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Milk has been consumed by humans for thousands of years for its nutritional properties. In recent years, raw milk demand has increased, valued for its authenticity and connection to local traditions. In Italy, the sale of raw milk is allowed exclusively through direct sale from the producing farm to the final consumer, either at the producing farm itself or through vending machines. However, the consumption of raw milk is not without risks. Among these, microbiological ones are relevant. These can lead to severe symptoms, particularly in vulnerable populations. For this reason, although consumers are advised to boil raw milk before consumption, producing farms in Italy are required to meet the microbiological criteria outlined in the Provision of 25 January 2007. In this retrospective study, the results of the analyses performed on 355 raw milk samples collected in Liguria between 2014 and 2023 for the detection of Campylobacter spp., Salmonella spp., Listeria monocytogenes , Staphylococcus aureus , and Escherichia coli O157 were analysed to better characterise the associated risk for consumers. The samples were collected during official controls by the local veterinary health services at vending machines of seven producing farms. Overall, six samples tested positive for C. jejuni , while only one sample tested positive for Salmonella enterica subsp. enterica , Serovar Veneziana. Listeria monocytogenes , S. aureus , and E. coli O157 were never responsible for non-compliances. Interestingly, three of the six samples positive for C. jejuni derived from the same producer. In farms where positive samples were detected, certain structural and/or operational non-compliances were identified. It can be concluded that, although the scenario in question does not present any cause for concern, it is nevertheless essential to implement a series of preventive measures in order to guarantee the safety of raw milk. These measures include the implementation of biosecurity practices, the maintenance of strict hygiene protocols during milking, and the adherence to the cold chain distribution protocol until the final stage of distribution.

The European subtype of tick-borne encephalitis virus (TBEV-Eur; species Orthoflavivirus encephalitidis , family Flaviviridae ) was the only tick-borne flavivirus present in central Europe known to cause neurologic disease in humans and several animal species. Here, we report a tick-borne flavivirus isolated from Alpine chamois ( Rupicapra rupicapra rupicapra ) with encephalitis and attached ticks, present over a wide area in the Alps. Cases were detected in 2017 in Salzburg, Austria, and 2023 in Lombardy and Piedmont, Italy. The virus strains exhibit 94.8-97.3% nucleotide identities to each other and are more closely related to Louping ill viruses (LIV; Orthoflavivirus loupingi ; 90-92% identities) than to TBEV-Eur (less than 88%). The chamois-derived virus strains, tentatively termed "Alpine chamois encephalitis virus", form a well-supported independent genetic clade with Spanish goat encephalitis virus, clearly separated from other LIV. This supports its designation as a new virus subtype with the proposed shared taxonomic name "Spanish goat and Alpine chamois encephalitis virus subtype" within the species Orthoflavivirus loupingi . The zoonotic potential of this newly identified virus subtype as well as its host range in other animal species including farm animals needs to be further investigated.

The presence of rare earth elements (REE) in the southern hemisphere, particularly marine ecosystems of Patagonia, have received little attention. The Magellanic penguin, which is also known as the Patagonian penguin, inhabits only in austral regions of South America. Although seabird feathers have been used extensively as a bio-monitoring tool, no studies have addressed the effect of age on REE accumulation in Magellanic penguins. In this study, the concentrations of REE were determined by ICP-MS to detect La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Sc, Lu and Y in the feathers of Magellanic penguins from Magdalena Island, an important rookery in the Strait of Magallan. Age-related differences were studied to investigate the different patterns of REE bioaccumulation between adults and juveniles. The data showed that juvenile penguins exhibit higher REE-levels than adult individuals (p < 0.05). Mean REE-values (μg g -1 d.w) differed several orders of magnitude, ranging from 0.002 for Lu in adults to 1.15 for Ce in juvenile individuals. The results are useful to understand the bioaccumulation of REE in fauna from remote and cold regions of the southern hemisphere., 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 Elsevier Inc. All rights reserved.)

Although wild and domestic carnivores share some haemotropic Mycoplasma species, information about the circulation of this pathogen in grey wolves (Canis lupus) populations is still very limited. Thus, a geographically broad-based investigation was performed for determining the occurrence and diversity of Mycoplasma spp. in three different wolf populations from southern Europe. Between 2001 and 2023, spleen samples from 285 grey wolves from Spain (n = 129), Italy (n = 113), and Portugal (n = 43) were collected. The presence of haemotropic Mycoplasma was assessed targeting the 16S rRNA gene using two PCR assays in parallel; in addition, the 16S-23S rRNA intergenic spacer was analysed for further identification of the positive samples. The influence of the sampling country, sex, and age of the animals on the prevalence of Mycoplasma spp. was also assessed by a generalized linear model analysis. The percentage of positive wolves was 13.3 % (38/285), and the occurrence was significantly higher in Spain (20.9 %) than in Italy (8.0 %) and Portugal (4.7 %). Mycoplasma haemocanis (10.5 %) and Candidatus M. haematoparvum (2.1 %), were identified; in addition, an uncultured Mycoplasma sp. was also detected (0.7 %). Our results confirm the circulation of potentially zoonotic Mycoplasma in wolf populations from southern Europe. To our knowledge, this is the first report of Ca. M. haematoparvum in wolves from Italy and Portugal. In addition, a Mycoplasma sp., previously found in dogs, has been detected for the first time in wolves. Further studies are needed to fully molecularly characterise haemotropic Mycoplasma spp., which will serve as a basis for the study of its ecoepidemiology., Competing Interests: Declaration of Competing Interest Authors declare that they have no competing interests., (Copyright © 2025 Elsevier B.V. All rights reserved.)

African swine fever (ASF), characterized by high mortality rates in infected animals, remains a significant global veterinary and economic concern, due to the widespread distribution of ASF virus (ASFV) genotype II across five continents. In this study, ASFV strains collected in Italy during 2022-2023 from two geographical clusters, North-West (Alessandria) and Calabria, were fully sequenced. In addition, an in vivo experiment in pigs was performed. Complete genomic sequencing of 30 strains revealed large-fragment deletions and translocations. In Alessandria, five samples showed two different deletions in the 5' genomic region: a ~4340 bp deletion (positions ~9020-13,356 in Georgia 2007/1) in four samples and a 2162 bp deletion (positions 17,837-19,998) in one sample. Another strain showed a truncation of 1950 bp at the 3' end. In Calabria, strains showed a 5137 bp deletion (positions 10,755-15,891) and ~2 kb truncations in the 3' region. Two strains showed a translocation from the region 1-2244 to positions 188,631-190,584. In vivo characterization of the deleted strain 22489.4&#95;2312/RC/2023 revealed identical disease progression to the wild-type strain, with severe ASF symptoms in inoculated pigs. This study is the first to report natural deleted strains of ASFV in Italy, revealing unique genomic deletions distinct from those in previously known strains.

Neurofilament light chain (Nf-L) is a biomarker for axonal damage in human neurology but is understudied in cattle. With this study we wanted to determine Nf-L stability at two different storage temperatures and Nf-L levels in healthy cattle and the relationship with age, evaluate whether Nf-L holds diagnostic potential for neurological disorders, and whether an association exists between Nf-L in serum and in cerebrospinal fluid (CSF). To do this, we measured Nf-L levels in CSF and serum samples from 49 healthy and 75 sick cattle. Storage at -80 °C or -20 °C had no impact on Nf-L concentration. Physiological median Nf-L levels were 6.3 pg/mL (serum) and 414 pg/mL (CSF) in calves and 5.5 pg/mL (serum) and 828 pg/mL (CSF) in adult cattle. There was no association between Nf-L levels in CSF and calf age (r 2 0.07, p = 0.13), while a weak association was found for Nf-L in serum (r 2 0.26, p = 0.01), and a significant association in adult cattle (CSF, r 2 0.69, p = 0.0001; serum, r 2 0.68, p = 0.0003). CSF Nf-L levels were higher in samples from animals with degenerative (median Nf-L 49971 pg/mL) and infectious central nervous system (CNS) disorders (median Nf-L, age < 2 months 8863 pg/mL; age 2-12 months 17474 pg/mL; age 1-6 years 3546 pg/mL), CNS anomalies and metabolic/toxic disorders. There was a significant association between CSF Nf-L and serum Nf-L in cattle with neurological disorders (r 2 0.2, p = 0.009). Taken together, these findings suggest the potential of Nf-L as a diagnostic tool in cattle neurology., Competing Interests: Declarations. Ethics approval and consent to participate: This study adhered to current animal welfare regulations (Directive 98/58/EC and Italian Decree Law 146/2001). All procedures were conducted as described by the institutional guidelines in accordance with national (D.L.26/2014) and international laws and policies (EEC Council Directive 63/2010). Ethical approval for the study was obtained from the Bioethics Committee of the University of Turin (protocol no. 0251347) and from the Italian Ministry of Health (Auth. no. 242/2020–PR). Owners provided written informed consent for veterinary assessment and treatment of their animals. Competing interests: The authors declare that they have no competing interests., (© 2025. The Author(s).)

Chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), is a significant threat to global amphibian populations, leading to widespread declines and extinctions. In the spring of 2023, Bd presence was detected in different amphibian species within two protected areas near Turin, Piedmont, Italy, following an unusual mortality event among the common toad ( Bufo bufo ). Histological and molecular analyses confirmed Bd infection in a deceased Pelophylax sp. specimen, prompting further investigation. Sampling of 166 individuals across seven amphibian taxa revealed an overall Bd occurrence of 38.6%, with Pelophylax sp. showing the highest detection rate (50.5% of 93 individuals). A marked difference in the positivity rate was observed between the two locations, with La Mandria (67.2% of 58) exhibiting significantly higher rates than Vauda (22.9% of 35). While Bd was identified in the sampled amphibians, the exact cause of the observed mortality remains unclear and may involve other pathogens or multifactorial causes, including but not limited to Bd. These findings represent the first documented case of Bd presence in Piedmont after an 18-year gap, highlighting the potential influence of local environmental factors on infection dynamics. The study emphasises the need for expanded, standardised field sampling and further investigation into the various factors affecting amphibian health to guide conservation efforts for vulnerable amphibian species.

Rose Bengal antigen and smooth lipopolysaccharide (s-LPS) were produced from a field strain of Brucella ceti ("homologous" antigens) and from the reference strain B. abortus S99 ("heterologous" antigens); they are currently used for the diagnosis of brucellosis in cattle, water buffaloes, sheep, goats, and pigs, as recommended in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (WOAH). "Homologous" and "heterologous" antigens were used in a rapid serum agglutination test (Rose Bengal test, RBT) and a competitive ELISA assay (c-ELISA) to test a panel of sera, blood, and other body fluids (cerebrospinal fluid, pericardial fluid, tracheal fluid, and aqueous humor) collected from 71 individuals belonging to five cetacean species ( Stenella coeruleoalba ; Tursiops truncatus ; Grampus griseus ; Globicephala melas ; and Ziphius cavirostris ), which were found stranded on the Italian coastline. Six animals were positive for Brucella spp. for bacterial isolation and/or PCR, and 55 animals were negative; for the remaining 10 animals, no PCR/isolation data were available. A total of 90 samples were tested. Results obtained from the two tests were compared in order to identify the most suitable antigen for the serological diagnosis of Brucella infection in cetaceans. The RBT performed with the "homologous" antigen showed the best results in comparison with the "heterologous" antigen: diagnostic sensitivity, specificity, and accuracy were 80.0%, 44.1%, and 46.9% for the "homologous" antigen and 80.0%, 17.0%, and 21.9% for the "heterologous" antigen. For the c-ELISA, "homologous" and "heterologous" s-LPS showed similar results (diagnostic sensitivity 66.7%, diagnostic specificity 97.3%, and diagnostic accuracy 95.0%). Therefore, the RBT using the "homologous" antigen and c-ELISA with "homologous" or "heterologous" s-LPS could be used in parallel for the detection of antibodies against Brucella spp. in cetaceans.

Understanding oxidative stress in high-mountain lake fish offers crucial insights into their health, resilience, and adaptation to extreme environmental changes. This study investigates the oxidative stress response of Cottus gobio in a high-mountain lake (Dimon Lake) located in the northeast Italy during the ice-free season, focusing on the relationship between oxidative stress biomarkers and physicochemical water parameters, as well as persistent and emerging contaminants. Significant seasonal variations were observed in water parameters, with lower oxygen, pH, conductivity, and phosphate levels in summer compared to autumn, while temperature, ammonium, and nitrate were higher in summer. Metal concentrations in C. gobio muscle were higher in autumn, with Zn showing the most significant increase. PAHs, NDL-PCBs, and pesticides were all below the limit of quantification in the fish muscle samples. No microplastics items were found in the gastrointestinal tracts of fish. Oxidative stress biomarkers revealed organ-specific and seasonal variations. The liver exhibited the highest activities of catalase (CAT), glutathione peroxidase (GPx), glutathione S-tranferase (GST), and glutathione reductase (GR), highlighting its central role in detoxification and metabolic processes. Superoxide dismutase (SOD) activity was notably higher in muscle tissue during summer, suggesting increased metabolic activity. A strong correlation was found between pH and the activities of SOD, CAT, GPx, GR, and metallothioneins (MTs), emphasizing the importance of water chemistry in influencing oxidative stress responses. This approach not only aids in understanding the local adaptations of these fish but also highlights the impacts of environmental stressors on high-mountain ecosystems. Continuous monitoring of water chemistry, particularly pH, is crucial for understanding and managing oxidative stress in aquatic organisms, especially in the context of global environmental changes., 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 Elsevier Inc. All rights reserved.)

Neonicotinoid insecticides (NNIs) are the fastest-growing class in agricultural protection. They target nicotinic acetylcholine receptors (nAChR) in pests, stimulating the nervous system at low doses and causing paralysis and death at higher concentrations. NNIs are used in crop protection, seed treatment, forestry, agriculture, and flea control in domestic cattle. Effective at lower concentrations and offering long-term control, NNIs are favoured for their systemic activity. However, due to their water solubility, mobility, and moderate persistence, NNIs easily contaminate adjacent aquatic environments via runoff, leaching, or spray drift. While less toxic to vertebrates, their widespread use poses threats to aquatic and terrestrial organisms, causing neurotoxicity, nephrotoxicity, cytotoxicity, genotoxicity, immunotoxicity, hepatotoxicity, endocrine disruption, and reproductive malformations. This review synthesizes research to address knowledge gaps on the environmental impact of NNIs and proposes policies to mitigate their harmful effects on aquatic non-target species., 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.)

Background and Objectives : Janus kinase inhibitors (JAKi) have significantly advanced the treatment of various dermatological conditions by modulating the JAK-STAT signalling pathway. While these inhibitors have proven effective, they also pose challenges due to associated increase in serum lipid levels and relative potential cardiovascular risks. This perspective work aims to discuss the implications of these lipid alterations proposing management strategies for patients with dermatological disorders under JAKi treatments. Materials and Methods : This manuscript reviews existing and recent literature on the metabolic effects of JAKi, particularly focusing on their impact on lipid profiles in patients treated for dermatological diseases. Results : JAK inhibitors are consistently associated with an increase in both LDL and HDL levels shortly after treatment initiation, which tend to stabilise over time. Despite these changes, there is no clear evidence linking these alterations to an increased risk of major adverse cardiovascular events (MACE), indicating a complex interaction between lipid metabolism and JAK-STAT signalling. Conclusions : Although JAKi may induce lipid changes in patients, raising concerns, especially in ones with existing cardiovascular risks, currently there is no proven link to increased MACE in this population. Monitoring lipid levels, alongside lifestyle modifications and possible statin use, can manage these effects without the need to stopping treatment.

Bivalve molluscs are filter-feeding organisms, capable of concentrating pathogenic microorganisms from the surrounding environment, thus contributing to the spread of viral pathogens, which they can transmit to humans, especially if eaten raw or undercooked. Although norovirus (NoV) and the hepatitis A virus (HAV) are considered the most common causes of foodborne infections, in recent years, other viruses with a zoonotic potential have been identified in shellfish, such as the hepatitis E virus (HEV), astrovirus (AsV), and aichi virus (AiV). The aim of the study was to investigate the presence of classical and emerging pathogenic enteric viruses in oysters ( Crassostrea gigas ) and mussels ( Mytilus galloprovincialis ) from a mollusc farming area in the northwest of Italy, between April 2022 and March 2023. In the period considered, a total of 168 samples (84 oysters and 84 mussels) were analysed. The prevalence of NoV was highest, with 32.7% (55/168) positive samples, followed by 18.4% (31/168) for AsV and 19.6% (33/168) for AiV. This study revealed, for the first time, the presence of AsV and AiV in molluscs farmed in this sea area. All the samples tested were negative for HAV and HEV. The emergence of new enteric viruses like AiV and AsV in bivalve molluscs underscores the importance of improving surveillance and environmental monitoring methods, particularly in shellfish production areas.

The acute and chronic toxicity of lead to Anabas testudineus was determined in this study using static replacement bioassay testing. During the chronic toxicity studies, an experiment on the bioremediation of lead toxicity using Ocimum sanctum leaf powder was conducted. The 96 h LC 50 values of lead for Anabas testudineus was 1.08 mg/L. Different biomarkers, such as the hepatosomatic index, gonadosomatic index, and fecundity, were significantly lower in fish subjected to 10% and 20% of the 96 h LC 50 values of lead, compared to controls. The 45-day chronic exposure of fish to lead concentrations of 0.2 mg/L and above significantly lowered the number of total RBC, hemoglobin content, HCT (%), plasma protein, and cholesterol while decreasing the level of total WBC, plasma glucose, creatinine, serum AST and serum ALT. The leaf powder of Ocimum sanctum plays a significant role in ameliorating lead toxicity., Competing Interests: The authors declare no conflicts of interest.

The research aimed to investigate the perinatal pathology of Alpine ibex ( Capra ibex ) through the study of four young subjects (at the age of 3 to 4 months) found dead in Valle d'Aosta, a region of northwestern Italy. The carcasses were submitted to necropsy followed by an examination of ecto- and endoparasites (ECP and ENP); samples from the gross lesions (in summary, cutaneous papilloma and crusts, ocular discharge, lobular haemorrhagic areas in the lungs, catarrhal-haemorrhagic enterocolitis) were analysed by bacteriological, histopathological, and biomolecular methods to define the etiological agent. The subjects, with various co-infection patterns, were affected by contagious ecthyma virus (ORFV) (agent of a highly diffusive pustular dermatitis transmissible to small ruminants and humans), Enteropathogenic Escherichia coli (EPEC) (major etiological agent of infantile diarrhoea especially in developing countries), Mycoplasma conjunctivae (MC) (cause of an ocular infection common to goats and sheep), various ECP (ticks and keds) and ENP (lung and intestinal nematodes, and coccidia). This study emphasises the potential role of the Alpine ibex in the transmission of infectious diseases to other animals such as to humans and, secondly, the need to apply diversified analytical approaches, with the commitment of various specialistic skills, in order to define, in detail, the various and frequently overlapping causes that led a free-ranging animal to the death.

In 2022, a novel parvovirus was identified from an outbreak of fatal enteritis in weaned European hedgehogs ( Erinaceus europaeus ) at a wildlife rescue center in Southern Italy. During sequence analysis, the strain was found to be closely related (90.4% nucleotide identity) to a chaphamaparvovirus (ChPV) discovered in Amur hedgehogs ( Erinaceus amurensis ) during a large metaviromic investigation in game animals in China. In this study, we investigated the presence of this novel ChPV in necropsied European hedgehogs from different areas of North-Western Italy. Duodenal and liver samples collected from 194 necropsied hedgehogs were screened by using a specific quantitative PCR. A total of 38/194 animals (19.6%) tested positive, with ChPV DNA being detected in the duodenum (9.3%, 18/194), liver (7.2%, 14/194) or in both (3.1%, 6/194) tissue samples, with comparable rates and mean viral loads. The nearly full-length genome of four hedgehog ChPV strains was reconstructed. During phylogenetic analysis based on the NS1 and partial VP aa sequences, the four strains detected in this study tightly clustered with the prototype ChPVs previously identified in Amur and European hedgehogs within a potential novel candidate species of the genus Chaphamaparvovirus .

Microplastics (MPs) have recently gained attention as emerging environmental contaminants, yet knowledge of their distribution, sources, and risks in freshwater lakes remains limited. This study examined the occurrence and risk of MPs in water and sediment samples from eight locations in Mohamaya Lake (Bangladesh) collected in April and May 2023. MPs were identified using stereomicroscopy and FTIR, revealing concentrations of 20-95 particles/L in water and 550-1900 particles/kg (d.w.) in sediment, with mean values of 50.62 ± 9.95 particles/L and 1068.75 ± 521.49 particles/kg (d.w.). Dominant MPs were blue fibers, 0-0.5 mm in size, with HDPE, PET and LDPE as the most common polymers. This study used four indices (nemerow pollution index-NPI, contamination factor-CF, pollution load index-PLI, and polymer hazard index-PHI) to assess MP pollution, revealing light to high contamination levels. While NPI indicated light pollution, CF, PLI, and PHI highlighted areas of moderate to high risk, with certain polymers showing high to extreme toxicity. This study deepens understanding of MP contamination in Bangladesh's freshwater lakes, underscoring the need for research on ecotoxicology, regulation, and associated challenges., Competing Interests: Declaration of competing interest The authors declare no personal or financial interests that may have influenced the work presented in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: Intestinal microeukaryote parasites are major contributors to the burden of diarrhoea in humans and domestic animals, but their epidemiology in wildlife is not fully understood. We investigated the frequency, genetic diversity and zoonotic potential of protists of animal and public health significance in free-ranging grey wolf (Canis lupus) populations in south-western Europe., Methods: Individually formed faecal samples collected from necropsied wolves or scat trails in Italy (n = 47), Portugal (n = 43) and Spain (n = 225) during the period 2011-2023 were retrospectively analysed using molecular (PCR and Sanger sequencing) methods. Complementary epidemiological data were gathered when available., Results: Giardia duodenalis was the most frequent microeukaryote found (40.3%, 127/315; 95% CI: 34.9-46.0), followed by Cryptosporidium spp. (3.5%, 11/315; 95% CI: 1.8-6.2), Enterocytozoon bieneusi and Encephalitozoon spp. (1.6%, 5/315; 95% CI: 0.5-3.7 each). Blastocystis was not identified in any of the faecal samples analysed. Sequence analyses confirmed the presence of canine-adapted assemblage D within G. duodenalis (n = 7). Three Cryptosporidium species were identified, namely canine-adapted C. canis (n = 9), zoonotic C. parvum (n = 1) and primarily anthroponotic C.hominis (n = 1). Genotyping tools enabled the identification of subtype family XXe2 within C. canis. Among microsporidia, the canine-adapted genotype PtEb IX was identified within E. bieneusi. Two samples were confirmed as Enc. intestinalis and three more as Enc. cuniculi genotype IV. This is the first record of Enc. intestinalis and Enc. cuniculi in the grey wolf globally., Conclusions: Silent carriage of intestinal microeukaryotes seems common in free-ranging grey wolves in southwestern Europe. Wolves can contribute to environmental contamination through the transmission stages (cysts, oocysts, spores) of species/genotypes potentially infective to humans. Individuals in close contact with wolf carcasses or their faecal material may be at potential risk of infection by microeukaryotic pathogens., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Canine vector-borne diseases (CVBDs) pose a global threat to both canine and public health. This study evaluates the prevalence of Anaplasma spp., Ehrlichia spp., Borrelia burgdorferi sensu lato (s.l.), Rickettsia conorii , and Dirofilaria immitis in domestic dogs in Liguria, north-west Italy, a region where data were lacking. From 2013 to 2023, 8584 blood samples from shelter (74%) and owned dogs (26%) were submitted to the Istituto Zooprofilattico Sperimentale of Piemonte, Liguria and Valle d'Aosta (IZSPLV) for serological testing (indirect immunofluorescence and/or rapid immunochromatographic tests). Overall, 18.8% (95% CI: 18.0-19.7) of the dogs tested positive for at least one pathogen, with positivity against R. conorii antigen being the most frequently recorded (24.4%, 95% CI: 23.3-25.5). Lower prevalence levels were observed for Anaplasma spp. (1.82%, 95% CI: 1.47-2.23), Ehrlichia spp., (1.25%, 95% CI: 0.97-1.60), B. burgdorferi s.l. (0.22%, 95% CI: 0.11-0.39), and D. immitis (0.84%, 95% CI: 0.65-1.06). Positive cases for all pathogens increased over time, with prevalence differing significantly between owned and shelter dogs. Topographical factors, land use, and monthly relative humidity appeared to influence the positivity in the dogs. These results update the epidemiology of the investigated CVBDs in Liguria, indicating a widespread exposure to Rickettsia spp. among local dogs.

Dengue virus circulation is on the rise globally, with increased epidemic activity in previously unaffected countries, including within Europe. In 2023, global dengue activity peaked, and Italy reported the highest number of dengue cases and local chains of transmission to date. By curating several sources of information, we introduce a novel data repository focused on dengue reporting in Italy. We integrate data from such a repository with other geographic, genomic and climatic spatiotemporal data to present an overview of transmission patterns of the past eight years related to circulating viral lineages, geographic distribution, hotspots of reporting, and the theoretical contribution of local climate. The novel data repository can contribute to a better understanding of an evolving epidemiological scenario in Italy, with the potential to inform reassessment and planning of adequate national and European public health strategies to manage the emergence of the dengue virus., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Swine are considered one of the most relevant large animal biomedical models since they share many immunological similarities with humans. Despite that, macrophage polarization has not comprehensively investigated in pigs. In this study, porcine monocyte-derived macrophages (moMΦ) were untreated or stimulated with IFN-γ + LPS (classical activation), or by different M2 polarizing stimuli: IL-4, IL-10, TGF-β, or dexamethasone. Expression of key cytokine genes (IL1B2, IL33, IL19, IL22, IL26, CCL17, CCL24, IFNA, IFNB) in macrophage subsets were investigated over time. Expression of the genes encoding the two main enzymes of the arginine pathway (ARG1, NOS2), and molecules related to alternative macrophage polarization in human and mice (MMP9, MRC1, FIZZ1, VEGFA) were also assessed. Stimulation with IFN-γ + LPS triggered up-regulation of IL1B2, IFNB, NOS2, whereas IL-4 triggered upregulation of CCL17, CCL24, CXCR2, and ARG1 expression. IL19 and IL22 expression was enhanced by stimulation with IFN-γ + LPS or TGF-β, but not IL-4, IL-10, or dexamethasone. Our data highlighted some peculiarities in swine, such as induced expression of IL33 after stimulation with IFN-γ + LPS, and no up-regulation of FIZZ1, VEGFA or MMP9 after exposure to any of the M2 polarizing stimuli. A better understanding of porcine macrophage polarization could benefit translational studies using this large animal model., Competing Interests: Declaration of Competing Interest The authors have nothing to declare., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Meagre ( Argyrosomus regius ) is one of the fast-growing species considered for sustainable aquaculture development along the Mediterranean and Eastern Atlantic coasts. The emergence of Systemic Granulomatosis (SG), a disease marked by multiple granulomas in various tissues, poses a significant challenge in meagre aquaculture. In the current study, we investigate the association of Mycobacterium spp. and SG in offshore aquaculture facilities in Sardinia, Italy. A total of 34 adult seemingly healthy meagre were arbitrarily collected and analyzed, combining histological, microbiological, molecular, metagenomics, and in situ techniques to investigate the presence of pathogens. Ziehl-Neelsen (ZN), periodic acid-schiff (PAS), and Giemsa stains were performed for the detection of acid-fast bacteria, common parasites, and fungi within granulomas, respectively. Granulomas were detected in 91% (31/34) of fish. The affected organs were kidney (88%), liver (47%), heart (41%), intestine (17.6%), and brain (5%). Acid-fast staining, along with Mycobacterium spp. specific quantitative PCR (qPCR), in situ hybridization (ISH) assay, and microbiological analyses showed negative results for the detection of Mycobacterium spp. and other bacteria implicated in granuloma formation. However, PCR amplification and sequencing of the 65-kDa heat shock protein gene revealed the presence of M. chelonae in 13% of both formalin-fixed and frozen liver tissues. Bacterial isolation failed to detect nontuberculous mycobacteria (NTM) and other bacteria typically associated with granulomas. Consistently, the use of an M. chelonae -specific probe in ISH failed to identify this bacterial species in granulomas. Collectively, results do not support the role of M. chelonae in the development of granulomas and suggest rejecting the hypothesis of a potential link between NTM and SG.

Cetacean brains are uniquely adapted to diving, but can be affected by diseases and exposure to toxins, triggering neurodegenerative processes that may cause stranding. Some species exhibit a significant post-reproductive lifespan (PRLS), increasing the likelihood of observing cumulative and age-related pathology. Immunohistochemistry against amyloid-β and hyperphosphorylated tau proteins is increasingly implemented to assess Alzheimer's Disease-like neuropathology in cetaceans, but comparisons between geographically distinct populations, animals of different age groups, sex, and with concomitant pathologies are lacking. We tested 43 cetaceans' (30 Tursiops truncatus; 13 Stenella coeruleoalba) parietal cortex, our most consistently archived cerebral tissue, in immunohistochemical analyses with amyloid-β oligomer 42 (Aβ-42) and hyperphosphorylated tau (pTau AT180 and AT8) antibodies. Aβ-42 antibody cross-reacted with plaques in three aged bottlenose and two aged striped dolphins, but was more often detected within neurons, glia, and blood vessels of all the dolphins. Histoscore comparisons between dolphins of different ages, sexes, and pathologies revealed significant correlations between older age, viral infections, and plaque presence. Protozoan cysts cross-reacted with Aβ-42 antibody. pTau signal was observed as single foci in neurons and neuropil in two young and two aged bottlenose dolphins. To our knowledge, this study is the first of its kind for the Mediterranean region and will help establish baseline understanding of physiological and pathological expression of proteins associated with human neurodegenerative disease in cetacean brains., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Orekhova et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Approximately 30 distinct Mycoplasma species have been isolated from cattle, but only a few are pathogenic and can cause serious respiratory diseases. Consequently, this study aimed to identify Mycoplasma spp. infections in cattle with bovine respiratory disease (BRD), considering factors such as animal demographics, concurrent infections with other pathogens, post-mortem clinical findings and histological examinations, and seasonality. A total of 326 samples were collected from 322 cattle that had died from BRD in Northwestern Italy. A total of 54 animals (16.8%) tested positive for Mycoplasma spp., and Mycoplasma bovis ( n = 22, 40.7%) and Mycoplasma dispar ( n = 13, 24.1%) were the most frequently detected species among the examined cattle. Among positive cattle, those aged five months or younger were approximately five times more likely to be infected by Mycoplasma dispar than by Mycoplasma bovis compared to those older than five months (proportional incidence ratio: 5.1, 95% CI 1.2-21.2). The main bacterial pathogens identified in cattle exhibiting co-infection was Pasteurella multocida , whereas the main viral pathogens were BRSV and BoHV-1. Histopathological investigations predominantly revealed catarrhal bronchopneumonia or purulent catarrhal bronchopneumonia among the examined cattle. Finally, Mycoplasma hyopharyngis , a species isolated from the pharyngeal and nasal cavities of pigs so far, was detected for the first time in the pneumonic lung of a bovine infected with BRD. Further investigations are necessary to thoroughly characterize its host range and pathogenic potential.

Lactococcus garvieae has been considered for a long time the only causal agent of lactococcosis. In recent years, different papers reported the involvement of other two bacterial species: Lactococcus petauri and Lactococcus formosensis . A different host tropism has been described for these species where L. garvieae and L. petauri are predominant species in fish and humans' infections, while L. formosensis in bovine. L. garvieae has been reported as rare infectious agent in dog. This paper represents the first isolation of L. petauri in two domesticated dog cases from urine and skin samples, respectively. The recovered L. petauri has been identified using PCR and sequencing based on Internal Transcribe Spacer (ITS) and phylogenetic analysis showed that it belongs to the L. petauri cluster with a 100% of identity with sequences previously reported from fish isolates while there were differences with L. petauri isolated from urinary tract infection from humans. L. petauri in human infection has been considered not necessarily deriving from the ingestion of contaminated food but rather as an opportunistic pathogen colonization intestinal tract. Differences among virulotypes have been reported for humans and dogs, and a comparison was also made between the virulotyping of L. petauri and L. garvieae in dogs. The antimicrobial pattern showed susceptibility for the election treatment molecules. These data contribute to our understanding of the host trophism of this species which was misclassified for long time and provide new data on its virulence factors and antimicrobial resistance.

Data collected by C. Re. Di. Ma over a 3-year period (2020-2022) were considered to assess anthropic pressure on cetaceans living in the Ligurian sea. Out of a total of 37 stranded cetaceans, a complete post mortem examination was performed on 23 cases. Of these, 14 were further selected considering at least one of these conditions: (i) confirmed, probable, or suspected interaction with fishing activities through the application of a standardized diagnostic framework (7/14; 50%), (ii) toxicological stress through the evaluation of OCs hazardous levels (14/14; 100%), and (iii) terrestrial pathogen-associated disease (systemic infection and/or associated lesions) (7/14; 50%). For 9 animals out of a total of 14 selected, the cause of death was classified as natural (6/14; 42,8%), anthropic (3/14; 21,4%), or not determined (5/14; 35,7%) based on gross and histological pathology and ancillary testing. These findings extend our knowledge of the anthropic pressure to which cetaceans stranded along the Ligurian coastline are subjected from a multidisciplinary point of view.

Non-front-fanged snakes (NFFS) have long been overlooked by snake venom research, likely due to most of them being considered non-medically relevant for humans. The paucity of information about composition and activities of NFFS venoms and oral secretions makes it difficult to assess whether a given species can inflict medically significant bites. Here, we provide a review of the information currently available about the symptoms/signs elicited by bites from European NFFS, aiming to offer a foundation for understanding the threat they pose in terms of snakebite. Despite an overall limited amount of available data for most of the considered taxa, the genus Malpolon is notable for its capacity to cause local and systemic envenoming, including neurotoxic symptoms. Bites by other genera like, Hemorrhois, Hierophis, Natrix, Platyceps, Telescopus, and Zamenis are mainly associated with local symptoms, but the extent of their medical significance remains unclear. Our findings suggest that, although bites from European NFFS generally cause only mild effects, the potential occurrence of systemic effects from some species cannot be ruled out. Considering the above, any bite by European NFFS should receive professional medical evaluation in order to ensure patient safety and appropriate management, as well as detailed documentation facilitating construction of an accurate medical risk profile for the species., 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 Elsevier Ltd. All rights reserved.)

Background: Knowledge of the distribution and abundance of disease-causing mosquito vectors is fundamental for assessing the risk of disease circulation and introduction. Aedes caspius (Pallas, 1771) and Aedes vexans (Meigen, 1830) have been implicated, to different extents, in the circulation of several arthropod-borne viruses (arboviruses). These two mosquitoes are vectors of Tahyna virus in Europe and are considered potential vectors of Rift Valley fever virus, a virus not present but at risk of introduction on the continent., Methods: In this work, we analysed abundance data collected during West Nile virus (WNV) surveillance in northern Italy (Po Plain) via 292 CO 2 -baited traps to evaluate the distribution and density of these two non-target mosquitoes. We modelled the distribution and abundance of these two mosquito species in the surveyed area using two distinct spatial analysis approaches (geostatistical and machine learning), which yielded congruent results., Results: Both species are more abundant close to the Po River than elsewhere, but Ae. caspius is present in the eastern and western parts of the plain, linked with the occurrence of rice fields and wetlands, while Ae. vexans is observed in the middle area of the plain., Conclusions: Presence and abundance data at the municipality level were obtained and made available through this work. This work demonstrates the importance of maintaining and improving entomological surveillance programs with an adequate sampling effort., (© 2024. The Author(s).)

Ticks represent a reservoir of zoonotic pathogens, and their numbers are increasing largely in wildlife. This work is aimed at producing maps of suitable habitats for ticks in Aosta Valley, Italy based on multitemporal EO data and veterinary datasets (tick species and distribution in wild hosts). EO data were processed in Google Earth Engine considering the following inputs: A) Growing Degree Ticks (GDT), B) NDVI from MOD09GA, C) NDVI entropy, D) distance from water bodies, E) topography, F) rainfalls from CHIRPS as monthly composites along the 2020, 2021 and 2022 years. Ticks were collected from hunted, injured, and found-dead wild animals ( Sus scrofa, Capreolus capreolus, Rupicapra rupicapra, Cervus elaphus); they were labeled at species level using taxonomic keys. Between September 2020 and December 2022, a total of 90 ticks were collected from 89 wild animals. Ixodes ricinus was the most prevalent tick species, followed by Dermacentor marginatus and Dermacentor spp. Molecular analyses demonstrated the presence of Anaplasma spp., B. burgdorferi sensu lato and Rickettsia spp. pathogens in infected ticks. To assess human population potential exposure to tick Meta® population dataset was used. In conclusion this study shows the potentialities of Remote sensing improving the technological transfer to the veterinarian sector.

Leptospirosis is a bacterial disease of worldwide distribution with relevant implications for animal and human health. Different large wild carnivore species can act as reservoirs of this zoonotic pathogen. This study aimed to evaluate the circulation of Leptospira spp. in free-ranging wolves (Canis lupus) from southern Europe. A total of 281 kidney samples of wolves from Spain and Italy were collected between 2017 and 2023. The presence of Leptospira DNA was analysed by real-time PCR and phylogenetic analyses were carried out using a Bayesian approach. The overall prevalence was 3.2 % (9/281; 95 %CI: 1.1-5.3). Leptospira DNA was detected in nine of the 180 wolves from Spain (5.0 %; 95 %CI: 1.8-8.2), but not in the Italian wolf population (0 %; 0/101). Molecular analyses revealed high homology between the sequences obtained in the present study and isolates of Leptospira interrogans and Leptospira borgpetersenii from different rodent and domestic ungulate species. Our results provide evidence of a low and spatially heterogeneous circulation of this pathogen in wolf populations of southern Europe. The detection of zoonotic Leptospira species in this survey supports the need to consider wolf populations in monitoring programs for leptospirosis with a One Health approach., Competing Interests: Declaration of Competing Interest None of the authors of this study has a financial or personal relationship with other people or organizations that could inappropriately influence or bias the content of the paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Lactococcosis is a common bacterial fish disease caused by Lactococcus garvieae, L. petauri and L. formosensis. Although there are different PCR-based techniques to identify the etiological agent, none of these can differentiate these two bacteria without sequencing PCR-amplified fragments. In the present study, we developed a multiplex PCR assay for simultaneous detection and differentiation of L. garvieae and L. petauri. The specificity of the primers was validated against the bacterial DNA of the targeted and non-targeted bacteria. The sizes of the PCR amplicons were obtained as 204 bp for the DUF1430 domain-containing protein gene of L. garvieae, 465 bp for the Lichenan permease IIC component gene of L. petauri, and 302 bp for the teichoic acid biosynthesis protein F gene of both L. garvieae and L. petauri. The PCR amplicons were clearly separated by agarose gel electrophoresis. The multiplex PCR assay did not produce any amplification products with the DNA of the non-targeted bacteria. The multiplex PCR detection limits for L. garvieae and L. petauri were 5 and 4 CFU in pure culture and 50 and 40 CFU/g in spiked tissue samples, respectively. It takes less than 2 h from plate-cultured bacteria and 3 h from tissue samples to get results. In conclusion, the developed multiplex PCR assay is a rapid, specific, accurate, and cost-effective method for the detection and differentiation of L. garvieae and L. petauri and is suitable to be used for routine laboratory diagnosis of L. garvieae and L. petauri., (© 2024 The Author(s). Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Bacteria-mediated treatments gained increasing attention as alternative therapies against tumors. An attenuated mutant strain of Salmonella enterica serovar Typhimurium (STMΔznuABC) has recently been considered as a potential new anti-cancer strategy. However, it is unclear whether this activity is tumor-induced or species-specific, and no data are available regarding STMΔznuABC on canine mammary tumors (CMTs). This study aimed to investigate the ability of STMΔznuABC in modulating the response of CMTs, focusing on cancer-associated fibroblasts. Four CMT cell lines (CF33, TM51, TM52 TM53) were treated with STMΔznuABC. Then, antiproliferative activity (MTT assay), bacterial invasion, and CMT cell lines gene expression analysis (RT-qPCR) of genes involved in immune response and cancer aggressiveness were evaluated. STMΔznuABC penetrated in TM51, TM52, TM53, and CF33 cell lines, causing a significant reduction of cell viability. Moreover, the expression of several genes was significantly modulated in all CMT cell lines: STMΔznuABC infection determined a significant up-regulation of CXCL8, IL18, IL10, TLR4 and RAD51, while CD14, IL6, CXCR4, P53, PTEN, STAT5, TLR5 and TGFB1 were downregulated in TM53. In CF33, CXCL8 and P53 were upregulated, while MYD88, MD2, IL18, TLR4,5, TGFB1 were downregulated. In TM52, CXCL8, CD44 and MD2 were upregulated and PTEN was downregulated, while in TM51 CXCL8, CD44 and ErbB2 were downregulated. We demonstrated the anti-proliferative and immuno-modulatory activity of STMΔznuABC in CMTs, paving the way for potential new anti-cancer treatments., Competing Interests: Declaration of competing interest The authors have no competing interests to declare. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Global fish and seafood consumption is increasing annually, frequently leading to the emergence of food fraud, mainly related to mislabeling and adulteration like, for example, the use of illicit/unauthorized food additives to mask or delay fish spoilage. Among the available diagnostic tools for control purposes, spectroscopic techniques have often been proposed to identify these kinds of illicit practices in fish and seafood products. The presented study aims to test two cheap and portable near infrared (NIR) spectrometers, a handheld MicroNIR and a pocket-sized SCiO, to uncover use of the illicit food additive Cafodos, a mixture of sodium citrate and hydrogen peroxide used to preserve some fish characteristics (like smell, color, na dtexture). The NIR spectroscopy in combination with chemometric approaches, allowed the successfully classification of (81-100%) samples of sea bass ( Dicentrarchus labrax ) treated with Cafodos. The study highlights the potential of this technique that, by not requiring pre-treatment of samples with further reagents, is cheaper and safer for the environment. In conclusion, the study confirmed the potential of portable devices for rapid NIR spectroscopy analysis to identify food fraud and ensure consumer safety.

Histamine is a biogenic amine and an indicator of fishery product freshness and hygienic quality. The European Regulation EC 2073/2005 sets the standards for fish sample collection and establishes quantitative levels of histamine in fishery products to ensure consumer health and safety. This retrospective study presents data on histamine monitoring in fish and fishery products collected in northern Italy between 2018 and 2022. A total of 138 samples were analysed via enzyme-linked immunosorbent assay (ELISA) and then confirmed by high-performance liquid chromatography with diode-array detection (HPLC-DAD). Four samples found positive contained histamine levels above the legal limit. Monitoring via ELISA and HPLC-DAD can efficiently detect histamine in fish and fishery products and protect consumers' health.