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.

Metabolomics Provide Sensitive Insights into the Impacts of Low Level Environmental Contamination on Fish Health-A Pilot Study.

This exploratory study aims to investigate the health of sand flathead (Platycephalus bassensis) sampled from five sites in Port Phillip Bay, Australia using gas chromatography-mass spectrometry (GC-MS) metabolomics approaches. Three of the sites were the recipients of industrial, agricultural, and urban run-off and were considered urban sites, while the remaining two sites were remote from contaminant inputs, and hence classed as rural sites. Morphological parameters as well as polar and free fatty acid metabolites were used to investigate inter-site differences in fish health. Significant differences in liver somatic index (LSI) and metabolite abundance were observed between the urban and rural sites. Differences included higher LSI, an increased abundance of amino acids and energy metabolites, and reduced abundance of free fatty acids at the urban sites compared to the rural sites. These differences might be related to the additional energy requirements needed to cope with low-level contaminant exposure through energy demanding processes such as detoxification and antioxidant responses as well as differences in diet between the sites. In this study, we demonstrate that metabolomics approaches can offer a greater level of sensitivity compared to traditional parameters such as physiological parameters or biochemical markers of fish health, most of which showed no or little inter-site differences in the present study. Moreover, the metabolite responses are more informative than traditional biomarkers in terms of biological significance as disturbances in specific metabolic pathways can be identified.

Superimposed microplastic pollution in a coastal metropolis.

The mitigation of microplastic pollution in the environment calls for a better understanding of the sources and transportation, especially from land sources to the open ocean. We conducted a large-scale investigation of microplastic pollution across the Greater Melbourne Area and the Western Port area, Australia, spanning gradients of land-use from un-developed catchments in conservation areas to more heavily-developed areas. Microplastics were detected in 94% of water samples and 96% of sediment samples, with abundances ranging from 0.06 to 2.5 items/L in water and 0.9 to 298.1 items/kg in sediment. The variation of microplastic abundance in sediments was closely related to that of the overlying waters. Fiber was the most abundant (89.1% and 68.6% of microplastics in water and sediment respectively), and polyester was the dominant polymer in water and sediment. The size of more than 40% of all total microplastics observed was less than 1 mm. Both light and dense polymers of different shapes were more abundant in sediments than those in water, indicating that there is microplastic accumulation in sediments. The abundance of microplastics was higher near coastal cities than at less densely-populated inland areas. A spatial analysis of the data suggests that the abundance of microplastics increases downstream in rivers and accumulates in estuaries and the lentic reaches of these rivers. Correlation and redundancy analysis were used to explore the associations between microplastic pollution and different land-use types. More microplastics and polymer types were found at areas with large amounts of commercial, industrial and transport activities. Microplastic abundances were also correlated with mean particle size. Microplastic hotspots within a coastal metropolis might be caused by a combination of natural accumulation via hydrological dynamics and contribution from increasing anthropogenic influences. Our results strongly suggest that coastal metropolis superimposed on increasing microplastic levels in waterbodies from inland areas to the estuaries and open oceans.

Dietary Uptake and Depuration Kinetics of Perfluorooctane Sulfonate, Perfluorooctanoic Acid, and Hexafluoropropylene Oxide Dimer Acid (GenX) in a Benthic Fish.

Per- and poly-fluoroalkyl substances (PFAS) are ubiquitously distributed throughout aquatic environments and can bioaccumulate in organisms. We examined dietary uptake and depuration of a mixture of 3 PFAS: perfluorooctanoic acid (PFOA; C8 HF15 O2 ), perfluorooctane sulfonate (PFOS; C8 HF17 SO3 ), and hexafluoropropylene oxide dimer acid (HPFO-DA; C6 HF11 O3 ; trade name GenX). Benthic fish (blue spot gobies, Pseudogobius sp.) were fed contaminated food (nominal dose 500 ng g-1 ) daily for a 21-d uptake period, followed by a 42-d depuration period. The compounds PFOA, linear-PFOS (linear PFOS), and total PFOS (sum of linear and branched PFOS) were detected in freeze-dried fish, whereas GenX was not, indicating either a lack of uptake or rapid elimination (<24 h). Depuration rates (d-1 ) were 0.150 (PFOA), 0.045 (linear-PFOS), and 0.042 (linear+branched-PFOS) with corresponding biological half-lives of 5.9, 15, and 16 d, respectively. The PFOS isomers were eliminated differently, resulting in enrichment of linear-PFOS (70-90%) throughout the depuration period. The present study is the first reported study of GenX dietary bioaccumulation potential in fish, and the first dietary study to investigate uptake and depuration of multiple PFASs simultaneously, allowing us to determine that whereas PFOA and PFOS accumulated as expected, GenX, administered in the same way, did not appear to bioaccumulate. Environ Toxicol Chem 2020;39:595-603. © 2019 SETAC.

A Re-Evaluation of Chironomid Deformities as an Environmental Stress Response: Avoiding Survivorship Bias and Testing Noncontaminant Biological Factors.

Larval deformities have been observed in chironomids, and are thought to be associated with aquatic contaminant exposure. However, in laboratory assays, deformities have not been linked with contaminants in the absence of potential confounding variables including mortality, which introduces a survivorship bias. There is also a paucity of data on noncontaminant causes. In addition, power analyses are rarely undertaken, meaning that effect sizes detectable are usually uncertain. We therefore aimed to clarify factors associated with deformities, by running survivorship bias-free (i.e., sublethal) assays, assessing contaminant (copper and imidacloprid) and noncontaminant (malnutrition) stressors, and considering natural biological (metamorphosis) factors in Chironomus tepperi. We included a posteriori power analyses for all tests. Our assays found no significant association between tested factors and deformity rate. Power analyses indicated that the stressor experiment had moderate power to detect deformity effects. The metamorphosis assay had relatively lower power (due to an unexpectedly high control deformity rate), highlighting the importance of power tests in these types of evaluations. These results, in conjunction with others recently published, raise doubts as to the causal effects of environmental stressors on deformity incidence. By avoiding survivorship bias, and by testing noncontaminant factors and statistical power, we present a more holistic methodology, to resolve ongoing uncertainty in this area. We also discuss possible future directions for chironomid deformity research, and concerns regarding survivorship bias in ecotoxicology. Environ Toxicol Chem 2019;38:1658-1667. © 2019 SETAC.

Toxicological effects of fungicide mixtures on the amphipod Austrochiltonia subtenuis.

Approaches to assess the toxicity of mixtures often use predictive models with acute mortality as an endpoint at relatively high concentrations. However, these approaches do not reflect realistic situations where organisms could be exposed to chemical mixtures over long periods at low concentrations at which no significant mortalities occur. The present study investigated chronic effects of 2 common fungicides, Filan® (active ingredient [a.i]) boscalid) and Systhane™ (a.i. myclobutanil), on the amphipod Austrochiltonia subtenuis at environmentally relevant concentrations under laboratory conditions. Sexually mature amphipods were exposed singly and in combination to Filan (1, 10, and 40 µg a.i./L) and Systhane (3 µg a.i./L) over 28 d. Survival, growth, a wide range of reproduction endpoints, and glutathione-S-transferase (GST) activity were measured at the end of the experiment. Both fungicides had significant independent effects on male growth, sex ratio, and juvenile size. Filan mainly affected female growth and the number of embryos per gravid female, whereas Systhane mainly affected the time for females to become gravid. The combined effects of these fungicides on numbers of gravid females and juveniles were antagonistic, causing a 61% reduction in the number of gravid females and a 77% reduction in the number of juveniles produced at the highest concentrations (40 µg a.i./L of boscalid and 3 µg a.i./L of myclobutanil) compared with the controls. There were no significant effects on survival or GST activity. The present study demonstrated that the effects of mixtures were endpoint dependent and that using a variety of endpoints should be considered for a comprehensive understanding of mixture effects. Also, chronic studies are more informative than acute studies for environmentally relevant fungicide concentrations. Environ Toxicol Chem 2017;36:2651-2659. © 2017 SETAC.

Effects of two commonly used fungicides on the amphipod Austrochiltonia subtenuis.

Fungicides are used widely in agriculture and have been detected in adjacent rivers and wetlands. However, relatively little is known about the potential effects of fungicides on aquatic organisms. The present study investigated the effects of 2 commonly used fungicides, the boscalid fungicide Filan® and the myclobutanil fungicide Systhane™ 400 WP, on life history traits (survival, growth, and reproduction) and energy reserves (lipid, protein, and glycogen content) of the amphipod Austrochiltonia subtenuis under laboratory conditions, at concentrations detected in aquatic environments. Amphipods were exposed to 3 concentrations of Filan (1 µg active ingredient [a.i.]/L, 10 µg a.i./L, and 40 µg a.i./L) and Systhane (0.3 µg a.i./L, 3 µg a.i./L, and 30 µg a.i./L) over 56 d. Both fungicides had similar effects on the amphipod at the organism level. Reproduction was the most sensitive endpoint, with offspring produced in controls but none produced in any of the fungicide treatments, and total numbers of gravid females in all fungicide treatments were reduced by up to 95%. Female amphipods were more sensitive than males in terms of growth. Systhane had significant effects on survival at all concentrations, whereas significant effects of Filan on survival were observed only at 10 µg a.i./L and 40 µg a.i./L. The effects of fungicides on energy reserves of the female amphipod were different. Filan significantly reduced amphipod protein content, whereas Systhane significantly reduced the lipid content. The present study demonstrates wide-ranging effects of 2 common fungicides on an ecologically important species that has a key role in trophic transfer and nutrient recycling in aquatic environments. These results emphasize the importance of considering the long-term effects of fungicides in the risk assessment of aquatic ecosystems. Environ Toxicol Chem 2017;36:720-726. © 2016 SETAC.

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.

Health Status of Sand Flathead (Platycephalus bassensis), Inhabiting an Industrialised and Urbanised Embayment, Port Phillip Bay, Victoria as Measured by Biomarkers of Exposure and Effects.

Port Phillip Bay, Australia, is a large semi-closed bay with over four million people living in its catchment basin. The Bay receives waters from the Yarra River which drains the city of Melbourne, as well as receiving the discharges of sewage treatment plants and petrochemical and agricultural chemicals. A 1999 study demonstrated that fish inhabiting Port Phillip Bay showed signs of effects related to pollutant exposure despite pollution management practices having been implemented for over a decade. To assess the current health status of the fish inhabiting the Bay, a follow up survey was conducted in 2015. A suite of biomarkers of exposure and effects were measured to determine the health status of Port Phillip Bay sand flathead (Platycephalus bassensis), namely ethoxyresorufin-O-deethylase (EROD) activity, polycyclic aromatic hydrocarbons (PAH) biliary metabolites, carboxylesterase activity (CbE) and DNA damage (8-oxo-dG). The reduction in EROD activity in the present study suggests a decline in the presence of EROD activity-inducing chemicals within the Bay since the 1990s. Fish collected in the most industrialised/urbanised sites did not display higher PAH metabolite levels than those in less developed areas of the Bay. Ratios of PAH biliary metabolite types were used to indicate PAH contaminant origin. Ratios indicated fish collected at Corio Bay and Hobsons Bay were subjected to increased low molecular weight hydrocarbons of petrogenic origin, likely attributed to the close proximity of these sites to oil refineries, compared to PAH biliary metabolites in fish from Geelong Arm and Mordialloc. Quantification of DNA damage indicated a localised effect of exposure to pollutants, with a 10-fold higher DNA damage level in fish sampled from the industrial site of Corio Bay relative to the less developed site of Sorrento. Overall, integration of biomarkers by multivariate analysis indicated that the health of fish collected in industrialised areas was compromised, with biologically significant biomarkers of effects (LSI, CF and DNA damage) discriminating between individuals collected in industrialised areas from observations made in fish collected in less developed areas of the Bay.

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.

Detecting long-term temporal trends in sediment-bound trace metals from urbanised catchments.

The shift from rural lifestyles to urban living has dramatically altered the way humans interact and live across the globe. With over 50% of the world's populations living within cities, and significant increases expected over the next 50 years, it is critical that changes to social, economic and environmental sustainability of cities globally be implicit. Protecting and enhancing aquatic ecosystems, which provide important ecosystem services, is challenging. A number of factors influence pollutants in urban waterways including changes in land-use, impervious area and stormwater discharges, with sediment-bound pollution a major issue worldwide. This work aimed to investigate the spatial and temporal distribution of trace metals in freshwater sediments from six urbanised catchment over a 30-year period. It provides an estimate of pollution using a geoaccumulation index and examines possible toxicity using a probable effect concentration quotient (mPECq). Results showed significant temporal changes in metal concentrations over time, with lead generally decreasing in all but one of the sites, attributed to significant changes in environmental policies and the active elimination of lead products. Temporal changes in other metals were variable and likely dependent on site-specific factors. While it is likely that diffuse pollution is driving changes in zinc, for metals such as lead, chromium and copper, it is likely that watershed landuse and/or point sources are more important. The results clearly indicated that changes to watershed landuse, environmental policy and pollution abatement programs are all driving changes in sediment quality, highlighting the utility of long-term sediment monitoring for assessment of urban watershed condition. While this study has demonstrated the utility of detecting long-term changes in metal concentrations, this approach could easily be adapted to detect and assess future trends in other hydrophobic contaminants and emerging chemicals of concern, such as synthetic pyrethroids, providing essential information for the protection of catchment.

Effects of Lumbriculus variegatus (Annelida, Oligochaete) bioturbation on zinc sediment chemistry and toxicity to the epi-benthic invertebrate Chironomus tepperi (Diptera: Chironomidae).

Classical laboratory-based single-species sediment bioassays do not account for modifications to toxicity from bioturbation by benthic organisms which may impact predictions of contaminated sediment risk to biota in the field. This study aims to determine the effects of bioturbation on the toxicity of zinc measured in a standard laboratory bioassay conducted with chironomid larvae (Chironomus tepperi). The epi-benthic chironomid larvae were exposed to two different levels of sediment contamination (1600 and 1980 mg/kg of dry weight zinc) in the presence or absence of annelid worms (Lumbriculus variegatus) which are known to be tolerant to metal and to have a large impact on sediment properties through bioturbation. Chironomids had 5-6x higher survival in the presence of L. variegatus which shows that bioturbation had a beneficial effect on the chironomid larvae. Chemical analyses showed that bioturbation induced a flux of zinc from the pore water into the water column, thereby reducing the bioavailability of zinc in pore water to the chironomid larvae. This also suggested that pore water was the major exposure path for the chironomids to metals in sediment. During the study, annelid worms (Oligochaetes) produced a thin layer of faecal pellets at the sediment surface, a process known to: (i) create additional adsorption sites for zinc, thus reducing its availability, (ii) increase the microbial abundance that in turn could represent an additional food source for opportunistic C. tepperi larvae, and (iii) modify the microbial community's structure and alter the biogeochemical processes it governs thus indirectly impact zinc toxicity. This study represents a contribution in recognising bioturbating organisms as "ecological engineers" as they directly and indirectly influence metal bioavailability and impact other sediment-inhabiting species. This is significant and should be considered in risk assessment of zinc levels (and other metals) in contaminated sediment when extrapolating from laboratory studies to the field.

Contaminant (PAHs, OCs, PCBs and trace metals) concentrations are declining in axial tissue of sand flathead (Platycephalus bassensis) collected from an urbanised catchment (Port Phillip Bay, Australia).

Concentrations of PAHs, OCs, PCBs and trace metals were determined in the white muscle of sand flathead Platycephalus bassensis collected at 6 locations in Port Phillip Bay during 2015. No PAHs, OCs or PCBs were detected in the white muscle of sand flathead at any of the locations, however measurable levels of As, Cu, Hg, Se and Zn were detected at all sites. Only As and Hg exhibited regional difference in white muscle concentrations, with As present only in a non-toxic organic form and Hg measured at levels that are comparable to levels reported in reference sites in other studies. All contaminants detected in the white muscle of sand flathead collected in Port Phillip Bay in 2015 were below Australian Food Standards guideline values, and by world standards, the Port Phillip Bay sand flathead population is considered minimally contaminated. Furthermore, tissue contaminant concentrations appear to be decreasing over time.

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.

Effects of the boscalid fungicide Filan® on the marine amphipod Allorchestes compressa at environmentally relevant concentrations.

Fungicides are widely used in agriculture to control fungal diseases. After application, fungicides can be transported offsite to surface and groundwater and ultimately enter estuarine and marine environments. The presence of fungicides in the marine environment may pose risks to marine organisms, but little is known about fungicide effects on these organisms, especially invertebrates. The present study investigated the effects of the commonly used boscalid fungicide Filan® on life history traits, feeding rate, and energy reserves (lipid, glycogen, and protein content) of the marine amphipod Allorchestes compressa over 6 wk under laboratory conditions. Amphipods were exposed to 3 concentrations of Filan (1 µg, 10 µg, and 40 µg active ingredient [a.i.]/L), with 5 replicates per treatment. Lipid content and reproduction were the most sensitive measures of effect, with lipid content reduced by 53.8% at the highest concentration. Survival, growth, and other energy reserves of amphipods were also negatively affected by Filan, and the effects were concentration dependent. Antennal deformities were incidentally observed on the amphipods at a concentration of 40 µg a.i./L. The results of the present study indicate comprehensive effects of the boscalid fungicide Filan on A. compressa at environmentally relevant concentrations. The decline or absence of A. compressa in marine ecosystems could impair the ecosystem function because of their important role in trophic transfer and nutrient recycling. The authors' results suggest that even though the use of fungicides is often regarded as posing only a minor risk to aquatic organisms, the assessment of their long-term effects is critical.

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.

A multi-platform metabolomics approach demonstrates changes in energy metabolism and the transsulfuration pathway in Chironomus tepperi following exposure to zinc.

Measuring biological responses in resident biota is a commonly used approach to monitoring polluted habitats. The challenge is to choose sensitive and, ideally, stressor-specific endpoints that reflect the responses of the ecosystem. Metabolomics is a potentially useful approach for identifying sensitive and consistent responses since it provides a holistic view to understanding the effects of exposure to chemicals upon the physiological functioning of organisms. In this study, we exposed the aquatic non-biting midge, Chironomus tepperi, to two concentrations of zinc chloride and measured global changes in polar metabolite levels using an untargeted gas chromatography-mass spectrometry (GC-MS) analysis and a targeted liquid chromatography-mass spectrometry (LC-MS) analysis of amine-containing metabolites. These data were correlated with changes in the expression of a number of target genes. Zinc exposure resulted in a reduction in levels of intermediates in carbohydrate metabolism (i.e., glucose 6-phosphate, fructose 6-phosphate and disaccharides) and an increase in a number of TCA cycle intermediates. Zinc exposure also resulted in decreases in concentrations of the amine containing metabolites, lanthionine, methionine and cystathionine, and an increase in metallothionein gene expression. Methionine and cystathionine are intermediates in the transsulfuration pathway which is involved in the conversion of methionine to cysteine. These responses provide an understanding of the pathways affected by zinc toxicity, and how these effects are different to other heavy metals such as cadmium and copper. The use of complementary metabolomics analytical approaches was particularly useful for understanding the effects of zinc exposure and importantly, identified a suite of candidate biomarkers of zinc exposure useful for the development of biomonitoring programs.

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.

Development of a passive sampler for Zinc(II) in urban pond waters using a polymer inclusion membrane.

The use of a polymer inclusion membrane (PIM) in a novel passive sampler to measure the time-weighted average concentration of Zn(II) in urban waters is described. The passive sampler consists of a compartment containing an acidic receiving solution, which is separated from the external source solution by a PIM consisting of 40 wt% di-2-(ethylhexyl) phosphoric acid as the extractant, and 60 wt% poly-(vinyl chloride) as the base polymer. Two laboratory passive sampling techniques were tested. One involved immersion of the passive sampler into a source solution ("dip-in" approach) for a predetermined period of time while in the other one the source solution was flown past the membrane of the sampler ("flow-through" approach). The latter approach was found to be more suitable for the calibration of the passive sampler under laboratory conditions. A successful application using the "dip-in" sampling approach in urban waters has been conducted for proof of concept.