Exposure to virgin and marine incubated microparticles of biodegradable and conventional polymers modulates the hepatopancreas transcriptome of Mytilus galloprovincialis.

Biodegradable polymers have been proposed as an alternative to conventional plastics to mitigate the impact of marine litter, but the research investigating their toxicity is still in its infancy. This study evaluates the potential ecotoxicological effects of both virgin and marine-incubated microparticles (MPs), at environmentally relevant concentration (0.1 mg/l), made of different biodegradable polymers (Polycaprolactone, Mater-Bi, cellulose) and conventional polymers (Polyethylene) on Mytilus galloprovincialis by using transcriptomics. This approach is increasingly being used to assess the effects of pollutants on organisms, obtaining data on numerous biological pathways simultaneously. Whole hepatopancreas de novo transcriptome sequencing was performed, individuating 972 genes differentially expressed across experimental groups compared to the control. Through the comparative transcriptomic profiling emerges that the preponderant effect is attributable to the marine incubation of MPs, especially for incubated polycaprolactone (731 DEGs). Mater-Bi and cellulose alter the smallest number of genes and biological processes in the mussel hepatopancreas. All microparticles, regardless of their polymeric composition, dysregulated innate immunity, and fatty acid metabolism biological processes. These findings highlight the necessity of considering the interactions of MPs with the environmental factors in the marine ecosystem when performing ecotoxicological evaluations. The results obtained contribute to fill current knowledge gaps regarding the potential environmental impacts of biodegradable polymers.

Spatial distribution and characteristics of plastic pollution in the salt marshes of Bahía Blanca Estuary, Argentina.

This study delves into the magnitude and attributes of plastic pollution in the salt marshes of the Bahía Blanca Estuary, Argentina, with a specific focus on its spatial distribution. The investigation included the evaluation of microplastics (1-5 mm), mesoplastics (5-25 mm) and macroplastics (25-100 mm), discovering elevated levels along the high salt marsh strandline compared to low salt marsh and mudflat areas. Notably, the abundance of plastic reached staggering levels, reaching up to 20,060 items/m2 in the vicinity of an illegal dumpsite. Microplastics, particularly in the 2-4 mm range, were dominant, and the main plastic components were high-density polyethylene (HDPE), low-density polyethylene (LDPE), polypropylene (PP), and polystyrene (PS). Plastic films emerged as the predominant plastic type, while the presence of pellets hinted at potential sources such as illegal dumping and port-related activities. This contamination could be largely attributed to inappropriate waste management practices and urban runoff, which pose a substantial ecological threat to these ecosystems. Urgent remedial action is essential to protect these marshes, underscoring the critical need for comprehensive wetland management and educational initiatives to ensure their long-term sustainability.

Oceanographic and anthropogenic variables driving marine litter distribution in Mediterranean protected areas: Extensive field data supported by forecasting modelling.

Marine litter concentration in the Mediterranean Sea is strongly influenced both by anthropogenic pressures and hydrodynamic factors that locally characterise the basin. Within the Plastic Busters MPAs (Marine Protected Areas) Interreg Mediterranean Project, a comprehensive assessment of floating macro- and microlitter in the Pelagos Sanctuary and the Tuscan Archipelago National Park was performed. An innovative multilevel experimental design has been planned ad-hoc according to a litter provisional distribution model, harmonising and implementing the current sampling methodologies. The simultaneous presence of floating macro- and microlitter items and the potential influences of environmental and anthropogenic factors affecting litter distribution have been evaluated to identify hotspot accumulation areas representing a major hazard for marine species. A total of 273 monitoring transects of floating macrolitter and 141 manta trawl samples were collected in the study areas to evaluate the abundance and composition of marine litter. High mean concentrations of floating macrolitter (399 items/km2) and microplastics (259,490 items/km2) have been found in the facing waters of the Gulf of La Spezia and Tuscan Archipelago National Park as well in the Genova canyon and Janua seamount area. Accordingly, strong litter inputs were identified to originate from the mainland and accumulate in coastal waters within 10-15 nautical miles. Harbours and riverine outfalls contribute significantly to plastic pollution representing the main sources of contamination as well as areas with warmer waters and weak oceanographic features that could facilitate its accumulation. The results achieved may indicate a potentially threatening trend of litter accumulation that may pose a serious risk to the Pelagos Sanctuary biodiversity and provide further indications for dealing with plastic pollution in protected areas, facilitating future management recommendations and mitigation actions in these fragile marines and coastal environments.

Monk seal faeces as a non-invasive technique to monitor the incidence of ingested microplastics and potential presence of plastic additives.

Anthropogenic debris, including plastics, has recently been identified as a major threat for marine mammals and the Marine Strategy Framework Directive aims to achieve the good environmental status of European waters by addressing among other criteria, the effects of marine litter on biota. This study implemented for the first time a non-invasive technique for collecting monk seal samples to assess microdebris ingestion in combination with identifying plastic additives and porphyrins biomarkers. A total of 12 samples of monk seal faeces were collected from marine caves in Zakynthos Island, Greece. A total of 166 microplastic particles were identified; 75 % of the particles were smaller than 3 mm. Nine phthalates and three porphyrins were detected. A strong correlation was found between the number of microplastics and the concentration of phthalates. The values of both phthalates and porphyrins were found lower than in other marine mammal tissues, suggesting that seals might not be impacted by them yet.

Monitoring the composition, sources and spatial distribution of seafloor litter in the Adriatic Sea (Mediterranean Sea) through Fishing for Litter initiatives.

Although the seafloor is an important sink for marine litter, its less accessible environment makes seafloor litter the least studied component. Nevertheless, detailed monitoring of its composition and spatial distribution is urgently needed to develop appropriate mitigation strategies in areas exposed to multiple anthropogenic pressures, such as the Adriatic Sea. Commercial fisheries such as bottom trawls can serve as an opportunistic platform for collecting data on seafloor litter and help researchers in addressing the lack of data through Fishing for Litter initiatives. In this study, the crews of twelve trawlers from two Italian Adriatic ports collected seafloor litter as part of their Fishing for Litter initiatives. In addition to the classical scheme, the collected litter was sorted on board into different bags, and speed, duration and geographical coordinates of each haul were recorded to allow an assessment of both densities and spatial distribution. More than 600 kg of litter was removed from the seafloor by the fishers in approximately 1 month and characterised by the researchers on the docks. Most of the found items were made of synthetic polymers and related to packaging, fisheries and aquaculture activities. Abandoned, lost or discarded fishing gear and mussel nets were identified as of particular concern in the maritime compartment of Chioggia and Civitanova Marche, respectively. Three hotspots for marine litter were identified: near the Venice lagoon, in the centre of the northern basin and at greater depth in the central Adriatic. This study provides a detailed picture of the spatial distribution and composition of seafloor litter in the areas studied and highlights the importance of increased cooperation between fishermen and scientists to improve the identification of hotspots and sources while removing marine litter from the seafloor and raising awareness of the problem.

Microplastic occurrence and phthalate ester levels in neuston samples and skin biopsies of filter-feeding megafauna from La Paz Bay (Mexico).

The impacts of microplastics on filter feeders megafauna have recently received increased attention. These organisms are potentially exposed to plastic ingestion and the release of added/sorbed contaminants during feeding activities. An assessment of microplastic abundance and the chemical impact of Phthalates esters (PAEs) were performed in neustonic samples and skin biopsies of Balaenoptera physalus and Rhincodon typus inhabiting the Gulf of California (Mexico). Sixty-eight percent of the net tows contained plastics with a maximum of 0.24 items/m3 mainly composed of polyethylene fragments. PAE levels were detected both in environmental and skin biopsy samples, with the highest values in the fin whale specimens (5291 ng/g d.w). Plasticizer fingerprint showed a similar distribution pattern between neustonic samples and filter-feeding species, with DEHP and MBP having the highest concentrations. The detection of PAE levels confirmed their potential role as plastic tracers and give preliminary information about the toxicological status of these species feeding in La Paz Bay.

Sorbed environmental contaminants increase the harmful effects of microplastics in adult zebrafish, Danio rerio.

Aquatic animals ingest Microplastics (MPs) which have the potential to affect the uptake and bioavailability of sorbed co-contaminants. However, the effects on living organisms still need to be properly understood. The present study was designed to assess the combined effects of MPs and environmental contaminants on zebrafish (Danio rerio) health and behavior. Adult specimens were fed according to three different protocols: 1) untreated food (Control group); 2) food supplemented with 0.4 mg/L pristine polyethylene-MPs (PE-MPs; 0.1-0.3 mm diameter) (PEv group); 3) food supplemented with 0.4 mg/L PE-MPs previously incubated (PEi group) for 2 months in seawater. Analysis of contaminants in PEi detected trace elements, such as lead and copper. After 15 days of exposure, zebrafish underwent behavioral analysis and were then dissected to sample gills and intestine for histology, and the latter also for microbiome analysis. Occurrence of PEv and PEi in the intestine and contaminants in the fish carcass were analyzed. Both PEv- and PEi-administered fish differed from controls in the assays performed, but PEi produced more harmful effects in most instances. Overall, MPs after environmental exposure revealed higher potential to alter fish health through combined effects (e.g. proportion of microplastics, pollutants and/or microorganisms).

Trophic niche influences ingestion of micro- and mesoplastics in pelagic and demersal fish from the Western Mediterranean Sea.

Plastic pollution has been extensively documented in the marine food web, but targeted studies focusing on the relationship between microplastic ingestion and fish trophic niches are still limited. In this study we investigated the frequency of occurrence and the abundance of micro- and mesoplastics (MMPs) in eight fish species with different feeding habits from the western Mediterranean Sea. Stable isotope analysis (δ13C and δ15N) was used to describe the trophic niche and its metrics for each species. A total of 139 plastic items were found in 98 out of the 396 fish analysed (25%). The bogue revealed the highest occurrence with 37% of individuals with MMPs in their gastrointestinal tract, followed by the European sardine (35%). We highlighted how some of the assessed trophic niche metrics seem to influence MMPs occurrence. Fish species with a wider isotopic niche and higher trophic diversity were more probable to ingest plastic particles in pelagic, benthopelagic and demersal habitats. Additionally, fish trophic habits, habitat and body condition influenced the abundance of ingested MMPs. A higher number of MMPs per individual was found in zooplanktivorous than in benthivore and piscivorous species. Similarly, our results show a higher plastic particles ingestion per individual in benthopelagic and pelagic species than in demersal species, which also resulted in lower body condition. Altogether, these results suggest that feeding habits and trophic niche descriptors can play a significant role in the ingestion of plastic particles in fish species.

Detection of anthropogenic fibres in marine organisms: Knowledge gaps and methodological issues.

Most studies examining the occurrence of plastics and microplastics in marine organisms have identified anthropogenic fibres, of natural and synthetic origin, as the most commonly occurring category. Anthropogenic fibres may have been chemically treated with additives making them more persistent and a potential threat to marine organisms. However, fibres have often been excluded from analytical data for the difficulties related to the sampling and analytical procedures, including potential overestimation of the results due to airborne contamination. This review aimed to collect and analyse all studies focusing on the interaction between anthropogenic fibres and marine organisms worldwide, highlighting critical issues that need to be overcome for the analysis fibres on marine organisms. Furthermore, emphasis was placed on the species studied in the Mediterranean Sea, which is particularly affected by this type of pollution. Overall, this review shows that fibre pollution is an underestimated threat to marine organisms and that a specific, harmonised protocol for the analysis of different anthropogenic fibres needs to be developed.

Gradients of Variation in the At-Vessel Mortality Rate between Twelve Species of Sharks and Skates Sampled through a Fishery-Independent Trawl Survey in the Asinara Gulf (NW Mediterranean Sea).

Elasmobranchs are priority species for conservation due to their rapid decline determined by the unbalanced struggle between a fragile bio-ecology and strong anthropogenic impacts, such as bycatch from professional fishing. In this context, measuring species resistance to catch of poorly selective gear is of paramount importance. During June-October 2022, five experimental fishing campaigns were carried out in the Asinara Gulf (northern Sardinia) through 35 geographically and bathymetrically representative hauls of an area between 30 and 600 m in depth. Skates prevailed over sharks in the number of species, with seven and five species, respectively. We first evaluated the status of each individual with respect to stress due to the trawl's catch using a three-graded scale. We also recorded individual biometrics (total and disk length, weight and sex, and maturity for males) on board by implementing the best practices in manipulating individuals for physiological recovery and release at sea. After capture, skates resulted in generally better conditions than sharks, although deepwater species of both groups exhibited a worse state than coastal species. The estimated vitality rates also depended on the size of the individuals. This work provides standardized data on the intermingled effect of size, species type, and inhabited depth on the resistance response of some elasmobranch species against capture by trawl fishery activities.

Chemical pollution and ecotoxicological effects of high-density polyethylene microplastics in Mytilus galloprovincialis from two Italian lagoon ecosystems.

Transitional water ecosystems have low water exchanges and can trap chemicals and microplastics (MPs). In this study, MPs, trace elements, polycyclic aromatic hydrocarbon-PHAs levels and the oxidative stress response were assessed in Mytilus galloprovincialis from two Italian lagoon ecosystems (Orbetello and Varano). In addition, the ecotoxicological effects induced by the exposure of M. galloprovincialis to high-density polyethylene-HDPE MPs were also determined. Levels of trace elements were almost always comparable among the sites, whereas MPs were found only in mussels from Orbetello. PAHs were always under the limit of quantification. Glutathione peroxidase and malondialdehyde levels were significantly higher in mussels from Varano. As regard the exposure test, it was found a significant effect of treatment, site and their interaction on mortality and biochemical biomarkers in both fed and unfed mussels. However, principal component analysis suggests similar effects of both color and nourishment condition on biochemical biomarkers. These findings warrant further investigation.

Microplastics in water surface and in the gastrointestinal tract of target marine organisms in Salento coastal seas (Italy, Southern Puglia).

Microplastic pollution is a major global environmental threat that has attracted increasing interest from the scientific community over the past decade. The semi-closed and highly urbanized Mediterranean Sea has been investigated since 2012, in several specific studies that have identified it as a target hotspot for microplastic contamination. The marine coastal zone of the Salento peninsula (Apulia, Italy) has peculiar geographical and hydrodynamic features, although there are few published data detailing the level of microplastics present in this area. The present manuscript contains both data on the concentration of microplastics in surface waters and the level of microplastics ingested by selected marine organisms in the Salento coastal zone. Microplastics floating on the water surface were monitored during Autumn 2020 and Spring 2021 using neuston Manta net at three different distances from the coasts (Lizzano, Gallipoli and Otranto). The level of microplastic ingestion was monitored in fish species (Sardina pilchardus, Boops boops, Mullus barbatus) and in mussels (Mytilus galloprovincialis). Episodic peaks of microplastic concentrations were found on the sea surface during transects performed in the 3 nautic miles from the seashore. High values of ingested microplastics were found in S. pilchardus. and B. boops (5.4 and 4.6 items/individual respectively). A higher concentration of microplastics was detected in the Adriatic Sea than in the Ionian Sea by comparing the gastrointestinal tract of S. pilchardus and B. boops, in the monitored areas. These results are correlated with the concentration of floating microplastics, although this last result is not validated by statistical analysis. These results support the effectiveness of S. pilchardus and B. boops used as targets in monitoring activity for these pollutants. Results show a worrying increase in the concentration of microplastics on the sea surface and in the gastrointestinal tract of the target species compared to data reported in the literature.

First record of microplastic in the environmental matrices of a Mediterranean marine cave (Bue Marino, Sardinia, Italy).

This study investigates for the first time the presence of microplastics in sediment, water, and benthic organisms (foraminifera) of a marine cave in the Gulf of Orosei (Sardinia, Italy). Microplastics were found in all water, and sediment samples with similar shapes, sizes, and compositions; identified items were mainly fragments and fibers constituted by PVC and polyethylene. Their provenance was supposed to be predominantly from the sea than from the seasonal freshwater supplies from the karst system. Foraminiferal assemblages were mainly constituted by calcareous hyaline taxa in the outer station, while in the inner ones, the agglutinated Eggerelloides advenus was dominant. FTIR analyses on agglutinated shells identified polyethylene. Microplastic items are collected by the foraminifers and sediment grains building the shell chambers. This is the first study providing evidence that marine caves may be collectors of microplastics and that, in these habitats, microplastics enter the biotic matrix at the protist's level.

Integrated biomarker responses in European seabass Dicentrarchus labrax (Linnaeus, 1758) chronically exposed to PVC microplastics.

Few studies evaluated long-term effects of polyvinyl chloride (PVC) microplastics (MPs) ingestion in fish. The present study aimed to investigate the integrated biomarker responses in the liver and blood of 162 European seabass, Dicentrarchus labrax, exposed for 90 days to control, virgin and marine incubated PVC enriched diets (0.1 % w/w) under controlled laboratory condition. Enzymatic and tissue alterations, oxidative stress, gene expression alterations and genotoxicity were examined. Additives and environmental contaminants levels in PVC-MPs, control feed matrices and in seabass muscles were also detected. The results showed that the chronic exposure at environmentally realistic PVC-MPs concentrations in seabass, cause early warning signs of toxicological harm in liver by induction of oxidative stress, the histopathological alterations and also by the modulation of the Peroxisome proliferator-activated receptors (PPARs) and Estrogen receptor alpha (ER-α) genes expression. A trend of increase of DNA alterations and the observation of some neoformations attributable to lipomas suggest also genotoxic and cancerogenic effects of PVC. This investigation provides important data to understand the regulatory biological processes affected by PVC-MPs ingestion in marine organisms and may also support the interpretation of results provided by studies on wild species.

Microplastic abundance and biodiversity richness overlap: Identification of sensitive areas in the Western Ionian Sea.

Plastic pollution in the Mediterranean Sea has been widely reported, but its impact on biodiversity has not been fully explored. Simultaneous sampling of microplastics (MP) with a manta net and surveys of large marine vertebrates were conducted along the coastal waters of Sicily (Western Ionian Sea). A total of 17 neustonic samples have been collected and 17 marine species (cetaceans, sea turtles, seabirds, and fish) have been sighted in the target area. Kernel density estimation was evaluated to highlight a possible overlap between the presence of large marine fauna and MP densities to provide a preliminary risk assessment. The highest biodiversity and MP concentration (0.197 ±â€¯0.130 items/m2) were observed in the southernmost part of the studied area. The overlap between biodiversity hotspots and the occurrence of MP, potential contribute to the identification of sensitive areas of exposure in a poorly studied region.

Analysis of the Gastro-Intestinal Tract of Marine Mammals: A Multidisciplinary Approach with a New Multi-Sieves Tool.

Organs and content of the gastro-intestinal tract (GIT) of marine mammals are relevant for a variety of investigations and provide data to researchers from different fields. Currently used protocols applied to the GIT for specific analysis limit the possibility to execute other investigations and important information could be lost. To ensure a proper sample collection and a multidisciplinary investigation of the GIT of marine mammals, a new multi-sieves tool and a specific protocol have been developed. This new device and approach allowed the simultaneous sampling of the GIT and its content for the main investigations concerned. The samples collected during these preliminary trials were suitable to perform all the different research procedures considered in this work. The obtained results show that with a few and easy procedural adjustments, a multidisciplinary sampling and evaluation of the GIT of marine mammals is possible. This will reduce the risk of losing important data aimed at understanding the cause of death of the animal, but also biology and ecology of marine mammals, and other important data for their conservation and habitats management.

Effects of microplastics on head kidney gene expression and enzymatic biomarkers in adult zebrafish.

Due to massive production, improper use, and disposal of plastics, microplastics have become global environmental pollutants affecting both freshwater and marine ecosystems. Several studies have documented the uptake of microplastics in wild species and the correlated biological effects, such as epithelial damage, inflammation, metabolic alterations, and neurotoxicity. However, the effects of microplastics are not fully understood yet. In this study, adult zebrafish have been exposed for twenty days to two concentrations of a mix of polystyrene and high-density polyethylene microplastics. The biological effects were investigated through the expression levels of a set of selected genes in head kidney samples and two enzymatic biomarkers, acetylcholinesterase and lactate dehydrogenase, in head and body homogenates respectively. The lowest microplastic concentration up-regulated genes involved in xenobiotics catabolic processes (cyp2p8), and adaptive immunity (tcra). Acetylcholinesterase activity was inhibited by the highest microplastics exposure, while a weaker and no significant inhibition was induced by the lowest concentration. No significant effects on lactate dehydrogenase activity were observed. The results presented in this study support the hypothesis that MPs exposure could induce the activation of an immune response and the xenobiotic metabolism, suggesting also that the cytochrome P450 enzyme cyp2p8 and acetylcholinesterase may be sensitive to MPs contamination.

Seafloor litter along the Italian coastal zone: An integrated approach to identify sources of marine litter.

A heterogeneous amount of waste of different origins is continuously generated along Italian coasts in the Mediterranean Sea. In this paper, using information regarding the quantities and the different types of marine litter based on Project AWARE's "Dive Against Debris" data, we try to identify the main sources of this waste. In particular, the methodological approach used associates the origin of marine litter sources with the dataset of geo-localised state-owned maritime concessions, which are loaded by the granting bodies (regions, municipalities, port authorities) on the State Property Information System (Sid). The sources of marine litter were evaluated using the matrix scoring technique (MST). Then, to assess the weight of each source in each station, the community-level weighted mean (CWM) was calculated. Single-use plastic accounted for the highest percentage (19.13%), followed by glass beverage bottles (10.90%), shopping bags (9.03%), aluminium beverage cans (4.91%), and cigarette butts (4.61%). Tourism and beach users contributed to 42.3% of the litter found, followed by fishing (15.7%) and yachting (11.3%). The overlapping of the data collected by scuba divers with the state concessions of activities from offshore and mainland areas was used to distinguish the anthropic pressures that impact the coast. Policy makers and the local administrator may use these results to define new methods of collection and reuse of anthropic waste through a more harmonised approach in the management of marine waste.

Interlaboratory comparison of microplastic extraction methods from marine biota tissues: A harmonization exercise of the Plastic Busters MPAs project.

In the framework of the Plastic Busters MPAs project, a harmonization exercise on two methods of microplastic extraction from biological samples i.e. 15% H2O2 digestion and 10% KOH digestion was carried out. The two methods were tested in four laboratories on fish gastrointestinal tracts and mussel tissues spiked with polyethylene, polypropylene and polyethylene terephthalate. The recovery percentage of microplastics for each method, species and polymer tested were overall similar among laboratories, and interlaboratory coefficient of variation was less than 11% for the majority of samples. Microplastic recovery rates for the two methods were similar for each sample tested, but overall mean interlaboratory recovery rate using KOH (96.67%) was higher than H2O2 (88.75%). Results validate the use of both methods for extracting microplastics from biota tissues. However, when comparing the two methods in terms of microplastic recovery rate, time consumed, technical difficulties and cost, digestion with 10% KOH is considered optimal.

Skin distress associated with xenobiotics exposure: An epigenetic study in the Mediterranean fin whale (Balaenoptera physalus).

The phenotypic plasticity of many organisms is mediated in part by epigenetics, the heritable changes in gene activity that occur without any alterations to DNA sequence. A major mechanism in epigenetics is the DNA methylation (DNAm). Hypo- and hyper-methylation are generalized responses to control gene expression however recent studies have demonstrated that classes of contaminants could mark specific DNAm signatures, that could usefully signal prior environmental exposure. We collected skin and blubber from 6 free-ranging fin whale (Balaenoptera physalus) individuals sampled as a part of a previous published study in the northern Mediterranean Sea. Genomic DNA extracted from the skin of the fin whales and levels of contaminants measured in the blubber of the same individuals were used for DNAm profiling through reduced representation bisulfite sequencing (RRBS). We tested the hypothesis that differences in the methylation patterns could be related to environmental exposure to contaminants and load in the whale tissues. The aims of this study were to determine the DNAm profiles of the methylation contexts (CpGs and non-CpGs) of differently contaminated groups using the RRBS, and to identify potential contaminant exposure related genes. Amount and proportion of methylcytosines in CpG and non-CpG regions (CHH and CHG) was very similar across the 6 samples. The proportion of methylcytosines sites in CpG was n = 32,682, the highest among all the sequence contexts (n = 3216 in CHH; n = 1743 in CHG). The majority of the methylcytosine occurred in the intron regions, followed by exon and promoter regions in CpG, CHH and CHG. Gene Ontology results indicated that DNAm affected genes that take place in cell differentiation and function in cutaneous, vascular and nervous systems. The identification of cellular response pathways allows a better understanding of the organism biological reaction to a specific environmental challenge and the development of sensitive tools based on the predictive responses. Eco-epigenetics analyses have an extraordinary potential to address growing issues on pollution biomonitoring, ecotoxicity assessment, conservation and management planning.