High Amounts of Halogenated Natural Products in Sperm Whales (Physeter macrocephalus) from Two Italian Regions in the Mediterranean Sea.

Halogenated natural products (HNPs) are considered to be emerging contaminants whose environmental distribution and fate are only incompletely known. Therefore, several persistent and bioaccumulative HNP groups, together with man-made polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), were quantified in the blubber of nine sperm whales (Physeter macrocephalus) stranded on the coast of the Mediterranean Sea in Italy. The naturally occurring polybrominated hexahydroxanthene derivatives (PBHDs; sum of TetraBHD and TriBHD) were the most prominent substance class with up to 77,000 ng/g blubber. The mean PBHD content (35,800 ng/g blubber) even exceeded the one of PCBs (28,400 ng/g blubber), although the region is known to be highly contaminated with man-made contaminants. Based on mean values, Q1 ∼ PBDEs > MeO-BDEs ∼ 2,2'-diMeO-BB 80 and several other HNPs followed with decreasing amounts. All blubber samples contained an abundant compound whose molecular formula (C16H19Br3O2) was verified using high-resolution mass spectrometry. The only plausible matching isomer was (2S,4'S,9R,9'S)-2,7-dibromo-4'-bromomethyl-1,1-dimethyl-2,3,4,4',9,9'-9,9'-hexahydro-1H-xanthen-9-ol (OH-TriBHD), a hydroxylated secondary metabolite previously detected together with TriBHD and TetraBHD in a sponge known to be a natural producer of PBHDs. The estimated mean amount of the presumed OH-TriBHD was 3000 ng/g blubber, which is unexpectedly high for hydroxylated compounds in the lipids of marine mammals.

Assessment of the impact of microplastic ingestion in striped red mullets from an Eastern Mediterranean coastal area (Zakynthos Island, Ionian Sea).

Monitoring microplastics (MPs) in the marine environment is an ongoing process, and our understanding of their impact on marine organisms is limited. The present study evaluates the effects of ingested MPs on the marine MP pollution bioindicator fish species Mullus surmuletus. The study follows a three-fold approach to assess the impact of MPs on marine organisms by investigating: 1) the ingestion of MPs, 2) the bioaccumulation of phthalate compounds as plastic additives, and 3) the evaluation of toxicological biochemical and cellular biomarkers. Striped red mullets were sampled in the marine protected area (MPA) of the National Marine Park of Zakynthos and coastal sites with high touristic pressure in Zakynthos Island in the Ionian Sea, Greece. Fewer ingested MPs and lower phthalate concentrations were found in fish inside the MPA compared to those sampled outside the marine park. However, no relationship was found between either phthalate concentrations or biomarker levels with the ingested MPs in the red striped mullets. Biomarker levels were influenced by season and site, but no effect could be attributed to the ingested MPs. The lack of association of biomarker responses and plasticizer bioaccumulation to MP ingestion can be explained by the low number of ingested MPs in the fish from Zakynthos coastal area as MP abundance ranged from 0.15 to 0.55 items per individual fish.

Novel ultrasound assisted extraction and d-SPE clean-up for the analysis of multiple legacy and emerging organic contaminants in edible fish.

Polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), novel brominated flame retardants (NBFRs), phthalate esters (PAEs) are pervasive environmental pollutants, posing threats to both ecosystems and human health. Although several analytical methods were developed for these compounds, they are not performed simultaneously. This study addresses the need for a sustainable, novel, analytical approach capable of simultaneously determining these diverse chemical classes in edible fish muscles. Employing ultrasound extraction coupled with dispersive solid-phase extraction (d-SPE) as a cleanup procedure, the method was compared to conventional techniques, revealing significant improvements. Analytical parameters were thoroughly assessed, and the innovative method demonstrated notable advantages, reducing extraction and purification times by approximately 74-80 % and solvent consumption by around 94-97 %. Applied to Mediterranean Sea fish samples, the results underscore the method's potential as a viable, sustainable alternative to traditional approaches, promising enhanced efficiency and reduced environmental impact.

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.

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.

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.

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).

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.

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.

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.

A multidisciplinary integrated approach using Pachygrapsus marmoratus to assess the impact of port activities on mediterranean marine protected areas.

The establishment of marine protected areas is considered the main global strategy to halt the loss of marine biodiversity. Since most of marine areas are open systems, this form of habitat protection cannot prevent their contamination due to human activities performed outside of their borders. Innovative approaches to assess the health status of protected marine habitats are therefore needed. Here we developed a multidisciplinary approach that combines ecological characteristics, bioaccumulation of inorganic and organic pollutants, cell damage (micronuclei frequency, nuclear alterations and LPO) and enzymatic (AChE, CAT, IDH, LDH, GST and CAT) markers focused on an intertidal brachyuran crab, Pachygrapsus marmoratus, to assess the impacts of contaminant exposure on Mediterranean coastal habitats. As study sites we selected two protected areas and two sites within industrial ports of the Ligurian Sea. Our results showed that the selected crab species is an excellent bioindicator. Individuals collected in sites with the highest levels of heavy metal pollution showed the highest signals of stress responses at both cellular and enzymatic levels, coupled with a high incidence of the parasite Sacculina carcini, a signal of impairment of their standard development and reproduction cycle. We could also prove that one of the selected marine protected areas showed the same intensity of impact as its adjacent port site. Our multidisciplinary approach proved to be a valuable tool to assess the environmental quality and health of protected and disturbed Mediterranean coastal environments and to inform efficient management and protection schemes for such habitats.

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.

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.

The Culturable Mycobiota of Sediments and Associated Microplastics: From a Harbor to a Marine Protected Area, a Comparative Study.

Fungi are an essential component of marine ecosystems, although little is known about their global distribution and underwater diversity, especially in sediments. Microplastics (MPs) are widespread contaminants worldwide and threaten the organisms present in the oceans. In this study, we investigated the fungal abundance and diversity in sediments, as well as the MPs, of three sites with different anthropogenic impacts in the Mediterranean Sea: the harbor of Livorno, the marine protected area "Secche della Meloria"; and an intermediate point, respectively. A total of 1526 isolates were cultured and identified using a polyphasic approach. For many of the fungal species this is the first record in a marine environment. A comparison with the mycobiota associated with the sediments and MPs underlined a "substrate specificity", highlighting the complexity of MP-associated fungal assemblages, potentially leading to altered microbial activities and hence changes in ecosystem functions. A further driving force that acts on the fungal communities associated with sediments and MPs is sampling sites with different anthropogenic impacts.

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.