Nanoplastics affect moulting and faecal pellet sinking in Antarctic krill (Euphausia superba) juveniles.

Plastic debris has been identified as a potential threat to Antarctic marine ecosystems, however, the impact of nanoplastics (<1 µm) is currently unexplored. Antarctic krill (Euphausia superba) is a keystone species of Southern Ocean pelagic ecosystems, which plays a central role in the Antarctic food webs and carbon (C) cycle. Krill has been shown to rapidly fragment microplastic beads through the digestive system, releasing nanoplastics with unknown toxicological effects. Here we exposed krill juveniles to carboxylic (COOH, anionic) and amino- (NH2, cationic) polystyrene nanoparticles (PS NPs) and we investigated lethal and sub-lethal endpoints after 48 h. The analysis of PS NP suspensions in Antarctic sea water (SW) media showed that PS-COOH formed large agglomerates (1043 ± 121 nm), while PS-NH2 kept their nominal size (56.8 ± 3 nm) during the exposure time. After 48 h, no mortality was found but increase in exuviae production (12.6 ± 1.3%) and reduced swimming activity were observed in juveniles exposed to PS-NH2. The microbial community composition in SW supports the release of krill moults upon PS NP exposure and stimulates further research on the pivotal role of krill in shaping Southern Ocean bacterial assemblages. The presence of fluorescent signal in krill faecal pellets (FPs) confirmed the waterborne ingestion and egestion of PS-COOH at 48 h of exposure. Changes in FP structure and properties were also associated to the incorporation of PS NPs regardless of their surface charge. The effects of PS NPs on krill FP properties were compared to Control 0 h as a reference for full FPs (plastic vs food) and Control 48 h as a reference for more empty-like FPs (plastic vs lack of food). Exposure to PS NPs led to a FP sinking rate comparable to Control 48 h, but significantly lower than Control 0 h (58.40 ± 23.60 m/d and 51.23 ± 28.60 m/d for PS-COOH and PS-NH2; 168.80 ± 74.58 m/d for Control 0 h). Considering the important role played by krill in the food web and C export in the Southern Ocean, the present study provides cues about the potential impact of nanoplastics on Antarctic pelagic ecosystems and their biogeochemical cycles.

Uptake and biological responses in land snail Cornu aspersum exposed to vaporized CdCl2.

The uptake of Cd and some biomarkers of exposure and effects have been investigated in specimens of land snail Cornu aspersum exposed to vaporized CdCl2 (10mg/L) for 7 days. The Cd levels quantified in snail's whole bodies confirmed Cd bioavailability trough vaporization and an higher accumulation in the midgut gland compared to the foot. Biological responses investigated showed a reduction of destabilization time of lysosomal membranes (NRRT) in hemocytes and an induction of catalase activities (CAT) in midgut gland. A further evidence of CdCl2 vaporized exposure was given by an increase in MT protein content as well as induction of Cd-MT gene expression, highlighting the central role of the midgut gland in Cd detoxification. These biomarkers can thus be considered as sensitive tools for the assessment of Cd contamination in the air using land snails as bioindicators. No changes in of GST activity and MDA were observed. From the overall results, the land snail, C. aspersum, could be used as good bioindicator of air quality for pollution monitoring purposes having shown clear signs of exposure and effects due Cd exposure by air.

Long-term toxicity of surface-charged polystyrene nanoplastics to marine planktonic species Dunaliella tertiolecta and Artemia franciscana.

Plastic pollution has been globally recognized as a critical issue for marine ecosystems and nanoplastics constitute one of the last unexplored areas to understand the magnitude of this threat. However, current difficulties in sampling and identifying nano-sized debris make hard to assess their occurrence in marine environment. Polystyrene nanoparticles (PS NPs) are largely used as nanoplastics in ecotoxicological studies and although acute exposures have been already investigated, long-term toxicity on marine organisms is unknown. Our study aims at evaluating the effects of 40nm PS anionic carboxylated (PS-COOH) and 50nm cationic amino-modified (PS-NH2) NPs in two planktonic species, the green microalga Dunaliella tertiolecta and the brine shrimp Artemia franciscana, respectively prey and predator. PS NP behaviour in exposure media was determined through DLS, while their toxicity to microalgae and brine shrimps evaluated through 72h growth inhibition test and 14 d long-term toxicity test respectively. Moreover, the expression of target genes (i.e. clap and cstb), having a role in brine shrimp larval growth and molting, was measured in 48h brine shrimp larvae. A different behaviour of the two PS NPs in exposure media as well as diverse toxicity to the two planktonic species was observed. PS-COOH formed micro-scale aggregates (Z-Average>1µm) and did not affect the growth of microalgae up to 50µg/ml or that of brine shrimps up to 10µg/ml. However, these negatively charged NPs were adsorbed on microalgae and accumulated (and excreted) in brine shrimps, suggesting a potential trophic transfer from prey to predator. On the opposite, PS-NH2-formed nano-scale aggregates (Z-Average<200nm), caused inhibition of algal growth (EC50=12.97µg/ml) and mortality in brine shrimps at 14 d (LC50=0.83µg/ml). Moreover, 1µg/ml PS-NH2 significantly induced clap and cstb genes, explaining the physiological alterations (e.g. increase in molting) previously observed in 48h larvae, but also suggesting an apoptotic pathway triggered by cathepsin L-like protease in brine shrimps upon PS-NH2 exposure. These findings provide a first insight into long-term toxicity of nanoplastics to marine plankton, underlining the role of the surface chemistry in determining the behaviour and effects of PS NPs, in terms of adsorption, growth inhibition, accumulation, gene modulation and mortality. The use of long-term end-point has been identified as valuable tool for assessing the impact of nanoplastics on marine planktonic species, being more predictable of real exposure scenarios for risk assessment purposes.