Interactions Between Heavy Metal Exposure and Blood Biochemistry in an Urban Population of the Black Swan (Cygnus atratus) in Australia.

There is growing recognition of the threat posed to wildlife by pollutants. Waterbirds are robust bioindicators of ecosystem health, and metal toxicity is a threat to these species in waterways worldwide. Urban waterbirds are likely to be at the highest risk of heavy metal exposure, but this issue has not been widely explored in Australia. Our aim was to estimate contemporary heavy metal exposure in a sedentary urban waterbird population: black swans (Cygnus atratus) inhabiting an inner-city wetland in one of Australia's largest cities, Melbourne. To investigate the physiological implications of legacy heavy metal exposure in these birds, we quantified blood biochemistry profiles and examined their relationships with metal concentrations in feathers. We caught 15 swans in 2021 and took feather samples to measure the concentration of eight heavy metals (chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), lead (Pb), and mercury (Hg)), and blood samples to measure the concentration of 13 plasma analytes. Multivariate regression analysis revealed few associations between heavy metals and biochemistry markers, and no differences between sexes or age classes. This study presents a baseline dataset of these contaminants and blood biochemical profiles of swans at this wetland that can be used for future monitoring and is an important step toward a better understanding of the threat posed by heavy metals to Australian urban waterbirds.

Exposure to Persistent Organic Pollutants in Australian Waterbirds.

There is growing worldwide recognition of the threat posed by persistent organic pollutants (POPs) to wildlife populations. We aimed to measure exposure levels to POPs in a Southern Hemisphere aquatic waterbird species, the nomadic gray teal (Anas gracilis), which is found across Australia. We collected wings from 39 ducks harvested by recreational hunters at two sites (one coastal, one inland) in Victoria, southeastern Australia, in 2021. We examined three groups of POPs: nine congeners of polychlorinated biphenyls (PCBs), 13 organochlorine pesticides (OCPs), and 12 polycyclic aromatic hydrocarbons (PAHs). The PCBs, OCPs, and PAHs were detected at quantifiable levels in 13%, 72%, and 100% of birds, respectively. Of the congeners we tested for in PCBs, OCPs, and PAHs, 33%, 38%, and 100% were detected at quantifiable levels, respectively. The highest levels of exposure to POPs that we found were to the PAH benzo[b]fluoranthene, occurring at a concentration range of 1.78 to 161.05 ng/g wet weight. There were some trends detected relating to differences between geographical sites, with higher levels of several PAHs at the coastal versus inland site. There were several strong, positive associations among PAHs found. We discuss potential sources for the POPs detected, including industrial and agricultural sources, and the likely role of large-scale forest fires in PAH levels. Our results confirm that while Australian waterbirds are exposed to a variety of POPs, exposure levels are currently relatively low. Additional future investigations are required to further characterize POPs within Australian waterbird species. Environ Toxicol Chem 2024;43:736-747. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

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

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

Environmentally relevant concentrations of chemically complex shale gas wastewater led to reduced fitness of water fleas (Daphnia carinata): Multiple lines of evidence approach.

Shale gas hydraulic fracturing generates flowback waters that pose a threat to aquatic organisms if released into the environment. In order to prevent adverse effects on aquatic ecosystems, multiple lines of evidence are needed to guide better decisions and management actions. This study employed a multi-disciplinary approach, combining direct toxicity assessment (DTA) on the water flea Daphnia carinata and LC-MS metabolomics analysis to determine the impact of a major ion salinity control (SC) and a cumulative flowback shale gas wastewater (SGW) from a well in the Beetaloo Sub-basin, Northern Territory, Australia. The exposures included a culture water control, simply further referred to as 'control', SC at 1% and 2% (v/v) and SGW at 0.125, 0.25, 0.5, 1% and 2% (v/v). The results showed that reproduction was significantly increased at SGW 0.5%, and significantly decreased when exposed to SC 2%. SGW 2% was found to be acutely toxic for the D. carinata (< 48-h). Second generation (F1) of D. carinata exposed to 0.125-1% SGW generally saw reduced activity in four oxidative biomarkers: glutathione S-transferase, lipid peroxidation, reactive oxygen species, and superoxide dismutase. At the metabolomics level, we observed significant changes in 103 metabolites in Daphnia exposed to both SGW and elevated salinity, in comparison to the control group. These changes indicate a range of metabolic disturbances induced by SGW and salinity, such as lipid metabolism, amino acid metabolism, nucleotide synthesis, energy production, and the biosynthesis of crucial molecules like hormones and pigments. These multiple lines of evidence approach not only highlights the complexities of SGW's impact on aquatic ecosystems but also underscores the importance of informed decision-making and management practices to safeguard the environment and its inhabitants.

Environmental organic contaminant body burdens and GC-MS based untargeted metabolomics in mediterranean mussels from Port Phillip Bay, Australia☆.

Mussels were collected from four coastal sites around Port Phillip Bay, Australia in Mar and Apr 2021). Body burdens of Polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and organochlorine pesticides (OCPs) were measured and the possible sources of toxicants discussed. In addition, a gas chromatography-mass spectrometry (GC-MS) based untargeted metabolomics analysis was performed using the mantle tissues of mussels. Correlations between the results of contaminant body burdens and metabolic variations were investigated. The results demonstrated that high accumulations of low-molecular-weight PAHs were found in mussels. High body burdens of PCBs and OCPs were only found at mussels from the site close to the river mouth. Some of the metabolic pathways were correlated with the accumulation of PAHs. No correlations were found between PCB and OCP accumulations and metabolic abundances. According to the food and environmental standards of the European Union (EU), the PAH, PCB, and OCP accumulation in mussels in this study are a serious food safety concern.

Changes in metabolic profiles of amphipods Allorchestes compressa after acute exposures to copper, pyrene, and their mixtures.

Amphipods are ideal indicators for biomonitoring and ecotoxicological studies of environmental contaminants because they are extensively distributed in aquatic environments, are easy to collect and are important in nutrient cycling. Marine amphipods (Allorchestes compressa) were exposed to two concentrations of copper and pyrene, and their mixtures, for 24 and 48 h. Changes in polar metabolites were assessed using Gas Chromatography Mass Spectrometry (GC-MS)-based untargeted metabolomics. Generally, limited metabolite changes were observed for copper and pyrene single exposures (eight and two significant metabolites, respectively), while 28 metabolites had changed following exposures to mixtures. Furthermore, changes were mainly observed after 24 h but had seemingly returned to control levels after 48 h. Multiple types of metabolites were affected including amino acids, Tricarboxylic acid (TCA) cycle intermediates, sugars, fatty acids, and hormones. This study highlights the sensitivity of metabolomics in assessing the impacts of low concentrations of chemicals compared to traditional ecotoxicological endpoints.

Expanding the use of portable XRF to monitor lead exposure in an Australian duck species two decades after a ban on lead shot.

There is growing worldwide recognition of the threat posed by toxic lead for wildlife and humans. Lead toxicity from ammunition has been shown to be a threat to waterbirds across the globe. Lead shot was banned for all waterfowl hunting in Victoria, Australia, in 2002. However, no assessments of lead exposure in Australian waterfowl have been published since the 1990s. Our aim was to estimate contemporary lead exposure via measuring bone lead concentrations in a harvested dabbling duck, the Pacific black duck (Anas superciliosa). We collected wings from 77 Pacific black ducks, spanning 2018 (n = 30) and 2021 (n = 47), from nine sites with long-term histories of regular waterfowl hunting. We sought to validate portable X-ray fluorescence (XRF) for this purpose by taking a piece of humerus bone from each bird, and measuring lead concentration (mg/kg), first via non-destructive XRF and then via destructive inductively coupled plasma mass spectrometry (ICP-MS) and validated the relationship via regression analysis. Portable XRF bone lead measurement demonstrated a strong correlation with ICP-MS results using root-transformed regression (R2 = 0.85). Greater than 92 % of ducks had only background lead exposure (<10 mg/kg). When compared to historical studies in the same species at similar field sites from the 1990s, lead exposure levels were considerably lower, with mean lead concentrations ∼2-fold lower (3.7 c.f. 7.7 mg/kg), and the frequency of birds with severe lead exposure (>20 mg/kg) ∼3-fold lower (2.6 c.f. 7.5 %). Our results confirm that portable XRF is a useful option for measurement of bone lead in Australasian waterbird species. Our findings also demonstrate that a ban on the use of lead shot around 20 years ago has been associated with a substantial reduction in lead exposure in at least one species of waterfowl.

Mixture Toxicity of Three Unconventional Gas Fracking Chemicals, Barium, O-Cresol, and Sodium Chloride, to the Freshwater Shrimp Paratya australiensis.

The 96-h acute toxicity of barium (Ba2+ ), o-cresol, and sodium chloride (NaCl) to Paratya australiensis was assessed in single, binary, and ternary combinations in addition to three biochemical assays: glutathione S-transferase, acetylcholinesterase, and sodium-potassium adenosine triphosphatase. The 96-h lethal concentrations that expressed 50% mortality (LC50) in the single-toxicant exposures were Ba2+ = 23.4 mg/L, o-cresol = 12.2 mg/L, and NaCl = 4198 mg/L. Mortality from o-cresol exposure occurred between 11 and 22 mg/L, whereas Ba2+ was more gradual across 10-105 mg/L, and most of the NaCl mortality occurred between 2050 and 4100 mg/L. Toxic units were used to assess the binary and ternary interactions of the toxicants. A more than additive effect was observed for most combinations in the binary chemical exposures, with the ternary combinations yielding highly synergistic interactions. Greater synergism was observed with the 96-h LC50 of o-cresol in combination with the three concentrations of NaCl (1025, 2050, and 3075 mg/L) compared with Ba2+ , with toxic units of 0.38, 0.48, and 0.10 (o-cresol) and 0.71, 0.67, and 0.50 (Ba2+ ). No notable enzyme activity trends were observed in the enzyme biomarker responses from both individual and mixture exposures. Although acute single-species toxicity tests tend to underestimate the effects of Ba2+ , o-cresol, and NaCl on populations, communities, and ecosystems in seminatural (e.g., mesocosms) and natural systems, there are currently no published acute toxicity data available for P. australiensis and the three toxicants used in the present study. The present study shows that chemicals with different toxicity mechanisms can potentially lead to more synergistic responses. Environ Toxicol Chem 2023;42:481-494. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

The Acute Toxicity of Salinity in Onshore Unconventional Gas Waters to Freshwater Invertebrates in Receiving Environments: A Systematic Review.

Industries such as unconventional natural gas have seen increased global expansion to meet the increasing energy needs of our increasing global population. Unconventional gas uses hydraulic fracturing that produces significant volumes of produced waters, which can be highly saline and pose a toxic threat to freshwater invertebrates if exposure via discharges, spills, leaks, or runoff were to occur. The primary aim of the present review was to determine the sodium (Na+ ) and chloride (Cl- ) content of these waters as an approximate measure of salinity and how these values compare to the NaCl or synthetic marine salt acute toxicity values of freshwater invertebrate taxa. Shale gas produced waters are much more saline with 78 900 ± 10 200 NaCl mg/L and total dissolved solids (TDS) of 83 200 ± 12 200 mg/L compared to coal bed methane (CBM) produced waters with 4300 ± 1100 NaCl mg/L and TDS of 5900 ± 1300 mg/L and pose a far greater toxicity risk from NaCl to freshwater invertebrates. In addition, the toxicity of other major ions (Ca2+ , K+ , Mg2+ , CO 3 2 - , HCO3 - , and SO 4 2 - ) and their influence on the toxicity of Na+ and Cl- were evaluated. Exposure of untreated and undiluted shale gas produced waters to freshwater invertebrates is likely to result in significant or complete mortality. Shale gas produced waters have higher concentrations of various metals compared with CBM produced waters and are more acidic. We recommend future research to increase the reporting and consistency of water quality parameters, metals, and particularly organics of produced waters to provide a better baseline and help in further investigations. Environ Toxicol Chem 2022;41:2928-2949. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Real-time automated behavioural monitoring of mussels during contaminant exposures using an improved microcontroller-based device.

An improved microcontroller-based device for real-time biomonitoring of mussel behaviour is introduced in this study. Open source Arduino platforms were used as processing cores and infra-red (IR) sensors (with transistor output) and Hall sensors to record the cardiac activities and valve movements of mussels. Compared to the devices described in previous studies, this device has low cost, high throughput, and high portability, and can be applied to conduct real-time preliminary automatic data processing. Mediterranean mussels were exposed to Cu2+ and microplastics and their cardiac activities and valve movements were recorded. The results demonstrated that Cu2+ exposure caused valve closure and a drop in the heart rate, similar to the behaviour during natural periods of bradycardia in mussels. The microplastic exposures tended to cause high oscillations (low concentration of microplastics) and slow decreases (high concentration of microplastics) of the maximum valve open positions. Such oscillations and decreases appeared to reset and restart after the bradycardia period. The device has potential to measure and establish behavioural responses of mussels and other bivalves, to the stress of exposure from environmental contaminants.

Trends in environmental and toxicity research on organic ultraviolet filters: A scientometric review.

In recent decades, the potential toxicological and environmental effects of organic ultraviolet filters (OUVF) have received growing attention. The number of studies in this area has increased; however, presently there is no scientometric perspective addressing this topic. The purpose of this study is to identify the intellectual base and research front using the visualization and analysis software, CiteSpace. We retrieved 453 articles, published in print or online as an early-access article between 2002 and 2020, from the Web of Science with a topic search related to OUVFs, environment, and toxicology. We then analysed synthesized networks of co-authorship (author, institution, country), co-citation (author, document, journal) and co-occurring keywords. The annual publication output has trended upwards since 2002. Authors based in China accounted for 29.4% of the total publications, followed by USA (17.4%); but overall publications from Switzerland and Spain were more influential. Major research themes identified included OUVF concentrations in aquatic environments, and hormonal effects. Emerging themes included improving the sensitivity of analytical detection methods for both OUVFs and their metabolites, consequences of OUVF transport to the marine environment, and concerns over prenatal exposure. Based on keyword analysis, benzophenone-3, 4-methylbenzylidene-camphor, 3-benzylidene camphor, and ethylhexyl-methoxycinnamate are the most studied OUVFs, and effects on estrogenic activity, gene expression, reproduction, and more recently, oxidative stress, have received most attention from a toxicological perspective. Other prominent topics were sources of environmental contamination and ecological risk assessments. This study maps the major research domains of OUVF environmental toxicology research; explanations and implications of the findings are discussed; and emerging trends highlighted.

Climate change impacts on pollutants mobilization and interactive effects of climate change and pollutants on toxicity and bioaccumulation of pollutants in estuarine and marine biota and linkage to seafood security.

This article provides an overview of the impacts of climate change stressors (temperature, ocean acidification, sea-level rise, and hypoxia) on estuarine and marine biota (algae, crustaceans, molluscs, corals, and fish). It also assessed possible/likely interactive impacts (combined impacts of climate change stressors and pollutants) on pollutants mobilization, pollutants toxicity (effects on growth, reproduction, mortality) and pollutants bioaccumulation in estuarine and marine biota. An increase in temperature and extreme events may enhance the release, degradation, transportation, and mobilization of both hydrophobic and hydrophilic pollutants in the estuarine and marine environments. Based on the available pollutants' toxicity trend data and information it reveals that the toxicity of several high-risk pollutants may increase with increasing levels of climate change stressors. It is likely that the interactive effects of climate change and pollutants may enhance the bioaccumulation of pollutants in seafood organisms. There is a paucity of literature relating to realistic interactive effects of climate change and pollutants. Therefore, future research should be directed towards the combined effects of climate change stressors and pollutants on estuarine and marine bota. A sustainable solution for pollution control caused by both greenhouse gas emissions (that cause climate change) and chemical pollutants would be required to safeguard the estuarine and marine biota.

Microplastics alter digestive enzyme activities in the marine bivalve, Mytilus galloprovincialis.

Microplastics are eaten by many invertebrates, particularly filter-feeding organisms like mussels. Since microplastics can be retained in the digestive system for extended periods, there is ample opportunity for them to interact with the functions of digestive enzymes. This study determined how the polymer type, size and concentration of ingested spherical microplastics affects the activities of seven key digestive enzymes in the digestive gland of Mytilus galloprovincialis, a common marine mussel. Polymer type significantly affected the activities of carbohydrase enzymes: polystyrene reduced amylase and xylanase activities, and increased cellulase activity. High concentrations of microplastics (5 × 104 microplastics L-1) caused a 2.5-fold increase in total protease activity. The activities of laminarinase, lipases and lipolytic esterases were unaffected by the polymer type, size or concentration of microplastics. Microplastics-induced changes to digestive enzyme activities can affect mussels' ability to acquire energy from food and reduce their energy reserves.

Legacy and emerging per- and polyfluoroalkyl substances (PFASs) in Australian biosolids.

Biosolids samples were collected from 19 Australian WWTPs during 2018 that cover a range of catchment types (urban, rural, industrial waste discharges) and treatment technologies. Samples were analysed for 44 PFAS using isotope dilution and alkaline extraction coupled with quantification with LC-MS/MS. The Σ44PFAS mean concentration was 260 ng/g dry weight (dw) and ranged between 4.2 and 910 ng/g dw. The dominant compound class detected were the di-substituted phosphate esters (Σ3PAPs mean 140 ng/g dw; range ND - 730 ng/g dw) which contributed 45% of the total mean Σ44PFAS mass, followed by perfluoroalkyl carboxylic acids (Σ11PFCAs mean 39 ng/g dw; range 2.3-120 ng/g dw) contributing 17%, and the perfluoroalkyl sulfonates (Σ8PFSAs mean 28 ng/g dw; range 0.9-220 ng/g) which contributed 16%. Using the population data supplied by the participating WWTPs, the mean annual estimated biosolids-associated PFAS contribution is 6 mg per person per year and ranged between 0.6 mg and 15 mg. A similar population normalised concentration regardless of WWTP, region or capacity suggests that the domestic environment provides the baseline PFAS loading. Statistically significant higher Σ44PFAS and PFOS concentrations were observed at urban locations. A weak correlation was observed between annual mass of PFAS associated with each individual WWTP and their percentage industrial waste contribution. This may be important for elevated PFAS concentrations observed in WWTPs with higher industrial waste inputs and requires further research.

A systematic review and ecological risk assessment for organic ultraviolet filters in aquatic environments.

Organic ultraviolet filters (OUVFs) are used in a wide range of manufactured products including personal care (e.g. sunscreens) and plastic items. This review summarizes the available data regarding the toxic effects of OUVFs on marine and freshwater organisms and generates the predicted no-effect concentration (PNEC) values necessary for assessing ecological risk. Through a systematic search of the literature, 89 studies were identified and ecotoxicological data extracted. Collectively, these studies described toxicity testing with 39 OUVF from 10 structural classes, with derivatives of benzophenones (49%) and camphors (16%) most studied. There was a bias towards selecting freshwater species (61%), and evaluating single OUVF effects (87%) rather than OUVF mixtures. Short-term (acute) experimentation (58%) was marginally more common than long-term (chronic) testing (42%). Reproductive, developmental, genetic, and neurological toxicity were the most commonly identified effects in aquatic organism, and were associated with molecular interactions with steroid receptors, DNA, or the production of reactive oxygen species. Species sensitivity distribution and/or assessment factors were used to calculate PNECs for 22 OUVFs and the risk quotients for 12 OUVFs. When using maximum concentrations, high risk was observed for six OUVFs in marine environments (4-methylbenzylidene-camphor, octocrylene, padimate-O, benzophenone-1, and oxybenzone, ethylhexyl-4-methoxycinnamate), and for four OUVFs in freshwater environments (ethylhexyl-4-methoxycinnamate, octocrylene, avobenzone and oxybenzone). When using median concentrations, a risk to marine environments was observed for oxybenzone. The results of this review underline that there is limited knowledge of the pathological effects of OUVFs and their metabolites in aquatic environments, and this inhibits the development of informed water-quality guidelines.

Population- and sex-specific sensitivity of the marine amphipod Allorchestes compressa to metal exposure.

The sensitivity to contaminants of natural populations varies greatly depending on their historical exposure and on the sex of the individual. These factors result in great uncertainty in ecotoxicological risk assessments and challenge the protection of marine biodiversity. This study investigated the role of background pollution in the environment in shaping the sensitivity of males and females of the common marine amphipod Allorchestes compressa to the common trace marine pollutant, copper (Cu). Female and male amphipods were collected from two sites: Geelong (the polluted site) and Clifton Springs (the clean site). Amphipods were exposed to Cu treatments of 0, 50, 100, and 250 µg/L for 10 days, followed by a 10-day recovery period. Cu-exposed males from Geelong showed a reduction in feeding rate at a higher Cu concentration than males from Clifton Springs, suggesting that they have a higher tolerance to Cu than males from Clifton Springs. This can be explained by their higher base level of metallothioneins (MTs) and glutathione-S-transferase (GST), the key physiological responses for detoxification and defence against damages from Cu toxicity. Males showed a higher tolerance to Cu than females. This pattern was similar in both populations, which may be associated with a higher level of GST. During the recovery period, only males from Geelong fully recovered to the control level. Our results emphasize the importance of considering population- and sex-specific sensitivity of invertebrates to contaminants in ecotoxicological risk assessments.

The PAH body burdens and biomarkers of wild mussels in Port Phillip Bay, Australia and their food safety implications.

Wild Mediterranean mussels (Mytilus galloprovincialis) were collected from 10 coastal sites of Port Phillip Bay, Australia, and 16 types of Polycyclic aromatic hydrocarbons (PAHs) in their bodies were measured. Also, mussels from 4 of the 10 sites were sampled and the activities of Metallothioneins (MTs), superoxide dismutase (SOD), glutathione S-transferase (GST), and Ethoxyresorufin -o- deethylase (EROD) in the gills were tested. The results demonstrated spatial variation of different PAHs in mussels, which differed from trace metal accumulations to mussels reported in previous studies. Besides, the results of SOD, GST, and EROD showed highly similar features among mussels from all the sites while the results of MTs appeared to be different. The estimation of possible excessive cancer risks caused by wild mussel consumption using the results in this study demonstrated that the PAHs in wild mussels in Port Phillip Bay is a moderate concern for consumers.

Invertebrate responses to microplastic ingestion: Reviewing the role of the antioxidant system.

Microplastic ingestion in invertebrates reduces somatic and reproductive growth. This could be caused by energy reserves being detracted from growth processes and redistributed to maintenance processes that preserve life. A potential sink for this diverted energy is the antioxidant system, which minimises oxidative damage and reinstates redox homeostasis following disturbances caused by exposure to pollution. Several microplastic studies have used genetic and molecular redox biomarkers to assess how microplastic ingestion affects the functioning of the antioxidant system. This systematic review synthesises the current understanding of redox biomarker responses in invertebrates that have ingested microplastics. We found that biomarker response information exists for only seven invertebrate taxa, and early life stages have received little scientific attention. The microplastics used by most studies were polystyrene (45% of studies), spherical (51% of studies), and were < 10 µm in diameter (31% of studies). We found multiple examples of microplastic ingestion posing an oxidative challenge to invertebrates, which required upregulation of antioxidant system components. However, the lack of systematic experiments prevented us from clearly identifying which characteristic of microplastics caused these responses. We identify several areas for consideration when investigating biomarker responses to microplastic ingestion and offer research priorities for future studies.

Spatial and temporal variations of trace metal body burdens of live mussels Mytilus galloprovincialis and field validation of the Artificial Mussels in Australian inshore marine environment.

The body burdens of eight trace metals: Cd, Pb, Cu, Zn, Cr, Se, Hg, and As, were measured in live mussels (LMs) Mytilus galloprovincialis at 14 coastal sites in Port Phillip Bay, Victoria, Australia in winter and summer between 2017 and 2018. The spatial and temporal variations of body burdens were evaluated. The results revealed significantly higher body burdens of Cd, Pb, Cu, Zn, Cr, and Hg in summer at the sites where the city centre and industries are located. Elevated levels of most trace metals including 3 toxic, non-essential metals (Cd, Pb, and Hg) were detected in LMs from the site of Geelong. The body burdens of Zn, Cr, Se, and As appeared higher at the sites from the Bellarine Peninsula and the mouth of Port Phillip Bay. Besides, the "Artificial Mussels" (AMs) were deployed at the same sites in summer for 28 d and retrieved when the LMs were collected. The accumulations of the eight metals were compared between AMs and LMs summer results. It indicated significant correlations for Pb, Cu, and Cr, lower correlations in Zn and Hg, and irregular correlations for Se and As. The AM results of Cd were below the detection limit of the analytical method. This study demonstrates that AMs are excellent replacement of LMs for the biomonitoring of multiple kinds of trace metals.

Foaming at the mouth: Ingestion of floral foam microplastics by aquatic animals.

Phenol-formaldehyde plastics are used globally as floral foam and generate microplastics that can enter the environment. This study is the first to describe how aquatic animals interact with this type of microplastic, and the resultant physiological responses. We analysed "regular foam" microplastics generated from petroleum-derived phenol-formaldehyde plastic, and "biofoam" microplastics generated from plant-derived phenol-formaldehyde plastic. Regular foam and biofoam microplastics showed similar FTIR spectra. Both types of microplastics were consumed by all six invertebrate species tested: the freshwater gastropod Physa acuta, the marine gastropod Bembicium nanum, the marine bivalve Mytilus galloprovincialis, adults and neonates of the freshwater crustacean Daphnia magna, the marine amphipod Allorchestes compressa, and nauplii of the marine crustacean Artemia sp. For all species, the occurrence of ingestion was similar for regular foam and biofoam microplastics. Biofoam microplastics leached more than twice as much phenolic compounds than regular foam microplastics. The leachates from regular foam and biofoam microplastics showed the same acute toxicity to Artemia nauplii (24-h LC50 = 27.4 mg mL-1 and 22.8 mg mL-1, respectively) and D. magna (48-h LC50 = 17.8 mg mL-1 and 15.3 mg mL-1, respectively). However, biofoam microplastic leachate was twice as toxic to embryos of the zebrafish, Danio rerio, compared with leachate from regular foam microplastic (96-h LC50 = 43.8 mg mL-1 vs 27.1 mg mL-1). Using M. galloprovincialis, we show that regular foam microplastic leachate and the physical presence of the microplastics exerted separate and cumulative effects on catalase (CAT) activity, glutathione-s-transferase (GST) activity and lipid peroxidation. Microplastic ingestion did not affect the activity of acetylcholinesterase (AChE). Taken together, these results show that phenol-formaldehyde microplastics can interact with a range of aquatic animals, and affect sublethal endpoints by leaching toxic compounds, and through the physical presence of the microplastics themselves.