Microplastic exposure and effects in aquatic organisms: A physiological perspective.

The impact of microplastics (MPs) on aquatic life, given their ubiquitous presence in the water compartment, represents a growing concern. Consistently, scientific knowledge is advancing rapidly, although evidence on actual adverse effects is still highly fragmented. This paper summarizes the recent literature on MP impacts on aquatic organisms in an attempt to link routes of uptake, possible alterations of physiological processes, and outcomes at different levels of biological organization. Animal feeding strategies and MP biodistribution is discussed, alongside with relevant effects at molecular, cellular, and systemic level. Pathways from animal exposure to apical physiological responses are examined to define the relevance of MPs for animal health, and to point out open questions and research gaps. Emphasis is given to emerging threats posed by leaching of plastic additives, many of which have endocrine disruption potential. The potential role of MPs as substrates for microorganism growth and vehicle for pathogen spreading is also addressed.

The Multixenobiotic resistance system as a possible protective response triggered by microplastic ingestion in Mediterranean mussels (Mytilus galloprovincialis): Larvae and adult stages.

The emerging paradigm on plastic pollution in marine environments is that microsize particles (MPs) have far more subtle effects than bigger fragments, given their size range overlapping with that of particles ingested by filter-feeders. The impacts include gut blockage, altered feeding and energy allocation, with knock-on effects on widespread physiological processes. This study investigated whether ingestion of polystyrene MPs (PS-MPs) triggers protective processes in marine mussels. The Multixenobiotic resistance (MXR) system is a cytoprotective mechanism acting as an active barrier against harmful xenobiotics and a route of metabolite detoxification. Both larvae and adults were employed in laboratory experiments with different concentrations of 3-µm PS-MPs (larvae), and 3-µm and 45-µm PS-MPs (adults) matching size range of planktonic food through the mussel lifecycle. Embryos grown in the presence of 3-µm PS-MPs showed significant reduction of MXR activity and down-regulation of ABCB and ABCC transcripts encoding the two main MXR-related transporters P-glycoprotein and the Multidrug resistance-related protein, respectively. In adults, effects of PS-MPs were assessed in haemocytes and gills, which showed different modulation of MXR activity and ABCB/ABCC expression according to MP size (haemocyte and gills) or particle concentration (haemocyte). These data showed that modulation of MXR activity is part of a generalized response triggered by particle ingestion.

Uptake and transcriptional effects of polystyrene microplastics in larval stages of the Mediterranean mussel Mytilus galloprovincialis.

The widespread occurrence of microplastics (MP) in the marine environment is cause of increasing concerns about the safety of the exposed ecosystems. Although the effects associated to the MP uptake have been studied in most marine taxa, the knowledge about their sub-lethal impacts on early life stages of marine species is still limited. Here, we investigated the uptake/retention of 3-µm polystyrene MP by early stages of the Mediterranean mussel Mytilus galloprovincialis, and the related effects on gut clearance, feeding efficiency, morphological and transcriptional parameters involved in embryo-larval development. Uptake measurements were performed on larvae at 48 h, 3, 6 and 9 days post fertilization (pf) after exposure to a range of 50-10,000 particles mL-1. At all tested pf periods, treatments resulted in a significant and linear increase of MP uptake with increasing concentrations, though levels measured at 48 h pf were significantly lower compared to 3-9 d pf. Ingested MP were retained up to 192 h in larvae's gut, suggesting a physical impact on digestive functions. No change was noted between the consumption of microalgae Nannochloropsis oculata by larvae when administered alone or in the presence of an identical concentration (2000 items mL-1) of MP. The exposure to 50-10,000 MP mL-1 did not alter the morphological development of mussel embryos; however, transcriptional alterations were observed at 50 and 500 MP mL-1, including the up-regulation of genes involved in shell biogenesis (extrapallial protein; carbonic anhydrase; chitin synthase) and immunomodulation (myticin C; mytilin B), and the inhibition of those coding for lysosomal enzymes (hexosaminidase; ß-glucorinidase; catepsin-L). In conclusion, though not highlighting morphological or feeding abnormalities, data from this study revealed the onset of physical and transcriptional impairments induced by MP in mussel larvae, indicating sub-lethal impacts which could increase their vulnerability toward further environmental stressors.

Impact of cationic polystyrene nanoparticles (PS-NH2) on early embryo development of Mytilus galloprovincialis: Effects on shell formation.

The potential release of nanoparticles (NPs) into aquatic environments represents a growing concern for their possible impact on aquatic organisms. In this light, exposure studies during early life stages, which can be highly sensitive to environmental perturbations, would greatly help identifying potential adverse effects of NPs. Although in the marine bivalve Mytilus spp. the effects of different types of NPs have been widely investigated, little is known on the effects of NPs on the developing embryo. In M. galloprovincialis, emerging contaminants were shown to affect gene expression profiles during early embryo development (from trocophorae-24 hpf to D-veligers-48 hpf). In this work, the effects of amino-modified polystyrene NPs (PS-NH2) on mussel embryos were investigated. PS-NH2 affected the development of normal D-shaped larvae at 48 hpf (EC50 = 0.142 mg/L). Higher concentrations (5-20 mg/L) resulted in high embryotoxicity/developmental arrest. At concentrations ≅ EC50, PS-NH2 affected shell formation, as shown by optical and polarized light microscopy. In these conditions, transcription of 12 genes involved in different biological processes were evaluated. PS-NH2 induced dysregulation of transcription of genes involved in early shell formation (Chitin synthase, Carbonic anhydrase, Extrapallial Protein) at both 24 and 48 hpf. Decreased mRNA levels for ABC transporter p-glycoprotein-ABCB and Lysozyme were also observed at 48 hpf. SEM observations confirmed developmental toxicity at higher concentrations (5 mg/L). These data underline the sensitivity of Mytilus early embryos to PS-NH2 and support the hypothesis that calcifying larvae of marine species are particularly vulnerable to abiotic stressors, including exposure to selected types of NPs.

Heavy metals in tissues of loggerhead turtles (Caretta caretta) from the northwestern Adriatic Sea.

Thirty-five specimens of Caretta caretta were collected dead along the Adriatic Sea coast from the Po Delta to the Reno mouth (Italy). Turtles were classified into four size categories ranging from 24.5 to 74 cm, by measuring the minimum straight-line carapace length (MSCL). Cd, Cu, Fe, Mn, Ni, and Zn levels were assessed in liver, lung, muscle and adipose tissue. Cd, Cu and Fe mainly accumulated in the liver (8.9, 23.7 and 1180 mg/kg dry mass [d.w.], respectively), and Mn in the lung (29.5 mg/kg d.w.). Levels of Ni were higher in adipose (22 mg/kg d.w.) than other tissues, while Zn concentrations were higher in muscle (about 140 mg/kg d.w.). Negative correlations with size were established for Zn in liver and Cu in adipose tissue, while positive correlations were observed for Mn and Ni in adipose tissue. Metal concentrations did not differ between males and females, nor between individuals found stranded and those victims of by-catch. On average, Cd, Cu, Mn and Ni concentrations in our specimens were higher than in loggerhead turtles and other species living in other areas. We hypothesize that trace metals could be used as "acquired markers" to help investigate migration routes of C. caretta.