Vito Tranquillo

Qualitative frameworks are widely employed to tackle urgent animal or public health issues when data are scarce and/or urgent decisions need to be made. In qualitative models, the degree of belief regarding the probabilities of the events occurring along the risk pathway(s) and the outcomes is described in nonnumerical terms, typically using words such as Low, Medium, or High. The main methodological challenge, intrinsic in qualitative models, relates to performing mathematical operations and adherence to the rule of probabilities when probabilities are nonnumerical. Although methods to obtain the qualitative probability from the conditional realization of n events are well‐established and consistent with the multiplication rule of probabilities, there is a lack of accepted methods for addressing situations where the probability of an event occurring can increase, and the rule of probability P(AUB) = P(A) + P(B) − P(A∩B) should apply. In this work, we propose a method based on the pairwise summation to fill this methodological gap. Our method was tested on two qualitative models and compared by means of scenario analysis to other approaches found in literature. The qualitative nature of the models prevented formal validation; however, when using the pairwise summation, results consistently appeared more coherent with probability rules. Even if the final qualitative estimate can only represent an approximation of the actual probability of the event occurring, qualitative models have proven to be effective in providing scientific‐based evidence to support decision‐making. The method proposed in this study contributes to reducing the subjectivity that characterizes qualitative models, improving transparency and reproducibility.

In countries endemic for foot-and-mouth disease (FMD), routine or emergency vaccinations are strategic tools to control the infection. According to the WOAH/FAO guidelines, a prior estimation of vaccine effectiveness is recommendable to optimize control programs. This study reports the results of a small-scale immunogenicity study performed in Transcaucasian Countries. Polyvalent vaccines, including FMDV serotypes O, A (two topotypes) and Asia1 from two different manufacturers, were evaluated in Georgia, Azerbaijan and Armenia. Naïve large and small ruminants were vaccinated once and a subgroup received a second booster dose. The titers of neutralizing antibodies in sera collected sequentially up to 180 DPV were determined through the Virus Neutralization Test versus homologous strains. This study led to the estimate that both the vaccines evaluated will not induce a protective and long-lasting population immunity, even after a second vaccination, stressing that consecutive administrations of both vaccines every three months are mandatory if one aspires to achieve protective herd immunity.

Intensive agricultural practices, such as pesticides use, may negatively affect bee health and hive products. Glyphosate is one of the most widely used polar pesticides applied in crops for weed control. In this study, honey samples, collected from beekeeping farms located in the Lombardy and Emilia-Romagna regions in Italy in the framework of regional monitoring plans activated from 2020 to 2022, were analyzed for the presence of residues of polar pesticides. The analytical method based on ion chromatography coupled to high-resolution mass spectrometry was applied to quantify glyphosate, glufosinate, ethephon, fosetyl aluminum, and their related metabolites. Residues of glyphosate were detected in around 28% of analyzed honey samples. Observations on the distribution of the honey-production-site locations suggest that honey samples originating from the provinces within the Lombardy region, where the agricultural sector is highly developed, were more affected by glyphosate contamination than the samples collected from the areas with low agricultural activity, where no glyphosate residues were detected over the three years of the monitoring program.

Growing attention is being given to the European hedgehog (Erinaceus europaeus) because of its synanthropic behaviour and its potential role in harbouring parasites, viruses, fungi and bacteria and disseminating them to several animals and humans. Salmonella are the most frequently detected zoonotic bacteria that hedgehogs could transmit through contaminating water and food sources with faeces. This study aimed to determine the prevalence and distribution of Salmonella spp. in wild hedgehogs in the Emilia-Romagna region (northern Italy). From 2019 to 2022, 212 European hedgehogs that died naturally were tested for Salmonella spp. through culture isolation. Positive samples were subjected to serological typing. A total of 82 samples tested positive for Salmonella spp., with the overall Bayesian posterior estimated prevalence ranging from 35% (95% CI: 23–47%) to a maximum of 45% (95% CI: 31–59%) during the years considered and with an overall prevalence calculated at 39% (95% CI: 33–45%). Salmonella enterica Enteritidis and Veneziana were the most prevalent detected serovars in 65% and 17% of the positive samples, respectively. Since 2021, S. Typhimurium, S. Typhimurium Monofasica, S. Zaiman, S. Hessarek, S. Muenster, S. Isangi serovars, S. enterica subsp. Diarizonae and S. enterica subsp. Houtenae have been detected. These findings show a high prevalence of Salmonella spp. in tested hedgehogs, suggesting an important role of this animal species in the epidemiology of potentially zoonotic serovars circulating in the Emilia-Romagna region.

Listeria monocytogenes is a widespread Gram-positive pathogenic bacterium that causes listeriosis, a rather rare but severe foodborne disease. Pregnant women, infants, the elderly, and immunocompromised individuals are considered particularly at risk. L. monocytogenes can contaminate food and food-processing environments. In particular, ready-to-eat (RTE) products are the most common source associated with listeriosis. L. monocytogenes virulence factors include internalin A (InlA), a surface protein known to facilitate bacterial uptake by human intestinal epithelial cells that express the E-cadherin receptor. Previous studies have demonstrated that the presence of premature stop codon (PMSC) mutations naturally occurring in inlA lead to the production of a truncated protein correlated with attenuate virulence. In this study, 849 L. monocytogenes isolates, collected from food, food-processing plants, and clinical cases in Italy, were typed and analyzed for the presence of PMSCs in the inlA gene using Sanger sequencing or whole-genome sequencing (WGS). PMSC mutations were found in 27% of the isolates, predominantly in those belonging to hypovirulent clones (ST9 and ST121). The presence of inlA PMSC mutations in food and environmental isolates was higher than that in clinical isolates. The results reveal the distribution of the virulence potential of L. monocytogenes circulating in Italy and could help to improve risk assessment approaches.

Salmonella is a pathogen of considerable health concern, given its zoonotic potential, and, in Italy, is the most frequently reported causative agent for foodborne outbreaks. Wild animals and in particular wild carnivores may be carriers of different Salmonella enterica subspecies and serotypes. Given their potential role as reservoirs, surveillance activities are necessary. This study aims to investigate the presence of different Salmonella subspecies and serotypes in wild carnivores in the Emilia-Romagna Region. A total of 718 fox (Vulpes vulpes), 182 badger (Meles meles) and 27 wolf (Canis lupus) carcasses, submitted between 2016–2022, were included for the present work. Gender and age data were collected along with geographical coordinates of carcass’ discovery site. Contents of the large intestine were sampled and cultured according to ISO 6579-1 and both serogroup and serotype identification were performed according to ISO/TR 6579-3:2014. Salmonella was retrieved from 42 foxes (6%), 21 badgers (12%) and 3 wolves (12%), respectively. Isolated Salmonella enterica strains belonged to 4 different subspecies and 25 different serotypes. S. veneziana and S. typhimurium were the most frequent serotypes found (11/67 and 10/67, respectively). In conclusion, zoonotic serotypes were found in all these species of wildlife, thus confirming their potential role in the ecology of Salmonella spp.

Leishmaniases are severe vector-borne diseases affecting humans and animals, caused by Leishmania protozoans. Immune polarization plays a major role in determining the outcome of Leishmania infections: hosts displaying M1-polarized macrophages are protected, while those biased on the M2 side acquire a chronic infection, that could develop into an overt and potentially deadly disease. The identification of the factors involved in M1 polarization is essential for the design of therapeutic and prophylactic interventions, including vaccines. Infection by the filarial nematode Dirofilaria immitis could be one of the factors that interfere with leishmaniasis in dogs. Indeed, filarial nematodes induce a partial skew of the immune response towards M1, likely caused by their bacterial endosymbionts, Wolbachia . Here we have examined the potential of Asaia ^WSP , a bacterium engineered for the expression of the Wolbachia surface protein (WSP), as an inductor of M1 macrophage activation and Leishmania killing. Macrophages stimulated with Asaia ^WSP displayed a strong leishmanicidal activity, comparable to that determined by the choice-drug amphotericin B. Additionally, Asaia ^WSP determined the expression of markers of classical macrophage activation, including M1 cytokines, ROS and NO, and an increase in phagocytosis activity. Asaia not expressing WSP also induced macrophage activation, although at a lower extent compared to Asaia ^WSP . In summary, our study, while providing a strong evidence for the immune-stimulating properties of Wolbachia , highlights the translational potential of Asaia ^WSP in the areas of the immune-prophylaxis and therapy of leishmaniases, as well as of other diseases that could be subverted by M1 macrophage activation.

Welfare of animals significantly depends on how stakeholders perceive their needs and behave in a way to favor production systems that promote better welfare outcomes. This study aimed at investigating stakeholders’ perception of the welfare of equines, small ruminants, and turkeys using text mining analysis. A survey composed by open-ended questions referring to different aspects of animal welfare was carried out. Text mining analysis was performed. A total of 270 surveys were filled out (horses = 122, sheep = 81, goats = 36, turkeys = 18, donkeys = 13). The respondents (41% veterinarians) came from 32 different countries. To describe welfare requirements, the words “feeding” and “water” were the most frequently used in all the species, meaning that respondents considered the welfare principle “good feeding” as the most relevant. The word “environment” was considered particularly important for turkeys, as well as the word “dry”, never mentioned for other species. Horses stakeholders also considered “exercise” and “proper training” important. Goat stakeholders’ concerns are often expressed by the word “space”, probably because goats are often intensively managed in industrialized countries. Although the sample was too small to be representative, text mining analysis seems to be a promising method to investigate stakeholders’ perception of animal welfare, as it emphasizes their real perception, without the constraints deriving by close-ended questions.