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.

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.

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.

Unsuitable use of DMSO for assessing behavioral endpoints in aquatic model species.

Dimethyl sulfoxide (DMSO) is a universally used aprotic solvent with the ability to permeate biological membranes and thus is commonly used to achieve appropriate biological availability of hydrophobic toxicants. While DMSO as a carrier medium has a reportedly low toxicity and is routinely employed in ecotoxicology, very little is known about its effect on dynamic behavioral parameters. This study presents a comparative analysis of the lethal and behavioral effects of exposures to DMSO concentrations of 0.1-10% on several test species such as: neonates of the freshwater crustacean Daphnia magna, nauplii of the marine crustacean Artemia franciscana, the marine crustacean Allorchestes compressa, embryos and larvae of the freshwater fish Danio rerio. The results demonstrated that DMSO did not cause statistically significant mortality even at concentrations close to 1% but induced clear and significant behavioral abnormalities in response to sublethal concentrations on all test species. These included hypoactivity syndrome in A. franciscana, A. compressa, D. magna and zebrafish larvae while a slight time-dependent hyperactivity response was observed in zebrafish embryos. For the majority of test species, behavioral changes such as moving distance, acceleration and burst movement were often observed during the first hours of exposure. These results indicate that caution should be exercised when using DMSO as a carrier solvent in experiments assessing behavioral endpoints.