Nicola Pozzato

Interfaces among humans, wildlife and domestic animals represent critical hotspots for the emergence and spread of infectious agents. The gray wolf (Canis lupus) and the golden jackal (Canis aureus) are expanding wild canid populations in Italy, whose increasing overlap with anthropized environments may facilitate pathogen transmission. This study investigated mammalian orthoreovirus (MRV), rotavirus A (RVA), canine coronavirus (CCoV), canine parvovirus (CPV2) and feline parvovirus (FPV) in wild canids from north-eastern Italy. The 101 carcasses (83 gray wolves and 18 golden jackals) obtained through passive wildlife surveillance were examined post-mortem. Tissue samples and swabs were screened using molecular assays, and positive samples were characterized and tested for virus isolation. MRV was detected in gray wolves (27.7%) and golden jackals (33.3%), representing the first report and successful isolation from both species. RVA prevalence was 60.2% in gray wolves and 61.1% in golden jackals. CCoV occurred exclusively in gray wolves (12.5%). CPV-2 was found in 10.8% of gray wolves and 35.3% of golden jackals, with FPV accounting for 66.7% of CPV-2-positive golden jackal samples. These findings provide valuable data on the circulation of MRV, RVA, CCoV, and CPV-2/FPV in free-ranging gray wolves and golden jackals and highlight the value of passive surveillance within a One Health framework overall.

Animal disease outbreaks at the wildlife-livestock interface threaten animal health, ecosystems, food systems, and, for zoonoses, human health, while veterinary authorities must maintain preparedness with limited resources. However, existing preparedness tools characterize capacity, exposure, or risk largely in isolation. This study developed a framework to assess whether preparedness is commensurate with disease-specific challenge. Questionnaire-derived Preparedness capacity for 26 European countries was estimated using Item Response Theory and adjusted for socioeconomic and governance context. Context-adjusted preparedness was then compared with disease-specific Challenge Indices combining livestock population, livestock density, animal imports, and wildlife reservoir abundance, for two diseases at the wildlife-livestock interface: avian influenza (AI) in domestic poultry and African swine fever (ASF) in domestic pigs. Countries were classified into four Preparedness-Challenge profiles and summarized regionally, with uncertainty propagated by Monte Carlo simulation and decomposed by variance-based sensitivity analysis. The Underprepared (high challenge) profile included 9 countries for AI and 10 for ASF and was the most common profile in Southern and Western Europe for both diseases, and in Central Europe for ASF. Livestock weighting sharpened this pattern, with this profile holding 66-91% of poultry and 79-97% of pigs in Central, Southern, and Western Europe. In Northern Europe, 76% of pigs were in the Prepared (high challenge) profile. Preparedness estimation was the largest source of uncertainty in country-level classifications and regional AI results, whereas Challenge Index weighting dominated regional ASF results. Countries with less preparedness relative to their challenge tended to have greater recorded outbreak burden (concordance probability: AI 0.63-0.70 for continuous burden measures, ASF 0.58-0.85 for all measures). Preparedness gaps were therefore disease-specific, and unweighted country summaries obscured where livestock exposure was concentrated. Disease-specific, livestock-weighted comparisons offer veterinary authorities a transparent basis for prioritizing preparedness investment, complementing established One Health tools.

Abnormal sudden mortality of 2 goats and 10 sheep occurred in a 27-head flock in Verona province (northeastern Italy). In the 24 h before death, animals had acute ataxia, lateral recumbency, spastic convulsions, and dyspnea. Autopsy of 4 animals was performed at the Verona Veterinary Diagnostic Laboratory of the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe; Italy). In the forestomachs, abundant fibrous material and numerous seeds were observed. Bacteriologic, parasitologic, and histologic investigations were carried out; samples of rumen contents and liver were analyzed by direct analysis real-time high-resolution mass spectrometry (DART-HRMS), which allows rapid screening of toxic substances. The combination of DART-HRMS and liquid chromatography high-resolution tandem MS (LC-HRMS/MS) confirmed the acute intoxication and provided insights into the clinicopathologic findings due to the ingestion of Chimonanthus praecox (wintersweet), a plant species that contains alkaloids including calycanthine, which is known to be toxic in several domestic species, including small ruminants.

Periodontal disease is a prevalent condition in companion animals. It is crucial to prevent the plaque and bacteria on tooth surfaces to avoid gingivitis and the more harmful periodontitis. The aim of the present study was to assess the impact of MLS laser treatment on the bacterial load by analyzing the gingival swabs of a total of 16 owned dogs with no history of dental disease that were selected from a cohort of patients admitted for plaque removal and dental hygiene procedures. Following each dental hygiene session, each dog received a single MLS laser therapy treatment (808–905 nm diode laser, frequency 36 Hz, and fluence 0.16 J/cm²). Swabs were collected from the two upper premolars before and after the laser treatment. These were submitted for mesophilic bacteria counts, and microbiological analysis was conducted on 10 positive cultures to evaluate the changes in the oral bacterial microbiota. MLS laser therapy statistically significantly reduced the mesophilic bacteria count by 1 log, with higher pre-laser treatment counts (n = 0.44; 5.77 ± 0.21 95%CI) in comparison to post-laser treatment counts (n = 0.73; 4.80 ± 0.346 95%CI). The MLS laser therapy was found to reduce the bacterial load in 80% of the subjects (p < 0.05). No significant differences pre- and post-laser treatment were observed in the bacteria species isolated from the microbiological cultures. MLS laser therapy appears to be a useful, non-invasive method for reducing the bacterial load in the treatment and prophylaxis of periodontal disease in dogs.

Background: The presence of carbapenemase-producing bacteria (CPB) in animal hosts and along the food chain may result in the development of reservoirs for human infections. Several CPB strains isolated from animals have been reported, suggesting that transmission and dissemination of the corresponding genes between humans and animals may occur. Animal and food samples have complex backgrounds that hinder the detection of CPB present in low concentrations by standard detection procedures. Methods: We evaluated the possibility of detecting bla_KPC, bla_VIM, and bla_OXA-48-like carbapenemases in 286 animal and food samples (faeces from farm and companion animals, raw meat, bivalve molluscs) by culture-based and standard molecular methods and by ddPCR. Results: The proposed ddPCR managed to detect the target genes, also in samples resulting negative to standard methods. While the presence of bla_KPC and bla_VIM was detected in few samples (~3%), one third of the samples (n = 94/283) carried different variants of bla_OXA-48-like genes. Conclusion: A specific and sensitive method such as ddPCR could be suitable to evaluate the current veterinarian and environmental situation and to assess the dynamic transmission and persistence of CPB between animals and humans and vice versa.

Laboratory tests provide essential support to the veterinary practitioner, and their use has grown exponentially. This growth is the result of several factors, such as the eradication of historical diseases, the occurrence of multifactorial diseases, and the obligation to control endemic and epidemic diseases. However, the introduction of novel techniques is counterbalanced by economic constraints, and the establishment of evidence- and consensus-based guidelines is essential to support the pathologist. Therefore, we developed standardized protocols, categorized by species, type of production, age, and syndrome at the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), a multicenter institution for animal health and food safety. We have 72 protocols in use for livestock, poultry, and pets, categorized as, for example, “bovine enteric calf”, “rabbit respiratory”, “broiler articular”. Each protocol consists of a panel of tests, divided into ‘mandatory’ and ‘ancillary’, to be selected by the pathologist in order to reach the final diagnosis. After autopsy, the case is categorized into a specific syndrome, subsequently referred to as a syndrome-specific panel of analyses. The activity of the laboratories is monitored through a web-based dynamic reporting system developed using a business intelligence product (QlikView) connected to the laboratory information management system (IZILAB). On a daily basis, reports become available at general, laboratory, and case levels, and are updated as needed. The reporting system highlights epidemiologic variations in the field and allows verification of compliance with the protocols within the organization. The diagnostic protocols are revised annually to increase system efficiency and to address stakeholder requests.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis [Johne's disease (JD)], a chronic disease that causes substantial economic losses in the dairy cattle industry. The long incubation period means clinical signs are visible in animals only after years, and some cases remain undetected because of the subclinical manifestation of the disease. Considering the complexity of JD pathogenesis, animals can be classified as infected, infectious, or affected. The major limitation of currently available diagnostic tests is their failure in detecting infected non-infectious animals. The present study aimed to identify metabolic markers associated with infected and infectious stages of JD. Direct analysis in real time coupled with high resolution mass spectrometry (DART-HRMS) was, hence, applied in a prospective study where cohorts of heifers and cows were followed up annually for 2–4 years. The animals' infectious status was assigned based on a positive result of both serum ELISA and fecal PCR, or culture. The same animals were retrospectively assigned to the status of infected at the previous sampling for which all JD tests were negative. Stored sera from 10 infected animals and 17 infectious animals were compared with sera from 20 negative animals from the same herds. Two extraction protocols and two (-/+) ionization modes were tested. The three most informative datasets out of the four were merged by a mid-level data fusion approach and submitted to partial least squares discriminant analysis (PLS-DA). Compared to the MAP negative subjects, metabolomic analysis revealed the m/z signals of isobutyrate, dimethylethanolamine, palmitic acid, and rhamnitol were more intense in infected animals. Both infected and infectious animals showed higher relative intensities of tryptamine and creatine/creatinine as well as lower relative abundances of urea, glutamic acid and/or pyroglutamic acid. These metabolic differences could indicate altered fat metabolism and reduced energy intake in both infected and infectious cattle. In conclusion, DART-HRMS coupled to a mid-level data fusion approach allowed the molecular features that identified preclinical stages of JD to be teased out.