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A number of marine species in the Mediterranean are threatened by the presence of several pressure factors, which include climate change, collisions with vessels, entanglement and ingestion of marine litter, especially plastic. Risk reduction policies can only be conceived starting from an accurate analysis of the exposure to such pressure factors. To estimate spatial abundance of both marine species and plastic litter and to assess the exposure risk, a two-stage analysis approach was applied, using aerial survey data from the ACCOBAMS Survey Initiative (ASI), in synergy with the Plastic Busters MPAs (PB MPAs) project. First, a detection function was fitted to observation data to obtain detection probabilities for individuals, then a Generalized Additive Model (GAM) was employed to estimate the spatial distribution of relative abundance, based on survey observations. A bivariate Local Indicator of Spatial Association (LISA) was then applied to the maps of relative abundance to derive risk maps of exposure of marine species to marine litter. The maps, obtained with a spatial resolution of about 10 km, allow us to identify areas with the highest neighboring abundance of taxa and marine litter, in particular for the MPAs studied by the PB MPAs project, which include the North-Western Mediterranean (Pelagos Sanctuary and Tuscan Archipelago), the Ionian and Aegean Sea (Zakynthos), and the Strait of Sicily (Cabrera Archipelago). [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

To evaluate the physiological state of the wild fish inhabiting the Biobío River in South-Central Chile, susceptible to the chemical contamination from different sources, biochemical and physiological biomarkers were applied to wild fish Percilia irwini and Trichomycterus areolatus in situ. Fish caught in the Biobío river in low, medium, and high anthropic impacts areas, with different pollution degrees along the river. Ethoxyresorufin O–O deethylase (EROD) activity was evaluated in fish liver. Length, weight, Gonad weight and Liver weight, Physiological Index, and gill and liver histopathology were conducted. Physicochemical parameters (pH, Temperature, Conductivity, and TDS) were measured at each sampling site. The results indicated a deteriorating condition in the biological parameters of both species in a high anthropic zone. Fishes show an increase in physiological indices and EROD liver activity, agreeing with previous studies supporting evidence of reproductive change development as we move downstream the river. Also, an increase in histopathological lesions towards the lower third stretch of the Biobío River. The Integrated Index of Physiological Biomarkers (IPBR) indicated that sites located in the high impact area (P. irwini: BC: 4.09; RC: 3.38; PC: 3.50; SJ: 2.34 and T. areolatus BC: 6.06, PC: 5.37; SJ: 5.42) have the most detrimental environmental quality, compared to reference area. The integrated biomarker analysis demonstrates that the alterations observed are related to the high anthropic activity levels downstream from the sites with the least intervention, demonstrating that the IPBR used is a complementary tool for studies of the Environmental Effects Monitoring approach. [ABSTRACT FROM AUTHOR]

The northern Gulf of Mexico has a long history of polycyclic aromatic hydrocarbon (PAH) contamination from anthropogenic activities, natural oil seepages, and the 2010 Deepwater Horizon explosion and oil spill. The continental shelf of the same area is a known breeding ground for sperm whales (Physeter macrocephalus). To evaluate PAH-DNA damage, a biomarker for potential cancer risk, we compared skin biopsies collected from Gulf of Mexico sperm whales in 2012 with skin biopsies collected from sperm whales in areas of the Pacific Ocean in 1999–2001. All samples were obtained by crossbow and comprised both epidermis and subcutaneous blubber. To evaluate exposure, 7 carcinogenic PAHs were analyzed in lipids extracted from Pacific Ocean sperm whale blubber, pooled by sex, and location. To evaluate PAH-DNA damage, portions of all tissue samples were formalin-fixed, paraffin-embedded, sectioned, and examined for PAH-DNA adducts by immunohistochemistry (IHC) using an antiserum elicited against benzo[ a ]pyrene-modified DNA, which crossreacts with several high molecular weight carcinogenic PAHs bound to DNA. The IHC showed widespread epidermal nuclear localization of PAH-DNA adducts in the Gulf of Mexico whales (n  = 15) but not in the Pacific Ocean whales (n  = 4). A standard semiquantitative scoring system revealed significantly higher PAH-DNA adducts in the Gulf of Mexico whales compared to the whales from the Pacific Ocean study (p  =  .0002). [ABSTRACT FROM AUTHOR]

The Cuvier's beaked whale (Ziphius cavirostris) is one of the least known cetacean species worldwide. The decreasing population trend and associated threats has led to the IUCN categorising the Mediterranean subpopulation as Vulnerable on the Red List of Threatened Species. This study aimed to investigate for the first time the ecotoxicological status of Cuvier's beaked whale in the NW Mediterranean Sea. The study sampled around the 20% of the individuals belonging to the Ligurian subpopulation, collecting skin biopsies from free-ranging specimens. The levels of polychlorinated biphenyl (PCBs), polybrominated diphenyl ethers (PBDEs) and induction of cytochrome's P450 (CYP1A1 and CYP2B isoforms) were evaluated. Results highlighted that the pattern of concentration for the target contaminants was PCBs > PBDEs and the accumulation values were linked to age and sex, with adult males showing significantly higher levels than juvenile. Concerns raised by the fact that 80% of the individuals had PCB levels above the toxicity threshold for negative physiological effects in marine mammals. Therefore, these findings shed light on this silent and serious threat never assessed in the Mediterranean Cuvier's beaked whale population, indicating that anthropogenic pressures, including chemical pollution, may represent menaces for the conservation of this species in the Mediterranean Sea. [ABSTRACT FROM AUTHOR]

Floating plastic debris tends to fragment into smaller pieces, termed microplastics, which may increase the likelihood of ingestion of plastics by marine organisms entering the food web. This study analyses the amount and spatial distribution of microplastics and zooplankton in an area near Asinara National Park (NW Sardinia) and overlapping the Pelagos Sanctuary (Mediterranean Sea). Analysis showed microplastics in 81 % of the 27 samples analysed, with a mean value of 0.17 ± 0.32 items m-3. From geographic information system processing of the data, microplastics appeared more abundant (by a factor of four) in the pelagic than in the neritic environment, and showed a size range of the same order as major zooplanktonic taxa determined in the area. These findings suggest a potential risk of mesozooplankton and species preying on plankton mistaking microplastics for food. Further functional and toxicological studies are therefore necessary to assess the hazard associated with microplastics in the marine food web. [ABSTRACT FROM AUTHOR]

The need for powerful new tools to detect the effects of chemical pollution, in particular of persistent organic pollutants (POPs) and polycyclic aromatic hydrocarbons (PAHs) on Mediterranean cetaceans led us to develop and apply a suite of sensitive biomarkers for integument biopsies of stranded and free-ranging animals. This multi-response ex vivo method has the aim to detect toxicological effects of contaminant mixtures. In the present study, we applied an ex vivo assay using skin biopsy and liver slices, combining molecular biomarkers [Western blot of Cytochrome P450 1A1 (CYP1A1) and Cytochrome P450 2B (CYP2B)] and gene expression biomarkers (Quantitative real-time PCR of CYP1A1, heat shock protein 70, estrogen receptor alpha and E2F transcription factor) in response to chemical exposure [organochlorines compounds (OCs), polybrominated diphenyl ethers (PBDEs), and PAHs] for stranded Mediterranean Stenella coeruleoalba. The main goal of this experiment was to identify the biomarker and/or a suite of biomarkers that could best detect the presence of a specific class of pollutants (OCs, PBDEs, and PAHs) or a mixture of them. This multi-response biomarker methodology revealed an high sensitivity and selectivity of responses (such as CYP1A and ER α mRNA variations after OCs and PAHs exposure) and could represent a valid future approach for the study of inter- and intra-species sensitivities to various classes of environmental contaminants. © 2013 Wiley Periodicals, Inc. Environ Toxicol 29: 1107-1121, 2014. [ABSTRACT FROM AUTHOR]

The possible existence of any cause–effect relationships between the concentrations of organochlorines (OCs) and the presence of Morbillivirus and Toxoplasma gondii infections was investigated in both free-living and stranded specimens of Stenella coeruleoalba, Tursiops truncatus, Globicephala melas, Balaenoptera physalus and Physeter macrocephalus from the Mediterranean Sea. High blubber concentrations of polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT) were recorded in free-ranging G. melas. Tissue concentrations of PCBs and DDT in stranded T. truncatus (367 lipid weight (l.w.) and 143.7 mg/kg l.w., respectively) and S. coeruleoalba (139.9 l.w.; 92.9 mg/kg l.w.) were beyond the PCB threshold value for the appearance of adverse effects in marine mammals. Evidence of T. gondii infection was molecularly detected in three S. coeruleoalba and six T. truncatus. [ABSTRACT FROM PUBLISHER]

BACKGROUND: Ocean pollution affects marine organisms and ecosystems as well as humans. The International Oceanographic Commission recommends ocean health monitoring programs to investigate the presence of marine contaminants and the health of threatened species and the use of multiple and early-warning biomarker approaches. OBJECTIVE: We explored the hypothesis that biomarker and contaminant analyses in skin biopsies of the threatened sperm whale (Physeter macrocephalus) could reveal geographical trends in exposure on an oceanwide scale. METHODS: We analyzed cytochrome P450 1A1 (CYP1A1) expression (by immunohistochemistry), stable nitrogen and carbon isotope ratios (as general indicators of trophic position and latitude, respectively), and contaminant burdens in skin biopsies to explore regional trends in the Pacific Ocean. RESULTS: Biomarker analyses revealed significant regional differences within the Pacific Ocean. CYP1A1 expression was highest in whales from the Galapagos, a United Nations Educational, Scientific, and Cultural Organization World Heritage marine reserve, and was lowest in the sampling sites farthest away from continents. We examined the possible influence of the whales' sex, diet, or range and other parameters on regional variation in CYP1A1 expression, but data were inconclusive. In general, CYP1A1 expression was not significantly correlated with contaminant burdens in blubber. However, small sample sizes precluded detailed chemical analyses, and power to detect significant associations was limited. CONCLUSIONS: Our large-scale monitoring study was successful at identifying regional differences in CYP1A1 expression, providing a baseline for this known biomarker of exposure to aryl hydrocarbon receptor agonists. However, we could not identify factors that explained this variation. Future oceanwide CYP1A1 expression profiles in cetacean skin biopsies are warranted and could reveal whether globally distributed chemicals occur at biochemically relevant concentrations on a global basis, which may provide a measure of ocean integrity. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the potential genotoxic effects of produced water (PW) from an Italian on-shore oil plant. Produced water is a complex mixture containing residual hydrocarbons, trace elements, naturally occurring radioactive material and potentially toxic treatment chemicals such as biocides, dispersants, detergents and scale inhibitors used in oil production. The test organism, mosquito fish (Gambusia affinis), was divided into male and female groups and exposed for 8 days in the laboratory to 50% concentrations of different produced waters: PW before treatment and after settling treatment. The fish were also exposed to lower concentrations (10%) of the same PW for 30 days. DNA damage was evaluated in erythrocytes by single cell gel electrophoresis (Comet assay) and micronucleus test, while an oxidative stress biomarker, was assessed. Polycyclic aromatic hydrocarbons (PAH) metabolites in bile were also evaluated. A higher sensitivity in biomarker responses was found in females in comparison to males. An increase in DNA strand breaks was observed in both genders after 30 days exposure and a statistically significant increase of micronucleated cells was found in females after 8 days exposure. A positive correlation between presence of micronucleated cells and PAH metabolites in bile was also observed. [ABSTRACT FROM AUTHOR]

. Zooplankton is an essential component of the marine and brackish food chains. The ecotoxicological risk of zooplanktonic communities, estimated by the modern methodological approach of biomarkers, can be used as an early warning signal of ecosystem health. The aim of this project is to estimate the potential use of several biomarkers (esterases, mixed function oxidases, porphyrins) in zooplanktonic organisms. Studies were carried out with different zooplanktonic crustaceans: the copepods Acartia margalefi and Acartia latisetosa collected in Ganzirri Lake (Messina); the mysid Siriella clausi collected in Faro Lake (Messina); the mysids Diamysis bahirensis, Siriella armata and Mysidopsis gibbosa collected in Stagnone di Marsala (Palermo); the Antarctic euphausiids Euphausia crystallorophias and Euphausia superba; the am-phipod Streetsia challengeri and the euphausiid Meganycthiphanes norvegica collected after a shore-stranding along Messina's Ionian coast. Moreover, experiments were carried out with the benthic decapods Eriphia verrucosa and Pachygrapsus marmoratus from a rocky shore of Messina's Ionian coast. Acetylcholinesterase activity (AChE) was determined in homogenates of whole organisms. The key result of this project concerns the different AChE activity basal values of different crustacean species. Particular attention should be paid to the difference in basal activity found between the Antarctic and the Mediterranean species. [ABSTRACT FROM AUTHOR]

As part of a programme to develop biomarker assays for polycyclic aromatic hydrocarbons (PAHs) in marine invertebrates, two species of crabs, Carcinus maenas and Carcinus aestuarii were exposed to benzo(a)pyrene (B(a)P) or crude oil. Microsomes were prepared from the midgut gland (hepatopancreas), examined by gel electrophoresis and Western blotting and assayed for B(a)P monooxygenase activity. In early experiments there was evidence of protein degradation and results were inconsistent and inconclusive. However, when steps were taken to minimize this in subsequent experiments, including the inclusion of four protease inhibitors in the homogenization buffer, there was consistent evidence for an increase of proteins of estimated molecular weight 45-60 kDa, and particularly of a distinct band at c. 48 kDa, following exposure to PAH at levels down to 0.1 ppm in ambient water. In C. aestuarii the increase in this band was found to coincide with an 8-12-fold increaseof B(a)P monooxygenase activity in midgut gland microsomes. These results suggest that one or more forms of cytochrome P450 may be induced by PAHs in these species. However, Western blotting using antibodies raised to vertebrate P450s, and representing four different gene families, failed to recognize any proteins in either the PAH-treated samples or in the controls. The isolation and characterization of induced protein, and the production of antibodies may provide the basis for a biomarker assay to measure a response to environmental PAHs in crabs. [ABSTRACT FROM AUTHOR]

Simple Summary: Currently procedures used to obtain samples from the gastro-intestinal tract (GIT) and protocols used to perform their respective analyses do not allow a multidisciplinary approach of this system. In fact, the investigations applied on the GIT, when performed singularly, could impair or limit the other analyses, because the currently procedures do not consider the needs of other disciplines. This means that the analyses to perform must be selected a priori, sacrificing the collection of other types of data and leads to the risk of losing important information, especially for wildlife species. To solve this conflict, we implement and standardize a new methodological approach to the GIT of marine mammals, which allow the collection of samples for different disciplines at the same time, performing the respective analyses, interpret and compare their results in a multidisciplinary way. The compatibility of multiple analyses allows the gaining of more information about the cause of death of stranded marine mammals and to enhance the knowledge of their biology and ecology. 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. [ABSTRACT FROM AUTHOR]

Microplastics have become pervasive environmental pollutants in both freshwater and marine ecosystems. The presence of microplastics have been recorded in the tissues of many wild fish species, and laboratory studies have demonstrated that microplastics can exert adverse health effects. To further investigate the biological mechanisms underlying microplastics toxicity we applied an integrated approach, analyzing the effects of microplastics at transcriptomic, histological and behavioral level. Adult zebrafish have been exposed to two concentrations of high-density polyethylene and polystyrene microplastics for twenty days. Transcriptomic results indicate alterations in the expression of immune system genes and the down-regulation of genes correlated with epithelium integrity and lipid metabolism. The transcriptomic findings are supported by tissue alterations and higher occurrence of neutrophils observed in gills and intestinal epithelium. Even the daily rhythm of activity of zebrafish appears to be affected, although the regular pattern of activity is recovered over time. Considering the transcriptomic and histological findings reported, we hypothesize that the effects on mucosal epithelium integrity and immune response could potentially reduce the organism defense against pathogens, and lead to a different utilization of energy stores. [ABSTRACT FROM AUTHOR]

Mass strandings of sperm whales (Physeter macrocephalus) are rare in the Mediterranean Sea. Nevertheless, in 2014 a pod of 7 specimens stranded alive along the Italian coast of the Central Adriatic Sea: 3 individuals died on the beach after a few hours due to internal damages induced by prolonged recumbency; the remaining 4 whales were refloated after great efforts. All the dead animals were genetically related females; one was pregnant. All the animals were infected by dolphin morbillivirus (DMV) and the pregnant whale was also affected by a severe nephropathy due to a large kidney stone. Other analyses ruled out other possible relevant factors related to weather conditions or human activities. The results of multidisciplinary post-mortem analyses revealed that the 7 sperm whales entered the Adriatic Sea encountering adverse weather conditions and then kept heading northward following the pregnant but sick leader of the pod, thereby reaching the stranding site. DMV infection most likely played a crucial role in impairing the health condition and orientation abilities of the whales. They did not steer back towards deeper waters, but eventually stranded along the Central Adriatic Sea coastline, a real trap for sperm whales. [ABSTRACT FROM AUTHOR]