Relevance of tributary inflows for driving molecular composition of dissolved organic matter (DOM) in a regulated river system.

River regulation by dams can alter flow regimes and organic matter dynamics, but less is known about how unregulated tributaries regulate organic matter composition and processing in the regulated river below the confluence. This study reports on water chemistry, especially dissolved organic matter (DOM) concentration and composition (dissolved organic carbon (DOC), organic nitrogen (DON), organic phosphorus (DOP) and combined amino acids (DCAA)) along the regulated Tumut and unregulated Goobarragandra (tributary) rivers under different flow conditions (base flow vs storm event) in south-east Australia. The tributary was significantly different from regulated and downstream sites during base flow conditions with higher temperature, pH, buffering capacity, DOC and nutrient concentrations (DON, DOP, DCAA). DOM characterisation by spectrometry and size exclusion chromatography revealed that the tributary contained a higher proportion of terrestrially derived humic-like and fulvic-like DOM. In contrast, regulated and downstream sites contained higher proportion of microbially derived DOM such as low molecular weight neutrals and protein-like components. Storm pulses of tributary flows into the regulated system, influenced both concentration and composition of DOM at the downstream site, which more strongly resembled the tributary site than the regulated site during the storm event. Additionally, we found that the tributary supplied fresh DOM, including small organic molecules to the regulated system during storm events. The presence of these different types of labile DOM can increase primary productivity and ecological functioning within regulated river reaches downstream of tributary junctions. This has important implications for the protection of unregulated tributary inflows within regulated river basins.

Two wild carnivores selectively forage for prey but not amino acids.

In nutritional ecology the intake target is the diet that maximises consumer fitness. A key hypothesis of nutritional ecology is that natural selection has acted upon the behavioural and physiological traits of consumers to result in them Selectively Consuming prey to match the Intake Target (SCIT). SCIT has been documented in some herbivores and omnivores, which experience strong heterogeneity in the nutritional quality of available foods. Although carnivores experience a prey community with a much more homogeneous nutrient composition, SCIT by carnivores has nevertheless been deemed highly likely by some researchers. Here we test for SCIT for micronutrients (amino acids) in two freshwater carnivores: the river blackfish and the two-spined blackfish. Although both blackfishes exhibited non-random consumption of prey from the environment, this resulted in non-random consumption of amino acids in only one species, the river blackfish. Non-random consumption of amino acids by river blackfish was not SCIT, but instead an artefact of habitat-specific foraging. We present hypotheses to explain why wild populations of freshwater carnivores may not exhibit SCIT for amino acids. Our work highlights the need for careful, critical tests of the hypotheses and assumptions of nutritional ecology and its application to wild populations.

Biomolecular modifications in the sacfry of Mogurnda adspersa in response to copper stress.

Copper (Cu) is one of the most harmful contaminants in fresh-water systems. Fish larvae such as sacfry are particularly vulnerable to metals such as copper (Cu) due to a less-developed excretory organ system and permeable skin that can absorb metals directly from the water. However, the sublethal effects of metals on this life stage are not well understood. This study assessed the sublethal toxicity of Cu on purple-spotted gudgeon sacfry (PSG, Mogurnda adspersa). For this purpose, 96 h Cu toxicity bioassays were performed and toxic effects of Cu on PSG were measured at different levels of biological organization, from the individual (loss of equilibrium, wet weight), to tissue (chemical changes in retinal tissue composition) and molecular responses (whole body amino acid (AA) profiles). The EC10 and EC50 (ECx: effect concentration that affected X% of test organisms) were found to be 12 (9 - 15) µg Cu L-1 and 22 (19 - 24) µg Cu L-1, respectively. Copper stress caused a decrease in total amino acid content and changed the AA profile of PSG compared to the controls. Proton-induced X-ray emission (PIXE) mapping techniques showed accumulation of Cu in the retinal tissues disturbing the distribution of other elements such as zinc, sulfur, phosphorus and potassium. Fourier-transform infrared (FTIR) microspectroscopy of control and Cu treated eye tissues revealed a change in protein secondary structure in retinal tissues in response to Cu accumulation, as well as decreased levels of the molecular retinal, consistent with the degradation of rhodopsin, a key protein in the visual sensory system. This is the first study to demonstrate the multi-level responses of PSG arising from exposure to environmentally realistic Cu concentrations and suggests that AA profiling can serve as a useful tool to assess the impacts of metals on fresh-water organisms.

Changes to the amino acid profile and proteome of the tropical freshwater microalga Chlorella sp. in response to copper stress.

Contamination of freshwaters is increasing globally, with microalgae considered one of the most sensitive taxa to metal pollution. Here, we used 72 h bioassays to explore the biochemical effects of copper (Cu) on the amino acid (AA) profile and proteome of Chlorella sp. and advance our understanding of the molecular changes that occur in algal cells during exposure to environmentally realistic Cu concentrations. The Cu concentrations required to inhibit algal growth rate by 10% (EC10) and 50% (EC50) were 1.0 (0.7-1.2) µg L-1 and 2.0 (1.9-2.4) µg L-1, respectively. The AA profile of Chlorella sp. showed increases in glycine and decreases in isoleucine, leucine, valine, and arginine, with increasing Cu. Proteomic analysis revealed the modulation of several proteins involved in energy production pathways, including: photosynthesis, carbon fixation, glycolysis, and oxidative phosphorylation, which likely assists in meeting increased energy demands under Cu-stressed conditions. Copper exposure also caused up-regulation of cellular processes and signalling proteins, and the down-regulation of proteins related to ribosomal structure and protein translation. These changes in biomolecular pathways have direct effects on the AA profile and total protein content and provide an explanation for the observed changes in amino acid profile, cell growth and morphology. This study shows the complex mode of action of Cu on Chlorella under environmentally realistic Cu concentrations and highlights several potential biomarkers for future investigations.

Methanotroph community structure and processes in an inland river affected by natural gas macro-seeps.

Methane availability in freshwaters is usually associated with spatial-temporal variation in methanogenesis. Unusually, however, natural gas macro-seeps occur along the Condamine River in eastern Australia which elevate ambient water-column methane concentrations more than 3,000 times. We quantified the spatial-temporal variation in methane oxidation rates and the total microbial and methanotroph community composition (through the amplification and sequencing of 16S rRNA and particulate methane monooxygenase (pmoA) genes), and the factors mediating this variation, in reaches with and without macro-seeps. Sediment methane oxidation rates were, on average, 29 times greater, and the abundance of methanotrophs significantly higher, in the vicinity of methane macro-seeps compared to non-seep sites. Methylocystis was the most abundant methanotroph group at all sites, but type Ib methanotrophs showed the steepest increase in abundance at seep sites. pmoA gene analysis identified these as clade 501, while 16S rRNA gene analysis identified these as the closely related genus Methylocaldum. Sediment methane oxidation rates and the relative abundance and composition of benthic microbial communities were primarily influenced by methane availability which was in turn related to variation in river discharge. Methane-derived carbon may be an important energy source for the aquatic food webs in reaches affected by natural gas macro-seeps.

2019-2020 Bushfire impacts on sediment and contaminant transport following rainfall in the Upper Murray River catchment.

During the 2019-2020 Australian bushfire season, large expanses (~47%) of agricultural and forested land in the Upper Murray River catchment of southeastern (SE) Australia were burned. Storm activity and rainfall following the fires increased sediment loads in rivers, resulting in localized fish kills and widespread water-quality deterioration. We collected water samples from the headwaters of the Murray River for sediment and contaminant analysis and assessed changes in water quality using long-term monitoring data. A robust runoff routing model was used to estimate the effect of fire on sediment loads in the Murray River. Peak turbidity in the Murray River reached values of up to 4200 nephelometric turbidity units (NTU), shown as pitch-black water coming down the river. The increase in suspended solids was accompanied by elevated nutrient concentrations during post-bushfire runoff events. The model simulations demonstrated that the sediment load could be five times greater in the first year after a bushfire than in the prefire condition. It was estimated that Lake Hume, a large reservoir downstream from fire-affected areas, would receive a maximum of 600 000 metric tonnes of sediment per month in the period immediately following the bushfire, depending on rainfall. Total zinc, arsenic, chromium, nickel, copper, and lead concentrations were above the 99% toxicant default guideline values (DGVs) for freshwater ecosystems. It is also likely that increased nutrient loads in Lake Hume will have ongoing implications for algal dynamics, in both the lake and the Murray River downstream. Information from this study provides a valuable basis for future research to support bushfire-related policy developments in fire-prone catchments and the mitigation of postfire water quality and aquatic ecosystem impacts. Integr Environ Assess Manag 2021;17:1203-1214. © 2021 Commonwealth of Australia. Integrated Environmental Assessment and Management © 2021 Society of Environmental Toxicology & Chemistry (SETAC).

Spatial variation in the amino acid profile of four macroinvertebrate taxa along a highly polluted river.

Acid mine drainage (AMD) is one of the major environmental problems impacting aquatic ecosystems globally. We studied changes in the community composition of macroinvertebrates and amino acid (AA) profiles of dominant taxa along an AMD contamination gradient within the Dee River, Queensland, Australia to understand how AMD can affect the biomolecular composition of macroinvertebrates. Taxa richness and community composition of macroinvertebrates changed widely along the AMD gradient with significantly lower taxa richness recorded at the polluted sites compared to upstream and downstream sites. The Dipteran families: Chironomidae and Ceratopogonidae, the Odonata family Gomphidae, and the Coleoptera family Dytiscidae were the only families found at all sampling sites and were used here for AA analysis. There were significant variations in the AA profiles among the studied taxa. The AA profile of each taxon also varied among upstream, polluted and downstream sites suggesting that contamination of a river system with acid mine drainage not only alters the overall macroinvertebrate community composition but also significantly influences the AA profile of organisms that are tolerant to AMD. This study highlights the potential of using AA profiling to study the response of aquatic organisms to contamination gradients such as those associated with AMD.

Connectivity, not short-range endemism, characterises the groundwater biota of a northern Australian karst system.

Groundwater ecosystems have a diverse and unique fauna, often dominated by Crustacea and generally characterised by short range endemics confined to single aquifers. Much of this knowledge has come from studies conducted either in fractured rock aquifers or alluvial aquifers. Karstic subterranean environments are present in the Cambrian Limestone Aquifer (CLA) in the Northern Territory, Australia, a freshwater aquifer which spans an area of ~28,000 km2. The presence of underground caverns and channels potentially allows extensive connectivity within this groundwater system. The emerging shale gas industry in the Beetaloo region, which underlies the CLA, provided the impetus to undertake the first survey of the potential existence of a stygofaunal community. Twenty-six groundwater wells (bores) and two springs were sampled in August and October 2019, across a distance of ~500 km, from the sub-tropical Mataranka region in the north to the semi-arid Barkly Tablelands in the south. Plankton nets and motorised pumps were used to collect water samples and conventional microscope-based morphological examinations in conjunction with environmental DNA (eDNA) were used to determine the presence of stygofauna. COI barcoding and 16S rRNA regions were also used for phylogenetic analysis. All stygofaunal communities were dominated by crustaceans, namely shrimps, amphipods, ostracods, copepods and syncarids. This fauna showed little affinity with the stygofauna recorded from more extensively sampled aquifers in north-western Australia, with new genera and species present in the CLA. eDNA analysis showed the presence of diverse biota at sites where direct water sampling for intact animals was difficult. COI and 16S analysis confirmed that a species of blind shrimp, Parisia unguis, occurred extensively throughout the aquifer, over a distance of at least ~300 km. The presence of Pa. unguis at widely separated sites across the CLA is consistent with substantial connectivity within the aquifer. This connectivity indicates that the risk of groundwater contamination from fracking chemicals needs to be adequately mitigated to prevent widespread effects.

Managed floodplain inundation maintains ecological function in lowland rivers.

Managed environmental flows are one mechanism by which managers may restore carbon dynamics, diversity and ecological function of rivers affected by anthropogenic activities. Empirical studies that quantify such interactions in detail are few, so we measured the amounts of dissolved organic carbon, nutrients, algae and invertebrates in the main river channel following a managed environmental flow that inundated an adjacent floodplain forest. Dissolved organic carbon (DOC), seston carbon, total nitrogen (TN), and chlorophyll-a (chl-a) concentrations were greatly increased downstream. The net yield of DOC, seston carbon, TN and chl-a from the floodplain peaked at approximately 100, 50, 5 and 0.1 t d-1, respectively during the major flow event. Total phosphorus mobilisation peaked at approximately 0.4 t d-1. Stable isotope analysis showed that allochthonously-derived carbon was rapidly incorporated into biofilm and grazing macroinvertebrates, persisting in riverine food webs for up to four months following the flood. During a subsequent smaller flow event, the floodplain either generated no further carbon or nutrients, or was a sink for carbon and nutrients. Our results provide empirical support for the River Wave Concept and show that allowing floodplain water to return to the river downstream of forests is important for maintaining ecological function within the river channel.

Dissolved organic matter and metabolic dynamics in dryland lowland rivers.

Dissolved organic matter (DOM) within freshwaters is essential for broad ecosystem function. The concentration and type of DOM within rivers depends on the relative contributions of allochthonous sources and the production and consumption of DOM by microbes. In this work we have examined the temporal patterns in DOM quality and productivity in three lowland rivers in dryland Australia using fluorescence excitation emission scans. We assessed the production and consumption of DOM within light and dark bottle assays to quantify the relative contribution of bacteria and algae to the DOM pool and simultaneously assessed whether the systems were autotrophic or heterotrophic. DOM varied temporally within the three river systems over the course of the study period. Characterisation of DOM within light and dark bottles following a 6-hour incubation revealed microbial consumption of a humic-like component and production of protein-like components similar in nature to the amino acids tryptophan and tyrosine. The lack of a significant difference in DOM quality between the light and dark bottles indicated that the protein-like DOM is likely derived from bacterial activity. Respiration was shown to be higher than gross primary production in both whole river and bottle assays, yielding negative net production values and demonstrating that these rivers were predominately heterotrophic. Our work suggests that bacterial metabolism of DOM may be a significant contributor to the production of protein-like components within heterotrophic freshwater systems.

Causal modelling applied to the risk assessment of a wastewater discharge.

Bayesian networks (BNs), or causal Bayesian networks, have become quite popular in ecological risk assessment and natural resource management because of their utility as a communication and decision-support tool. Since their development in the field of artificial intelligence in the 1980s, however, Bayesian networks have evolved and merged with structural equation modelling (SEM). Unlike BNs, which are constrained to encode causal knowledge in conditional probability tables, SEMs encode this knowledge in structural equations, which is thought to be a more natural language for expressing causal information. This merger has clarified the causal content of SEMs and generalised the method such that it can now be performed using standard statistical techniques. As it was with BNs, the utility of this new generation of SEM in ecological risk assessment will need to be demonstrated with examples to foster an understanding and acceptance of the method. Here, we applied SEM to the risk assessment of a wastewater discharge to a stream, with a particular focus on the process of translating a causal diagram (conceptual model) into a statistical model which might then be used in the decision-making and evaluation stages of the risk assessment. The process of building and testing a spatial causal model is demonstrated using data from a spatial sampling design, and the implications of the resulting model are discussed in terms of the risk assessment. It is argued that a spatiotemporal causal model would have greater external validity than the spatial model, enabling broader generalisations to be made regarding the impact of a discharge, and greater value as a tool for evaluating the effects of potential treatment plant upgrades. Suggestions are made on how the causal model could be augmented to include temporal as well as spatial information, including suggestions for appropriate statistical models and analyses.

Characterisation of adaptive genetic diversity in environmentally contrasted populations of Eucalyptus camaldulensis Dehnh. (river red gum).

As an increasing number of ecosystems face departures from long standing environmental conditions under climate change, our understanding of the capacity of species to adapt will become important for directing conservation and management of biodiversity. Insights into the potential for genetic adaptation might be gained by assessing genomic signatures of adaptation to historic or prevailing environmental conditions. The river red gum (Eucalyptus camaldulensis Dehnh.) is a widespread Australian eucalypt inhabiting riverine and floodplain habitats which spans strong environmental gradients. We investigated the effects of adaptation to environment on population level genetic diversity of E. camaldulensis, examining SNP variation in candidate gene loci sampled across 20 climatically diverse populations approximating the species natural distribution. Genetic differentiation among populations was high (F(ST) = 17%), exceeding previous estimates based on neutral markers. Complementary statistical approaches identified 6 SNP loci in four genes (COMT, Dehydrin, ERECTA and PIP2) which, after accounting for demographic effects, exhibited higher than expected levels of genetic differentiation among populations and whose allelic variation was associated with local environment. While this study employs but a small proportion of available diversity in the eucalyptus genome, it draws our attention to the potential for application of wide spread eucalypt species to test adaptive hypotheses.

Comparing activated sludge fungal community population diversity using denaturing gradient gel electrophoresis and terminal restriction fragment length polymorphism.

We compared the relative values of denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) for profiling fungal communities in wastewater treatment plants using both ITS and 18S rRNA gene fragments as phylogenetic markers. A similar number of fungal ribotypes was obtained with both methods for the same treatment plant when the ITS primer set was used, while a greater number of ribotypes was obtained with T-RFLP compared to DGGE with the 18S rRNA primer set. Non-metric multi-dimensional scaling of presence/absence data and analysis of similarity showed that both methods could distinguish between the different plant communities at a statistically significant level (p < 0.05), regardless of which phylogenetic marker was used. The data suggest that both methods can be used preferably together to profile activated sludge fungal communities. A comparison of profiles generated with both these phylogenetic markers based on the number of ribotypes/bands, suggests that the 18S rRNA region is more discriminatory than the ITS region. Detected differences in fungal community compositions between plants probably reflect differences in their influent compositions and operational parameters.

High spatial resolution infrared micro-spectroscopy reveals the mechanism of leaf lignin decomposition by aquatic fungi.

Organic carbon is a critical component of aquatic systems, providing energy storage and transfer between organisms. Fungi are a major decomposer group in the aquatic carbon cycle, and are one of few groups thought to be capable of breaking down woody (lignified) tissue. In this work we have used high spatial resolution (synchrotron light source) infrared micro-spectroscopy to study the interaction between aquatic fungi and lignified leaf vein material (xylem) from River Redgum trees (E. camaldulensis) endemic to the lowland rivers of South-Eastern Australia. The work provides spatially explicit evidence that fungal colonisation of leaf litter involves the oxidative breakdown of lignin immediately adjacent to the fungal tissue and depletion of the lignin-bound cellulose. Cellulose depletion occurs over relatively short length scales (5-15 µm) and highlights the likely importance of mechanical breakdown in accessing the carbohydrate content of this resource. Low bioavailability compounds (oxidized lignin and polyphenols of plant origin) remain in colonised leaves, even after fungal activity diminishes, and suggests a possible pathway for the sequestration of carbon in wetlands. The work shows that fungi likely have a critical role in the partitioning of lignified material into a biodegradable fraction that can re-enter the aquatic carbon cycle, and a recalcitrant fraction that enters long-term storage in sediments or contribute to the formation of dissolved organic carbon in the water column.

Impacts of inundation and drought on eukaryote biodiversity in semi-arid floodplain soils.

Floodplain ecosystems are characterized by alternating wet and dry phases and periodic inundation defines their ecological character. Climate change, river regulation and the construction of levees have substantially altered natural flooding and drying regimes worldwide with uncertain effects on key biotic groups. In southern Australia, we hypothesized that soil eukaryotic communities in climate change affected areas of a semi-arid floodplain would transition towards comprising mainly dry-soil specialist species with increasing drought severity. Here, we used 18S rRNA amplicon pyrosequencing to measure the eukaryote community composition in soils that had been depleted of water to varying degrees to confirm that reproducible transitional changes occur in eukaryotic biodiversity on this floodplain. Interflood community structures (3 years post-flood) were dominated by persistent rather than either aquatic or dry-specialist organisms. Only 2% of taxa were unique to dry locations by 8 years post-flood, and 10% were restricted to wet locations (inundated a year to 2 weeks post-flood). Almost half (48%) of the total soil biota were detected in both these environments. The discovery of a large suite of organisms able to survive nearly a decade of drought, and up to a year submerged supports the concept of inherent resilience of Australian semi-arid floodplain soil communities under increasing pressure from climatic induced changes in water availability.

Provisioning of bioavailable carbon between the wet and dry phases in a semi-arid floodplain.

Ecosystem functioning on arid and semi-arid floodplains may be described by two alternate traditional paradigms. The pulse-reserve model suggests that rainfall is the main driver of plant growth and subsequent carbon and energy reserve formation in the soil of arid and semi-arid regions. The flood pulse concept suggests that periodic flooding facilitates the two-way transfer of materials between a river and its adjacent floodplain, but focuses mainly on the period when the floodplain is inundated. We compared the effects of both rainfall and flooding on soil moisture and carbon in a semi-arid floodplain to determine the relative importance of each for soil moisture recharge and the generation of a bioavailable organic carbon reserve that can potentially be utilised during the dry phase. Flooding, not rainfall, made a substantial contribution to moisture in the soil profile. Furthermore, the growth of aquatic macrophytes during the wet phase produced at least an order of magnitude more organic material than rainfall-induced pulse-reserve responses during the dry phase, and remained as recognizable soil carbon for years following flood recession. These observations have led us to extend existing paradigms to encompass the reciprocal provisioning of carbon between the wet and dry phases on the floodplain, whereby, in addition to carbon fixed during the dry phase being important for driving biogeochemical transformations upon return of the next wet phase, aquatic macrophyte carbon fixed during the wet phase is recognized as an important source of energy for the dry phase. Reciprocal provisioning presents a conceptual framework on which to formulate questions about the resistance and ecosystem resilience of arid and semi-arid floodplains in the face of threats like climate change and alterations to flood regimes.

Rehabilitation options for inland waterways impacted by sulfidic sediments--field trials in a south-eastern Australian wetland.

The accumulation of significant pools of sulfidic sediments in inland wetlands and creeks is an emerging risk for the management of inland waterways. We used replicated plot trials to appraise the viability of various strategies for neutralizing oxidized, acidified sulfidic sediments in a highly degraded wetland. Of the twenty different treatments trialed only addition of calcium hydroxide or calcium carbonate, burning of wood, and planting of Phragmites australis, Typha domingensis and Atriplex nummularia into beds prepared with CaCO3 or P. australis and T. domingensis into beds of sediment and mulch, decreased total actual acidity (TAA) in the top 5 cm of sediment in the first two weeks following treatment. Only the calcium hydroxide treatments and planting of P. australis, T. domingensis and A. nummularia into beds prepared with CaCO3 decreased TAA for a longer period of time (6 months). None of the treatments, except the planting of P. australis into beds prepared with lime, decreased TAA in the 5-30 cm layer of sediments. Therefore, the only effective treatment appears to be the application of highly alkaline ameliorants which need to be transported to the site. A survey of the wetland was undertaken to estimate the total amount of actual and potential acidity stored in the wetland's sediment and overlying water and showed that up to 1200 tonnes of calcium carbonate would be required to neutralise all of the actual and potential acidity in the 10 ha wetland. However, neutralisation of the remaining water in the wetland (about 12.5 ML) would produce approximately 2750 m3 of metal rich sludge (approximately 100 tonnes dry weight) that would require separate disposal.

Acidification and buffering mechanisms in acid sulfate soil wetlands of the Murray-Darling Basin, Australia.

The acid generation mechanisms and neutralizing capacities of sulfidic sediments from two inland wetlands have been studied in order to understand the response of these types of systems to drying events. The two systems show vastly different responses to oxidation, with one (Bottle Bend (BB) lagoon) having virtually no acid neutralizing capacity (ANC) and the other (Psyche Bend (PB) lagoon) an ANC that is an order of magnitude greater than the acid generation potential. While BB strongly acidifies during oxidation the free acid generation is less than that expected from the measured proton production and consumption processes, with additional proton consumption attributed to the formation of an acid-anion (chloride) FeIII (oxyhydr)oxide product, similar to akaganéite (Fe(OH)2.7Cl0.3). While such products can partially attenuate the acidification of these systems, resilience to acidification is primarily imparted by sediment ANC.

Urban stormwater runoff drives denitrifying community composition through changes in sediment texture and carbon content.

The export of nitrogen from urban catchments is a global problem, and denitrifying bacteria in stream ecosystems are critical for reducing in-stream N. However, the environmental factors that control the composition of denitrifying communities in streams are not well understood. We determined whether denitrifying community composition in sediments of nine streams on the eastern fringe of Melbourne, Australia was correlated with two measures of catchment urban impact: effective imperviousness (EI, the proportion of a catchment covered by impervious surfaces with direct connection to streams) or septic tank density (which affects stream water chemistry, particularly stream N concentrations). Denitrifying community structure was examined by comparing terminal restriction fragment length polymorphisms of nosZ genes in the sediments, as the nosZ gene codes for nitrous oxide reductase, the last step in the denitrification pathway. We also determined the chemical and physical characteristics of the streams that were best correlated with denitrifying community composition. EI was strongly correlated with community composition and sediment physical and chemical properties, while septic tank density was not. Sites with high EI were sandier, with less fine sediment and lower organic carbon content, higher sediment cations (calcium, sodium and magnesium) and water filterable reactive phosphorus concentrations. These were also the best small-scale environmental variables that explained denitrifying community composition. Among our study streams, which differed in the degree of urban stormwater impact, sediment grain size and carbon content are the most likely drivers of change in community composition. Denitrifying community composition is another in a long list of ecological indicators that suggest the profound degradation of streams is caused by urban stormwater runoff. While the relationships between denitrifying community composition and denitrification rates are yet to be unequivocally established, landscape-scale indices of environmental impact such as EI may prove to be useful indicators of change in microbial communities.

Sulfide formation in freshwater sediments, by sulfate-reducing microorganisms with diverse tolerance to salt.

Understanding how sulfate-reducing microbes in freshwater systems respond to added salt, and therefore sulfate, is becoming increasingly important in inland systems where the threat from salinisation is increasing. To address this knowledge gap, we carried out mesocosm studies to determine how the sulfate-reducing microbial community in sediments from a freshwater wetland would respond to salinisation. The levels of inorganic mineral sulfides produced after 6months incubation were measured to determine whether they were in sufficient quantity to be harmful if re-oxidized. Comparative sequence analysis of the dissimilatory sulfite reductase (DSR) gene was used to compare the sulfate-reducing community structure in mesocosms without salt and those incubated with moderate levels of salt. The amount of total S, acid volatile sulfide or chromium-reducible sulfide produced in sediments with 0, 1 or 5gL(-1) added salt were not significantly different. Sediments subjected to 15gL(-1) salt contained significantly higher total S and acid volatile sulfide, and levels were above trigger values for potential harm if re-oxidation occurred. The overall community structure of the sulfate-reducing microbiota (SRM) was explained by the level of salt added to sediments. However, a group of sulfate reducers were identified that occurred in both the high salt and freshwater treatments. These results demonstrate that freshwater sediments contain sulfate reducers with diverse abilities to respond to salt and can respond rapidly to increasing salinity, explaining the observation that harmful levels of acid volatile sulfides can form rapidly in sediments with no history of exposure to salt.