Are we even close? Five years marine litter ingestion monitoring in loggerhead turtles along Italian coast reveals how far we are from the Good Environmental Status.

The loggerhead sea turtle Caretta caretta has been chosen as bioindicator to monitor the amount of litter ingested by marine animals within the European Marine Strategy Framework Directive and the Barcelona Regional Sea Convention. European Member States and Contracting Parties are committed to achieve the Good Environmental Status (GES), which is reached when the quantity of ingested litter does not adversely affect the health of the species concerned. Although the monitoring strategy has been outlined for more than a decade, to date no threshold values have been adopted to verify GES achievement. After five years of extensive monitoring along the Italian coasts, this study evaluates the suitability of five different GES scenarios and proposes a new threshold value (i.e., "there should be less than 33% of sea turtles having more than 0.05 g of ingested plastic in the GI") for its implementation in the European seas and the Mediterranean basin.

Phthalate levels in common sea anemone Actinia equina and Anemonia viridis: A proxy of short-term microplastic interaction?

Phthalates are widely employed plasticizers blended to plastic polymers that, during plastic aging and weathering are prone to leach in the surrounding environment. Thus, phthalates were proposed to indirectly evaluate MPs contamination in marine environments, with still uncertain and scarce data, particularly for wildlife. This study investigates simultaneously microplastics (MPs) and phthalates (PAEs) occurrence in wild Actinia equina and Anemonia viridis, two common and edible sea anemone species. Both species had a 100 % frequency of MPs occurrence, with similar average concentrations. PAEs were detected in 70 % of samples, with concentrations up to 150 ng/g in A. equina and 144.3 ng/g for A. viridis. MPs and PAEs present in sea anemone tissues appear to reflect seawater plastic contamination conditions in the study area. Given the rapid biodegradation of PAEs, occurrence and concentrations of both these additives and their metabolites could be useful tracers of short-term plastic debris-biota interactions.

Tracing the route: Using stable isotope analysis to understand microplastic pathways through the pelagic-neritic food web of the Tyrrhenian Sea (Western Mediterranean).

This study provides a pilot investigation of the relationship between microplastic ingestion and the trophic ecology of three pelagic fish species (Engraulis encrasicolus, Scomber scombrus, and Trachurus trachurus) from Anzio coast, Tyrrhenian Sea (Western Mediterranean). Stable isotope analysis has been performed to determine the trophic position and the isotopic niche of the three species. Then, data on the occurrence, abundance, and diversity of ingested microplastics have been analyzed considering the observed foraging patterns. The detected differences in the estimated trophic position (E. encrasicolus = 3.08 ± 0.18; S. scombrus = 3.57 ± 0.21; T. trachurus = 4.07 ± 0.21), together with the absence of overlap in the isotopic niches confirm that the three examined species cover different ecological roles within the coastal-pelagic food web. Results from the analysis of ingested microplastics show that the trophic position has no remarkable effects on the incidence of microplastic ingestion, with no significant differences detected in terms of both frequency of occurrence and number of ingested microplastics per individual. However, differences among species emerge when considering the diversity of ingested microplastic types in terms of shape, size, color, and polymer composition. Species at higher trophic levels have shown to ingest a greater diversity of microplastics, including a significant increase in the size of the ingested particles (median surface area: 0.011 mm2 in E. encrasicolus; 0.021 mm2 in S. scombrus; 0.036 mm2 in T. trachurus). The ingestion of larger microplastics might be due to the larger gape sizes but also to active selection mechanisms, likely stimulated by the similarity of these particles to natural or potential prey of both S. scombrus and T. trachurus. Overall, this study suggests that microplastic ingestion can be affected by the different trophic position of fish species, providing new insights about the impact of microplastic contamination on the pelagic community.

From inshore to offshore: distribution of microplastics in three Italian seawaters.

A comprehensive understanding of the concentration of microplastics (MPs) in seawaters is essential to implement monitoring programs and understand the impacts on ecosystems, as required by the European legislation to protect the marine environment. In this context, the purpose of this study is to investigate the composition, quantity, and spatial distribution of microplastics from coastal to offshore areas in three Italian seawaters. In addition, the distribution of microplastics between surface and subsurface water layers was analyzed in order to better understand the dynamics of MPs in the upper layers of the water column. A total number of 6069 MPs (mean total concentration of 0.029 microplastics · m-2) were found to be heterogeneous in type, shape, and color. In general, MPs concentrations decrease with coastal distance, except when environmental forcings are predominant (such as sea currents). Moreover, the amount of surface MPs was almost four times that of subsurface microplastics, which consisted mostly of fibers. In light of these results, it becomes clear how critical it is to plan remediation actions and programs to minimize microplastic accumulations in the sea.

Image processing tools in the study of environmental contamination by microplastics: reliability and perspectives.

Microplastic pollution is one of the greatest environmental concerns for contemporary times and the future. In the last years, the number of publications about microplastic contamination has increased rapidly and the list is daily updated. However, the lack of standard analytical approaches might generate data inconsistencies, reducing the comparability among different studies. The present study investigates the potential of two image processing tools (namely the shapeR package for R and ImageJ 1.52v) in providing an accurate characterization of the shape of microplastics using a restricted set of shape descriptors. To ascertain that the selected tools can measure small shape differences, we perform an experiment to verify the detection of pre-post variations in the shape of different microplastic types (i.e., nylon [NY], polyethylene [PE], polyethylene terephthalate [PET], polypropylene [PP], polystyrene [PS], and polyvinylchloride [PVC]) treated with mildly corrosive chemicals (i.e., 10% KOH at 60 °C, 30% H2O2 at 50 °C, and 15% H2O2 + 5% HNO3 at 40 °C; incubation time ≈ 12 h). Analysis of surface area variations returns results about the vulnerability of plastic polymers to digestive solutions that are aligned with most of the acquired knowledge. The largest decrease in surface area occurs for KOH-treated PET particles, while NY results in the most susceptible polymer to the 30% H2O2 treatment, followed by PVC and PS. PE and PP are the most resistant polymers to all the used treatments. The adopted methods to characterize microplastics seem reliable tools for detecting small differences in the shape and size of these particles. Then, the analytic perspectives that can be developed using such widely accessible and low-cost equipment are discussed.

One is not enough: Monitoring microplastic ingestion by fish needs a multispecies approach.

The development of monitoring programs based on bioindicators is crucial for assessing the impact of microplastic ingestion on marine organisms. This study presents results from an Italian pilot action aimed at investigating the suitability of a monitoring strategy based on a multispecies approach. The benthic-feeder Mullus barbatus, the demersal species Merluccius merluccius, and the pelagic-feeder species of the genus Scomber were used to assess the environmental contamination by microplastics in three different marine areas, namely Ancona (Adriatic Sea), Anzio (Tyrrhenian Sea), and Oristano (Western Sardinia). Microplastic ingestion frequencies were higher in samples from Anzio (26.7 %) and Ancona (25.0 %) than Oristano (14.4 %), suggesting a relationship between microplastic bioavailability and the proximity to urban settlements and river flows. Furthermore, microplastic ingestion was affected by the feeding habits of the examined species. The detected differences reinforce the hypothesis that a multispecies approach is needed to evaluate microplastic ingestion by marine animals.

Distribution of seafloor litter and its interaction with benthic organisms in deep waters of the Ligurian Sea (Northwestern Mediterranean).

The Mediterranean Sea is one of the most polluted marine basins and currently serves as a hotspot for marine litter. The seafloor represents the ultimate sink for most litter worldwide. Nevertheless, the knowledge about litter distribution and its interactions with benthic organisms in deep water is poorly understood. In 2018, we investigated spatial patterns of macro- and micro-litter distribution, and their effects on benthic communities in the Ligurian Sea. An oceanographic survey was carried out with a remotely operated vehicle and a multibeam echosounder on seven seamounts and canyons, at depths ranging from 350 to 2200 m. High litter accumulations were discovered at the mouth of the Monaco canyon, where estimated densities of up to 3.8 × 104 items km-2 were found at 2200 m depth. The highest abundance of urban litter items was found on the soft substrate, at the bottom of the deeper parts of the submarine canyons, which seem to act as conduits carrying litter from the shelf towards deeper areas. In contrast, fishing-related items were most abundant in the upper layer of the seamounts (300-600 m depths). Furthermore, more than 10% of the observed deep gorgonian colonies were entangled by lost longlines, indicating the detrimental effects of this fishing gear on benthic habitats. The discovery of new litter hotspots and the evaluation of how deep-sea species interact with litter contribute to increasing the knowledge about litter distribution and its effects on the deep ecosystem of the Mediterranean basin. All the observations recorded in this study showed substantial and irreversible changes in the deep and remote areas of marine environments, and these changes were found to be caused by humans. Our findings further stress the need for urgent and specific measures for the management of deep-sea pollution and the reduction of litter inputs in the environment.

Using Boops boops (osteichthyes) to assess microplastic ingestion in the Mediterranean Sea.

This study assesses microplastic ingestion in Boops boops at different geographical areas in the Mediterranean Sea. A total of 884 fish were caught at 20 coastal sites in Spain, France, Italy and Greece and analyzed using a common methodological protocol. Microplastics were found in 46.8% of the sampled fish, with an average number of items per individual of 1.17 ± 0.07. Filaments were the predominant shape type, while polyethylene and polypropylene were indicated by FTIR as the most common polymer types of ingested microplastics. The frequency of occurrence, as well as the abundance and proportion of types (size, shape, color and polymer) of ingested microplastics, varied among geographical areas. The spatial heterogeneity of the abundance of ingested microplastics was mainly related to the degree of coastal anthropogenic pressure at the sampling sites. Our findings further support the suitability of B. boops as bioindicator of microplastic pollution in the Mediterranean Sea.

Macro-litter ingestion in deep-water habitats: is an underestimation occurring?

Marine litter affects marine organisms, posing threats to biodiversity conservation and to ecosystem structure and functioning. Providing a suitable assessment of marine litter effects on marine life through bioindicator species is crucial to drive an effective waste management policy. However, to date no standardized tool has been developed to describe and monitor the impact of marine macro-litter on marine life within deep-water habitats. Modifying the protocol proposed to monitor macro-litter ingestion by the loggerhead sea turtle Caretta caretta, we perform a preliminary investigation on the suitability of different elasmobranch species for monitoring macro-litter ingestion in deep-sea. A total of 122 specimens representing 7 elasmobranch species were collected and examined. External visual inspections documented no clear evidences of disease due to marine litter entanglement. A total of 7 ingested litter items were found in the stomach or in the esophagus of 6 specimens of 4 different species (frequency of occurrence = 4.9%), with a maximum number of 2 items per specimen. No litter items were found in the intestinal contents, as well as no evidence of gastrointestinal blockages due to litter items were detected. The low number of collected litter items suggests that the number of samples required is too large for a feasible monitoring program. Moreover, we observed that the anatomy of the intestinal spiral valve may represent an obstacle to the transit of macro-litter items, which could be spontaneously regurgitated as it happens in the selective elimination of undigested remains, such as bones and scales. Therefore, macro-litter retention time in the gastrointestinal tract of elasmobranchs could be brief and shorter than retention time of food. This may lead to an underestimation of macro-litter ingestion frequency by deep-water elasmobranchs. This study reports the first evidence of plastic ingestion by the kitefin shark Dalatias licha.

Food preference determines the best suitable digestion protocol for analysing microplastic ingestion by fish.

Microplastic presence in the marine environment has generated considerable concern. Many procedures for microplastics detection in fish gastrointestinal tract have been recently developed. In this study, we compared efficiencies of two common procedures applied for the digestion of organic matter (10% KOH; 15% H2O2) with a new proposal (mixture of 5% HNO3 and 15% H2O2). We considered ecological diversity among species and differences in their diet compositions as factors that could affect the efficiency and feasibility of analytical approaches. Our aim was to understand whether either one of the three protocols might be suitable for all species or it might be more advisable to select a method according to the gut content determined by different food preferences. The results showed that the trophic level and feeding habits should be considered for protocol selection. Finally, we applied the best protocols on samples from the Tyrrhenian sea.

Spatial variability and influence of biological parameters on microplastic ingestion by Boops boops (L.) along the Italian coasts (Western Mediterranean Sea).

Recently, many studies focus on the ingestion of microplastics by marine biota. Fish exploit almost every kind of marine environment, occupy many ecological niches and are an important food source for human populations worldwide. For these reasons, they seem to represent very appropriate biological indicators of microplastic ingestion. UNEP/MAP SPA/RAC (2018) identified the bogue, Boops boops (Linnaeus, 1758), as a possible target species for monitoring microplastic ingestion in fish populations. This study provides the first report of microplastic ingestion by B. boops from the Tyrrhenian and the Ligurian Seas (Western Mediterranean Sea). Generalized Linear Mixed Models were used to analyse the relationship among biological parameters and environmental factors. A total of 379 bogues were collected in three Italian regions, subject to different anthropogenic pressures (river input, human population, shipping lanes and distance from the coast). Microplastics were detected in the gastrointestinal tract of most individuals (56%) with a mean of 1.8 (±0.2) microplastics per individual. Our study further confirms that this species is able to highlight differences in the ingestion of microplastics according to local anthropization, resulting Latium region to be the most polluted. Fish with lower physical condition are more likely to ingest microplastics, suggesting a relationship with the level of local environmental contamination. Finally, the ingestion of microplastics might be influenced by behavioural differences between sexes. According to our results, males ingest significantly more microplastics than females (p < 0.05). Our research confirms that an extensive knowledge on the biology of a bioindicator species is a priority for developing a valid monitoring strategy, such as the Marine Strategy Framework Directive for European waters.

Exploring microplastic ingestion by three deep-water elasmobranch species: A case study from the Tyrrhenian Sea.

This study analyzes microplastic ingestion by three deep-water elasmobranch species (Galeus melastomus, Scyliorhinus canicula and Etmopterus spinax) from the Tyrrhenian Sea, discriminating between stomach and intestine contents. The absence of significant differences in frequency and abundance of plastic items into stomachs seems to suggest that ecological diversity among the three sharks does not strongly influence the probability of plastic ingestion in the study area. On the other hand, the detected differences in the microplastic content into the intestine might be due to a different retention time of microplastics, suggesting how feeding habits could influence metabolic features, and therefore affect the recovery of ingested plastic items. This information would improve the future development of marine micro-litter monitoring systems, following the MSFD requirements. Moreover, this study shows that all the three examined elasmobranch species can give important information even with relatively small sample sizes (N ≈ 30), and they could be used as target species for monitoring micro-litter ingestion in deep-water habitats.