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.

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.

Drivers of litter ingestion by sea turtles: Three decades of empirical data collected in Atlantic Europe and the Mediterranean.

Sea turtles are considered as bio-indicators for monitoring the efficiency of restoration measures to reduce marine litter impacts on health. However, the lack of extended and standardised empirical data has prevented the accurate analysis of the factors influencing litter ingestion and the relationships with individual health. Historic data collected from 1988 and standard data collected from 2016 were harmonised to enable such analyses on necropsied loggerhead turtles (Caretta caretta) in eight Mediterranean and North-East Atlantic countries. Litter was found in 69.24 % of the 1121 individuals, mostly single-use and fishing-related plastics. Spatial location, sex and life history stage explained a minor part of litter ingestion. While no relationships with health could be detected, indicating that all individuals can be integrated as bio-indicators, the mechanistic models published in literature suggest that the high proportion of plastics in the digestive contents (38.77 % per individual) could have long-term repercussions on population dynamics.

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.

Phthalate esters (PAEs) concentration pattern reflects dietary habitats (δ13C) in blood of Mediterranean loggerhead turtles (Caretta caretta).

Phthalic acid esters (PAEs) are classified as endocrine disruptors, but it remains unclear if they can enter the marine food-web and result in severe health effects for organisms. Loggerhead turtles (Caretta caretta) can be chronically exposed to PAEs by ingesting plastic debris, but no information is available about PAEs levels in blood, and how these concentrations are related to diet during different life stages. This paper investigated, for the first time, six PAEs in blood of 18 wild-caught Mediterranean loggerhead turtles throughout solid-phase extraction coupled with gas chromatography-ion trap/mass spectrometry. Stable isotope analyses of carbon and nitrogen were also performed to assess the resource use pattern of loggerhead turtles. DEHP (12-63 ng mL-1) and DBP (6-57 ng mL-1) were the most frequently represented PAEs, followed by DiBP, DMP, DEP and DOP. The total PAEs concentration was highest in three turtles (124-260 ng mL-1) whereas three other turtles had concentrations below the detection limit. PAEs were clustered in three groups according to concentration in all samples: DEHP in the first group, DBP, DEP, and DiBP in the second group, and DOP and DMP in the third group. The total phthalates concentration did not differ between large-sized (96.3 ± 86.0 ng mL-1) and small-sized (67.1 ± 34.2 ng mL-1) turtles (p < 0.001). However, DMP and DEP were found only in large-sized turtles and DiBP and DBP had higher concentrations in large-sized turtles. On the other hand, DEHP and DOP were found in both small- and large-sized turtles with similar concentrations, i.e. ~ 21.0/32.0 ng mL-1 and ~ 7.1/9.9 ng mL-1, respectively. Winsored robust models indicated that δ13C is a good predictor for DBP and DiBP concentrations (significant Akaike Information criterion weight, AICwt). Our results indicate that blood is a good matrix to evaluate acute exposure to PAEs in marine turtles. Moreover, this approach is here suggested as a useful tool to explain the internal dose of PAEs in term of dietary habits (δ13C), suggesting that all marine species at high trophic levels may be particularly exposed to PAEs, despite their different dietary habitats and levels of exposure.

Polymer composition assessment suggests prevalence of single-use plastics among items ingested by loggerhead sea turtles in the western mediterranean sub-region.

The ingestion of plastic is becoming a major concern for various species and particularly for marine turtles across the globe. The loggerhead sea turtle (Caretta caretta) was recently chosen by the European Commission as a bio-indicator for plastic pollution within the Mediterranean basin. We further investigated which items this key species is more prone to ingest, following the standardised Marine Strategy Framework Directive protocols. Moreover, we integrated to this protocol the Fourier Transform Infrared Spectroscopy, which allowed us to determine the polymer type of each item. We analysed samples from 226 sea turtles from 2008 to 2017 in two areas of the western Mediterranean sub-region (sensu MSFD). In the Lazio area we found a frequency of occurrence of plastic ingestion of 78.33%, while in Sardinia 41.79%. The analysis of the litter categories, among all individuals, highlights a prevalence of user-sheet (Use-She; 69.13%) and user-fragment plastics (Use-Fra; 20.84%). In addition, the polymer analysis showed a dominance of polyethylene (65.98%) and polypropylene (26.23%). As a result, by looking at other works that have investigated polymer types and items sources, we are able to infer that 77.25% of the objects ingested by the C. caretta individuals are attributable to disposable daily-life objects managed in an improper way. Therefore, C. caretta apart from being an efficient bio-indicator for plastic pollution, highlighting spatial and temporal concentration differences, it could also be used to verify the effectiveness of the Single-use Plastic Directive (EU 2019/904).

A novel approach based on multiple fish species and water column compartments in assessing vertical microlitter distribution and composition.

The assessment of the distribution and composition of microlitter in the sea is a great challenge. Biological indicators can be an irreplaceable tool since they measure microlitter levels in their environments in a way that is virtually impossible to replicate by direct physical measurements. Furthermore, trends can provide policymakers with statistically robust analysis. We looked into the capacity of multiple fish species to describe the distribution and composition of microlitter vertically across different compartments of the water column. A total of 502 individuals from six selected species (Scomber scombrus, Oblada melanura, Spicara smaris, Boops boops, Merluccius merluccius and Mullus barbatus) were collected on the western side of Sardinia island and allocated to three compartments: surface, mid-water and bottom. The species of the surface exhibited a higher frequency of occurrence (41.89%) of microlitter ingestion, compared to those of the mid-water and bottom (19.60%; 22.58%). A significant difference in the average number of ingested microlitter was found between the surface and the bottom compartment. All the microlitter fragments found were analysed through Fourier Transform Infrared Spectroscopy (FTIR). The comparison of the expected buoyancies of the polymers identified puth faith in the allocation of the species to the respective compartments. Therefore, considering the Marine Strategy Framework Directive objective, this approach could be useful in assessing microlitter distribution and composition vertically across the water column.

Fishery management in a marine protected area with compliance gaps: Socio-economic and biological insights as a first step on the path of sustainability.

Overfishing is one of the main impacts on the marine environment and multiple-use Marine Protected Areas (MPAs) could be a useful tool to conserve biodiversity and promote sustainable resource exploitation. However, ensuring a high level of protection on the ground is a difficult task. This work contributes to the analysis of the causes at the root of MPAs' ineffectiveness by examining the management of Paracentrotus lividus fishery in an Italian MPA, employing a multidisciplinary approach built on biological and socio-economic competences. This sea urchin species has a determinant ecological role in structuring infralittoral benthic assemblages and is the most exploited echinoid in Europe. From 2010 to 2018, underwater sampling was conducted over 39 monitoring sites to define P. lividus spatial and temporal trends. Declared catches and semi-structured interviews with local stakeholders were used to define the socio-economical context, underline existing conflicts among them, as well as to trace the historical evolution of sea urchin fishery. The results show that the management of sea urchin fishery is not sustainable, primarily because of the stakeholders' non-compliance with the rules. P. lividus stock is progressively declining (-73% in 9 years), showing no difference between MPA (0.5 ± 0.15 ind./m2) and control sites (0.3 ± 0.04 ind./m2). Moreover, fishermen dominate the social arena while scientists, civil society and local press have little relevance. Additionally, the untruthfulness of catch declarations was proved, the IUU fishery is relevant and the black market is hiding the actual economic value. This work offers management solutions that may be useful in other areas that show similar compliance issues.

Data Collection on Marine Litter Ingestion in Sea Turtles and Thresholds for Good Environmental Status.

The following protocol is intended to respond to the requirements set by the European Union's Marine Strategy Framework Directives (MSFD) for the D10C3 Criteria reported in the Commission Decision (EU), related to the amount of litter ingested by marine animals. Standardized methodologies for extracting litter items ingested from dead sea turtles along with guidelines on data analysis are provided. The protocol starts with the collection of dead sea turtles and classification of samples according to the decomposition status. Turtle necropsy must be performed in authorized centers and the protocol described here explains the best procedure for gastrointestinal (GI) tract isolation. The three parts of the GI (esophagus, stomach, intestine) should be separated, opened lengthways and contents filtered using a 1 mm mesh sieve. The article describes the classification and quantification of ingested litter, classifying GI contents into seven different categories of marine litter and two categories of natural remains. The quantity of ingested litter should be reported as total dry mass (weight in grams, with two decimal places) and abundance (number of items). The protocol proposes two possible scenarios to achieve the Good Environmental Status (GES). First: "There should be less than X% of sea turtles having Y g or more plastic in the GI in samples of 50-100 dead turtles from each sub-region", where Y is the average weight of plastic ingested and X% is the percentage of sea turtles with more weight (in grams) of plastic than Y. The second one, which considers the food remain versus plastic as a proxy of individual health, is: "There should be less than X% of sea turtles having more weight of plastic (in grams) than food remains in the GI in samples of 50-100 dead turtles from each sub-region".

Self-organisation in striped seagrass meadows affects the distributional pattern of the sessile bivalve Pinna nobilis.

Striped seagrass meadows are formed by narrow ribbons which are elevated over the seabed and separated by channels. Limited information on the genesis and development of this morphological pattern, including the adaptive responses of associated biota, is preventing holistic insight into the functioning of such protected ecosystems. This paper assessed the structural dynamics of a Posidonia oceanica striped meadow and the distribution and 3D orientation of the associated bivalve Pinna nobilis. Our analysis of the interaction between bedforms, bottom currents, and the distribution of P. nobilis revealed that the striped seascape is the result of a self-organisation process driven by feedback interactions among seagrass growth, sediment deposition, and hydrodynamics. The results suggest that the ribbon wall is the most suitable sub-habitat for this species, because it supports the highest density of P. nobilis, compared to the meadow top and bottom. Here, specimens can take advantage of the resuspension induced by hydrodynamics and open their shells towards the current, thus enhancing food intake. Therefore, our results show that self-organisation in striped seagrass meadow affects the distributional pattern of P. nobilis, providing new insights into the autoecology of this species beyond the conservation implications for its habitat.

Determining suitable fish to monitor plastic ingestion trends in the Mediterranean Sea.

The presence of marine litter is a complex, yet persistent, threat to the health and biodiversity of the marine environment, and plastic is the most abundant, and ubiquitous type of marine litter. To monitor the level of plastic waste in an area, and the prospect of it entering the food chain, bioindicator species are used extensively throughout Northern European Seas, however due to their distribution ranges many are not applicable to the Mediterranean Sea. Guidance published for the Marine Strategy Framework Directive suggests that the contents of fish stomachs may be analyzed to determine trends of marine plastic ingestion. In order to equate transnational trends in marine plastic ingestion, the use of standardized fish species that widely occur throughout the basin is favoured, however for the Mediterranean Sea, specific species are not listed. Here we propose a methodology to assess how effective Mediterranean fish species, that are known to have ingested marine plastic, are as bioindicators. A new Bioindicator Index (BI) was established by incorporating several parameters considered important for bioindicators. These parameters included species distribution throughout the Mediterranean basin, several life history traits, the commercial value of each species, and the occurrence of marine litter in their gut contents. By collecting existing data for Mediterranean fish, ranked scores were assigned to each trait and an average value (BI value) was calculated for each species. Based on their habitat preferences, Engraulis encrasicolus (pelagic), Boops boops (benthopelagic), three species of Myctophidae (Hygophum benoiti, Myctophum punctatum and Electrona risso) (mesopelagic), Mullus barbatus barbartus (demersal) and Chelidonichthys lucerna (benthic), were identified as currently, the most suitable fish for monitoring the ingestion of marine plastics throughout the Mediterranean basin. The use of standardized indicator species will ensure coherence in the reporting of marine litter ingestion trends throughout the Mediterranean Sea.

Pressure and impact of anthropogenic litter on marine and estuarine reptiles: an updated "blacklist" highlighting gaps of evidence.

We report an arrangement on the effect of anthropogenic litter on marine and estuarine reptiles, checking for evidence about different types of impact (ingestion vs. entanglement) and pressure (three size-based categories). From 1976 to 2018, we obtained a "blacklist" of 11 species impacted by marine litter (about 13% of 85 species of marine and estuarine reptiles), belonging to three orders (Testudines, Squamata, and Crocodilia). We obtained only occasional evidence of an impact for Squamata (Hidrophis elegans, Disteira major) and Crocodilia (Crocodylus porosus). Regarding the different types of pressure, the highest number of evidence has been obtained for macro-litter (10 species) and the lowest for micro-litter (4 species, all Chelonidae). Among Testudines, Lepidochelys kempii and Natator depressus evidenced a lack of data for micro-plastic. In Squamata, information is lacking for micro-plastic with only occasional references for meso-plastic (in Hydrophis elegans) and macro-plastic (Disteira major and Crocodylus porosus). We obtained a direct correlation between the research effort and the number of citations regarding different types of pressure and impact of marine litter: therefore, our blacklist of impacted species could be increased, carrying out further research focused on other poorly studied marine and estuarine reptiles. We suggest the use of a standardized nomenclature to reduce the amount of lost information.

Loggerhead sea turtles (Caretta caretta): A target species for monitoring litter ingested by marine organisms in the Mediterranean Sea.

Marine litter is any persistent, manufactured or processed solid material discarded, disposed of or abandoned in the marine and coastal environment. Ingestion of marine litter can have lethal and sub-lethal effects on wildlife that accidentally ingests it, and sea turtles are particularly susceptible to this threat. The European Commission drafted the 2008/56/EC Marine Strategy Framework Directive with the aim to achieve a Good Environmental Status (GES), and the loggerhead sea turtle (Caretta caretta, Linnaeus 1758) was selected for monitoring the amount and composition of litter ingested by marine animals. An analogous decision has been made under the UNEP/MAP Barcelona Convention for the protection of the Mediterranean Sea, following the Ecosystem Approach. This work provides for the first time, two possible scenarios for the Marine Strategy Framework Directive GES, both related to "Trends in the amount and composition of litter ingested by marine animals" in the Mediterranean Sea. The study validates the use of the loggerhead turtle as target indicator for monitoring the impact of litter on marine biota and calls for immediate use of this protocol throughout the Mediterranean basin and European Region. Both GES scenarios are relevant worldwide, where sea turtles and marine litter are present, for measuring the impact of ingested plastics and developing policy strategies to reduce it. In the period between 2011 and 2014, 150 loggerhead sea turtles, found dead, were collected from the Italian Coast, West Mediterranean Sea Sub-Region. The presence of marine litter was investigated using a standardized protocol for necropsies and lab analysis. The collected items were subdivided into 4 main categories, namely, IND-Industrial plastic, USE-User plastic, RUB-Non plastic rubbish, POL-Pollutants and 14 sub-categories, to detect local diversity. Eighty-five percent of the individuals considered (n = 120) were found to have ingested an average of 1.3 ± 0.2 g of litter (dry mass) or 16 ± 3 items.

Recovery Trends of Commercial Fish: The Case of an Underperforming Mediterranean Marine Protected Area.

Temporal trends in the recovery of exploited species in marine protected areas (MPAs) are useful for a proper assessment of the efficacy of protection measures. The effects of protection on the fish assemblages of the sublittoral rocky reefs in the "Penisola del Sinis-Isola di Mal di Ventre" MPA (W. Sardinia, Italy) were evaluated using a multi-year series of data. Four surveys, conducted 7, 10, 13 and 15 years after the area was designated as an MPA and carried out in the period spanning June and July, were used to estimate the abundance and biomass of commercial species. The surveys were carried out in zones with decreasing levels of fishing restrictions within the MPA (zones A, B, C) and in unprotected zones (OUT1 and OUT2), and underwater video visual census techniques were used. Protected zones only occasionally showed higher levels of abundance or biomass, and the trajectories of those metrics were not consistent across the years. In addition, the zone with the highest level of protection (zone A) never presented levels of abundance and biomass higher than those in zones B and C. This study shows that even 15 years after designation, protection has had no appreciable effect in the MPA studied. It is argued that this is emblematic of several shortcomings in the planning, regulation and enforcement frameworks of the MPA.

Amount and distribution of neustonic micro-plastic off the western Sardinian coast (Central-Western Mediterranean Sea).

A plethora of different sampling methodologies has been used to document the presence of micro-plastic fragments in sea water. European Marine Strategy suggests to improve standard techniques to make future data comparable. We use Manta Trawl sampling technique to quantify abundance and distribution of micro-plastic fragments in Sardinian Sea (Western Mediterranean), and their relation with phthalates and organoclorine in the neustonic habitat. Our results highlight a quite high average plastic abundance value (0.15 items/m(3)), comparable to the levels detected in other areas of the Mediterranean. "Site" is the only factor that significantly explains the differences observed in micro-plastic densities. Contaminant levels show high spatial and temporal variation. In every station, HCB is the contaminant with the lowest concentration while PCBs shows the highest levels. This work, in line with Marine Strategy directives, represents a preliminary study for the analysis of plastic impact on marine environment of Sardinia.

Interaction between loggerhead sea turtles (Caretta caretta) and marine litter in Sardinia (Western Mediterranean Sea).

Anthropogenic debris in the environment affects many species that accidentally ingest it. The aim of this study is to evaluate the quantity and composition of marine litter ingested by loggerheads in Sardinia, thus supplying for the lack of data in the existing literature for this area. Seventeen of the 121 (14.04%) monitored turtles presented debris in their digestive tracts. Litter in faecal pellet of alive individuals (n = 91) and in gastro-intestinal contents of dead ones (n = 30) was categorized, counted and weighed. User plastic was the main category of ingested debris with a frequency of occurrence of 13.22% of the total sample, while sheet (12.39%) and fragments (9.09%) were the most relevant sub-categories. This study highlights for the first time the incidence of litter in alive turtles in Sardinia. This contribution improves the knowledge about marine litter interaction on Caretta caretta as bio-indicator. Results will be useful for the Marine Strategy implementation.