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Anthropogenic threats impacting ecological targets should be mitigated and solved using fast and schematic tools useful in conservation strategies. Herein, we suggest a mixed and quick approach implementing coarse-grained (and expert-based) threat analysis with the fine-grained (and analytical) DPSIR (driving forces, pressure, status, impact, and response) framework of indicators, all included in a single causal chain. Both approaches are largely used in conservation but never combined. A simulated example of the application of the set of indicators (status, pressure, impact, and response) on dune ecosystems (and nested targets represented by halo-psammophilous plants) has been included. Due to its schematic format, values as targets, pressures as threats, and responses as conservation strategies have been unified in a single conceptual framework. This synthetic framework can also be used to communicate to academic students the complexities of socio-ecological systems on the conservation front lines using a simplified cause–effect chain.

The monitoring of stranded marine mammals represents a strategic method to assess their health, conservation status, and ecological role in the marine ecosystem. Networks worldwide track stranding events for the passive monitoring of mortality patterns, emerging and reemerging pathogens, climate change, and environmental degradation from a One Health perspective. This study summarizes pathogen prevalence data from the Italian Stranding Network (ISN) derived from post-mortem investigations on cetaceans found dead stranded along the Italian coastline between 2015 and 2020. The decomposition of the carcasses and logistics limited the post-mortem examination to 585 individuals, out of 1236 single-stranding reports. The most relevant pathogens identified were Cetacean Morbillivirus, Herpesvirus, Brucella spp., and Toxoplasma gondii, whose roles as environmental stressors are well known, despite their real impact still needing to be investigated in depth. Statistical analysis showed that age and sex seem to be positively related to the presence of pathogens. This study represents the first step in harmonizing post-mortem investigations, which is crucial for evidence-based conservation efforts. Implementing diagnostic and forensic frameworks could offer an indirect insight into the systematic monitoring of diseases to improve the identification of regional and temporal hotspots in which to target specific mitigation, management, and conservation strategies.

Since 2013, loggerhead sea turtle (Caretta caretta, Linnaeus 1758) nesting has been observed further north along the Italian coast, reaching the Tuscan coast (NW Mediterranean Sea). The four nesting events that occurred in Tuscany in the summer of 2019 spurred the scientific community to monitor these occurrences more carefully, following them from egg deposition to hatching. This provided an opportunity to collect samples for conducting multidisciplinary investigations, including the toxicological investigations of the biological material collected from the four nests. The aim of this study was to conduct an initial assessment of persistent organic pollutants in the eggs laid in Tuscany, aiming to establish a baseline on this topic for subsequent nesting events that have occurred until today. Organochlorine compounds (OCs), specifically polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolites, and hexachlorobenzene (HCB), were analyzed and detected in unhatched eggs, embryos, and chorio-allantoic membranes (CAMs). OCs were detected in all samples, with PCBs > DDTs ≫ HCB. A significant spatial variation in pollutant levels and profiles among sea turtle nesting locations was found. Embryos showed higher levels of contamination than egg contents regardless of the developmental stages. Depth of the laying chamber and egg mass were not significant factors in OC bioaccumulation. For the first time in the Mediterranean Sea, this study assessed the role of CAM in the transfer of contaminants to the embryo. Overall, the OC levels found were lower compared to the results from other studies conducted worldwide on loggerhead sea turtle biological material. This was the first assessment of nest biological material for the North-Western coast of Italy.

Abstract Dolphin–fishery interaction is a worldwide issue affecting dolphins through bycatch and fishers through catch or gear damages. Concerning the Mediterranean Sea, problematic interactions mainly occur between common bottlenose dolphin and small-scale fisheries. Acoustic Deterrent Devices such as pingers, are one of the most widespread measures used in attempts to face this issue. Therefore, the efficiency of interactive pingers (DiD01) in protecting the trammel nets from dolphin interactions was assessed in the Northern Tyrrhenian Sea. From March to October 2021, a total of 139 fishing trials using nets with pingers (TEST) and without pingers (CTRL), respectively n = 97 and n = 42, were carried out. Non-parametric statistic of the Catch per Unit Effort, comparing control and test nets, was not significantly different (p > 0.05) using catches weights (CPUEW) while it was significant (p

Due to their late maturation, extreme longevity, low fecundity and slow growth rates, deep-sea Chondrichthyes are extremely vulnerable to human impacts. Moreover, assessing the impact of deep-sea fisheries is difficult, as many species (including sharks) are part of the bycatch and are often discarded at sea, and/or landed under generic commercial-species codes. The lack of this information on fishery data sets and the limited availability of species-specific life history data make challenging the management of deep-sea Chondrichthyes. The kitefin shark Dalatias licha is a cosmopolitan elasmobranch, mainly found on continental and insular shelf-breaks and slopes in warm-temperate and tropical waters. This species is a common by-catch of the deep-sea trawling, considered as “Endangered” by the IUCN Red List for all European waters, Mediterranean Sea included. Here we present the results of a study based on a total of 78 specimens of kitefin shark collected over 3 years in the Ligurian Sea (NW Mediterranean) as by-catch from deep-water fisheries. Total length ranged from 380 to 1164 mm, and individual weight ranged from 198 to 8000 g. Immature and mature individuals showed a sex ratio dominated by males. Adult males were observed throughout the year, while mature females were observed only in spring-summer. These data lead to hypothesise a spatial segregation between genders. The kitefin shark diet was dominated by bony fish (mainly Macrouridae) and other small sharks (e.g., Galeus melastomus and Etmopterus spinax), but their gut included plastic items and parasites. Data reported here underline the rarity, complex ecology and the threat for this shark species and support the urgency of promoting initiatives for their monitoring and conservation.

Information regarding bacterial diseases in Cuvier’s beaked whale (CBW, Ziphius cavirostris) is scattered and mostly incomplete. This report describes a case of septicemia by Morganella morganii in a juvenile male CBW with concurrent renal crassicaudiasis. The animal stranded along the Ligurian coastline (Italy) and underwent a systematic post-mortem examination to determine the cause of death. Histopathology showed lesions consistent with a septicemic infection, severe meningoencephalitis, and renal crassicaudiasis. An M. morganii alpha-hemolytic strain was isolated in pure culture from liver, lung, prescapular lymph node, spleen, hepatic and renal abscesses, and central nervous system (CNS). The antimicrobial susceptibility profile of the strain was evaluated with the minimum inhibitory concentrations (MICs) method and reduced susceptibility to Trimethoprim-Sulfamethoxazole is reported. Crassicauda sp. nematodes were retrieved from both kidneys. No other pathogens were detected by immunohistochemistry, serology, or biomolecular analyses. Toxicological investigations detected high concentrations of immunosuppressant pollutants in the blubber. The chronic parasitic infestation and the toxic effects of xenobiotics likely compromised the animal's health, predisposing it to an opportunistic bacterial infection. To our knowledge, this is the first description of M. morganii septicemia with CNS involvement in a wild cetacean.

Seas and oceans are contaminated by persistent organic pollutants (POPs), which are released into the environment by human activities. The chemical-physical properties of POPs induce high persistence and toxicity in marine organisms from the lowest to the highest trophic levels. Phyto- and zooplankton are at the base of the food chain, and they can adsorb and accumulate these xenobiotic compounds. Therefore, all planktophagous species, including the whale shark (Rhincodon typus), are susceptible to ingesting these contaminants during feeding. From October to December, whale sharks migrate along the north-west coast of Madagascar in search of dense patches of plankton. During scientific expeditions to the whale sharks’ foraging areas in the waters of the island of Nosy Be (which is in the north-west of Madagascar), plankton samples were taken. In these samples, the presence and levels of some chlorinated xenobiotics (HCB, DDT and its metabolites, and PCBs) were evaluated in order to estimate the possible impact of whale shark diet on organochlorine (OC) accumulation. The fresh plankton biomass sampled from this region did not seem to be sufficient for the sustenance of the animals, which suggests that the daily contamination input of Rhincodon typus individuals, depending on their plankton diet, is minimal.

Environmental pollution, particularly in the marine environment, has become a significant concern due to the increasing presence of pollutants and their adverse effects on ecosystems and human health. This study focuses on the bioaccumulation of trace elements in the muscle tissue of the blackmouth catshark (Galeus melastomus) from different areas in the Mediterranean Sea. Trace elements are of interest due to their persistence, toxicity, and potential for bioaccumulation. This research aims to assess the distribution and accumulation of trace elements in the muscle tissue of G. melastomus and investigate their potential impact on the deep-sea environment of the Mediterranean. The focused areas include the Ligurian Sea, the northern and central Tyrrhenian Sea, the southern Tyrrhenian Sea, the Ionian Sea, the Pantelleria Waters, and the Gela Waters. Samples were collected following established protocols, and trace element analysis was conducted using inductively coupled plasma mass spectrometry. The study provides data on the concentrations of 17 trace elements, namely aluminum, arsenic, cadmium, cobalt, copper, manganese, molybdenum, nickel, zinc, selenium, strontium, lead, chromium, iron, barium, bismuth, and uranium. The findings contribute to a better understanding of trace element bioaccumulation patterns in elasmobranch species, specifically G. melastomus, and highlight the potential risks associated with chemical contamination in the Mediterranean Sea. This research emphasizes the importance of studying the impacts of pollutants on marine organisms, particularly those occupying key ecological roles, like sharks, to support effective conservation and management strategies.

Between 2007 and 2022, 112 specimens of newborn and young pelagic sharks were recorded in the waters of Tuscany Region, in the South Ligurian–North Tyrrhenian Seas (north-western Mediterranean basin). The sharks belonged to the Carcharhinus plumbeus (n = 14), Prionace glauca (n = 66), Isurus oxyrinchus (n = 16), Mobula mobular (n = 5) Alopias vulpinus (n = 7) and Hexanchus griseus (n = 4) species. Each animal was correctly identified thanks to the photographs or videos collected. All specimens were incidentally captured with set nets in inshore shallow waters, except bluntnose six-gill sharks, which were bycatch of deep-water bottom-trawl fishery. Body mass, sex, total length and biometric measurements were recorded in 34 baby sharks following the Mediterranean Large Elasmobranches Monitoring (MEDLEM) protocol. The presence of very evident and often non-healed umbilical scar confirmed that some of the sample specimens were newborn. Further confirmation came from the comparison between the total length observed and the size at birth known for the sampled species as reported in the literature. Some baby sharks were preserved in the Museums of Natural History of Pisa and Florence University collections. The importance of the coastal area studied as a possible shark nursery is discussed.

Between 2019 and 2021, 21 bottlenose dolphins were found stranded dead along the Ligurian Pelagos Sanctuary coast (Italy). For 11 animals, out of a total of 14 recovered, the cause of death was classified as natural, anthropic, or not determined based on gross and histological pathology and ancillary testing. Hexachlorobenzene (HCB), dichlorodiphenyltrichloroethanes (DDTs), and polychlorinated biphenyls (PCBs) were examined in their blubber, and results were discussed according to their toxicological properties. All specimens showed the following trend: PCBs > DDTs >> HCB, and the levels of cancerogenic, mutagenic, and teratogenic organochlorine compounds (T-OCs) were >50% of total OCs. Immunosuppressant organochlorine compound (IS-OC) levels in 10 out of 11 animals were above 50% of the total OCs. PCB levels always exceeded the threshold of 17 mg/kg lipid weight for PCB-induced adverse health effects. The results suggest that bottlenose dolphins living in the Pelagos Sanctuary undergo a high level of exposure to pathogens and OCs, betraying the designation of the Cetacean Sanctuary and, consequently, of a region created for their conservation. Immune dysfunction and infectious disease susceptibility appear to be highly connected with high levels of OC xenobiotics. These data are useful to understand health and mortality trends in cetacean populations, as well as for developing policies for cetacean conservation and management in this important protected area of the Mediterranean.

Galeus melastomus is the most common Pentanchidae in the Mediterranean Sea. A scavenger and opportunistic feeder, and despite the wide distribution, little is known about its feeding habits in Italian waters. The main purpose of this study was to investigate the diet of the blackmouth catshark by analysing the stomach contents. The specimens analysed were obtained from five populations of the Tyrrhenian and of the Ionian Seas, collected from a depth between 40 and 700 m. A total of 259 stomachs were analysed. The stomach contents were grouped into macro-categories and identified to the lowest taxonomic level possible. Crustaceans such as Parapenaeus longirostris, the Cephalopods Heteroteuthis dispar and Onychoteuthis banksii, and Osteichthyes, mostly Myctophidae, were identified. Plastic debris was also found among the stomach contents and classified according to its colour and shape. Osteichthyes represent the most abundant item (44%), above all the Myctophidae family, except for the catshark population from Tuscany, in which the most frequent species were Cephalopods, such as Abralia veranyi and Heteroteuthis dispar. Differences in the plastic debris contents were also observed between the Tuscany population and other populations. These could be explained as a probable consequence of the different depths at which the blackmouth catshark populations were sampled.

Among killer whale forms, type C is a fish-eating form and is the most common in the Ross Sea. In the austral summer 2015, a study was conducted to evaluate the toxicological hazard these marine mammals face in the Antarctic ecosystem. Seven biopsy samples were collected from adult individuals (five males and two females) in the surroundings of the Italian Research Station Mario Zucchelli, Terra Nova Bay, by remote dart sampling from the pack ice. The accumulation levels of persistent organic pollutants (POPs) such as legacy (DDTs, PCBs, and HCB) and emerging (PBDEs and DP) were measured. Moreover, the protein expression of cytochrome P450 (CYP1A1 and 2B) and the mRNA level variations of the peroxisome proliferator-activated receptors α and γ (PPARα-γ) and the estrogen receptor α (ERα), aryl hydrocarbon receptor (AhR), and Cyp1a were evaluated. Twenty PCB congeners, six DDTs, HCB, three HCHs, and fourteen brominated BDEs and DP-syn and anti-isomers were analyzed on freeze-dried blubber biopsy samples by GC-MS. The protein expression was evaluated by Western Blot and the mRNA levels were quantified by quantitative real-time PCR. The average abundance pattern for the contaminants was DDTs > PCBs > HCB > HCHs ≈ PBDEs >> DP. Contaminant levels resulted to be lower when compared to the existing data from the Antarctic type C killer whales from the McMurdo Sound (Ross Sea) and those reported for fish-eating killer whales worldwide. The mRNA levels of the five target genes were successfully quantified, but no statistical correlation was found with POP levels, suggesting that either the low levels of quantified POPs in blubber may not significantly affect the biological responses investigated, or that other stressors could contribute to the alterations of the molecular biomarkers. Although the results showed a lower risk related to contamination compared to more impacted areas, this study provides baseline data for the conservation of this species in an area with high ecological value, recently declared as the largest Marine Protected Area in Antarctica, where pollutants should remain at minimum levels despite increasing multiple stresses existing in the region.

Human activities are at the origin of anthropogenic threats altering ecosystems at any hierarchical level. To mitigate them, environmental managers develop projects to obtain effective outcomes on biological targets of conservation concern. Here, we carried out two new approaches (TAN = Threat Analysis and TRA = Threat Reduction Assessment) aimed at assessing the effectiveness of conservation actions on two threatened beach-nesting landbird species, the Kentish Plover (Charadrius alexandrinus) and the Little Ringed Plover (C. dubius), breeding along a coastal beach of central Italy. Using a score-based evaluation (TAN approach), a panel of experts assessed the extent, intensity, and magnitude of a set of species-specific threats, ranking them from more to less impacting. Domestic dogs, dune trampling, and synanthropic predators appeared as the threats with the most significant magnitudes. Using the TRA approach, experts obtained a rank of threats that were more urgent to solve: i.e., domestic dogs and dune trampling. To contrast with these threats, in 2021, we carried out a conservation project with specific measures that were aimed at reducing the threat magnitude on birds. They included: dune borders demarcation, anti-predatory cages on plover nests, the removal of beach-stranded fishing lines and hooks, field surveillance by volunteers, dog control, social- and mass-media communication, and alliances with stakeholders and institutions. After the project, mechanical beach grooming (>80%), dune trampling, and synanthropic predators (both >60%) showed the highest percentage of impact reduction. The project showed a medium–high level of effectiveness in reducing the total threat magnitude (TRA-I index = 63.08%). The Threat Analysis should be routinely used to arrange a causal chain that is useful for defining the relationships among human-induced threats and ecological targets, selecting the threats with the highest magnitudes. After the projects, the Threat Reduction Assessment may assess the level of threat reduction, suggesting measures for adaptive management.

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.

Interactions between fishing and dolphins can be detrimental, since on one hand dolphins can be lethally entangled by nets and trawls, and on the other dolphins can predate fish caught by nets. For dolphins, this interaction can be dangerous as they can be wounded or accidentally killed; for fishers, the predation of their catch results in economic losses due to reduced quantity and/or quality of catches and damage to fishing gear. During July and November 2020, we surveyed the “dolphin–fisheries conflict” through compiling 209 fisher interviews from nine locations in Italy and Croatia. Fishers mentioned the common bottlenose dolphin (Tursiops truncatus) as the species primarily interacting with fishing, with the major issue being catch damage by predation. The interaction probability varied among gears and seasons, with some fishing activities (e.g., passive nets) more affected than others (e.g., bottom trawls), especially in terms of economic loss (1000–10,000 €/year on average). More than 70% of the fishers claimed that dolphin populations have increased over the last 10 years, in different degrees and based on different areas. Dolphin bycatch rates are generally low; however, 34.6% of respondents reported having captured at least one dolphin during their career. The fishers’ attitude towards acoustic deterrents (“pingers”) as a mitigation measure revealed that few of them were aware of these devices or were using them.

Cetaceans can be considered good natural samplers of biodiversity due to the different hunting strategies they adopt. In this study, the stomach contents of 28 Tursiops truncatus (15 females and 13 males), stranded along Tuscany coasts, NW Mediterranean, between 2008 and 2021, were analyzed. The prey items were identified at the lowest taxonomic level possible, and assessed in terms of abundance, weight, and frequency of occurrence. The index of relative importance (IRI) was also computed. Overall, 2201 bony fishes and 406 cephalopods were identified. The trophic spectrum resulted in high diversity (69 taxa) and the prey species, 53 fishes and 16 cephalopods, live at different levels of the water column. Predation was mainly based on European hake, Merluccius merluccius (%IRI 26.9), and conger eel, Conger conger (%IRI 25.1). The abundant presence of nocturnal species, such as Conger and Ophidion, indicates the nocturnal hunting activity of the bottlenose dolphin. Furthermore, evidence is presented of the dolphins’ ability to capture fish at night, taking advantage of the sound produced by these fish to locate them. Diet did not show any statistical differences among sexes, except that females preyed upon a significantly higher quantity of octopods than males.

We applied two recent approaches largely used in biological conservation: Threat Analysis (TAN) and Threat Reduction Assessments (TRAs), assessing the effectiveness of a project focused on two water-related bird species (common tern, Sterna hirundo and little tern, Sternula albifrons), commonly breeding in some wetlands of Italy. We used the IUCN standardized lexicon for the classification of threats, utilizing a panel of experts to assess a set of regime attributes (extent, severity and magnitude) of each human-induced disturbance. Our aims were: (i) through the TAN approach, to carry out an arrangement and quantification of the main threats acting on our focal species and select the priority ones; (ii) through the TRA approach, to test the effectiveness of an operational project focused on mitigating the threats and improving the breeding success of species (i.e., building rafts and floating islands to encourage their nesting). Using the TAN approach, experts identified the following human-induced threats (IUCN code): 6.1—Generic disturbance; 7.2—Water stress; 7.3—Salinization; 8.8—Vagrant dogs; 8.8—Mediterranean gulls; 8.8—Wild boars, all significantly different in their magnitude. Among them, wild boars and Mediterranean gulls appeared the priority threats with the greatest extent, intensity and magnitude. Using the TRA approach, after the project, we assessed an overall decrease in the threat magnitude of 23.08% (21.42% when considering only the threats directly affected by our project). These data suggest that further efforts should be devoted to achieving greater effectiveness of conservation actions focused on our target species. With limited time and resources to quantify threats, expert-based approaches could be useful for rapidly assessing the effectiveness of small conservation projects by providing a range of scores obtained following an analytical procedure. In this regard, Threat Analysis and Threat Reduction Assessment could be considered useful tools to support adaptive management in project management cycles.

Considering the rapid growth of tourism in recent years and the acknowledgement that exposure to solar UV radiation may cause skin cancer, sunscreens have been widely used by beachgoers in recent decades. UV filters contained in sunscreens, however, were recently identified as emerging pollutants in coastal waters since they accumulate in the marine environment with different adverse effects. In fact, exposure to these components was proven to be toxic to most invertebrate and vertebrate marine species. Some UV filters are linked to the production of significant amounts of reactive oxygen species (ROS), such as hydrogen peroxide, and the release of inorganic micronutrients that may alter the status of coastal habitats. Bioaccumulation and biomagnification have not yet been fully addressed. This review highlights recent progress in research and provides a comprehensive overview of the toxicological and ecotoxicological effects of the most used UV filters both on the abiotic and biotic compartments in different types of coastal areas, to gain a better understanding of the impacts on coastal biodiversity.

The aim of this study was to evaluate the expression of Major histocompatibility complex (MHC) class I chain-related protein A (MICA) in fibroblast cell cultures of cetaceans (skin biopsies of free-ranging specimens and skin samples of freshly stranded cetaceans) by an immunofluorescence technique and to outline possible variations in MICA expression linked to different ecological and biological factors, while also investigating MICA expression after in vitro treatments with different contaminants. Free-ranging or stranded specimens of cetaceans were sampled in the Sea of Cortez (Mexico) (Balaenoptera edeni, Delphinus capensis, and Orcinus orca) and in the Mediterranean Sea (Balaenoptera physalus, Physeter macrocephalus, Tursiops truncatus, and Stenella coeruleoalba). Cell cultures were treated with an OC mixture, flame retardants, PAHs, MeHg, and BPA. The three species from the Sea of Cortez showed higher basal activity of MICA and lower levels of DDTs and PCBs than the Mediterranean species. A Pearson's linear coefficient equal to −0.45 also confirmed this tendency to have high levels of MICA and low total OC levels. Treatment of cultured fibroblasts with different contaminants mostly resulted in the upregulation of MICA protein expression by at least one treatment dose; downregulation was also found in some species or treatments. MICA alteration indicates a state of stress of the organism and a modification of the immune system's response and can be proposed as a non-invasive immunological marker that can be measured in skin biopsy samples, thus offering a good alternative to blood measurements.

Immunology of marine mammals is a relatively understudied field and its monitoring plays an important role in the individual and group management of these animals, along with an increasing value as an environmental health indicator. This study was aimed at implementing the knowledge on the immune response in cetaceans stranded along the Italian coastline to provide a baseline useful for assessing the immune status of bottlenose (Tursiops truncatus) and striped (Stenella coeruleoalba) dolphins. In particular, since the Mediterranean Sea is considered a heavily polluted basin, a comparison with animals living in open waters such as the Atlantic Ocean was made. Formalin-fixed, paraffin-embedded spleen, thymus, and lymph node tissues from 16 animals stranded along Italian and 11 cetaceans from the Canary Island shores were sampled within 48 h from death. Information regarding stranding sites, gender, and age as well as virologic, microbiological, and parasitological investigations, and the cause and/or the death mechanism were also collected in order to carry out statistical analyses. Selected tissues were routinely stained with hematoxylin-eosin (H&E) and with immunohistochemical techniques (IHC). For IHC analysis, anti-human CD5 monoclonal mouse antibody to identify T lymphocytes, CD20 monoclonal mouse antibody for the identification of mature B lymphocytes and HLA-DR antigen (alpha-chain) monoclonal mouse antibody for the identification of the major histocompatibility complex type II were previously validated for both species by Western-blotting technique. T-test method applied to quantitative evaluation of IHC positive cells showed a significant relationship between the number of (expression) of CD20 stained lymphocytes and normal and hypoplastic lymph nodes, respectively. No other significant correlations were noticed. Analyses for organochlorines (OC) compounds were performed in animals (n°5) having frozen blubber tissue available. A simple linear regression was calculated to predict if the amount of OCs could influence the number of inflammatory cell subpopulations and a moderate negative correlation was found between the presence of high quantity of contaminants and the number of T lymphocytes. Future analysis should be aimed to understand the effect of the major immunomodulatory pathogens on sub-populations of B and T cells.

Two striped dolphins (SD1, SD2), stranded along the Ligurian coast of Italy, were diagnosed with a nonsuppurative meningoencephalitis associated with previously undescribed protozoan tissue cysts. As tissue cysts were morphologically different from those of Toxoplasma gondii, additional histopathological, immunohistochemical, ultrastructural, and biomolecular investigations were performed, aiming to fully characterize the organism. Histopathology revealed the presence of large Sarcocystis-like tissue cysts, associated with limited inflammatory lesions in all CNS areas studied. IHC was inconclusive, as positive staining with polyclonal antisera did not preclude cross-reaction with other Sarcocystidae coccidia. Applied to each animal, 11 different PCR protocols precluded a neural infection by Sarcocystis neurona, Sarcocystis falcatula, Hammondia hammondi, and Neospora caninum. T. gondii coinfection was confirmed only in dolphin SD2. Sarcocystis sp. sequences, showing the highest homology to species infecting the Bovidae family, were amplified from SD1 myocardium and SD2 skeletal muscle. The present study represents the first report of Sarcocystis-like tissue cysts in the brain of stranded cetaceans along with the first description of Sarcocystis sp. infection in muscle tissue of dolphins from the Mediterranean basin.

Tattoo skin disease (TSD) is a poxviral disease typical of cetaceans. Two juvenile and well-preserved male striped dolphins (Stenella coeruleoalba), found stranded along the Tuscany and Latium coasts of Italy in 2015 and 2016, respectively, showed typical skin lesions ascribable to TSD. Histological, ultrastructural and biomolecular investigations confirmed a poxviral aetiology for the aforementioned skin lesions. To our knowledge, this should be the first report of TSD in cetaceans stranded along the Italian coastline. As organochlorines like PCBs and DDTs are known to be highly immunotoxic, the tissue loads of these contaminants were evaluated, in order to increase our knowledge on their potential role as well as on the relationships between the level of exposure to these pollutants and poxviral infection's occurrence.

The Mediterranean Sea is greatly affected by marine litter. In this area, research on the impact of plastic debris (including microplastics) on biota, particularly large filter-feeding species such as the fin whale (Balaenoptera physalus), is still in its infancy. We investigated the possible overlap between microplastic, mesoplastic and macrolitter accumulation areas and the fin whale feeding grounds in in a pelagic Specially Protected Area of Mediterranean Importance (SPAMI): the Pelagos Sanctuary. Models of ocean circulation and fin whale potential habitat were merged to compare marine litter accumulation with the presence of whales. Additionally, field data on microplastics, mesoplastics, and macrolitter abundance and cetacean presence were simultaneously collected. The resulting data were compared, as a multi-layer, with the simulated distribution of plastic concentration and the whale habitat model. These data showed a high occurrence of microplastics (mean: 0.082 items/m2, STD ± 0.079 items/m2) spatial distribution agreed with our modeling results. Areas with high microplastic density significantly overlapped with areas of high macroplastic density. The most abundant polymer detected in all the sampling sites was polyethylene (PE), suggesting fragmentation of larger packaging items as the primary source. To our knowledge, this is the first study in the Pelagos Sanctuary in which the simulated microplastic distribution has been confirmed by field observations. The overlap between the fin whale feeding habitat and the microplastic hot spots is an important contribution for risk assessment of fin whale exposure to microplastics.

This paper summarizes more than four decades of cetacean research data collected by the Museo Civico di Storia Naturale di Milano and, between 1986 and 2003, by the Centro Studi Cetacei of the Società Italiana di Scienze Naturali. It is the result of the collaboration among scientists of several Italian museums. A detailed analysis concerning the amount and taxonomic representative- ness of the extant cetacean collections in Italian naturalistic museums and analogous institutions up until and including 2007 has been carried out. Adequately preserved and anatomically most significant specimens only have been taken into consideration. On the whole 1033 specimens representing 41 species are considered herein. They are housed in 53 institutions, of which the following ones hold the most important collections: the museums of Calci (Pisa), Genova, Firenze, Milano, Roma Zoology, Siena, Comiso and Napoli. All the surveyed institutions are listed, with a short introduction about their material and an inven- tory of it. Specimens are then arranged in systematic order and some further data are provided in a table for each species. Such tables report the items of each museum with regard to quantity, preserva- tion techniques and, whenever possible, collecting data. Finally, a comparative analysis of the results is presented under multiple profiles: historical, preservation techniques, suitability of the specimens for research, place of origin, and the importance of the Italian cetacean collections for research and education.

BackgroundMass strandings of sperm whales (Physeter macrocephalus) remain peculiar and rather unexplained events, which rarely occur in the Mediterranean Sea. Solar cycles and related changes in the geomagnetic field, variations in water temperature and weather conditions, coast geographical features and human activities have been proposed as possible causes. In December 2009, a pod of seven male sperm whales stranded along the Adriatic coast of Southern Italy. This is the sixth instance from 1555 in this basin.Methodology/principal findingsComplete necropsies were performed on three whales whose bodies were in good condition, carrying out on sampled tissues histopathology, virology, bacteriology, parasitology, and screening of veins looking for gas emboli. Furthermore, samples for age determination, genetic studies, gastric content evaluation, stable isotopes and toxicology were taken from all the seven specimens. The animals were part of the same group and determined by genetic and photo-identification to be part of the Mediterranean population. Causes of death did not include biological agents, or the "gas and fat embolic syndrome", associated with direct sonar exposure. Environmental pollutant tissue concentrations were relatively high, in particular organochlorinated xenobiotics. Gastric content and morphologic tissue examinations showed a prolonged starvation, which likely caused, at its turn, the mobilization of lipophilic contaminants from the adipose tissue. Chemical compounds subsequently entered the blood circulation and may have impaired immune and nervous functions.Conclusions/significanceA multi-factorial cause underlying this sperm whales' mass stranding is proposed herein based upon the results of postmortem investigations as well as of the detailed analyses of the geographical and historical background. The seven sperm whales took the same "wrong way" into the Adriatic Sea, a potentially dangerous trap for Mediterranean sperm whales. Seismic surveys should be also regarded as potential co-factors, even if no evidence of direct impact has been detected.