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.

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.

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.

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.

Effects of perfluorooctanoic acid (PFOA) on the thyroid status, vitellogenin, and oxidant-antioxidant balance in the Murray River rainbowfish.

Perfluorooctanoic acid's (PFOA) widespread use, presence and persistence in the aquatic environment has led to an increasing number of studies focusing on its toxicological effects. In Australia, PFOA has been detected in the aquatic environment, however its effects on Australian native fauna are unknown. In this study, male Australian native fish Murray River rainbowfish (Melanotaenia fluviatilis) were exposed to four different concentrations of PFOA (0.01, 0.1, 1 and 10 mg L-1). Variations in thyroid hormones (Triiodothyronine (T3)/Thyroxine (T4)) and the presence of vitellogenin were determined in plasma. Oxidative stress responses were evaluated in gills and liver. Exposure of male fish to PFOA resulted in altered T3/T4 ratios and the presence of vitellogenin in the plasma. Activities of catalase (CAT) and glutathione- S-transferase (GST) were significantly increased in the gills and significantly reduced in the liver. Lipid peroxidation was observed in both tissues showing that vital organs could not neutralize the peroxides generated by oxidative stress resulting from exposure to PFOA. In natural populations exposed to PFOA, such hormonal disturbances can have negative effects, notably through altered capacity to respond to changes in environmental conditions.

Trace metal biomonitoring in the east Gippsland Lakes estuary using the barnacle Amphibalanus variegatus and mussel Mytilus edulis.

The Gippsland Lakes estuary, a Ramsar listed wetland, in Victoria, Australia, is an area of potential concern for metal pollution due to influxes of human population and associated anthropogenic activities. A biomonitoring exercise was undertaken where the concentrations of 9 metals (Cr, Fe, Cu, Zn, As, Se, Ag, Cd and Hg) were analysed in the soft tissue of two common sessile invertebrates: the mussel Mytilus edulis and the barnacle Amphibalanus variegatus from 6 locations on two different occasions throughout the Gippsland Lakes estuary. A salinity gradient exists in the Lakes, from seawater at Lakes Entrance in the east, decreasing down to < 10 PSU in the west at Lake Wellington during times of rainfall, which is a major factor governing the growth and distribution of both species. Dissolved metal levels in general were low; however, Cu at most sites exceeded the 90% trigger values, while all Zn concentrations exceeded the lowest 80% trigger values of the ANZECC marine water quality guidelines for environmental health. Elevated levels of Cu and Zn were found particularly in barnacles at some sites with environmental contamination due to leaching from anti fouling paints and sacrificial zinc anodes. Elevated levels of Ag and Cd were found in mussels at the Hollands Landing site, which is immediately adjacent to a boat ramp, and Cd and Ag at this site are suspected to originate from inland anthropogenic sources. Concentrations of As in M. edulis across all 6 sites in both sampling periods had mean wet weight As concentrations exceeding the maximum level stated in the FSANZ guidelines. A. variegatus contained elevated levels of Hg especially at the North Arm site with a maximum of 13.6 µg Hg/g dry wt., while A. variegatus also showed temporal changes in Hg concentrations across sites. The maximum Hg concentration found in Mytilus edulis was 1.49 µg Hg/g dry wt. at the Hollands Landing site. Previous contaminant studies of biota in the Lakes have targeted sampling of singular predatory or migratory species, such as Black Bream (Acanthopagrus butcheri) and the Burrunan dolphin (Tursiops australis). This is the first biomonitoring study conducted on sessile organisms to assess metal contamination in the system.

The Toxicity of Nonaged and Aged Coated Silver Nanoparticles to Freshwater Alga Raphidocelis subcapitata.

The transformation of coated silver nanoparticles (AgNPs) and their impacts on aquatic organisms require further study. The present study investigated the role of aging on the transformation of differently coated AgNPs and their sublethal effects on the freshwater alga Raphidocelis subcapitata. The stability of AgNPs was evaluated over 32 d, and the results indicated that transformation of AgNPs occurred during the incubation; however, coating-specific effects were observed. Fresh AgNPs increased reactive oxygen species (ROS) formation, whereas aged AgNPs induced excessive ROS generation compared with their fresh counterparts. Increased ROS levels caused increased lipid peroxidation (LPO) in treatment groups exposed to both fresh and aged NPs, although LPO was comparatively higher in algae exposed to aged AgNPs. The observed increase in catalase (CAT) activity of algal cells was attributed to early stress responses induced by excessive intracellular ROS generation, and CAT levels were higher in the aged NP treatment groups. In conclusion, AgNPs increased ROS levels and LPO in algae and caused the activation of antioxidant enzymes such as CAT. Overall, the results suggest that aging and coating of AgNPs have major impacts on AgNP transformation in media and their effects on algae. Environ Toxicol Chem 2019;38:2371-2382. © 2019 SETAC.

Additives migrating from 3D-printed plastic induce developmental toxicity and neuro-behavioural alterations in early life zebrafish (Danio rerio).

The environmental impact of exposure to 3D-printed plastics as well as potential migration of toxic chemicals from 3D-printed plastics remains largely unexplored. In this work we applied leachates from plastics fabricated using a stereolithography (SLA) process to early developmental stages of zebrafish (Danio rerio) to investigate developmental toxicity and neurotoxicity. Migration of unpolymerized photoinitiator, 1-hydroxycyclohexyl phenyl ketone (1-HCHPK) from a plastic solid phase to aqueous media at up to 200 mg/L in the first 24 h was detected using gas chromatography-mass spectrometry. Both plastic extracts (LC50 22.25% v/v) and 1-HCHPK (LC50 60 mg/L) induced mortality and teratogenicity within 48 h of exposure. Developmental toxicity correlated with in situ generation of reactive oxygen species (ROS), an increase in lipid peroxidation and protein carbonylation markers and enhanced activity of superoxide dismutase (SOD) and glutathione-S-transferase (GST) in embryos exposed to concentrations as low as 20% v/v for plastic extracts and 16 mg/L for 1-HCHPK. ROS-induced cellular damage led to induction of caspase-dependent apoptosis which could be pharmacologically inhibited with both antioxidant ascorbic acid and a pan-caspase inhibitor. Neuro-behavioral analysis showed that exposure to plastic leachates reduced spontaneous embryonic movement in 24-36 hpf embryos. Plastic extracts in concentrations above 20% v/v induced rapid retardation of locomotion, changes in photomotor response and habituation to photic stimuli with progressive paralysis in 120 hpf larvae. Significantly decreased acetylcholinesterase (AChE) activity with lack of any CNS-specific apoptotic phenotypes as well as lack of changes in motor neuron density, axonal growth, muscle segment integrity or presence of myoseptal defects were detected upon exposure to plastic extracts during embryogenesis. Considering implications of the results for environmental risk assessment and the growing usage of 3D-printing technologies, we speculate that some 3D-printed plastic waste may represent a significant and yet very poorly uncharacterized environmental hazard that merits further investigation on a range of aquatic and terrestrial species.