ECHIDNA (Emerging CHemIcals Database for National Awareness): a framework to prioritise contaminants of emerging concern in water.

The widespread presence of contaminants of emerging concern (CEC) in surface waters, treated wastewater and drinking water is an ongoing issue for the water industry. The absence of regulatory guidance and limited occurrence, toxicity and removal data are defining criteria of CEC and make it difficult to prioritise which CEC pose the greatest risk. The online Emerging CHemIcals Database for National Awareness (ECHIDNA) aims to classify and prioritise CEC based on their potential risk, with the information presented in an easily accessible and intuitive manner. A candidate list of almost 1,800 potential CEC, including pesticides, pharmaceuticals and industrial compounds, was compiled using both Australian and international resources. These were ranked based on in silico assessment of their persistent, bioaccumulative and toxic (PBT) properties, as well as potential chronic toxicity hazard, yielding 247 CEC for further prioritisation. Risk Quotients (RQ) identified between 5 and 87 CEC posing a risk to human and ecosystem health, respectively, across drinking water, surface water, treated wastewater and raw wastewater. While the ability of the water industry to effectively prioritise CEC is limited by candidate identification and data availability, ECHIDNA can provide valuable information for better decision-making surrounding CEC management.

Functional and hepatic metabolite changes in aquatic turtle hatchlings exposed to the anti-androgenic fungicide vinclozolin.

Many man-made chemicals that are released into water bodies in agricultural landscapes have been identified as endocrine disruptors and can cause serious impacts on the growth and survival of aquatic species living in these environments. However, very little attention has been paid to their toxicological effects in cultured non-fish species, such as aquatic turtles. We exposed hatchlings of the Chinese soft-shelled turtle (Pelodiscus sinensis) to different concentrations of vinclozolin (0, 5, 50 and 500 µg/L) for 60 days to assess physiological and metabolic impacts of this fungicide. Despite no death occurrence, hatchling turtles exposed to the highest concentration of vinclozolin consumed less food, grew more slowly (resulting in smaller body size after exposure) and performed more poorly in behavioral swimming tests than controls and turtles exposed to lower concentrations. Hepatic metabolite profiles acquired via liquid chromatography-mass spectrometry (LC-MS) revealed multiple metabolic perturbations related to amino acid, lipid, and fatty acid metabolism in animals exposed to environmentally relevant concentrations. Specifically, many critical metabolites involved in energy-related metabolic pathways (such as some intermediates in the tricarboxylic acid cycle, lactate, and some amino acids) were present in livers of hatchling turtles exposed vinclozolin, though at lower concentrations, reflecting energy metabolism dysregulation induced by exposure to this fungicide. Overall, our results suggest that the changes in growth and behavioral performances caused by chronic vinclozolin exposure may be associated with internal physiological and metabolic disorders mediated at the biochemical level.

The Role of Behavioral Ecotoxicology in Environmental Protection.

For decades, we have known that chemicals affect human and wildlife behavior. Moreover, due to recent technological and computational advances, scientists are now increasingly aware that a wide variety of contaminants and other environmental stressors adversely affect organismal behavior and subsequent ecological outcomes in terrestrial and aquatic ecosystems. There is also a groundswell of concern that regulatory ecotoxicology does not adequately consider behavior, primarily due to a lack of standardized toxicity methods. This has, in turn, led to the exclusion of many behavioral ecotoxicology studies from chemical risk assessments. To improve understanding of the challenges and opportunities for behavioral ecotoxicology within regulatory toxicology/risk assessment, a unique workshop with international representatives from the fields of behavioral ecology, ecotoxicology, regulatory (eco)toxicology, neurotoxicology, test standardization, and risk assessment resulted in the formation of consensus perspectives and recommendations, which promise to serve as a roadmap to advance interfaces among the basic and translational sciences, and regulatory practices.

Sex-Specific Differences in Lysine, 3-Hydroxybutyric Acid and Acetic Acid in Offspring Exposed to Maternal and Postnatal High Linoleic Acid Diet, Independent of Diet.

BACKGROUND: Linoleic acid (LA) is an essential polyunsaturated fatty acid (PUFA) that is required for foetal growth and development. Excess intake of LA can be detrimental for metabolic health due to its pro-inflammatory properties; however, the effect of a diet high in LA on offspring metabolites is unknown. In this study, we aimed to determine the role of maternal or postnatal high linoleic acid (HLA) diet on plasma metabolites in adult offspring. METHODS: Female Wistar Kyoto (WKY) rats were fed with either low LA (LLA) or HLA diet for 10 weeks prior to conception and during gestation/lactation. Offspring were weaned at postnatal day 25 (PN25), treated with either LLA or HLA diets and sacrificed at PN180. Metabolite analysis was performed in plasma samples using Nuclear Magnetic Resonance. RESULTS: Maternal and postnatal HLA diet did not alter plasma metabolites in male and female adult offspring. There was no specific clustering among different treatment groups as demonstrated by principal component analysis. Interestingly, there was clustering among male and female offspring independent of maternal and postnatal dietary intervention. Lysine was higher in female offspring, while 3-hydroxybutyric acid and acetic acid were significantly higher in male offspring. CONCLUSION: In summary, maternal or postnatal HLA diet did not alter the plasma metabolites in the adult rat offspring; however, differences in metabolites between male and female offspring occurred independently of dietary intervention.

Bioaccumulation and Biodistribution of Selenium in Metamorphosing Tadpoles.

Selenium is an important macronutrient with a very narrow margin between essentiality and toxicity. Amphibians are hypothesized to be particularly sensitive due to the potential for metamorphosis-driven mobilization, which could transfer or concentrate contaminant burdens within specific organs. We explored the potential role of tissue degeneration and remodeling during anuran metamorphosis as a mechanism for altering tissue-specific Se burdens. Limnodynastes peronii tadpoles were exposed to dissolved 75Se (as selenite) for 7 days and depurated until completion of metamorphosis. Bioaccumulation and retention kinetics were assessed in whole tadpoles and excised tissues using gamma spectroscopy, and temporal changes in biodistribution were assessed using autoradiography. Tadpoles retained Se throughout metamorphosis, and partitioned the element predominantly within digestive and excretory tissues, including livers > mesonephros > guts > gallbladder. Importantly, our results demonstrate that Se biodistribution varies significantly throughout development. This is indicative of tissue transference, and particularly in tissues developing de novo after depuration. To the best of our knowledge, this is the first study demonstrating Se transference during metamorphic tissue remodelling. Further research is warranted to explore the fate and metabolism of Se (and other metal and metalloids) during anuran development and the implications of transference for influencing toxicity.

Uptake and tissue distributions of cadmium, selenium and zinc in striped marsh frog tadpoles exposed during early post-embryonic development.

Metals and metalloids released through anthropogenic activities can accumulate in aquatic organisms, resulting in adverse effects in sensitive species. We investigated the influence of feeding regime and exposure complexity (i.e., mixture) on bioaccumulation kinetics and body distribution of common metal(loid) pollutants in Limnodynastes peronii during early post-embryonic development. Tadpoles were exposed to radiolabelled 109Cd, 75Se and 65Zn alone and in a mixture for 4 days, followed by 3 days depuration in clean water. One group was fed directly in exposure aquaria, whereas a second group was transferred to clean water for feeding, to investigate the potential influence of sorption to food on uptake. Bioconcentration factor and retention was observed to be greatest for Se. Results demonstrate that tadpoles accumulated and retained half the amount of Cd when exposed in mixture, suggesting that Se and/or Zn may have antagonistic effects against Cd uptake. Additionally, tadpoles fed directly in exposure water accumulated 2-3-times more Cd and Zn compared to tadpoles fed in clean water, indicating that the presence of food particles is an important factor that may influence uptake. Interestingly, this had a negligible impact on Se uptake. The study reveals how exposure conditions can influence the bioaccumulation of metal(loid)s, highlighting experimental factors as important considerations for both controlled toxicity experiments and for understanding exposure risks for amphibian populations.

Individual and mixture toxicity of pharmaceuticals naproxen, carbamazepine, and sulfamethoxazole to Australian striped marsh frog tadpoles (Limnodynastes peronii).

Nonsteroidal human pharmaceuticals are prevalent in domestic wastewater and may find their way into the environment at low concentrations. Since most pharmaceuticals are designed to be biologically active at low concentrations, there is a risk that these compounds may affect aquatic wildlife. Of particular concern is the occurrence of pharmaceutical mixtures, which may lead to increased adverse effects compared to individual compounds. Interactive effects were previously demonstrated for amphibians exposed to pesticide mixtures, but no such studies investigating responses of amphibians to pharmaceutical mixtures are apparently available. Results demonstrated increased toxicity (loss of tactile response) of striped marsh frog (Limnodynastes peronii) tadpoles exposed to a mixture of naproxen, carbamazepine, and sulfamethoxazole, compared to exposures to the individual compounds. Significant time × treatment interactions were observed for tadpole development following chronic exposures to 10 or 100 µg/L of each compound and the mixture; however, responses were weak and main treatment effects were not significant. Despite minor effects at low exposure concentrations, results demonstrated a potential for mixtures of nonsteroidal pharmaceuticals commonly occurring in wastewater to influence amphibian development. With the vast numbers of pharmaceuticals that exist and are found in the environment, this work highlights a need for further research into mixtures of pharmaceutically active wastewater contaminants. Further, since pharmaceuticals exert extremely varied biological actions, it is suggested that future investigations would benefit from inclusion of endpoints that are indicative of physiological or metabolic performance, as well as assessment of sensitive behavioral responses.

Metabolomic response of striped marsh frog (Limnodynastes peronii) tadpoles exposed to the fire retardant Phos-Chek LC95W.

Climate change and other factors have contributed to an increased frequency and intensity of global wildfires in recent years. Ammonium-based fire retardants are widely used to suppress or delay the spread of fire and have generally been regarded as presenting a low risk of acute toxicity to fauna. However, studies have raised concerns about their potential to cause indirect or sub-lethal effects, and toxicity information regarding the potential for such impacts in aquatic species is limited. To address these knowledge gaps, we used an untargeted metabolomics approach to evaluate the sub-lethal physiological and metabolic responses of striped marsh frog (Limnodynastes peronii) tadpoles exposed to a concentration gradient of the ammonium polyphosphate (APP)-based fire retardant Phos-Chek LC95W (PC). Acute exposure (96 h) to PC significantly altered the relative abundance of 14 metabolites in whole tadpoles. The overall metabolic response pattern was consistent with effects reported for ammonia toxicity and also suggestive of energy dysregulation and osmotic stress associated with alterations to physicochemical water quality parameters in the PC treatments. Results suggest that run-off or accidental application of this formulation into waterways can have significant sub-lethal consequences on the biochemical profiles (i.e., the metabolome) of aquatic organisms and may be a concern for frog species that breed and develop in small, often ephemeral, waterbodies. Our study highlights the benefits of integrating untargeted metabolomics with other ecological and toxicological endpoints to provide a more holistic characterisation of the sub-lethal impacts associated with bushfire-fighting chemicals and with environmental contaminants more broadly.

Application of in vitro bioassays to monitor pharmaceuticals in water: A synthesis of chronological analysis, mode of action, and practical insights.

Pharmaceutical compounds in wastewater have emerged as a significant concern for the aquatic environment. The use of in vitro bioassays represents a sustainable and cost-effective approach for assessing the potential toxicological risks of these biologically active compounds in wastewater and aligns with ethical considerations in research. It facilitates high-throughput analysis, captures mixture effects, integrates impacts of both known and unknown chemicals, and reduces reliance on animal testing. The core aim of the current review was to explore the practical application of in vitro bioassays in evaluating the environmental impacts of pharmaceuticals in wastewater. This comprehensive review strives to achieve several key objectives. First, it provides a summary categorisation of pharmaceuticals based on their mode of action, providing a structured framework for understanding their ecological significance. Second, a chronological analysis of pharmaceutical research aims to document their prevalence and trends over time, shedding light on evolving environmental challenges. Third, the review critically analyses existing bioassay applications in wastewater, while also examining bioassay coverage of representative compounds within major pharmaceutical classes. Finally, it explores the potential for developing innovative bioassays tailored for water quality monitoring of pharmaceuticals, paving the way for more robust environmental monitoring and risk assessment. Overall, adopting effect-based methods for pharmaceutical monitoring in water holds significant promise. It encompasses a broad spectrum of biological impacts, promotes standardized protocols, and supports a bioassay test battery approach indicative of different endpoints, thereby enhancing the effectiveness of environmental risk assessment.

Polyethylene microplastics induced lipidomic responses in Chironomus tepperi: A two-generational exploration.

Microplastics (MPs) accumulating in freshwater sediment have raised concerns about potential risks to benthic dwelling organisms, yet few studies have examined the long-term impacts caused by MP exposure. This study investigated alterations to lipid profiles in an Australian freshwater invertebrate, Chironomus tepperi, induced by polyethylene MP fragments (1-45 µm) at environmentally relevant concentrations (125, 250, 500 and 1000 MPs/kg sediment), using a two-generational experimental design. In the parental generation, the relative abundance of triacylglycerols, total fatty acids and unsaturated fatty acids exhibited apparent hormetic patterns, with low-concentration stimulation and high-concentration inhibition observed. The overall trend in these lipid classes is consistent with previously observed changes to polar metabolite profiles, indicating that ingestion of MPs could inhibit nutrient assimilation from food leading to disruption of energy availability. In the first filial generation continuously exposed to MPs, however, abundance of cholesterol and total fatty acids increased with increasing exposure concentrations, suggesting different effects on energy metabolism between the parental generation and offspring. No differences in the lipidome were observed in first filial larvae that were not exposed, implying that MPs pose negligible carry-over effects. Overall, the combined results of this study together with a preceding metabolomics study provide evidence of a physical effect of MPs with subsequent impacts to bioenergetics. Nevertheless, future research is required to explore the potential long-term impacts caused by MPs, and to unravel the impacts of the surfactant control as a potential contributor to the observed hormetic response, particularly for studies exploring sub-lethal effects of MP exposure using sensitive omics techniques.

Minor metabolomic disturbances induced by glyphosate-isopropylammonium exposure at environmentally relevant concentrations in an aquatic turtle, Pelodiscus sinensis.

The ecotoxicological and environmental impacts of glyphosate-based herbicides have received considerable attention due to their extensive use globally. However, the potential for adverse effects in cultured non-fish vertebrate species are commonly ignored. In this study, effects on growth, indicators of functional performance, gut microbial diversity, liver antioxidant responses and metabolite profiles were evaluated in soft-shelled turtle hatchlings (Pelodiscus sinensis) exposed to different concentrations of glyphosate-isopropylammonium (0, 0.02, 0.2, 2 and 20 mg/L). No significant changes in growth or functional performance (food intake, swimming speed), gut microbiota, and liver antioxidant responses (SOD and CAT activities, MDA content) were observed in exposed turtles. However, hepatic metabolite profiles revealed distinct perturbations that primarily involved amino acid metabolism in turtles exposed to environmentally relevant concentrations. Overall, our results suggested that metabolite profiles may be more sensitive than phenotypic or general physiological endpoints and gut microbiota profiling, and indicate a potential mechanism of hepatotoxicity caused by glyphosate-isopropylammonium based on untargeted metabolomics analysis. Furthermore, the toxicity of glyphosate at environmentally relevant concentrations might be relatively minor in aquatic turtle species.

Influence of chemical dose and exposure duration on protein synthesis in green sea turtle primary cells.

Understanding the impacts of chemical exposure in marine wildlife is challenging, due to practical and ethical constraints that preclude traditional toxicology research on these animals. This study addressed some of these limitations by presenting an ethical and high throughput cell-based approach to elucidate molecular-level effects of contaminants on sea turtles. The experimental design addressed basic questions of cell-based toxicology, including chemical dose and exposure time. Primary green turtle skin cells were exposed to polychlorinated biphenyl (PCB) 153 and perfluorononanoic acid (PFNA) for 24 and 48 h, at three sub-lethal, environmentally relevant concentrations (1, 10 and 100 µg/L). Sequential window acquisition of all theoretical mass spectra (SWATH-MS) identified over 1000 differentially abundant proteins within the 1% false discovery rate (FDR) threshold. The 24 h exposure resulted in a greater number of differentially abundant proteins, compared to 48 h exposure, for both contaminants. However, there were no statistically significant dose-response relationships for the number of differentially synthesised proteins, nor differences in the proportion of increased vs decreased proteins between or within exposure times. Known in vivo markers of contaminant exposure, superoxide dismutase and glutathione S-transferase, were differentially abundant following exposure to PCB153 and PFNA. SIGNIFICANCE: Cell-based (in vitro) proteomics provides an ethical and high throughput approach to understanding the impacts of chemical contamination on sea turtles. Through investigating effects of chemical dose and exposure duration on unique protein abundance in vitro, this study provides an optimised framework for conducting cell-based studies in wildlife proteomics, and highlights that proteins detected in vitro could act as biomarkers of chemical exposure and effect in vivo.

Metabolomic responses in freshwater benthic invertebrate, Chironomus tepperi, exposed to polyethylene microplastics: A two-generational investigation.

The accumulation of microplastics (MPs) in sediments could pose risks to benthic organisms and their progeny. Here, we examined effects on traditional apical endpoints along with changes to whole body metabolite profiles induced by irregular shaped polyethylene MPs (1-45 µm) at environmentally relevant concentrations (125, 250, 500 and 1000 MPs/kg sediment) in Chironomus tepperi using a two-generation exposure regime. Survival and emergence of C. tepperi were negatively affected in the parental generation at the two highest concentrations, whereas endpoints associated with growth were only impacted at 1000 MPs/kg sediment. Metabolites associated with several amino acid and energy metabolism pathways were present at lower abundances at the highest exposure concentration suggesting an overall impact on bioenergetics which relates to the inhibition of food acquisition or nutrient assimilation caused by ingestion of MPs, rather than a traditional receptor-mediated toxicity response. In contrast, no significant effects on apical endpoints were observed in the continuous exposure of first filial generation, and lactic acid was the only metabolite that differed significantly between groups. Larvae in unexposed conditions showed no differences in survival or metabolite profiles suggesting that effects in the parental generation do not carry over to the next filial generation. The findings provide evidence on the underlying impacts of MP ingestion and potential adaption to MP exposure of C. tepperi.

Tadpole mortality varies across experimental venues: do laboratory populations predict responses in nature?

Laboratory experiments are widely used to study how populations in nature might respond to various biological interactions, but the relevance of experiments in artificial venues is not known. We compiled mortality and growth data from 424 anuran populations carried out under laboratory, mesocosm, field enclosure, and field settings to determine if major differences exist amongst experimental venues and how this might influence experimental responses of tadpoles amongst venues. Our results show that there are fundamental differences in survival amongst venues, with the highest mortality occurring in field populations and the lowest in laboratory populations. Separation of mesocosm and field enclosure data based on the possibility of predatory interactions indicates that predation is an important factor leading to increased mortality in natural populations. Comparisons of size distributions across venues (although size data were limited for field populations) suggest that variation in tadpole size is low in natural populations compared to populations in artificial venues. We infer from this that mortality has a homogenizing effect on size in nature, resulting in natural populations that are not a random sample of hatched individuals. This finding suggests that populations reared under controlled laboratory conditions in the absence of predation (and other selective pressures) may not be representative of natural populations.

Toxicity of naphthenic acids to wood frog tadpoles (Lithobates sylvaticus).

Increased incidences of mortality and adverse effects have been described for wildlife exposed to oil-sands-process-affected waters (OSPW). Naphthenic acids (NA) were identified as a primary toxic component of OSPW, yet little information exists regarding NA-induced toxicity in aquatic vertebrates. Amphibian larvae may be particularly susceptible to exposure to OSPW in groundwater surrounding oil sands regions, and increased frequency of mortality and adverse developmental effects were noted in exposed tadpoles. Despite this, there are no published studies investigating the effects of NA exposure on developing tadpoles. LC50 values of 4.76 mg/L NA were found for tadpoles at an early developmental stage (Gosner stage 28), and even greater toxicity with more developed tadpoles at 96 h, with an LC50 value of 3.04 mg/L in Gosner stage 36 tadpoles. These values are well below NA concentrations found in OSPW tailing ponds and similar to levels identified in groundwater in the Athabasca Oil Sands region.

Field-scale monitoring of green sea turtles (Chelonia mydas): Influence of site characteristics and capture technique on the blood metabolome.

Given their threatened status, there is considerable interest in establishing monitoring techniques that can be used to evaluate the health of sea turtles in the wild. The present study represents a methodological contribution towards field-scale metabolomic assessment of sea turtles, by exploring differences in blood biochemistry associated with site characteristics and capture technique. We compared the metabolome of blood from animals at three locations (two coastal and one reefal), collected from turtles that were either resting or active, and sampled across multiple seasons at one location. Our results show clear differences in the metabolome of turtles from the three locations, some of which are likely attributable to differences in diet or forage quality and others which may reflect differences in other factors (e.g., occurrence of land-based contaminants or other biotic and/or abiotic stressors) between coastal and reefal sites. Our analysis also revealed the influence of capture technique on metabolite profiles, with numerous markers of physical exertion in animals captured while active that were absent in turtles sampled while resting. We observed a modest potential for temporal differences in the metabolome, but controlling for sampling time did not change the overall conclusions of our study. This suggests that temporal differences in the metabolome warrant consideration when designing studies to evaluate the status of sea turtles in the wild, but that site characteristics and capture technique are bigger drivers. However, sample size for this comparison was relatively small and further investigation of seasonal differences in the metabolome are warranted. Research exploring each of these factors more closely will further contribute towards achieving robust metabolomics analysis of sea turtles across large spatial and temporal scales.

Estrogenic mixtures induce alterations in lipidomic profiles in the gonads of female oysters.

This study aimed to reveal possible alterations to lipidomic profiles in Sydney rock oysters, Saccostrea glomerata, exposed to estrogenic mixtures (i.e., estrone, E1; 17ß-estradiol, E2; estriol, E3; 17α-ethinylestradiol, EE2; bisphenol A, BPA; 4-t-octylphenol, 4-t-OP; and 4-nonylphenol, 4-NP) at "low" and "high" concentrations, typical of those detected in Australian and global receiving waters. A seven-day acute exposure window exhibited significantly lower abundances of many non-polar metabolites in digestive gland, gills, and gonads. Overall, there was a strong effect of the carrier solvent ethanol (despite a low exposure of 0.0002%), with all solvent containing treatments exhibiting lower abundances of lipidic metabolites, especially in the gill and digestive gland. No significant changes of the lipidome were exhibited in the male gonad by estrogenic exposure. However, in the female gonad, significant reductions of phospholipids and phosphatidylcholine were associated with exposure to high estrogenic mixtures. We hypothesise that the decreases in these phospholipids in the female gonad may be attributable to 1) lower algal consumption and thus lower uptake of lipidic building blocks; 2) a reduction of available substrates for phospholipid and phosphatidylcholine synthesis; and/or 3) induction of reactive oxygen species via estrogen metabolism, which may cause lipid peroxidation and lower abundance of phospholipids.

Ocean acidification impairs the physiology of symbiotic phyllosoma larvae of the lobster Thenus australiensis and their ability to detect cues from jellyfish.

Ocean acidification (OA) can alter the behaviour and physiology of marine fauna and impair their ability to interact with other species, including those in symbiotic and predatory relationships. Phyllosoma larvae of lobsters are symbionts to many invertebrates and often ride and feed on jellyfish, however OA may threaten interactions between phyllosomas and jellyfish. Here, we tested whether OA predicted for surface mid-shelf waters of Great Barrier Reef, Australia, under ∆ pH = -0.1 (pH ~7.9) and ∆pH = -0.3 (pH ~7.7) relative to the present pH (~8.0) (P) impaired the survival, moulting, respiration, and metabolite profiles of phyllosoma larvae of the slipper lobster Thenus australiensis, and the ability of phyllosomas to detect chemical cues of fresh jellyfish tissue. We discovered that OA was detrimental to survival of phyllosomas with only 20% survival under ∆pH = -0.3 compared to 49.2% and 45.3% in the P and ∆pH = -0.1 treatments, respectively. The numbers of phyllosomas that moulted in the P and ∆pH = -0.1 treatments were 40% and 34% higher, respectively, than those in the ∆pH = -0.3 treatment. Respiration rates varied between pH treatments, but were not consistent through time. Respiration rates in the ∆pH = -0.3 and ∆pH = -0.1 treatments were initially 40% and 22% higher, respectively, than in the P treatment on Day 2 and then rates varied to become 26% lower (∆pH = -0.3) and 17% (∆pH = -0.1) higher towards the end of the experiment. Larvae were attracted to jellyfish tissue in treatments P and ∆pH = -0.1 but avoided jellyfish at ∆pH = -0.3. Moreover, OA conditions under ∆pH = -0.1 and ∆pH = -0.3 levels reduced the relative abundances of 22 of the 34 metabolites detected in phyllosomas via Nuclear Magnetic Resonance (NMR) spectroscopy. Our study demonstrates that the physiology and ability to detect jellyfish tissue by phyllosomas of the lobster T. australiensis may be impaired under ∆pH = -0.3 relative to the present conditions, with potential negative consequences for adult populations of this commercially important species.

Chronic pesticide exposure elicits a subtle carry-over effect on the metabolome of Aurelia coerulea ephyrae.

Chemical pollutants, such as pesticides, often leach into aquatic environments and impact non-target organisms. Marine invertebrates have complex life cycles with multiple life-history stages. Exposure to pesticides during one life-history stage potentially influences subsequent stages; a process known as a carry-over effect. Here, we investigated carry-over effects on the jellyfish Aurelia coerulea. We exposed polyps to individual and combined concentrations of atrazine (2.5 µg/L) and chlorpyrifos (0.04 µg/L) for four weeks, after which they were induced to strobilate. The resultant ephyrae were then redistributed and exposed to either the same conditions as their parent-polyps or to filtered seawater to track potential carry-over effects. The percentage of deformities, ephyrae size, pulsation and respiration rates, as well as the metabolic profile of the ephyrae, were measured. We detected a subtle carry-over effect in two metabolites, acetoacetate and glycerophosphocholine, which are precursors of the neurotransmitter acetylcholine, important for energy metabolism and osmoregulation of the ephyrae. Although these carry-over effects were not reflected in the other response variables in the short-term, a persistent reduction of these two metabolites could have negative physiological consequences on A. coerulea jellyfish in the long-term. Our results highlight the importance of considering more than one life-history stage in ecotoxicology, and measuring a range of variables with different sensitivities to detect sub-lethal effects caused by anthropogenic stressors. Furthermore, since we identified few effects when using pesticides concentrations corresponding to Australian water quality guidelines, we suggest that future studies consider concentrations detected in the environment, which are higher than the water quality guidelines, to obtain a more realistic scenario by possible risk from pesticide exposure.

Changes in global protein expression in sea turtle cells exposed to common contaminants indicates new biomarkers of chemical exposure.

Non-targeted protein expression at the cellular level can provide insights into mechanistic effects of contaminants in wildlife, and hence new and potentially more accurate biomarkers of exposure and effect. However, this technique has been relatively unexplored in the realm of in vitro biomarker discovery in threatened wildlife, despite the vulnerability of this group of animals to adverse sublethal effects of contaminant exposure. Here we examined the usefulness of non-targeted protein expression for biomarker discovery in green sea turtles (Chelonia mydas) by investigating differences in the response of primary cells from five different tissue types that were exposed to three contaminants known to accumulate in this species. Cells derived from C. mydas skin, liver, kidney, ovary and small intestine were exposed to 100 µg/L of either polychlorinated biphenyl 153 (PCB153), perfluorononanoic acid (PFNA) or phenanthrene for 24 h. The global protein expression was then quantitatively evaluated using sequential window acquisition of all theoretical mass spectra (SWATH-MS). Comparison of the global protein profiles revealed that, while a majority of proteins were mutually expressed in controls of all tissue types (~90%), the response to exposure in terms of protein expression strength was significantly different between tissue types. Furthermore, a comparison to known markers of chemical exposure in sea turtles from the literature indicated that in vitro response can reflect known in vivo responses. In particular, markers such as heat shock protein (HSP) 60, glutathione S-transferases (GSTs) and superoxide dismutases (SODs), cytochrome P450 and catalase were dysregulated in response to exposure. Furthermore, potential new markers of exposure were discovered such as annexin, an important protein in cell signalling processes. While this methodology proved promising further studies are required to confirm the accuracy of in vitro protein expression as a tool for biomarker discovery in wildlife.