Frederic D.L. Leusch

Benchmarking Organic Micropollutants in Wastewater, Recycled Water and Drinking Water with In Vitro Bioassays

Thousands of organic micropollutants and their transformation products occur in water. Although often present at low concentrations, individual compounds contribute to mixture effects. Cell-based bioassays that target health-relevant biological endpoints may therefore complement chemical analysis for water quality assessment. The objective of this study was to evaluate cell-based bioassays for their suitability to benchmark water quality and to assess efficacy of water treatment processes. The selected bioassays cover relevant steps in the toxicity pathways including induction of xenobiotic metabolism, specific and reactive modes of toxic action, activation of adaptive stress response pathways and system responses. Twenty laboratories applied 103 unique in vitro bioassays to a common set of 10 water samples collected in Australia, including wastewater treatment plant effluent, two types of recycled water (reverse osmosis and ozonation/activated carbon filtration), stormwater, surface water, and drinking water. Sixty-five bioassays (63%) showed positive results in at least one sample, typically in wastewater treatment plant effluent, and only five (5%) were positive in the control (ultrapure water). Each water type had a characteristic bioanalytical profile with particular groups of toxicity pathways either consistently responsive or not responsive across test systems. The most responsive health-relevant endpoints were related to xenobiotic metabolism (pregnane X and aryl hydrocarbon receptors), hormone-mediated modes of action (mainly related to the estrogen, glucocorticoid, and antiandrogen activities), reactive modes of action (genotoxicity) and adaptive stress response pathway (oxidative stress response). This study has demonstrated that selected cell-based bioassays are suitable to benchmark water quality and it is recommended to use a purpose-tailored panel of bioassays for routine monitoring.

Comparison of five in vitro bioassays to measure estrogenic activity in environmental waters

Bioassays are well established in the pharmaceutical industry and single compound analysis, but there is still uncertainty about their usefulness in environmental monitoring. We compared the responses of five bioassays designed to measure estrogenic activity (the yeast estrogen screen, ER-CALUX, MELN, T47D-KBluc, and E-SCREEN assays) and chemical analysis on extracts from four different water sources (groundwater, raw sewage, treated sewage, and river water). All five bioassays displayed similar trends and there was good agreement with analytical chemistry results. The data from the ER-CALUX and E-SCREEN bioassays were robust and predictable, and well-correlated with predictions from chemical analysis. The T47D-KBluc appeared likewise promising, but with a more limited sample size it was less compelling. The YES assay was less sensitive than the other assays by an order of magnitude, which resulted in a larger number of nondetects. The MELN assay was less predictable, although the possibility that this was due to laboratory-specific difficulties cannot be discounted. With standardized bioassay data analysis and consistency of operating protocols, bioanalytical tools are a promising advance in the development of a tiered approach to environmental water quality monitoring.

Bioanalytical tools in water quality assessment.

Bioanalytical Tools in Water Quality Assessment reviews the application of bioanalytical tools for assessment of water quality including surveillance monitoring. The types of water included range from wastewater to drinking water, including recycled water, as well as treatment processes and advanced water treatment. Bioanalytical Tools in Water Quality Assessment not only demonstrates applications but also fills in the background knowledge in toxicology/ecotoxicology needed to appreciate these applications. Each chapter summarises fundamental material in a targeted way so that information can be applied to better understand the use of bioanalytical tools in water quality assessment. Bioanalytical tools in Water Quality Assessment can be used by lecturers teaching academic and professional courses and also by risk assessors, regulators, experts, consultants, researchers and managers working in the water sector. It can also be a reference manual for environmental engineers, analytical chemists, and toxicologists. ISBN: 9781843393689 (Print) ISBN: 9781780400778 (eBook)

Effects of a phytosterol mixture on male fish plasma lipoprotein fractions and testis P450scc activity

Plant sterols (phytosterols) have been identified as one potential source of reproductive effects in fish living downstream of pulp mills. beta-Sitosterol, the predominant plant sterol in pulp mill effluent, has previously been shown to decrease plasma sex steroid and cholesterol levels and in vitro gonadal steroid production in fish. In this study, male brook trout (Salvelinus fontinalis) and goldfish (Carassius auratus) were exposed to a phytosterol mixture (72% beta-sitosterol) via Silastic intraperitoneal implants to help elucidate the mechanisms of action of phytosterols on steroid depression. As cholesterol is exogenously supplied for gonadal steroidogenesis, changes in plasma cholesterol fractions were examined. In male brook trout, low-density lipoprotein cholesterol and triglyceride levels decreased significantly, 43 and 50%, respectively, in phytosterol-treated fish. It is improbable, however, that these decreases are linked to depressed gonadal steroidogenesis in fish. The activity of P450scc, which converts cholesterol to pregnenolone (the first step in the steroidogenic pathway), was not affected in testis mitochondria isolated from brook trout or goldfish. Further investigation of the mechanisms of action of phytosterols is required.

Removal of trace organic contaminants by an MBR comprising a mixed culture of bacteria and white-rot fungi

The degradation of 30 trace organic contaminants (TrOC) by a white-rot fungus-augmented membrane bioreactor (MBR) was investigated. The results show that white-rot fungal enzyme (laccase), coupled with a redox mediator (1-hydroxy benzotriazole, HBT), could degrade TrOC that are resistant to bacterial degradation (e.g. diclofenac, triclosan, naproxen and atrazine) but achieved low removal of compounds (e.g. ibuprofen, gemfibrozil and amitriptyline) that are well removed by conventional activated sludge treatment. Overall, the fungus-augmented MBR showed better TrOC removal compared to a system containing conventional activated sludge. The major role of biodegradation in removal by the MBR was noted. Continuous mediator dosing to MBR may potentially enhance its performance, although not as effectively as for mediator-enhanced batch laccase systems. A ToxScreen3 assay revealed no significant increase in the toxicity of the effluent during MBR treatment of the synthetic wastewater comprising TrOC, confirming that no toxic by-products were produced.

In vivo implants of β-sitosterol cause reductions of reactive cholesterol pools in mitochondria isolated from gonads of male goldfish (Carassius auratus)

beta-Sitosterol, a phytosterol found in high concentrations in pulp mill effluents, has been proposed as one of the causative agents for steroid depressions observed in fish exposed to pulp mill effluents. Previous studies have suggested a cholesterol-mediated mechanism; however, it is unknown how beta-sitosterol depresses gonadal steroidogenesis. In this study, adult male goldfish (Carassius auratus) were exposed for 24-31 days to beta-sitosterol (55% of a phytosterol mixture or 96% pure; 150 microg/g; Silastic implant) after which gonadal mitochondria were isolated. Pregnenolone production, an indicator of the size of the pool of reactive cholesterol, was then measured in the isolated mitochondria. Sterol exposure did not affect P450 side-chain cleavage enzyme (converts cholesterol to pregnenolone) activity but did decrease the size of the mitochondrial pool of reactive cholesterol, suggesting beta-sitosterol is impeding cholesterol transfer across the mitochondrial membrane. This finding is supported by the observation that 25-hydroxycholesterol, which passes through mitochondrial membranes without need for a membrane transporter, restores beta-sitosterol-induced reductions in pregnenolone production.

Assessing granular media filtration for the removal of chemical contaminants from wastewater

Granular media filtration was evaluated for the removal of a suite of chemical contaminants that can be found in wastewater. Laboratory- and pilot-scale sand and granular activated carbon (GAC) filters were trialled for their ability to remove atrazine, estrone (E1), 17α-ethynylestradiol (EE2), N-nitrosodimethylamine (NDMA), N-nitrosomorpholine (NMOR) and N-nitrosodiethylamine (NDEA). In general, sand filtration was ineffective in removing the contaminants from a tertiary treated wastewater, with the exception of E1 and EE2, where efficient removals were observed after approximately 150 d. Batch degradation experiments confirmed that the removal of E1 was through biological activity, with a pseudo-first-order degradation rate constant of 7.4 × 10(-3) h(-1). GAC filtration was initially able to effectively remove all contaminants; although removals decreased over time due to competition with other organics present in the water. The only exception was atrazine where removal remained consistently high throughout the experiment. Previously unreported differences were observed in the adsorption of the three nitrosamines, with the ease of removal following the trend, NDEA > NMOR > NDMA, consistent with their hydrophobic character. In most instances the removals from the pilot-scale filters were generally in agreement with the laboratory-scale filter, suggesting that there is potential in using laboratory-scale filters as monitoring tools to evaluate the performance of pilot- and possibly full-scale sand and GAC filters at wastewater treatment plants.

The effects of mediator and granular activated carbon addition on degradation of trace organic contaminants by an enzymatic membrane reactor

The removal of four recalcitrant trace organic contaminants (TrOCs), namely carbamazepine, diclofenac, sulfamethoxazole and atrazine by laccase in an enzymatic membrane reactor (EMR) was studied. Laccases are not effective for degrading non-phenolic compounds; nevertheless, 22-55% removal of these four TrOCs was achieved by the laccase EMR. Addition of the redox-mediator syringaldehyde (SA) to the EMR resulted in a notable dose-dependent improvement (15-45%) of TrOC removal affected by inherent TrOC properties and loading rates. However, SA addition resulted in a concomitant increase in the toxicity of the treated effluent. A further 14-25% improvement in aqueous phase removal of the TrOCs was consistently observed following a one-off dosing of 3g/L granular activated carbon (GAC). Mass balance analysis reveals that this improvement was not due solely to adsorption but also enhanced biodegradation. GAC addition also reduced membrane fouling and the SA-induced toxicity of the effluent.

Extreme weather events: Should drinking water quality management systems adapt to changing risk profiles?

Among the most widely predicted and accepted consequences of global climate change are increases in both the frequency and severity of a variety of extreme weather events. Such weather events include heavy rainfall and floods, cyclones, droughts, heatwaves, extreme cold, and wildfires, each of which can potentially impact drinking water quality by affecting water catchments, storage reservoirs, the performance of water treatment processes or the integrity of distribution systems. Drinking water guidelines, such as the Australian Drinking Water Guidelines and the World Health Organization Guidelines for Drinking-water Quality, provide guidance for the safe management of drinking water. These documents present principles and strategies for managing risks that may be posed to drinking water quality. While these principles and strategies are applicable to all types of water quality risks, very little specific attention has been paid to the management of extreme weather events. We present a review of recent literature on water quality impacts of extreme weather events and consider practical opportunities for improved guidance for water managers. We conclude that there is a case for an enhanced focus on the management of water quality impacts from extreme weather events in future revisions of water quality guidance documents.

In vitro bioassays to evaluate complex chemical mixtures in recycled water.

With burgeoning population and diminishing availability of freshwater resources, the world continues to expand the use of alternative water resources for drinking, and the quality of these sources has been a great concern for the public as well as public health professionals. In vitro bioassays are increasingly being used to enable rapid, relatively inexpensive toxicity screening that can be used in conjunction with analytical chemistry data to evaluate water quality and the effectiveness of water treatment. In this study, a comprehensive bioassay battery consisting of 36 bioassays covering 18 biological endpoints was applied to screen the bioactivity of waters of varying qualities with parallel treatments. Samples include wastewater effluent, ultraviolet light (UV) and/or ozone advanced oxidation processed (AOP) recycled water, and infiltrated recycled groundwater. Based on assay sensitivity and detection frequency in the samples, several endpoints were highlighted in the battery, including assays for genotoxicity, mutagenicity, estrogenic activity, glucocorticoid activity, arylhydrocarbon receptor activity, oxidative stress response, and cytotoxicity. Attenuation of bioactivity was found to be dependent on the treatment process and bioassay endpoint. For instance, ozone technology significantly removed oxidative stress activity, while UV based technologies were most efficient for the attenuation of glucocorticoid activity. Chlorination partially attenuated genotoxicity and greatly decreased herbicidal activity, while groundwater infiltration efficiently attenuated most of the evaluated bioactivity with the exception of genotoxicity. In some cases, bioactivity (e.g., mutagenicity, genotoxicity, and arylhydrocarbon receptor) increased following water treatment, indicating that transformation products of water treatment may be a concern. Furthermore, several types of bioassays with the same endpoint were compared in this study, which could help guide the selection of optimized methods in future studies. Overall, this research indicates that a battery of bioassays can be used to support decision-making on the application of advanced water treatment processes for removal of bioactivity.