Members of the Coronaviridae family infect humans as well as domestic and wild animals. Over the past three decades, three members of this family, all with zoonotic origins, have caused significant epidemics or pandemics (SARS, MERS, and COVID-19). Despite the spread of SARS-CoV-2 being primarily driven by human-to-human transmission, various animal species are susceptible to infection and may contribute to viral circulation. Aim of this work was to monitor coronavirus (CoV) infections in wild mammals in the Emilia-Romagna region (RER), Italy, using a combined approach of molecular screening for viral RNA detection and serological testing for anti-SARS-CoV-2 antibodies. Respiratory and gastrointestinal tissue samples were collected from wild animal carcasses between 2022 and 2024. Samples were tested for SARS-CoV-2 using two RT-qPCR assays targeting the E and N genes, and for other CoVs using a nested pan-coronavirus RT-PCR followed by Sanger sequencing of positive samples. Additionally, serum samples obtained from blood, cardiac clot, or thoracic exudate were screened for antibodies against the SARS-CoV-2 nucleocapsid (N) protein, with positive samples subsequently confirmed by an ELISA targeting antibodies to the receptor-binding domain (RBD) of the Spike (S) protein, focused on variants circulating during the study period. Molecular analyses were performed on 2,238 animals, all of which tested negative for SARS-CoV-2, while 90 (79

Marine litter is now recognized as one of the greatest environmental challenges of our century, with plastics accounting for almost 80

Poisoning caused by Taxus baccata is a well-known cause of sudden death in domestic animals due to the cardiotoxic effects of taxine alkaloids. This study describes two cases of fatal intoxication in donkeys (Equus africanus asinus) and demonstrates a multidisciplinary diagnostic approach combining pathology, botanical identification, and toxicology. Two animals were found dead without prior clinical signs on a farm in north-eastern Italy. Necropsies were performed, and samples were collected for further investigations. Histopathological findings were limited and non-specific, consistent with the hyperacute course typical of yew poisoning. Fragments of plant material resembling yew needles and twigs were identified in the gastric contents. Toxicological analysis using liquid chromatography-electrospray ionization mass spectrometry confirmed the presence of taxane alkaloids, supporting the diagnosis of yew poisoning. These data highlight the importance of integrating necropsy results with botanical examination and targeted toxicological analyses in cases of suspected plant poisoning. This multidisciplinary approach provides a reliable diagnostic framework for confirming yew poisoning in veterinary investigations.

Thanks to its ethological and physiological characteristics, the hedgehog is a synanthropic species of particular importance for the maintenance and possible spread of pathogens, some of which are zoonotic. Among these, we can include the mammalian orthoreovirus (MRV), which is characterized by respiratory, gastrointestinal, and neurological symptoms in both animals and humans. MRV is characterized by a high capacity for genetic reassortment and intragenic rearrangement, and the ability to infect a wide range of mammals. This work aims to investigate the presence of MRVs and its genomic characterization in hedgehogs. During the two-year period from 2022 to 2023, the intestine and lungs were collected from 293 hedgehogs and subjected to real-time PCR to detect the L1 gene. Positive samples were subjected to a typing RT-PCR targeting a portion of the S1 gene and then to sequencing. A total of 38 hedgehogs tested positive by real-time PCR (p = 13%). Typing RT-PCR demonstrated the positivity of 25 samples for serotype 3. Four samples, representative of the main groups recognized during the phylogenetic analysis, underwent whole genome sequencing, revealing the presence of reassortment phenomena between strains related to bats, chamois, and human MRVs.

Monitoring stranded cetaceans represents a strategic method to assess their health, conservation status, and ecological role in the marine ecosystem. Efficient stranding networks and standardized protocols are essential to monitor this phenomenon and investigate its causes. This study assesses the evidence of fishery interaction on stranded cetacean carcasses found along the Italian coastline from 1986 to 2023. Evidence assessment and post-mortem investigation methods evolved over three macro-periods, from non-standardized reporting (1986-2014) to an integrated national stranding network (2015-2019), and finally to the creation of a new standardized, evidence-based diagnostic framework under the EU-funded LIFE DELFI project (LIFE18 NAT/IT/000942) (2020-2023). A total of 5355 cases were selected for this analysis. A literature review and evidence of interaction on stranded carcasses supported the categorization of findings, ranging from case history to pathological observations, allowing the assessment of temporal variation, demographic parameters, geographical distribution, and fishing gear identification. Evidence of fishery interaction was found in 12.89% of the cases (690/5355), with an annual average of 18.15 affected animals, and fishery interaction was identified as the likely cause of death in 10.32% of the cases. The most frequently reported species were Stenella coeruleoalba and Tursiops truncatus, showing significant differences in fishery interactions, particularly in relation to sex, age class, and geographical distribution. Adult male Tursiops truncatus exhibited a higher susceptibility to gillnet interaction in the Adriatic Sea. The results of this study emphasize the importance of standardized post-mortem investigations and long-term monitoring to identify risk hotspots, implement species- and region-specific mitigation strategies, and establish threshold values for cetacean conservation.

The Adriatic Sea is a critical neritic habitat for juvenile and adult female loggerhead sea turtles (Caretta caretta), where intense anthropogenic pressures and environmental stressors may influence their reproductive biology. Knowledge on the onset of puberty in this population is limited by scarce information on the sub-adult stage, a transitional phase in which reproductive competence is acquired. This study integrated histological analysis and Fourier-transform infrared (FTIR) imaging spectroscopy to provide both structural and biochemical characterization of folliculogenesis, with emphasis on vitellogenesis, in C. caretta from the north-central Adriatic Sea. Histological analysis determined the progression of follicle development, while FTIR imaging, a label-free and spatially resolved technique, mapped the distribution of proteins, lipids, and nucleic acids across ovarian compartments. Logistic regression estimated the size at which 50% of females are sexually mature (L50) at 58.54 cm Curved Carapace Length (CCL). Based on this value, 60% of sub-adult females were already mature, indicating earlier puberty than previously inferred from macroscopic criteria. These preliminary results, along with reports of sporadic nesting in the Adriatic, raise the question of whether this basin may host further nesting events in the future. FTIR imaging proved to be a powerful tool for reproductive biology in non-model marine vertebrates.

Trichinella spp. are zoonotic nematodes with a global distribution, primarily maintained through wildlife reservoirs which complicates eradication efforts. In Europe, four species-T. spiralis, T. britovi, T. nativa, and T. pseudospiralis-circulate in wild carnivores and omnivores, with T. britovi and T. spiralis being the most prevalent in wild and domestic hosts. This study aimed to assess the presence and species distribution of Trichinella spp. in wild mammals. This study presents the results of five years of wildlife surveillance carried out within the long-standing monitoring program implemented in the Emilia-Romagna region, northern Italy, which has been in place since 2006. Between 2020 and 2024, a total of 104,338 wild mammals, including red foxes, wolves, and wild boar, were tested using the magnetic stirrer digestion method in accordance with EU regulations. A total of 12 animals (0.011%) were found to be infected with Trichinella larvae. Molecular identification of positive samples, performed at the European Union Reference Laboratory for Parasites, showed the presence of T. britovi in 11 animals, while one isolate remained unidentified. T. britovi was detected primarily in wolves and red foxes. No infections were identified in wild boar during the study period. Overall, the low prevalence observed confirms that Trichinella spp. infections in wildlife in Emilia-Romagna are sporadic but persistently present. Only T. britovi was detected, supporting its role as the predominant species in this area. These findings highlight the importance of long-term surveillance for early detection and risk assessment within a One Health framework.

Marine litter represents a growing threat to marine biodiversity, particularly to cetaceans, yet its impacts on these sentinel species remain insufficiently quantified. This study provides the first comprehensive, transboundary assessment of litter ingestion in stranded cetaceans along the Italian coastline and across the wider Adriatic basin, including Croatia and Slovenia, between 2009 and 2023. Through harmonized post-mortem examinations, and focusing on the period of consistent data collection and analysis (2009-2023), this study documented plastic litter ingestion in 2.9% of necropsied cetaceans in Italy and 3.7% in the broader Adriatic subregion, with sperm whales (Physeter macrocephalus) showing the highest frequency (50% FO) and susceptibility. In 11 cases, ingestion was associated with health deterioration and mortality. The most commonly ingested items were plastic sheets and fragments. The Italian Adriatic subregion emerged as a hotspot for plastic interactions, reflecting regional hydrodynamics and anthropogenic pressures. Applying criteria from regional and international frameworks, the results showed that 60% of P. macrocephalus had ingested more than 1 kg of plastic, with 40% exhibiting harmful effects. These data provide baseline values that can serve as reference points for proposing thresholds to achieve Good Environmental Status under the Marine Strategy Framework Directive. Despite these results supporting the use of sperm whales as suitable sentinel species for monitoring macroplastic pollution, the available data are limited to Italy and influenced by distributional patterns and unusual mortality events. A combined approach, where T. truncatus is monitored for its broad spatial representativeness and P. macrocephalus for its ecological susceptibility, may be a useful strategy to guide further research and inform management measures in the future. These findings underscore the need for standardized monitoring protocols, enhanced cross-border data sharing, and policy measures to mitigate plastic impacts. This work provides crucial baseline knowledge for conservation planning and reinforces the role of cetaceans as indicators of ecosystem health in the Mediterranean.

The detection of tetrodotoxins (TTXs) in European shellfish led the European Union to request a risk assessment from the European Food Safety Authority (EFSA). EFSA set a reference limit of 44 µg TTX equivalents kg−1 and called for more data on TTX occurrence, especially in gastropods, which can accumulate in TTXs but remain poorly studied. Recently, preliminary monitoring has revealed the recurrent presence of TTXs in mussels in three areas along the North–Central Adriatic coast of Italy, while research on non-bivalve organisms has not yet been carried out. This study presents a preliminary survey, conducted from January 2023 to March 2025, on the presence of TTXs in gastropods, echinoderms, and arthropods collected from this area. A method in Hydrophilic Interaction Liquid Chromatography coupled with tandem Mass Spectrometry (HILIC-MS/MS) for detecting TTXs in bivalve mollusks was first tested through an international proficiency test, then optimized for the other invertebrates, the object of this study. TTX levels in all gastropods and arthropod samples were undetectable, while traces (~5 µg kg−1) were found in one echinoderm sample (Paracentrotus lividus), marking the first reported occurrence of TTX in this species. Sea urchins are widely consumed in Italy; therefore, this finding is of particular importance from a public health perspective and deserves further investigation. Some gastropod genera or species sampled (e.g., Nassarius, Rapana venosa) have been identified as TTX carriers in other regions; therefore, the negative results obtained in this study may be related to seasonal or geographic variability. These results provide valuable data to EFSA’s call for monitoring emerging risks, particularly as climate change may increase TTX prevalence in European waters as well as worldwide.

Plastic marine litter is a critical issue that threatens marine ecosystems. This study investigated microplastics (MPs) contamination in the Italian seas, involving regions significantly affected by pollution from urban, industrial and agricultural sources. The research, conducted in collaborations between 10 different Experimental Zooprophylactic Institutes throughout Italy, analyzed Mytilus galloprovincialis (common mussels) for its filtration capacity and suitability as a bioindicator. Using data from two projects funded by the Italian Ministry of Health, MPs were detected from 7% to 13% of mussel samples, mainly polypropylene and polystyrene fragments and fibers. These findings align with previous studies highlighting the pervasive presence of MPs and their potential risks as mussels are consumed whole, allowing MPs to be ingested. The study underscores the need for standardized detection methods and coordinated policies to mitigate plastic pollution. Public awareness campaigns and improved waste management practices are key to addressing the environmental and health impacts of MPs. Further research on the long-term effects of MPs on marine ecosystems and human health is essential to developing comprehensive mitigation strategies.

Oleander (Nerium oleander) contains more than 30 different toxic cardiac glycosides, including oleandrin. Intoxication can occur through ingestion or inhalation in several species, including cattle. A total of 76 of 205 beef cattle died within 30 h due to the burning of oleander mowing. The burning area was located approximately 20 meters from the animal housing facility. The clinical signs before death were depression, sternal decubitus, and dyspnea. Macroscopic examination revealed cavitary effusions, diffuse edema, and hemorrhagic changes. Histopathological examination confirmed hemorrhagic and edematous findings, minimal neutrophilic infiltration and hemorrhage in the heart, centrilobular hepatic necrosis, multifocal myopathy and necrosis, and chronic bronchitis. Oleandrin was detected in the lungs, kidneys, intracardiac clot, ruminal content, and liver. The concentration of oleandrin differed among tissues and animals, supporting the hypothesis that 74 of the 76 animals died from inhalation intoxication caused by oleander. The remaining two animals, which survived the hyperacute phase, later died due to secondary oleandrin intoxication via ingestion, which aggravated their pre-existing health conditions. To the best of the authors’ knowledge, this case represents the first reported instance of inhalation intoxication by oleander in cattle.

Second-generation Anticoagulant Rodenticides (ARs) can be critical for carnivores, due to their widespread use and impacts. However, although many studies explored the impacts of ARs on small and mesocarnivores, none assessed the extent to which they could contaminate large carnivores in anthropized landscapes. We filled this gap by exploring spatiotemporal trends in grey wolf (Canis lupus) exposure to ARs in central and northern Italy, by subjecting a large sample of dead wolves (n = 186) to the LC-MS/MS method. Most wolves (n = 115/186, 61.8 %) tested positive for ARs (1 compound, n = 36; 2 compounds, n = 47; 3 compounds, n = 16; 4 or more compounds, n = 16). Bromadiolone, Brodifacoum and Difenacoum, were the most common compounds, with Brodifacoum andBromadiolone being the ARs that co-occurred the most (n = 61). Both the probability of testing positive for multiple ARs and the concentration of Brodifacoum, and Bromadiolone in the liver, systematically increased in wolves that were found at more anthropized sites. Moreover, wolves became more likely to test positive for ARs through time, particularly after 2020. Our results underline that rodent control, based on ARs, increases the risks of unintentional poisoning of non-target wildlife. However, this risk does not only involve small and mesocarnivores, but also large carnivores at the top of the food chain, such as wolves. Therefore, rodent control is adding one further conservation threat to endangered large carnivores in anthropized landscapes of Europe, whose severity could increase over time and be far higher than previously thought. Large-scale monitoring schemes for ARs in European large carnivores should be devised as soon as possible.

Abstract Background Benzalkonium chloride (BAC) is a quaternary ammonium compound (QAC), that can be found in a wide variety of household products–from disinfectants to medicaments and home fragrances–but also professional products. In pets, cats have long been reported as more sensitive than dogs to QACs; in fact, signs of irritation such as oral ulcerations, stomatitis and pharyngitis can be observed after contact with concentrations of 2% or lower. In a review of 245 cases of BAC exposure in cats, reported by the Veterinary Poisons Information Service (United Kingdom) only 1.2% of the cases died or were euthanized. Nevertheless, BAC toxidromes in cats can result in transitory CNS and respiratory distress, as well as severe mucosal and cutaneous lesions. Currently, only a few reports are available concerning BAC poisoning in this species. Case presentation A 4 month-old kitten presented with severe glossitis, lameness in the hindlimbs and episodes of vomiting and diarrhoea. The cause was unknown until the owners reported use of a BAC-containing mould remover (5%) 4 days later. The patient developed severe oral burns requiring a pharyngeal tube for feeding and severe cutaneous chemical burns. The kitten was managed with supportive therapy and required hospitalization for 10 days. The symptoms disappeared completely 3 weeks after exposure. Conclusions BAC is a very common compound contained in several household and professional products but, to the best of our knowledge, no previous case had been reported in Italy. We hope that this report will help raise awareness on the hazards of BAC products for cats in both domestic and work contexts.

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.

Along the Italian coasts, toxins of algal origin in wild and cultivated shellfish have been reported since the 1970s. In this study, we used data gathered by the Veterinary Public Health Institutes (IZS) and the Italian Environmental Health Protection Agencies (ARPA) from 2006 to 2019 to investigate toxicity events along the Italian coasts and relate them to the distribution of potentially toxic species. Among the detected toxins (OA and analogs, YTXs, PTXs, STXs, DAs, AZAs), OA and YTX were those most frequently reported. Levels exceeding regulatory limits in the case of OA (<= 2,448 mu g equivalent kg(-1)) were associated with high abundances of Dinophysis spp., and in the case of YTXs (<= 22 mg equivalent kg(-1)) with blooms of Gonyaulax spinifera, Lingulodinium polyedra, and Protoceratium reticulatum. Seasonal blooms of Pseudo-nitzschia spp. occur all along the Italian coast, but DA has only occasionally been detected in shellfish at concentrations always below the regulatory limit (<= 18 mg kg(-1)). Alexandrium spp. were recorded in several areas, although STXs (<= 13,782 mu g equivalent kg(-1)) rarely and only in few sites exceeded the regulatory limit in shellfish. Azadinium spp. have been sporadically recorded, and AZAs have been sometimes detected but always in low concentrations (<= 7 mu g equivalent kg(-1)). Among the emerging toxins, PLTX-like toxins (<= 971 mu g kg(-1) OVTX-a) have often been detected mainly in wild mussels and sea urchins from rocky shores due to the presence of Ostreopsis cf. ovata. Overall, Italian coastal waters harbour a high number of potentially toxic species, with a few HAB hotspots mainly related to DSP toxins. Nevertheless, rare cases of intoxications have occurred so far, reflecting the whole Mediterranean Sea conditions.

This study reports the first detection of the marine neurotoxin pinnatoxin-G (PnTX-G) in clams collected in the northwestern Adriatic Sea (Italy). It also represents the first report of the potential toxin-producing dinoflagellate, Vulcanodinium rugosum, in Italian seas. This result, from the coasts of the Emilia-Romagna Region, indicates a successful colonization process, reflecting conditions in France where V. rugosum was initially documented. In this case, the concentration of PnTXs was very low, making further sampling necessary to fully understand the extent of the phenomenon. Discussions on the need to obtain more data to support a proper risk assessment and the need to implement a monitoring program that includes emerging marine biotoxins are also included.

Animals, including wildlife, are part of One-Health concept since many infectious diseases can affect both humans and animals. In this study, 126 red foxes (Vulpes vulpes) from Northern Italy in 2022–2023 were tested by molecular assays for Protoparvovirus carnivoran 1 (PPVC-1), Canine adenovirus type 1 and 2 (CAdV-1 and CAdV-2), Circovirus canine (CanineCV), Canine distemper virus (CDV), and Leptospira spp. A total of 39 of 126 (30.9%) red foxes were infected with at least one pathogen and five of these were coinfected: 20/126 (15.9%) red foxes tested positive for PPVC-1, 3/126 (2.4%) for CAdV, 20/126 (15.9%) for CanineCV, and 2/126 (1.6%) for Leptospira spp. DNA. No foxes tested positive for CDV RNA. The pathogens identified were genetically analysed. New findings were reported such as a fox with multiple feline panleukopenia virus (FPV) and canine parvovirus type 2b (CPV-2b) infection associated with quasispecies dynamics, typical genetic characteristics of the identified CanineCV, and the first detection in red foxes of Leptospira ST198 related to L. interrogans serogroup Australis. Further studies are necessary to investigate the transmission between domestic animals and wildlife and to understand the role of red foxes in the maintenance of these pathogens not only in the wild but also in urban and peri-urban environments.

A precise and accurate method for enumeration of Escherichia coli is critical to classify production and relaying areas for live bivalve molluscs. Paired comparison of most probable number and direct plating count of E. coli was performed on a total of 918 samples of Manila clams (Ruditapes philippinarum) collected from official monitoring and from a regional shellfish monitoring program conducted from 2011 to 2017 in Emilia Romagna region, Italy. The records of E. coli enumeration resulting from both MPN reference method and alternative TBX count method were compared with McNemar's test using three cut-off values, namely 230, 700 and 4.600 MPN/100g, based on the requirements laid down in Regulation (EU) 2019/627. The relative trueness was calculated. Significant differences were observed between the two methods for each investigated cut-off: these incongruities concerned a total of 129 (14%), 131 (14%) and 56 (6%) samples observed as negative with TBX but positive by MPN (respectively using 230, 700 and 4.600 E. coli as cut-off), as well as, quite the opposite, a total of 20 (2,2%), 8 (0,9%) and 5 (0,5%) samples observed negative by MPN but positive with TBX method. A negative bias is reported between the two methods. Applying the outcomes of the two different methods to a real scenario, the Veterinary Competent Authority could classify production and relaying areas in the same area of classification only for 69% of samples. This high degree of disagreement between MPN and direct plating outcomes clearly shows the different and not negligible impact of the two methods on the specific E. coli requirements for the classification of production and relaying areas for R. philippinarum. Analytical methods, either official or validated, if used officially by Veterinary Authority must lead to the same classification of production and relaying areas for live bivalve molluscs.

Listeria monocytogenes is an ubiquitous pathogen isolated from different host species including fish, crustaceans, and molluscs, but it is rarely a pathogenic microorganism to marine reptiles. In particular, only two cases of fatal disseminated listeriosis have been described in the loggerhead sea turtle (Caretta caretta). In this study, we describe a lethal case of L. monocytogenes infection in a loggerhead sea turtle. The turtle was found alive, stranded on a beach in North-eastern Italy, but perished soon after being rescued. The autoptic examination revealed that heart, lung, liver, spleen, and urinary bladder were disseminated with multiple, firm, 0.1-0.5 mm sized, nodular, white-green lesions. Microscopically, these lesions corresponded with heterophilic granulomas with Gram+ bacteria within the necrotic center. Furthermore, the Ziehl-Neelsen stain was negative for acid-fast organisms. Colonies isolated from heart and liver were tested through MALDI-TOF for species identification, revealing the presence of L. monocytogenes. Whole Genome Sequencing on L. monocytogenes isolates was performed and the subsequent in silico genotyping revealed the belonging to Sequence Type 6 (ST 6); the virulence profile was evaluated, showing the presence of pathogenicity islands commonly observed in ST 6. Our results further confirm that L. monocytogenes should be posed in differential diagnosis in case of nodular lesions of loggerhead sea turtles; thus, given the zoonotic potential of the microorganism, animals should be treated with particular caution. In addition, wildlife animals can play an active role as carriers of possibly pathogenetic and virulent strains and contribute to the distribution of L. monocytogenes in the environment.