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Marine litter, particularly microplastics, is a growing threat to the Mediterranean Sea, impacting biodiversity and ecosystem health. However, most studies conducted in the Mediterranean Sea have focused on monitoring of only specific environmental compartments, and rarely have highlighted the overall impacts affecting an area. Therefore, using a new multi-compartment monitoring approach and a standardized methodology, this study investigates the abundance, distribution, composition and impact of marine litter on beaches, surface waters, fish and mussels in a coastal area of Tuscany (Italy). Concerning beach macro litter values, significant differences were found among the three beaches analysed, with the highest amount in the Feniglia beach (mean value = 1245 items/100 m). The top items found are cotton bud sticks (32.3 %), which in the winter survey at Feniglia beach, reached a remarkable density of 1983 items/100 m. Microlitter (1-5 mm), was detected in the beach sediments with a mean abundance of 130 items/m 2 . Regarding floating macrolitter, the transect with the highest values was the Feniglia site at 3 nautical miles in autumn (1083 items/km 2 ) while for floating microlitter the highest concentration (832,683 MPs/km 2 ) was found in front of Scarlino. Of the 234 fish analysed from 5 different species, 67 contained microplastics in the gastrointestinal tract (28 %) with a mean value of 0.4 items per individual. The species with the highest occurrence was the European anchovy (Engraulis encrasicolus) (53 %) followed by the bogue (Boops boops) (40 %). Through the application of the Marine Litter Impact Index (MLII), considering all the compartments analysed, the Feniglia area emerges as the ecosystem most impacted by marine litter (mean MLII = 3.5, high). This study highlights how a multi-compartment monitoring approach is crucial for understanding the complex interactions between land, sea, and biota., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:, (Copyright © 2024. Published by Elsevier Ltd.)

Agroecosystems provide habitats, food, and water for many pollinators and insects, but they are also heavily exposed to threats from the widespread use of pesticides and fertilizers. Managed honeybees and wild bees encounter pesticides in vineyards by collecting morning dew from vine leaves and accessing sugars from grapes, particularly during dry periods. This study assessed the toxicological effects of the commercial fungicide formulations Fantic FNCWG ® and Ramedit combi ® , both individually and in combination, on honeybees. Using a multi-biomarker approach, we evaluated neurotoxicity, metabolic disturbances, phase II detoxification processes, and immune system function. Our findings revealed that commercial fungicide mixtures with multiple active ingredients affect bees differently than single active compounds. Biomarker responses highlighted how these complex mixtures disrupt various enzymatic pathways; including immune function; altering critical enzyme kinetics involved in detoxification and potentially impairing essential bee functions. This study emphasizes the need for more comprehensive research into the sublethal effects of commercial pesticides, particularly those used in vineyards, which are understudied compared to pesticides used in orchards.

Sea turtles can host a wide range of parasitic taxa, some of which may lead to severe diseases, weakening and endangering their health. Assessing free-ranging sea turtles' responses to these potential pathogens using biomarkers can provide valuable insights into the cellular and molecular impacts of parasites. This information can serve as a crucial tool for conservation efforts. During rehabilitation, hematological, genotoxic and innate immune parameters were evaluated in 70 free-ranging sea turtles rescued along the Southern Adriatic coast (Eastern Mediterranean Sea) from 2021 to 2023, 24 of which were found to have parasitic infections. Hapalotrema mistroides, Sulcascaris sulcata and Neospirorchis sp. were identified by coprological and molecular techniques. Some significant differences for biomarkers were associated with these parasites. These findings highlight the importance of considering the infection status of free-ranging sea turtles when evaluating biomarker results, as major pathogens like parasites can influence certain parameters., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest and that they have no actual or potential competing financial interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Agrochemicals play a vital role in protecting crops and enhancing agricultural production by reducing threats from pests, pathogens and weeds. The toxicological status of honey bees can be influenced by a number of factors, including pesticides. While extensive research has focused on the lethal and sublethal effects of insecticides on individual bees and colonies, it is important to recognise that fungicides and herbicides can also affect bees' health. Unfortunately, in the field, honey bees are exposed to mixtures of compounds rather than single substances. This study aimed to evaluate the effects of a commercial fungicide and a commercial herbicide, both individually and in combination, on honey bees. Mortality assays, biomarkers and learning and memory tests were performed, and the results were integrated to assess the toxicological status of honey bees. Neurotoxicity (acetylcholinesterase and carboxylesterase activities), detoxification and metabolic processes (glutathione S-transferase and alkaline phosphatase activities), immune system function (lysozyme activity and haemocytes count) and genotoxicity biomarkers (Nuclear Abnormalities assay) were assessed. The fungicide Sakura® was found to activate detoxification enzymes and affect alkaline phosphatase activity. The herbicide Elegant 2FD and the combination of both pesticides showed neurotoxic effects and induced detoxification processes. Exposure to the herbicide/fungicide mixture impaired learning and memory in honey bees. This study represents a significant advance in understanding the toxicological effects of commonly used commercial pesticides in agriculture and contributes to the development of effective strategies to mitigate their adverse effects on non-target insects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Sea turtles, with their global distribution and complex life cycle, often accumulate pollutants such as metals and metalloids due to their extended lifespan and feeding habits. However, there are limited studies exploring the impact of metal pollution on the reproductive health of female sea turtles, specifically focusing on the quality of their eggs, which has significant implications for the future generations of these charismatic animals. São Tomé Island, a crucial nesting and feeding habitat for green sea turtles, underscores the urgent need for comprehensive research in this ecologically significant area. This study aimed to investigate whether metals and metalloids in the blood of nesting female green sea turtles induce genotoxic effects in their erythrocytes and affect their egg morphometric characteristics and the composition of related compartments. Additionally, this study aimed to evaluate whether the quality of energetic reserves for embryo development (fatty acids in yolk's polar and neutral lipids) is influenced by the contamination status of their predecessors. Results revealed correlations between Cu and Hg levels and increased "lobed" erythrocytes, while As and Cu negatively influenced shell thickness. In terms of energy reserves, both polar and neutral lipid fractions contained primarily saturated and monounsaturated fatty acids, with prevalent 18:1n-9, 18:0, 16:0, 14:0, and 12:0 fatty acids in yolk samples. The yolk polar fraction was more susceptible to contaminant levels in female sea turtles, showing consistent negative correlations between pollution load index and essential n3 fatty acids, including linolenic, eicosatrienoic, eicosapentaenoic, and docosapentaenoic acids, crucial for embryonic development. These metals accumulation, coupled with the reduced availability of these key fatty acids, may disrupt the eicosanoid and other important pathways, affecting reproductive development. This study reveals a negative correlation between metal contamination in female sea turtles' blood and egg lipid reserves, raising concerns about embryonic development and the species' future generations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The increase of urbanization and agricultural activities is causing a dramatic reduction of natural environments. As a consequence, animals need to physiologically adjust to these novel environments, in order to exploit them for foraging and breeding. The aim of this work was to compare the physiological status among nestling common kestrels (Falco tinnunculus) that were raised in nest-boxes located in more natural, rural, or urban areas in a landscape with a mosaic of land uses around Rome in Central Italy. A blood-based multi-biomarker approach was applied to evaluate physiological responses at multiple levels, including antioxidant concentrations, immunological functions, genotoxicity, and neurotoxicity. We found lower concentrations of glutathione and GSH:GSSG ratio values and higher proportions of monocytes in urban birds compared to the other areas. We also found higher DNA damage in rural compared to urban and natural krestels and inhibition of butyrylcholinesterase activity in urban and natural birds compared to rural area. Finally, we found similar values among study areas for respiratory burst, complement system, bactericidal capacity, and plasma non-enzymatic antioxidant capacity. These results suggest that (i) city life does not necessarily cause physiological alterations in kestrels compared to life in other habitats, and (ii) environmental pressures are likely to differ in typology and intensity across habitats requiring specific responses that a multi-biomarker approach can help to detect. Further studies are needed to assess which factors are responsible for the physiological differences among city, rural, and natural birds, and whether these differences are consistent across time and space., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Modern agriculture is mainly based on the use of pesticides to protect crops but their efficiency is very low, in fact, most of them reach water or soil ecosystems causing pollution and health hazards to non-target organisms. Fungicide triazoles and strobilurins based are the most widely used and require a specific effort to investigate toxicological effects on non-target species. This study evaluates the toxic effects of four commercial fungicides Prosaro® (tebuconazole and prothioconazole), Amistar®Xtra (azoxystrobin and cyproconazole), Mirador® (azoxystrobin) and Icarus® (Tebuconazole) on Eisenia fetida using several biomarkers: lipid peroxidation (LPO), catalase activity (CAT), glutathione S-transferase (GST), total glutathione (GSHt), DNA fragmentation (comet assay) and lysozyme activity tested for the first time in E. fetida. The exposure to Mirador® and AmistarXtra® caused an imbalance of ROS species, leading to the inhibition of the immune system. AmistarXtra® and Prosaro®, composed of two active ingredients, induced significant DNA alteration, indicating genotoxic effects. This study broadened our knowledge of the effects of pesticide product formulations on earthworms and showed the need for improvement in the evaluation of toxicological risk deriving from the changing of physicochemical and toxicological properties that occur when a commercial formulation contains more than one active ingredient and several unknown co-formulants., (© 2024. The Author(s).)

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/km 2 ) and microplastics (259,490 items/km 2 ) 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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Anthropogenic activities have caused a steady decline of common kestrel (Falco tinnunculus) since the 1980 s. Effects, especially sublethal effects of contaminants, need to be investigated to ensure the conservation of this species. Data about countries, biological material, contaminants classes, and methodological approaches were collected from scientific publications to highlight gaps on common kestrel toxicology and ecotoxicology. We found that most studies have been conducted in Europe and in the field, underlining a lack of in vitro studies. The studies investigated mainly contaminant levels, while sublethal effects, evaluation of emerging contaminants and use of non-invasive or low-invasive samples were scarce. This work shows important gaps on toxicological status of the common kestrel, highlighting the importance of developing a non-lethal approach that combines responses at different levels of biological organization, as well as data on chemical contamination and on the environment in which the different populations inhabit., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Brodifacoum is the most common rodenticide used for the eradication of invasive rodents from islands. It blocks the vitamin K cycle, resulting in hemorrhages in target mammals. Non-target species may be incidentally exposed to brodifacoum, including marine species. A case study conducted on the Italian Marine Protected Area of Tavolara Island was reported after a rodent eradication using the aerial broadcast of a brodifacoum pellet. Brodifacoum presence and effects on non-target marine organisms were investigated. Different fish species were sampled, and a set of analyses was conducted to determine vitamin K and vitamin K epoxide reductase concentrations, prothrombin time, and erythrocytic nuclear abnormalities (ENA) assay. In all the examined organisms, brodifacoum was not detected. The results obtained showed differences in vitamin K and vitamin K epoxide concentrations among the samples studied, with a positive correlation for three species between vitamin K, vitamin K epoxide, and fish weight. The prothrombin time assay showed a good blood clotting capacity in the fish. Higher abnormality values were recorded for four species. The results of this study suggest that it is possible to hypothesize that the sampled fish were not likely to have been exposed to brodifacoum and that consequently there are no negative issues concerning human consumption.