The Mediterranean basin is the second largest global biodiversity hotspot in the world, which coexists with a plethora of anthropogenic stress. This study examines the risks that marine litter poses to Mediterranean biodiversity, using the Special Protection Area of Mediterranean Importance (SPAMI) "Pelagos Sanctuary" as a case study. In this paper, a new survey method and data analysis strategies to assess the impact of marine litter, including microplastics, on Mediterranean organisms, is proposed. A total of 23 species, from invertebrates to cetaceans, were analysed using two main monitoring approaches. In the first approach, the core density distribution areas of 11 megafauna species (elasmobranchs, sea turtles, seabirds, and cetaceans) and 4 invertebrate species were evaluated through an extended field survey. Simultaneously, monitoring of floating macro- and micro-litter was performed to be overlapped with the species distribution and abundance. The second monitoring approach assessed the ingestion of marine litter, microplastics and the levels of plastic additives in 10 stranded megafauna species and in 7 invertebrate and fish species. The final data processing, merging the data on biodiversity and marine litter abundance and distribution in the environment, with the data of ingested marine litter, microplastics, and plastic additives in sentinel species, allowed to: a) create risk maps for the species inhabiting the Pelagos Sanctuary, identifying critical areas for biodiversity conservation; b) identify the most exposed species to the risk of marine litter by an "exposure score"; c) to select sentinel species for pelagic environment.
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