals, radionuclides, and organics. While accidental releases from storage tanks with flowback water are low-probability events if multiple containment barriers are put in place, they cannot be entirely excluded. Here the natural attenuation potential of deep unsaturated zones and groundwater was explored using predictive modelling involving a hypothetical leak from a storage tank. Actual chemical concentrations from flowback water at two shale gas wells with contrasting salinity (12,300 and 105,000 ppm TDS) in the Beetaloo Sub-basin (Northern Territory, Australia) served as input to the one-dimensional HYDRUS model for simulating chemical transport through the unsaturated zone, with groundwater at 50 and 100 m depth, respectively. Subsequent chemical transport in groundwater involved the use of a three-dimensional analytical transport model. For a total of 63 chemicals the long-term attenuation from dilution and dispersion in unsaturated sediments and groundwater was calculated. Predicted environmental concentrations for aquatic receptors were compared with no-effect levels of individual chemicals to derive risk quotients (RQ) and identify chemicals of no concern to ecosystem health (i.e. RQ <1). Except for salinity and radium-228 in one of the two wells, RQ < 1 for all other chemicals. The initial approach considered testing of toxicity to individual chemicals only. When direct toxicity assessments (DTAs) were used to account for effects of chemical mixtures, the required DTA-derived safe dilution factor for 95% species protection was 1.8 to 2.5 times higher than the dilution factor accounting for dispersion and dilution only. Accounting for biodegradation, sorption and radioactive decay decreased chemical concentrations in unsaturated sediments to safe levels using the DTA for all chemicals. The study highlighted the importance of incorporating DTA in chemical risk assessments involving complex chemical mixtures. Improved understanding of fate and transport of flowback chemicals will help effectively manage water-quality risks associated with shale gas extraction.

Microcystis aeruginosa is a widespread cyanobacteria capable of producing hepatotoxic microcystins. Understanding the environmental factors that influence its growth and toxin production is essential to managing the negative effects on freshwater systems. Some micronutrients are important cofactors in cyanobacterial proteins and can influence cyanobacterial growth when availability is limited. However, micronutrient requirements are often species specific, and can be influenced by substitution between metals or by luxury uptake. In this study, M. aeruginosa was grown in modified growth media that individually excluded some micronutrients (cobalt, copper, iron, manganese, molybdenum) to assess the effect on growth, toxin production, cell morphology and iron accumulation. M. aeruginosa growth was limited when iron, cobalt and manganese were excluded from the growth media, whereas the exclusion of copper and molybdenum had no effect on growth. Intracellular microcystin-LR concentrations were variable and were at times elevated in treatments undergoing growth limitation by cobalt. Intracellular iron was notably higher in treatments grown in cobalt-deplete media compared to other treatments possibly due to inhibition or competition for transporters, or due to irons role in detoxifying reactive oxygen species (ROS).

Environmental context Base Mine Lake (BML) is a demonstration pilot pit lake for long term oil sands tailings reclamation in Alberta, Canada. This study quantified BML water cap sulfur mass balance and speciation to help understand potential risks to oxygen levels during its early-stage development. Results provide important insights for the adaptive management of water-capped oil sands tailings reclamation. Rationale Sulfur cycling is crucial to the persistence of oxygen in the water cap of Base Mine Lake (BML), the first demonstration oil sands pit lake for water capped tailings technology (WCTT) in Alberta, Canada. Methodology Here, we report on the first investigation of sulfur mass balance and sulfur speciation (SO4 2−, SO3 2−, S2O3 2−, S0 and ƩH2S) over seasonal, annual and spatial scales in BML. Results and discussion High aqueous total sulfur concentration (1.7–2 mM), dominated by sulfate (>75%), decreased over the study period (2015–2021), due to the consolidation of fluid fine tailings (FFT) and operational pump-in and pump-out activities. Expanded BML water cap S biogeochemical cycling occurred after a 2016 alum amendment. Late summer hypolimnetic anoxia emerged post-alum (2017–2021), coincident with detectable total sulfide (ƩH2S) and elemental sulfur (S0) concentrations and expanded sulfur-reducing bacteria activity in anoxic bottom waters. Post spring turnover resuspended FFT and particle settling rates also likely increased post-alum, supported by the observed migration of epilimnetic highest sulfite concentration (pre-alum) to metalimnetic waters (post-alum). These sulfide containing particles are likely the primary reduced S substrate for spring–summer sulfur-oxidising bacteria activity, as winter aqueous reactive S species (ƩH2S, S0, S2O3 2 − and SO3 2−) were non-detectable across years. Concentrations of reactive S species reached up to 200 µM, posing risks to BML O2 levels (maximum 300–350 µM). Results of this study establish the interactive effects of physical and biogeochemical processes, as well as operational activities in emergent S risks to water cap oxygen levels, a key criterion for success of this reclamation tailings technology.

Micronutrients play key roles in numerous metabolic processes in cyanobacteria. However, our understanding of whether the micronutrient cobalt influences the productivity of freshwater systems or the occurrence of cyanobacterial blooms is limited. This study aimed to quantify the concentration of Co necessary for optimal cyanobacterial growth by exposing Microcystis aeruginosa to a range of Co concentrations under culture conditions. Extended exposure to concentrations below similar to 0.06 mu g center dot L-1 resulted in notable inhibition of M. aeruginosa growth. A clear negative relationship was observed between Co concentration in solution and intracellular Fe quota of M. aeruginosa, possibly due to decreased transport of Fe at higher Co concentrations. Cyanocobalamin and any Co within the structure of cyanocobalamin appears to be non-bioavailable to M. aeruginosa, instead they likely rely on the synthesis of a structural variant - pseudocobalamin, which may have implications for the wider algal community as the variants of cobalamin are not necessarily functionally exchangeable. To evaluate the likelihood of Co limitation of cyanobacterial growth under field conditions, a survey of 10 freshwater reservoirs in South-Eastern Australia was conducted. Four of the ten sites had dissolved Co concentrations below the 0.06 mu g center dot L-1 threshold value. All four of these sites rarely undergo cyanobacterial blooms, strengthening evidence of the potential for Co to limit growth, perhaps either alone or in combination with phosphorus.

The role of trace metal micronutrients in limiting cyanobacterial growth and structuring the phytoplankton community is becoming more evident. However, little is known regarding the extent of micronutrient limitation in freshwaters or which micronutrient conditions favour potentially-toxic cyanobacteria. To assess how freshwater phytoplankton respond to micronutrient and macronutrient additions, we conducted nutrient amendment bioassays at seven sites across South Eastern-Australia. Sites were variable in cyanobacterial cell densities and phytoplankton community compositions. At two sites, Mannus Lake and Burrendong Dam, micronutrient additions (iron, cobalt, copper, manganese, molybdenum and zinc) increased cyanobacterial growth, indicating micronutrient limitation. Both sites had cyanobacterial blooms present at the onset of the experiment, dominated by Chrysosporum ovalisporum at Mannus Lake and Microcystis aeruginosa at Burrendong Dam. This suggests that micronutrients may be an important regulator of the severity of cyanobacterial blooms and may become limiting when there is high competition for nutrient resources. The addition of the micronutrient mixture resulted in a higher proportion of cyanobacteria compared to the control and a lower diversity community compared to phosphorus additions, indicating that micronutrients can not only influence cyanobacterial biovolume but also their ability to dominate the phytoplankton community. This reinforces that micronutrient requirements of phytoplankton are often species specific. As micronutrient enrichment is often overlooked when assessing nutrient-constraints on cyanobacterial growth, this study provides valuable insight into the conditions that may influence cyanobacterial blooms and the potential contribution of micronutrients to eutrophication.

Successful decommissioning of offshore oil and gas infrastructure requires an effective and safe approach to assessing and managing chemical and radiological residues. Scale frequently accumulates on the interior surfaces of pipes and other structures and may persist long after extraction operations have ceased. Scale materials can contain a range of metal contaminants (including mercury), as well as naturally occurring radioactive materials. In newer or more accessible infrastructure, the scale is routinely removed, and becomes a waste product. The persistent nature of scale contaminants can result in a radiological dose to the organisms living on, or near an intact pipeline. Eventually, infrastructure corrosion following insitu decommissioning (abandonment) could lead to metal and radionuclide contaminants being accessible to the surrounding seafloor environment, where bioaccumulation and subsequent ecotoxicological effects from the chemical and radiological properties of the scale could occur. The paper describes a tiered approach to assess the ecological impacts of pipeline scale in order to assist operators with their plans for decommissioning offshore infrastructure, especially when considering ‘leave in place’ options.

Acidification of freshwater in mining impacted areas is a major global environmental problem catalyzed by sulfur-oxidizing bacteria (SOB). To date, little is known about the active bacteria in mine tailings impoundments and their environmental niches. Here, biological sulfur oxidation was investigated over four years in a mine tailings impoundment, integrating sulfur geochemistry, genome-resolved metagenomics and metatranscriptomics. We demonstrated oxygen driven niche partitioning of SOB and their metabolic pathways that explain acidity generation and thiosulfate persistence. Four chemolithoautotrophic SOB, Halothiobacillus, Thiobacillus, Sulfuricurvum and Sediminibacterium comprised 37% to 73% of the analyzed communities. The impoundment waters alternated between the dominance of Halothiobacillus versus a Thiobacillus, Halothiobacillus, Sulfuricurvum and Sediminibacterium consortia. Halothiobacillus dominance was associated with lower pH values (∼4.3), higher [H + ]/[SO 4 2- ] and lower [S 2 O 3 2- ], collectively indicative of extensive sulfur oxidation. Halothiobacillus , which couple sulfur oxidation via the Sox pathway to aerobic respiration or NO 2 - reduction, were present throughout the depth profile, yet their expression of sox genes occurred only in upper highly oxygenated waters. Conversely, when consortia of Thiobacillus, Halothiobacillus, Sulfuricurvum and Sediminibacterium dominated, recycling/disproportionating reactions were more prevalent. Thiobacillus, which dominated deeper micro-oxic/anoxic waters, oxidized sulfur primarily through the rDSR pathway, coupled to NO 3 - /NO 2 - reduction, resulting in lower [H + ]/[SO 4 2- ] and higher [S 2 O 3 2- ] relative to upper waters. These field results mirror the Sox/rDSR-geochemical patterns of experimental SOB enrichments and reveal opportunities for biological treatments of recalcitrant reduced sulfur compounds, as well as gene-based monitoring and in situ RNA detection to predict the onset of problematic geochemistry.

Environmental contextMany trace metals, including copper, are only sparingly soluble in seawater and may exist in both dissolved and particulate forms (e.g. as precipitates). Aquatic organisms may experience different toxic effects from exposure to dissolved and particulate trace metals. This study investigates how concentration, reaction time and changes to precipitate composition/mineral formation affect copper solubility in seawater, thus influencing metal bioavailability and toxicity in the field and laboratory. AbstractA lack of knowledge on the solubility of metals such as copper affects the ability to predict the forms (dissolved and particulate) that organisms are exposed to in field and laboratory waters. Laboratory tests were conducted where copper (total concentrations of 0.5 to 20mg L−1) was added to natural and artificial seawater (pH 8.15, 22°C), equilibrated for 28 days and dissolved copper monitored at periodic intervals. At 0.5mg L−1, dissolved copper concentrations remained stable over 28 days and no precipitates were detected. However, at higher total copper concentrations, an initial rapid precipitation phase was followed by the establishment of a metastable equilibrium that persisted for periods of days to weeks, and whose solubility concentrations and duration were influenced by the total copper concentration and typically in the range 0.6 to 0.9mg L−1. After 5 to 15 days, a step change decrease in dissolved copper concentration followed by a slow decline was observed in the >2mg L−1 total copper treatments. The minimum solubility measured after 28 days was 0.053mg L−1. Elemental and X-ray diffraction analyses indicated that the copper precipitates comprised similar proportions of amorphous copper hydroxycarbonate and amorphous dicopper trihydroxide chloride after 1 day and transformed to predominantly mineralised dicopper trihydroxide chloride in the clinoatacamite polymorph form after 28 days. These observations have particular relevance for toxicity tests of less sensitive organisms and highlight the need to consider metal solubility, exposure to precipitates and changes in precipitate mineral phases.