Environmental DNA Metabarcoding Supporting Community Assessment of Environmental Stressors in a Field-Based Sediment Microcosm Study.

Conventional ecological risk assessment on toxic stressors in sediment is limited to a small and selected fraction of benthic communities. Ecogenomic approaches provide unprecedented capacity to monitor the changes of biodiversity and community composition in the field, but how to utilize it to assess ecological impact by contaminates remains largely unexplored. Here, an environmental DNA (eDNA) metabarcoding approach was used to assess the effect of copper on changes in biodiversity and community composition across the tree of life (including bacteria, protists, algae, fungi, and metazoa) in a field-based microcosm. Many microorganisms across a broad range of taxa groups changed their relative abundance in response to increased copper concentrations in sediments. Changes in community structure of microbiota appeared to be more sensitive to copper than survival of laboratory-bred organisms and indigenous macroinvertebrates. Copper caused a significant shift in prokaryotic community composition via substitution of dominant species. Network heterogeneity and Shannon diversity of the bacterial community decreased in the high copper treatments. eDNA metabarcoding assessed the effects of copper-contaminated sediment with less effort than manually processing samples. Our study highlighted the value of community profiling by an eDNA-based approach in prospective and retrospective risk assessment of environmental stressors.

Improvements and cost-effective measures to the automated intermittent water renewal system for toxicity testing with sediments.

The push to make bioassays more sensitive has meant an increased duration of testing to look at more chronic endpoints. To conduct these longer bioassays through the use of traditional bioassay methods can be difficult, as many traditional bioassays have employed manual water changes, which take considerable time and effort. To that end, static-renewal systems were designed to provide researchers a technique to ease the manual water change burden. One of the most well-known static-renewal designs, the static intermittent renewal system (STIR) was produced by the United States Environmental Protection Agency in 1993. This system is still being used in laboratories across the globe today. However, these initial designs have become rather dated as new technologies and methods have been developed that make these systems easier to build and operate. The following information details changes to the initial design and a proof of concept experiment with the benthic invertebrate, Chironomus tepperi, to validate the modifications to the original system.

Bifenthrin Causes Toxicity in Urban Stormwater Wetlands: Field and Laboratory Assessment Using Austrochiltonia (Amphipoda).

Stormwater wetlands are engineered to accumulate sediment and pollutants from stormwater and provide environmental value to urban environments. Therefore, contaminated sediment risks causing toxicity to aquatic fauna. This research identifies contaminants of concern in urban wetland sediments by assessing sediment toxicity using the amphipod Austrochiltonia subtenuis. Sediments from 98 wetlands were analyzed for contaminants, and laboratory bioassays were performed with A. subtenuis. Wild Austrochiltonia spp. were also collected from wetlands to assess field populations. Random forest modeling was used to identify the most important variables predicting survival, growth, and field absence of Austrochiltonia spp. Bifenthrin was the most frequently detected pesticide and also the most important predictor of Austrochiltonia spp. responses. Copper, permethrin, chromium, triclosan, and lead were also important. The median lethal effect concentration (LC50) of bifenthrin to laboratory-based A. subtenuis (1.09 (±0.08) µg/gOC) exposed to wetland sediments was supported by a bifenthrin-spiked sediment experiment, indicating A. subtenuis is a suitable test species. Furthermore, Austrochiltonia spp. were absent from all sites that exceeded the calculated bifenthrin LC50, demonstrating the impact of this contaminant on wild populations. This research demonstrates the sensitivity of Austrochiltonia spp. to urban sediment contamination and identifies bifenthrin as a contaminant of concern in urban wetlands.

A Meta-Analysis Evaluating the Relationship between Aquatic Contaminants and Chironomid Larval Deformities in Laboratory Studies.

Chironomid larval deformities have been widely investigated as an aquatic pollution toxicity end point. Field chironomid surveys often show a spatial association between contaminants and deformities, suggesting contaminants cause deformities. However, over 40 years of laboratory assays have not been able to confirm this causality. We therefore conducted a review of the literature and meta-analysis, in order to (A) assess whether trends across assays indicated dose-response effects, (B) characterize the consistency of results, and (C) investigate whether experimental issues and publication bias were contributing to inconsistency and/or reducing confidence in results. The experimental issues we investigated were extraneous nonchemical laboratory stressors (which may mask or interact with chemical effects), and mortality (which can confound deformity results). Our meta-analysis of the most commonly tested chemicals suggested dose-response effects for copper, but not lead or zinc. However, we also found substantial inconsistency across studies. Both mortality and extraneous stressors were potentially contributing to this inconsistency, reducing confidence in most published data. We observed no evidence of publication bias. We conclude that any causal link between contaminants and deformities remains uncertain, and suggest improved experimental and data reporting procedures to better assess this relationship.

Development of a passive sampler based on a polymer inclusion membrane for total ammonia monitoring in freshwaters.

A passive sampler for determining the time-weighted average total ammonia (i.e. molecular ammonia and the ammonium cation) concentration (C TWA) in freshwaters, which incorporated a polymer inclusion membrane (PIM) as a semi-permeable barrier separating the aqueous source solution from the receiving solution (i.e. 0.8 mol L(-1) HCl), was developed for the first time. The PIM was composed of dinonylnaphthalene sulfonic acid (DNNS) as a carrier, poly (vinyl chloride) (PVC) as a base polymer and 1-tetradecanol as a modifier. Its optimal composition was found to be 35 wt% commercial DNNS, 55 wt% PVC and 10 wt% 1-tetradecanol. The effect of environmental variables such as the water matrix, pH and temperature were also studied using synthetic freshwaters. The passive sampler was calibrated under laboratory conditions using synthetic freshwaters and exhibited a linear response within the concentration range 0.59-2.8 mg L(-1) NH4(+) (0.46-2.1 mg N L(-1)) at 20 °C. The performance of the sampler was further investigated under field conditions over 7 days. A strong correlation between spot sampling and passive sampling was achieved, thus providing a proof-of-concept for the passive sampler for reliably measuring the C(TWA) of total ammonia in freshwaters, which can be used as an indicator in tracking sources of faecal contamination in stormwater drains.

No evidence of exposure to environmental estrogens in two feral fish species sampled from the Yarra River, Australia: A comparison with Northern Hemisphere studies.

Environmental estrogens originate from a variety of sources including sewage treatment plant (STP) effluents and adverse physiological effects (endocrine disruption) have been observed in several fish species sampled downstream of STP discharges. In this study we examined common carp (Cyprinus carpio) and roach (Rutilis rutilis) for signs of exposure to environmental estrogens in the iconic Yarra River, Melbourne, Australia. The Yarra River flows through the city of Melbourne and more than 2 million people live within the catchment. Two STPs discharge water into the Yarra River within the middle reaches, and the areas immediately downstream of these discharge locations were the focus of this study. Carp and roach were chosen as test species since both have been utilised extensively for endocrine disruption research throughout Europe, North America and Asia, and data from various international studies was used for comparison with the results of the present study. Neither species showed evidence of exposure to environmental estrogens, with no elevation of plasma vitellogenin levels in males and no incidence of intersex gonads. Most physiological endpoints in both species from this study were within ranges reported in carp and roach from reference sites in other studies, however some degenerative histological changes in both male and female gonads were observed. Surface water samples showed no estrogenic activity (measured by the yeast-estrogen screen, YES), but did display strong anti-estrogenic and weak androgenic activity (measured by the yeast-androgen screen, YAS). Whilst the results show no evidence of impacts from environmental estrogens in the Yarra River, the presence of both anti-estrogenic and androgenic activity in water samples, as well as some gonadal changes in carp is concerning and indicates that our focus needs to broaden, in order to look for biological impacts in resident fauna that might be due to environmental pollutants other than environmental estrogens.

Comparing the impacts of sediment-bound bifenthrin on aquatic macroinvertebrates in laboratory bioassays and field microcosms.

We conducted two laboratory bioassays and two field microcosm exposures with bifenthrin (a synthetic pyrethroid) in order to evaluate the capacity of single-species laboratory bioassays to predict lethal and sublethal impacts on aquatic invertebrates in microcosms. For the laboratory species, Chironomus tepperi, larval survival was reduced by 24% at 53.66µg/g OC, while adult emergence was reduced at concentrations of 33.33µg/g OC and higher, with a 61% decrease at 77.78µg/g OC and no emergence at 126.67µg/g OC. The abundance of several other microcosm taxa was reduced in the microcosms at a similar concentration range (33.33µg/g OC and above), however there was no impact on the abundance of the congeneric species, Chironomus oppositus. The differences in impacts between test systems were potentially due to both differing species sensitivity and the interaction of ambient temperature with bifenthrin toxicity. Bifenthrin also was associated with early emergence of Chironomus sp. in both test systems, at concentrations of 10µg/g OC and higher (laboratory) and 43.90µg/g OC (microcosm), and with a significant decrease in the proportion of C. oppositus males in a microcosm. These findings indicate that while laboratory bioassays accurately predict many impacts in the field, there are some limitations to the predictive capacity of these tests.

The Parthenogenetic Cosmopolitan Chironomid, Paratanytarsus grimmii, as a New Standard Test Species for Ecotoxicology: Culturing Methodology and Sensitivity to Aqueous Pollutants.

Chironomids from the genus Chironomus are widely used in laboratory ecotoxicology, but are prone to inbreeding depression, which can compromise test results. The standard Chironomus test species (C. riparius, C. dilutus and C. yoshimatsui) are also not cosmopolitan, making it difficult to compare results between geographic regions. In contrast, the chironomid Paratanytarsus grimmii is cosmopolitan, and not susceptible to inbreeding depression because it reproduces asexually by apomictic parthenogenesis. However, there is no standardised culturing methodology for P. grimmii, and a lack of acute toxicity data for common pollutants (metals and pesticides). In this study, we developed a reliable culturing methodology for P. grimmii. We also determined 24-h first instar LC50s for the metals Cu, Pb, Zn, Cd and the insecticide imidacloprid. By developing this culturing methodology and generating the first acute metal and imidacloprid LC50s for P. grimmii, we provide a basis for using P. grimmii in routine ecotoxicological testing.

Transgenerational effects of parental nutritional status on offspring development time, survival, fecundity, and sensitivity to zinc in Chironomus tepperi midges.

Environmental stimuli can induce plastic changes in life history traits, and stimuli experienced by parents can be transmitted to the next generation ("transgenerational") through the inheritance of factors unrelated to changes in DNA sequences. Transgenerational effects are common in species living in habitats subjected to recurrent stressful events, such as fluctuating resource availability. In a previous study, the nutritional status of the midge Chironomus tepperi has been reported to influence life history traits of the offspring. In this study we investigated whether they also alter sensitivity of offspring to zinc. Offspring of parents reared under low food conditions had a shorter development time and lower reproductive output compared to offspring of parents raised under excess food. While zinc exposure decreased the survival of offspring generally, the interaction between parental food level and zinc exposure did not influence the relative sensitivity of offspring toward zinc. Parental nutritional stress therefore triggered transgenerational effects, potentially acting as confounding factors in ecotoxicological studies, but they did not directly affect the susceptibility of offspring to zinc.

A national survey of trace organic contaminants in Australian rivers.

Trace organic contaminant (TrOC) studies in Australia have, to date, focused on wastewater effluents, leaving a knowledge gap of their occurrence and risk in freshwater environments. This study measured 42 TrOCs including industrial compounds, pesticides, and pharmaceuticals and personal care products by liquid chromatography tandem mass spectrometry at 73 river sites across Australia quarterly for 1 yr. Trace organic contaminants were found in 92% of samples, with a median of three compounds detected per sample (maximum 18). The five most commonly detected TrOCs were the pharmaceuticals salicylic acid (82%, maximum = 1530 ng/L), paracetamol (also known as acetaminophen; 45%, maximum = 7150 ng/L), and carbamazepine (27%, maximum = 682 ng/L), caffeine (65%, maximum = 3770 ng/L), and the flame retardant (2-chloroethyl) phosphate (44%, maximum = 184 ng/L). Pesticides were detected in 28% of the samples. To determine the risk posed by the detected TrOCs to the aquatic environment, hazard quotients were calculated by dividing the maximum concentration detected for each compound by the predicted no-effect concentrations. Three of the 42 compounds monitored (the pharmaceuticals carbamazepine and sulfamethoxazole and the herbicide simazine) had a hazard quotient >1, suggesting that they may be causing adverse effects at the most polluted sites. A further 10 compounds had hazard quotients >0.1, indicating a potential risk; these included four pharmaceuticals, three personal care products, and three pesticides. Most compounds had hazard quotients significantly <0.1. The number of TrOCs measured in this study was limited and further investigations are required to fully assess the risk posed by complex mixtures of TrOCs on exposed biota.

Investigation of 10 herbicides in surface waters of a horticultural production catchment in southeastern Australia.

Herbicides are regularly applied in horticultural production systems and may migrate off-site, potentially posing an ecological risk to surface waterways. However, few studies have investigated the levels and potential ecotoxicological impact of herbicides in horticultural catchments in southern Australia. This study investigated the presence of 10 herbicides at 18 sites during a 5-month period in horticulturally important areas of the Yarra Valley in southeastern Australia. Seven of the 10 herbicides were detected in the streams, in 39 % of spot water samples, in 25 % of surface sediment samples, and in >70 % of the passive sampler systems deployed. Few samples contained residues of ≥2 herbicides. Simazine was the herbicide most frequently detected in water, sediment, and passive sampler samples and had the highest concentrations in water (0.67 µg/L) and sediment (260 µg/kg dry weight). Generally the concentrations of the herbicides detected were several orders of magnitude lower than reported ecotoxicological effect values, including those for aquatic plants and algae, suggesting that concentrations of individual chemicals in the catchment were unlikely to pose an ecological risk. However, little is known about the combined effects of simultaneous, low-level exposure of multiple herbicides of the same mode of action on Australian aquatic organisms nor their contribution when found in mixtures with other pesticides. Further research is required to adequately assess the risk of pesticides in Victorian aquatic environments.

Background concentrations and spatial distribution of PFAS in surface waters and sediments of the greater Melbourne area, Australia.

A survey of per- and polyfluoroalkyl substances (PFAS) was conducted in Melbourne, Australia to determine background concentrations in residential, industrial, municipal wastewater treatment plants, and rural land uses. Surface water and sediment samples collected from 65 sites with different catchment land uses were analysed for thirty-three PFAS. Twenty-two out of thirty-three targeted PFAS were detected, with at least one PFAS species was detected in 98% water samples and 8% sediment samples. One site was determined to have point-source pollution from an airport (surface water Σ33PFAS = 4261 ng/L) and was excluded from statistical analyses. The median Σ33PFAS concentration in surface water was 63.5 ng/L and the average was 78.6 ng/L (range < DL-526 ng/L). PFAS species with the highest median concentrations were PFBA (11.3 ng/L), PFHxA (9.2 ng/L), PFOA (8.3 ng/L), PFOS (8.0 ng/L), PFPeA (7.5 ng/L), PFHpA (3.2 ng/L), and PFHxS (2.9 ng/L). The average Σ4PFAS in sediments was 0.35 ng/g d.w. (range =

Environmental monitoring using next generation sequencing: rapid identification of macroinvertebrate bioindicator species.

INTRODUCTION: Invertebrate communities are central to many environmental monitoring programs. In freshwater ecosystems, aquatic macroinvertebrates are collected, identified and then used to infer ecosystem condition. Yet the key step of species identification is often not taken, as it requires a high level of taxonomic expertise, which is lacking in most organizations, or species cannot be identified as they are morphologically cryptic or represent little known groups. Identifying species using DNA sequences can overcome many of these issues; with the power of next generation sequencing (NGS), using DNA sequences for routine monitoring becomes feasible. RESULTS: In this study, we test if NGS can be used to identify species from field-collected samples in an important bioindicator group, the Chironomidae. We show that Cytochrome oxidase I (COI) and Cytochrome B (CytB) sequences provide accurate DNA barcodes for chironomid species. We then develop a NGS analysis pipeline to identifying species using megablast searches of high quality sequences generated using 454 pyrosequencing against comprehensive reference libraries of Sanger-sequenced voucher specimens. We find that 454 generated COI sequences successfully identified up to 96% of species in samples, but this increased up to 99% when combined with CytB sequences. Accurate identification depends on having at least five sequences for a species; below this level species not expected in samples were detected. Incorrect incorporation of some multiplex identifiers (MID's) used to tag samples was a likely cause, and most errors could be detected when using MID tags on forward and reverse primers. We also found a strong quantitative relationship between the number of 454 sequences and individuals showing that it may be possible to estimate the abundance of species from 454 pyrosequencing data. CONCLUSIONS: Next generation sequencing using two genes was successful for identifying chironomid species. However, when detecting species from 454 pyrosequencing data sets it was critical to include known individuals for quality control and to establish thresholds for detecting species. The NGS approach developed here can lead to routine species-level diagnostic monitoring of aquatic ecosystems.

Structural changes in a macrozoobenthos assemblage after imidacloprid pulses in aquatic field-based microcosms.

A field-based microcosm experiment was performed to investigate the effects of repeated pulses of the neonicotinoid insecticide imidacloprid on a lentic benthos assemblage. This specific microcosm method was chosen because it allows for both testing of a wide range of organisms under natural conditions and as well as gaining insight into intraspecific and interspecific interactions. The macrozoobenthos that colonised the microcosms was exposed to three pulses each 1 week apart at nominal concentrations ranging from 0.6 to 40 µg/L. Imidacloprid underwent fast aqueous photolysis due to optimal sunlight conditions during the test phase (half-life = 28 ± 8 h [monitored for 21 days]). Nonetheless, decreased abundance and emergence of Ephemeroptera and decreased survival of chironomid species of the subfamilies Tanypodinae and Orthocladiinae were observed at time-weighted average concentrations of 2.3 µg/L. In contrast, the gastropod Radix sp. became dominant at high imidacloprid concentrations, probably due to decreased competition for food with sensitive species. The results of this study show that repeated short-term contamination of imidacloprid at low concentration levels may affect aquatic ecosystems even under optimal conditions for photodegradation. The microcosm approach, with its simple and field-relevant design, proved to be a useful tool for assessing the effects of imidacloprid contamination.

Seasonal estimation of groundwater vulnerability.

Index-based methods estimate a fixed value of groundwater vulnerability (GWV); however, the effects of time variations on this estimation have not been comprehensively studied. It is imperative to estimate a time-variant vulnerability that accounts for climatic changes. In this study, we used a Pesticide DRASTICL method separating hydrogeological factors into dynamic and static groups followed by correspondence analysis. The dynamic group is composed of depth and recharge, and the static group is composed of aquifer media, soil media, topography slope, impact of vadose zone, aquifer conductivity and land use. The model results were 42.25-179.89, 33.93-159.81, 34.08-168.74, and 45.56-205.20 for spring, summer, autumn, and winter, respectively. The results showed a moderate correlation between the model predictions and observed nitrogen concentrations with R2 = 0.568 and a high correlation for phosphorus concentrations with R2 = 0.706. Our results suggest that the time-variant GWV model provides a robust yet flexible method for investigating seasonal changes in GWV. This model is an improvement to the standard index-based methods, making them sensitive to climatic changes and portraying a true vulnerability estimation. Finally, the correction of the rating scale value fixes the problem of overestimation in standard models.

Food limitation in Chironomus tepperi: effects on survival, sex ratios and development across two generations.

Species from the Dipteran family Chironomidae are widely used in laboratory and field studies to identify toxicity in freshwater environments. However, toxicity assessments can be influenced by food availability, which can alter endpoints in assays including viability, sex ratios and development time. The aim of this study was to determine if food limitation affected the endpoints used in toxicity tests with the Australian model organism, Chironomus tepperi, including responses in offspring. First instar larvae were subjected to food treatments with larval density controlled and offspring were either raised under the same food conditions as their parents or under standard conditions. In lower food density treatments adults in the F0 generation experienced delayed emergence and females produced fewer egg masses. F0 diet affected the performance of F1 to continued exposure and there was evidence that the quality of the offspring was compromised. Although sex ratios were not skewed, males and females responded differently to food limitation, especially in the F1 generation where female development was more delayed. These results demonstrate that endpoints used in toxicity evaluation in C. tepperi also respond to food availability, highlighting the need to control for food in both laboratory and field toxicity studies. Multiple generations should ideally be exposed to stressors under laboratory conditions to elucidate likely long term effects in the field.

Environmental fate of fungicides in surface waters of a horticultural-production catchment in southeastern Australia.

Fungicides are regularly applied in horticultural production systems and may migrate off-site, potentially posing an ecological risk to surface waterways. However, few studies have investigated the fate of fungicides in horticultural catchments. This study investigated the presence of 24 fungicides at 18 sites during a 5-month period within a horticultural catchment in southeastern Australia. Seventeen of the 24 fungicides were detected in the waterways, with fungicides detected in 63% of spot water samples, 44% of surface sediment samples, and 44% of the passive sampler systems deployed. One third of the water samples contained residues of two or more fungicides. Myclobutanil, trifloxystrobin, pyrimethanil, difenoconazole, and metalaxyl were the fungicides most frequently detected, being present in 16-38% of the spot water samples. Iprodione, myclobutanil, pyrimethanil, cyproconazole, trifloxystrobin, and fenarimol were found at the highest concentrations in the water samples (> 0.2 µg/l). Relatively high concentrations of myclobutanil and pyrimethanil (≥ 120 µg/kg dry weight) were detected in the sediment samples. Generally the concentrations of the fungicides detected were several orders of magnitude lower than reported ecotoxicological effect values, suggesting that concentrations of individual fungicides in the catchment were unlikely to pose an ecological risk. However, there is little information on the effects of fungicides, especially fungi and microbes, on aquatic ecosystems. There is also little known about the combined effects of simultaneous low-level exposure of multiple fungicides to aquatic organisms. Further research is required to adequately assess the risk of fungicides in aquatic environments.

Household herbicide use as a source of simazine contamination in urban surface waters.

Contamination of urban surface waters by herbicides is an increasing concern; however, sources of contamination are poorly understood, hindering the development of mitigation and regulatory strategies. Impervious surfaces, such as concrete in driveways and paths are considered an important facilitator for herbicide runoff to urban surface waters following applications by residential homeowners. This study assessed the transferability of a herbicide from concrete pavers treated with an off-the-shelf product, containing simazine as the active herbicide, marketed for residential homeowner application to impervious surfaces. Commercially available pavers were treated according to label directions and the effects of exposure time prior to irrigation, repeated irrigations, and dry time between irrigations on transferability of simazine to runoff were assessed. Simazine transferability was greatest when receiving an initial irrigation 1 h after application, with concentrations in runoff reduced by half when exposure times prior to the first irrigation were >2 days. Concentrations remained stable for repeated irrigations up to 320 days and exposures to outdoor conditions of 180 days prior to a first irrigation. Dry time between irrigations significantly influenced simazine transfer to runoff. Dry periods of 140 days resulted in approximately a 4-times increase in simazine transferability to runoff. These results suggest that herbicides used by homeowners, or any other users, on impervious surfaces are available to contaminate runoff for prolonged time periods following application at concentrations that may pose risks to aquatic life and for reuse of harvested runoff on parks and gardens. Regulators should consider the potential of hard surfaces to act as reservoirs for herbicides when developing policies and labelling products.

Effects of pesticides monitored with three sampling methods in 24 sites on macroinvertebrates and microorganisms.

Grab water samples, sediment samples, and 2,2,4-trimethylpentane passive samplers (TRIMPS) were used to determine the exposure to 97 pesticides in 24 southeast Australian stream sites over 5 months. Macroinvertebrate communities and selected microorganisms (bacteria, flagellates, ciliates, amoebas, nematodes, and gastrotrichs) were sampled to detect relationships with pesticide toxicity. Sediment samples had the highest estimated toxicities in terms of toxic units (TU) for Daphnia magna (TUDM) and for Selenastrum capricornutum (TUSC). The pesticide-selective SPEARpesticides and the general SIGNAL index for macroinvertebrates exhibited negative linear relationships (r(2) = 0.67 and 0.36, respectively) with pesticide contamination in terms of log maximum TUDM (log mTUDM), suggesting macroinvertebrate community change due to pesticide exposure. Pesticide contamination was the only measured variable explaining variation in ecological quality. Variation in the densities of several microbial groups was best explained by environmental variables other than log TUs. The log mTUDM values derived from sediment concentrations were most important to establish a link with effects on macroinvertebrates, whereas log mTUDM of grab water samples had only minor contribution. Current-use insecticides and fungicides can affect macroinvertebrate communities and monitoring of sediment and continuous water sampling is needed to detect these effects.

Potamopyrgus antipodarum has the potential to detect effects from various land use activities on a freshwater ecosystem.

Identifying risks to ecosystems from contaminants needs a diversity of bioindicators, to understand the effects of these contaminants on a range of taxa. Molluscs are an ideal bioindicator because they are one of the largest phyla with extremely high ecological and economic importance. The aim of this study was to evaluate if laboratory bred Potamopyrgus antipodarum has the potential to show the impact of contaminants from various land use activities and degree of pollution on a freshwater ecosystem. We assessed the impact of contaminants arising from runoff and direct discharges in Merri Creek by measuring organism level responses (survival, growth, and reproduction), and sub-organism level responses (glutathione S-transferase (GST) activity, lipid peroxidation (LPO) activity and catalase (CAT) activity) in snails after 28-d of deployment at nine sites in Merri Creek and one site in Cardinia Creek. In Merri Creek, the top two sites were reference sites (with low impact from human activities), while the rest were impact sites (impacted by various anthropogenic land uses). Cardinia Creek (an additional reference site) had lower human activity. High concentrations of heavy metals, nutrients, and/or synthetic pyrethroids (bifenthrin) dominated these sites, which are likely to have contributed towards the negative responses observed in the snails. There was little influence from environmental conditions and site location on the endpoints because we found a similar response at an additional reference site compared to the reference sites in Merri Creek. At the organism level, reproduction increased and/or reduced, while CAT was affected at the sub-organism level. Potamopyrgus antipodarum has the potential to be a sensitive bioindicator for Australian conditions because the snails responded to varying concentrations of contaminants across different land use activities and showed similar sensitivity to P. antipodarum found in other regions of the globe and other bioindicators.