Demonstrating removal credits for contaminants of emerging concern in recycled water through a reverse osmosis barrier-A predictive framework.

The performance of individual reverse osmosis (RO) systems varies significantly with different contaminants of emerging concern (CECs). As such, log reduction values (LRVs) of the concentration of these chemicals cannot be arbitrarily credited in water treatment and water recycling. This study looks to present an approach to the management of chemical risks by providing a systematic validation of RO barrier performance with respect to LRV credits for various classes of CECs. In this work, a one-off sampling campaign across five treatment barriers (strainer filtration, ultrafiltration, RO, ion exchange, chlorination) of a full-scale water recycling plant was conducted, followed by a systematic sampling campaign for a period of six weeks across just the RO barrier. The CECs screening methodology used GC-MS for quantification of 948 trace organic chemicals along with specific 44 per- and polyfluoroalkyl substances (PFAS) screening using LC-MS/MS to demonstrate the removal credits of the RO barrier to a wide spectrum of CECs. The work was used to validate an LRV barrier credit framework so as to predict the performance of a polyamide RO membrane for removal of a range of chemical classes, under typical operational conditions. Conductivity was validated as an efficient surrogate for membrane integrity and RO performance, along with specified operational conditions associated with permeate flux and recovery rate. A bioassay method (photobacterium test) showed good potential to be used as a quick measure to indicate the general toxicity of a sample caused by chemical contamination, because of its high detection sensitivity and time and cost efficiency.

Sub-ppm determination of perfluorinated carboxylic acids in solution by UV-vis high-performance liquid chromatography through solid phase extraction.

This study investigated a novel and sensitive analytical method based on a simple heat-based derivatization using 3-bromoacetyl coumarin as the reagent and analysis with a HPLC-UV system or just a UV-vis spectrometer to allow the sub-ppm determination of PFCAs in water solution with the potential for utilization in simple laboratories and field laboratory scenarios. A Strata-X-AW cartridge was used for the solid phase extraction (SPE) procedure and higher than 98% recoveries were obtained. The derivatization condition showed that a high efficiency of peak separation was obtained with obviously different retention time among various PFCAs derivatives using HPLC-UV analysis. The derivatization stability and repeatability showed favorable results with stable derivatized analytes for ≤12 h and a relative standard deviation (RSD) of <2% for all repetitions. The limit of detection for the HPLC-UV analysis was between 0.1 ppm and 0.5 ppm. A satisfactory linearity response was found with R2 >0.998 for all individual PFCA compounds. The limit of detection for simple UV-Vis analysis was <0.0003 ppm to measure the presence of PFCAs. Contamination of standards with humic substances and measurement of industrial samples in a complex wastewater matrix showed no adverse effects on the accuracy of PFCA determination by using the developed methodology.

A chemical credit framework to predict the removal performance of organic chemicals of concern from water through an ozonation process.

Ozonation is an effective barrier for removing a wide spectrum of organic Chemicals of Concern (CoC) in a water treatment process. In this study, bench- and full-scale tests were conducted on a secondary treated effluent at the Eastern Treatment Plant (ETP) of Melbourne Water to probe the performance of ozonation in removing CoC in a wastewater discharge. From the secondary treated effluent as the feed to the ozone process, 58 organic chemicals were measured out of a possible 949 compounds by using the AIQS-DB analytical method. A chemical credit framework for the ozonation process has been established according to the bench-scale results. Chemical classifications based on the chemical structures (aromatics, aliphatic and halogenated aliphatic compounds) and reaction rate constants with O3 (KO3) and the ∙OH radical (K∙OH) and a combined O3/TOC ratio and O3 CT value as operating parameters were confirmed to be useful and important in determining whether a chemical would be removed by ozone. It is shown that an O3/TOC ratio of >0.404 was shown to be necessary to overcome the instantaneous ozone demand (IOD) to guarantee enough ozone to oxidise CoC. For CoC with KO3 >105M - 1s - 1 and K∙OH >109M - 1s - 1, an O3/TOC ratio of ≥0.461 or a measurable O3 CT of ≥0.063 mg min/L can achieve log reduction values (LRVs) of ≥1, these are chemicals with aromatic structures; For CoC with low KO3 and high KOH, a combined O3/TOC ratio and O3 CT value inclusive of a chemical structure classification is indicated as necessary criteria to evaluate the removal. UV254 and TOC were demonstrated to be good online surrogates of ozone barrier performance in the absence of continuous O3/TOC ratio and O3 CT value measurements. Full-scale operational results confirm the effective predictions of the chemical credit framework, which highlights the necessity and importance of monitoring both the O3/TOC ratio and O3 CT values to predict the removal efficiency of a given chemical with a known response to O3 or a known chemical structure.

Transcriptome-wide changes associated with the reproductive behaviour of male guppies exposed to 17α-ethinyl estradiol.

Although many pharmaceutical compounds (and their metabolites) can induce harmful impacts at the molecular, physiological and behavioural levels, their underlying mechanistic associations have remained largely unexplored. Here, we utilized RNA-Seq to build a whole brain transcriptome profile to examine the impact of a common endocrine disrupting pharmaceutical (17α-ethinyl estradiol, EE2) on reproductive behaviour in wild guppies (Poecilia reticulata). Specifically, we annotated 16,791 coding transcripts in whole brain tissue in relation to the courtship behaviour (i.e. sigmoid display) of EE2 exposed (at environmentally relevant concentration of 8 ng/L for 28-days) and unexposed guppies. Further, we obtained 10,960 assembled transcripts matching in the non-coding orthologous genomes. Behavioural responses were assessed using a standard mate choice experiment, which allowed us to disentangle chemical cues from visual cues. We found that a high proportion of the RNAseq reads aligned back to our de novo assembled transcriptome with 80.59% mapping rate. Behavioural experiments showed that when males were presented only with female visual cues, there was a significant interaction between male treatment and female treatment in the time spent in the preference zone. This is one of the first studies to show that transcriptome-wide changes are associated with the reproductive behaviour of fish: EE2 exposed male guppies that performed high levels of courtship had a gene profile that deviated the most from the other treatment groups, while both non-courting EE2 and control males had similar gene signatures. Using Gene Ontology pathway analysis, our study shows that EE2-exposed males had gene transcripts enriched for pathways associated with altered immunity, starvation, altered metabolism and spermatogenesis. Our study demonstrates that multiple gene networks orchestrate courting behaviour, emphasizing the importance of investigating impacts of pharmaceuticals on gene networks instead of single genes.

Glyphosate and aminomethylphosphonic acid (AMPA) are commonly found in urban streams and wetlands of Melbourne, Australia.

Glyphosate is the most widely-used pesticide for weed control in agriculture and in urban and residential areas. This is the first study to quantify glyphosate and AMPA levels in surface water in Australia from different land uses. Glyphosate and AMPA was measured in surface water from 10 rural streams, 30 urban stormwater wetlands and 9 urban streams located in and around the city of Melbourne, Australia on five occasions between October 2017 and February 2018. Glyphosate and AMPA were present in most of the urban surface water samples. The frequency of detection of glyphosate was 77% in wetlands and 79% in urban streams, whereas it was only detected in 4% of the rural streams. Similarly, AMPA detection was 91% in wetlands and 97% in urban streams, whereas it was only present in 6% of the rural stream samples. In both urban streams and wetlands, the highest average glyphosate concentrations occurred in November (1.8 ±â€¯2.2 µg L-1). Overall, wetlands and streams associated with urban land use are vulnerable to glyphosate contamination. These results highlight the importance of screening for contaminants in urban stormwater to identify the source of pollutants that may end up in aquatic ecosystems and the risks therefrom.

Occurrence of perfluoroalkyl substances in selected Victorian rivers and estuaries: An historical snapshot.

This reconnaissance study was undertaken in 2012 to examine the occurrence of common perfluoroalkyl substances (PFAS), including perfluoroalkyl sulphonic acids and perfluoroalkyl carboxylic acids in rivers and estuaries in Port Philip Bay, Victoria, Australia. In total, 19 PFAS were screened in grab samples of water using a combination of solid phase extraction and liquid chromatography - mass spectrometry measurement techniques. Eighteen of the PFAS screened were observed in samples. The highest level of PFOS observed at a freshwater site was 0.045 µg/L; this concentration is approximately half the draft Australian 95% species protection level for total PFOS. The highest level of PFOA in the study (0.014 µg/L) was some four orders of magnitude lower than the draft Australian trigger value for PFOA (220 µg/L). However, none of the PFAS observed at the freshwater sites had research quotient (RQ) or toxicity unit (TU) values above 1 or -3, respectively. The highest concentration of PFOS observed at an estuarine site was 0.075 µg/L; the highest level of PFOA, 0.09 µg/L). There are no Australian marine water quality trigger values for PFAS, so potential risk was assessed using the European environment quality standards (EQS) adopted in EU Directive 2013/39/EU, RQ and TU methods. In that context, none of the PFAS observed at estuary sites had concentrations higher than the EU standards, or RQ above 1 or Log 10 TU above -3. Together these assessments suggest none of the PFAS screened would have posed an acute risk to organisms in the fresh or estuary waters studied at the time of sampling on an individual or collective basis. However, the detection of these PFAS in Victorian estuaries highlights that the issue is not just an issue for more densely populated countries in the northern hemisphere, but also potentially of concern in Australia. And, in that context, more sampling campaigns in Port Philip Bay are of paramount importance to assess the potential risk pose by these compounds to aquatic ecosystems.

A critical control point approach to the removal of chemicals of concern from water for reuse.

The reuse of water in a range of potable and non-potable applications is an important factor in the augmentation of water supply and in improving water security and productivity worldwide. A key hindrance to the reuse of water is the cost of compliance testing and process validation associated with ensuring that pathogen and chemicals in the feedwater are removed to a level that ensures no acute or chronic health and/or environmental effects. The critical control point (CCP) approach is well established and widely adopted by water utilities to provide an operational and risk management framework for the removal of pathogens in the treatment system. The application of a CCP approach to barriers in a treatment system for the removal of chemicals is presented. The application exemplar is to a small community wastewater treatment system that aims to produce potable quality water from a secondary treated wastewater effluent, however, the concepts presented are generic. The example used seven treatment barriers, five of which were designed and operated as CCP barriers for pathogens. The work demonstrates a method and risk management framework by which three of the seven barriers could also include a CCP approach for the removal of chemicals. Analogous to a CCP approach for pathogens, the potential is to reduce the use of chemical analysis as a routine determinant of performance criteria. The operational deployment of a CCP approach for chemicals was augmented with the development of a decision tree encompassing the classification of chemicals and the total removal credits across the treatment train in terms of the mechanistic removal of chemicals for each barrier. Validation of the approach is shown for an activated sludge, ozone and reverse osmosis barrier.

The endocrine disruptor, 17α-ethinyl estradiol, alters male mate choice in a freshwater fish.

Among the handful of studies on the behavioural effects of endocrine disrupting chemicals (EDCs), only a few have set out to disentangle the mechanisms underpinning behavioural changes. In fish, previous studies have shown that both visual and chemical cues play an important role in mate choice. As such, contaminant-induced changes in either transmission or perception of mate choice cues could have direct implications for individual's fitness. One widespread contaminant of environmental concern is 17α-ethinyl estradiol (EE2), a synthetic estrogen used in the contraceptive pill. Here, we investigated the impacts of EE2 exposure (28 days; measured concentration 14 ng/L) on visual and chemical communication in wild guppies (Poecilia reticulata). Using a standard dichotomous mate choice assay, we first gave individual males (either control or EE2-exposed) the opportunity to court two size-matched females (one control and one EE2-exposed) using only visual cues. We then introduced chemical cues of females (control and EE2-exposed) to the trial tank. We found that there was no significant effect of EE2-treatment on total time males spent associating with the females, when given only visual cues. There was, however, a significant effect on male courtship behaviour, with both control and EE2-exposed males spending more time performing 'sigmoid' displays towards the visual cues of control females compared to EE2-exposed females. When males were presented with both visual and chemical female cues simultaneously, we found that males spent more time courting control females that were paired with EE2-chemical cues. Not only does our study uncover a previously unknown behavioural impact of EE2-exposure on chemical cues, but demonstrates that EE2-exposure can exert complex effects on visual and chemical communication in a mate choice context. Finally, we contribute to the discussion of intraspecific variability by providing data on the potential trade-offs underpinning contaminant-induced behavioural changes.

Occurrence and assessment of the risk of ultraviolet filters and light stabilizers in Victorian estuaries.

This reconnaissance study was undertaken to examine the occurrence of common ultraviolet filters (UVF) and light stabilizers (UVLS), and preservatives in four different estuaries in Port Philip Bay, Victoria, for the first time. In total, 11 UV filters, 10 UV stabilizers, 12 preservatives and a metabolite, and one fragrance were screened in grab samples of water and sediment using a combination of solid phase extraction and gas and liquid chromatography mass spectrometry measurement techniques. In that context, 16 of the UVF and UVLS and 5 of the preservatives screened were observed in water and/or sediment samples. There are no marine water quality guideline values for any of the fragrances, preservatives and UV filters and light stabilizers in Australia's current national water quality guidelines, so potential risk was assessed using the risk quotient (RQ) and toxic unit (TU) concepts. In that context, only two chemicals (OC and EHMC) had both an RQ above 1 and a log10TU above - 3, suggesting that few of the screened chemicals would have posed an individual, short-term risk to organisms in the waters studied at the time of sampling. However, the detection of common UV filters, such as 4MBC, EHMC, OC and the common preservatives 2-PE, MP, and PB in these Victorian estuaries highlights that the existence of personal care products in the environment is not just an issue for more densley populated countries in the northern hemisphere, but also potentially of concern in Australia. And, in that context, more sampling campaigns in Port Philip Bay are of paramount importance to assess the potential risk posed by these compounds to aquatic ecosystems.

The agricultural contaminant 17ß-trenbolone disrupts male-male competition in the guppy (Poecilia reticulata).

Despite a growing literature highlighting the potential impact of human-induced environmental change on mechanisms of sexual selection, relatively little is known about the effects of chemical pollutants on male-male competition. One class of environmental pollutant likely to impact male competitive interactions is the endocrine-disrupting chemicals (EDCs), a large and heterogeneous group of chemical contaminants with the potential to influence morphology, physiology and behaviour at minute concentrations. One EDC of increasing concern is the synthetic, androgenic steroid 17ß-trenbolone, which is used globally to promote growth in beef cattle. Although 17ß-trenbolone has been found to cause severe morphological and behavioural abnormalities in fish, its potential impact on male-male competition has yet to be investigated. To address this, we exposed wild male guppies (Poecilia reticulata) to an environmentally realistic concentration of 17ß-trenbolone (average measured concentration: 8 ng/L) for 21 days using a flow-through system. We found that, in the presence of a competitor, 17ß-trenbolone-exposed males carried out more frequent aggressive behaviours towards rival males than did unexposed males, as well as performing less courting behaviour and more sneak (i.e., coercive) mating attempts towards females. Considering that, by influencing mating outcomes, male-male competition has important consequences for population dynamics and broader evolutionary processes, this study highlights the need for greater understanding of the potential impact of EDCs on the mechanisms of sexual selection.

Characterisation of the transcriptome of male and female wild-type guppy brains with RNA-Seq and consequences of exposure to the pharmaceutical pollutant, 17α-ethinyl estradiol.

Waterways are increasingly being contaminated by chemical compounds that can disrupt the endocrinology of organisms. One such compound is 17α-ethinyl estradiol (EE2), a synthetic estrogen used in the contraceptive pill. Despite considerable research interest in the effects of EE2 on reproduction and gene expression, surprisingly, only a few studies have capitalised on technologies, such as next-generation sequencing (NGS), to uncover the molecular pathways related to EE2 exposure. Accordingly, using high-throughput sequencing technologies, the aim of our study was to explore the effects of EE2 on brain transcriptome in wild-type male and female guppy (Poecilia reticulata). We conducted two sets of experiments, where fish were exposed to EE2 (measured concentrations: 8ng/L and 38ng/L) in a flow-through system for 21days. The effects on the brain transcriptome on both males and females were assessed using Illumina sequencing (MiSeq and HiSeq) platform followed by bioinformatics analysis (edgeR, DESeq2). Here, we report that exposure to EE2 caused both up- and downregulation of specific transcript abundances, and affected transcript abundance in a sex-specific manner. Specifically, we found 773 transcripts, of which 60 were male-specific, 61 female-specific and 285 treatment-specific. EE2 affected expression of 165 transcripts in males, with 88 downregulated and 77 upregulated, while in females, 120 transcripts were affected with 62 downregulated and 58 upregulated. Finally, RT-qPCR validation demonstrated that expression of transcripts related to transposable elements, neuroserpin and heat shock protein were significantly affected by EE2-exposure. Our study is the first to report brain transcriptome libraries for guppies exposed to EE2. Not only does our study provide a valuable resource, it offers insights into the mechanisms underlying the feminizing effects on the brains of organisms exposed to environmentally realistic concentrations of EE2.

Herbicides and trace metals in urban waters in Melbourne, Australia (2011-12): concentrations and potential impact.

Urban stormwater samples were collected from five aquatic systems in Melbourne, Australia, on six occasions between October 2011 and March 2012 and tested for 30 herbicides and 14 trace metals. Nineteen different herbicides were observed in one or more water samples from the five sites; chemicals observed at more than 40% of sites were simazine (100%), MCPA (83%), diuron (63%) and atrazine (53%). Using the toxicity unit (TU) concept to assess potential risk to aquatic ecosystems, none of the detected herbicides were considered to pose an individual, group or collective short-term risk to fish or zooplankton in the waters studied. However, 13 herbicides had TU values suggesting they might have posed an individual risk to primary producers at the time of sampling. Water quality guideline levels were exceeded on many occasions for Cd, Cu, Cr, Pb and Zn. Similarly, RQmed and RQmax exceeded 1 for Cd, Cr, Cu, Mn, Ni, Pb, V and Zn. Almost all the metals screened exceeded a log10TU of -3 for every trophic level, suggesting that there may have been some impact on aquatic organisms in the studied waterbodies. Our data indicate that Melbourne's urban aquatic environments may be being impacted by approved domestic, industrial and sporting application of herbicides and that stormwater quality needs to be carefully assessed prior to reuse. Further research is required to understand the performance of different urban stormwater wetland designs in removing pesticides and trace metals. Applying the precautionary principle to herbicide regulation is important to ensure there is more research and assessment of the long-term 'performance' standard of all herbicides and throughout their 'life cycle'. Implementing such an approach will also ensure government, regulators, decision makers, researchers, policy makers and industry have the best possible information available to improve the management of chemicals, from manufacture to use.

Exposure to an agricultural contaminant, 17ß-trenbolone, impairs female mate choice in a freshwater fish.

Despite the pivotal role sexual selection plays in population dynamics and broader evolutionary processes, the impact of chemical pollution on female mate choice is poorly understood. One group of chemical contaminants with the potential to disrupt the mechanisms of female mate choice is endocrine disrupting chemicals (EDCs); a broad class of environmental pollutants that can interfere with the endocrinology of organisms at extremely low concentrations. Recent research has revealed that estrogenic EDCs can affect female mate choice in fish, but the impact of androgenic EDC exposure is yet to be studied. To address this, we investigated the effects of an environmentally relevant concentration of trenbolone - an androgenic steroid used as a growth promoter in the cattle industry - on female mate choice in wild-caught guppies (Poecilia reticulata). We exposed male and female guppies to 17ß-trenbolone for 21 days (measured concentration 4ng/L) via a flow-through system, and found that trenbolone-exposed female guppies spent less time associating with males, and were less choosy, compared to unexposed females. In contrast, trenbolone had no impact on male reproductive behavior or morphology. This is the first study to show that androgenic EDC exposure can disrupt female mate choice, highlighting the need for studies to investigate the behavioral impacts of environmental contaminants on both sexes.

Pesticide and trace metals in surface waters and sediments of rivers entering the Corner Inlet Marine National Park, Victoria, Australia.

Water and sediment samples were collected from up to 17 sites in waterways entering the Corner Inlet Marine National Park monthly between November 2009 and April 2010, with the Chemcatcher passive sampler system deployed at these sites in November 2009 and March 2010. Trace metal concentrations were low, with none occurring at concentrations with the potential for adverse ecological effects. The agrochemical residues data showed the presence of a small number of pesticides at very low concentration (ng/L) in the surface waters of streams entering the Corner Inlet, and as widespread, but still limited contamination of sediments. Concentrations of pesticides detected were relatively low and several orders of magnitude below reported ecotoxicological effect and hazardous concentration values. The low levels of pesticides detected in this study indicate that agricultural industries were responsible agrochemical users. This research project is a rarity in aligning both agrochemical usage data obtained from chemical resellers in the target catchment with residue analysis of environmental samples. Based on frequency of detection and concentrations, prometryn is the priority chemical of concern for both the water and sediments studied, but this chemical was not listed in reseller data. Consequently, the risks may be greater than the field data would suggest, and priorities for monitoring different since some commonly used herbicides (such as glyphosate, phenoxy acid herbicides, and sulfonyl urea herbicides) were not screened. Therefore, researchers, academia, industry, and government need to identify ways to achieve a more coordinated land use approach for obtaining information on the use of chemicals in a catchment, their presence in waterways, and the longer term performance of chemicals, particularly where they are used multiple times in a year.

POPs monitoring in Australia and New Zealand using plastic resin pellets, and International Pellet Watch as a tool for education and raising public awareness on plastic debris and POPs.

Persistent organic pollutants (i.e. PCBs, DDTs, and HCHs) were analyzed along Australia and New Zealand North Island coastlines. PCB concentrations were high in urban areas (107-294 ng/g-pellet), with Sydney Harbour the most polluted. Hepta-chlorinated PCB was abundant, with ~30% in urban areas suggesting legacy pollution. DDT concentrations showed similar pattern except in rural agricultural sites, Taupo Bay and Ahipara, New Zealand (23 and 47 ng/g-pellet). p,p'-DDE predominance at these 2 sites suggested historical input; they also had high HCH concentrations (17 and 29 ng/g-pellet). The role of International Pellet Watch (IPW) in science communication was studied through feedbacks from IPW volunteers, case studies and examples. IPW data were categorized into understandable terms and tailored reports based on volunteers' backgrounds complemented with pollution maps. The effectiveness of IPW science communication has led to its use in awareness and education activities focusing on both POPs and plastic debris issues.

Combining Passive Sampling with Recombinant Receptor-Reporter Gene Bioassays to Assess the Receptor Activity of Victorian Rivers.

This pilot study was initiated to provide new information on the 'hormonal' activity of Victorian rivers. Chemcatcher™ passive sampler systems containing Empore™ C18FF disks were deployed at eight riverine sites near Melbourne. Little estrogenic activity [<0.4-1.8 ng estradiol equivalents (EQ)/disk] and no retinoic acid activity (RAR, all samples <0.8 ng trans-retinoic acid EQ/disk) was observed. Almost all sample extracts showed aryl hydrocarbon receptor activity (from <4 to 29 ng ß-naphthoflavone EQ/disk). Overall, the disk extracts were eminently compatible with the bioassay screening technology, enabling the relative levels of 'hormonal activity' to be observed in the surface waters in and around Melbourne. From a practical perspective, the in situ sampling and pre-concentration provided by passive sampling reduces the manual handling risks associated with sample transport, and the number of laboratory operations required to obtain assay-ready solutions for analysis.

Sex in troubled waters: Widespread agricultural contaminant disrupts reproductive behaviour in fish.

Chemical pollution is a pervasive and insidious agent of environmental change. One class of chemical pollutant threatening ecosystems globally is the endocrine disrupting chemicals (EDCs). The capacity of EDCs to disrupt development and reproduction is well established, but their effects on behaviour have received far less attention. Here, we investigate the impact of a widespread androgenic EDC on reproductive behaviour in the guppy, Poecilia reticulata. We found that short-term exposure of male guppies to an environmentally relevant concentration of 17ß-trenbolone-a common environmental pollutant associated with livestock production-influenced the amount of male courtship and forced copulatory behaviour (sneaking) performed toward females, as well as the receptivity of females toward exposed males. Exposure to 17ß-trenbolone was also associated with greater male mass. However, no effect of female exposure to 17ß-trenbolone was detected on female reproductive behaviour, indicating sex-specific vulnerability at this dosage. Our study is the first to show altered male reproductive behaviour following exposure to an environmentally realistic concentration of 17ß-trenbolone, demonstrating the possibility of widespread disruption of mating systems of aquatic organisms by common agricultural contaminants.

Pesticide and trace metal occurrence and aquatic benchmark exceedances in surface waters and sediments of urban wetlands and retention ponds in Melbourne, Australia.

Samples of water and sediments were collected from 24 urban wetlands in Melbourne, Australia, in April 2010, and tested for more than 90 pesticides using a range of gas chromatographic (GC) and liquid chromatographic (LC) techniques, sample 'hormonal' activity using yeast-based recombinant receptor-reporter gene bioassays, and trace metals using spectroscopic techniques. At the time of sampling, there was almost no estrogenic activity in the water column. Twenty-three different pesticide residues were observed in one or more water samples from the 24 wetlands; chemicals observed at more than 40% of sites were simazine (100%), atrazine (79%), and metalaxyl and terbutryn (46%). Using the toxicity unit (TU) concept, less than 15% of the detected pesticides were considered to pose an individual, short-term risk to fish or zooplankton in the ponds and wetlands. However, one pesticide (fenvalerate) may have posed a possible short-term risk to fish (log10TUf > -3), and three pesticides (azoxystrobin, fenamiphos and fenvalerate) may have posed a risk to zooplankton (logTUzp between -2 and -3); all the photosystem II (PSII) inhibiting herbicides may have posed a risk to primary producers in the ponds and wetlands (log10TUap and/or log10TUalg > -3). The wetland sediments were contaminated with 16 different pesticides; no chemicals were observed at more than one third of sites, but based on frequency of detection and concentrations, bifenthrin (33%, maximum 59 µg/kg) is the priority insecticide of concern for the sediments studied. Five sites returned a TU greater than the possible effect threshold (i.e. log10TU > 1) as a result of bifenthrin contamination of their sediments. Most sediments did not exceed Australian sediment quality guideline levels for trace metals. However, more than half of the sites had threshold effect concentration quotients (TECQ) values >1 for Cu (58%), Pb (50%), Ni (67%) and Zn (63%), and 75% of sites had mean probable effect concentration quotients (PECQ) >0.2, suggesting that the collected sediments may have been having some impact on sediment-dwelling organisms.

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.

Altered reproductive behaviours in male mosquitofish living downstream from a sewage treatment plant.

Freshwater environments are common repositories for the discharge of large volumes of domestic and industrial waste, particularly through wastewater effluent. One common group of chemical pollutants present in wastewater are endocrine disrupting chemicals (EDCs), which can induce morphological and behavioural changes in aquatic organisms. The aim of this study was to compare the reproductive behaviour and morphology of a freshwater fish, the mosquitofish (Gambusia holbrooki), collected from two sites (wastewater treatment plant (WWTP) and a putative pristine site). The mosquitofish is a sexually dimorphic livebearer with a coercive mating system. Males inseminate females using their modified anal fin as an intromittent organ. Despite this, females are able to exert some control over the success of male mating attempts by selectively associating with, or avoiding, certain males over others. Using standard laboratory assays of reproductive behaviour, we found that mosquitofish males living in close proximity to WWTP showed increased mating activity compared to those inhabiting a pristine site. More specifically, during behavioural trials in which males were allowed to interact with females separated by a transparent divider, we found that WWTP-males spent more time associating with females. Concordant with this, when males and females were subsequently allowed to interact freely, WWTP-males also spent more time chasing and orienting towards the females. As a result, females from both sites showed more interest towards the WWTP-site males. Male anal fin morphology, however, did not differ between sites. Our study illustrates that lifetime exposure to WWTP-effluents can greatly affect male behaviour. The results underscore the importance of behaviour as a potential tool for investigating unknown contaminants in the environment.