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Abstract Antimicrobial resistance is a global threat, and pet-associated strains may pose a risk to human health. Equine veterinarians are at high risk of carrying methicillin-resistant staphylococci (MRS), but specific risk factors remain elusive, and few data are available for other personnel involved in the horse industry. The prevalence, characteristics, and risk factors for nasal carriage of MRS in horses and their caregivers were studied in northwestern Italy. Nasal swabs from 110 asymptomatic horses housed at 21 barns and 34 human caregivers were collected. Data on barns, horses, and personnel were acquired through questionnaires. The samples were incubated in selective media, and the bacterial isolates were identified by mass spectrometry. Risk factors were investigated by Poisson regression. MRS were isolated from 33 horses (30%), 11 humans (32.4%) and 3 environmental samples (14.2%). Most isolates were multidrug resistant (MDRS). The prevalence of MRS and MDRS was greater in racehorses and their personnel than in pleasurable and jumping/dressing horses. MRS carriage in caregivers was associated with an increased prevalence of MRS carriage in horses. The frequency of antimicrobial treatments administered in the barn during the last 12 months was a risk factor for MRS carriage in horses [prevalence ratio (PR) 3.97, 95% CI 1.11, 14.13] and caregivers (PR 2.00, 95% CI 1.05, 3.82), whereas a good ventilation index of the horse tabling environment was a protective factor (PR 0.43, 95% CI 0.20, 0.92). Our data reveal relevant interactions occurring between bacterial communities of horses and humans that share the same environment, suggesting that One Health surveillance programs should be implemented.

Abstract Background In this study, a probiotic mixture (Honeybeeotic) consisting of seven bacterial strains isolated from a unique population of honeybees (Apis mellifera ligustica) was used. That honeybee population was located in the Roti Abbey locality of the Marche Region in Italy, an area isolated from human activities, and genetic contamination from other honeybee populations. The aim was to investigate the effects of this probiotic mixture on the innate immunity and intestinal microbiome of healthy common honeybees in two hives of the same apiary. Hive A received a diet of 50% glucose syrup, while hive B received the same syrup supplemented with the probiotics, both administered daily for 1 month. To determine whether the probiotic altered the immune response, phenoloxidase activity and hemolymph cellular subtype count were investigated. Additionally, metagenomic approaches were used to analyze the effects on gut microbiota composition and function, considering the critical role the gut microbiota plays in modulating host physiology. Results The results revealed differences in hemocyte populations between the two hives, as hive A exhibited higher counts of oenocytoids and granulocytes. These findings indicated that the dietary supplementation with the probiotic mixture was safe and well-tolerated. Furthermore, phenoloxidase activity significantly decreased in hive B (1.75 ± 0.19 U/mg) compared to hive A (3.62 ± 0.44 U/mg, p

A semi-quantitative risk assessment was developed to classify pig farms in terms of the probability of introduction of African swine fever virus (ASFV). Following on-farm data collection via a specific checklist, we applied a modified failure mode and effect analysis (FMEA) to calculate the risk priority codes (RPC's), indicating increasing risk levels ranging from 1 to 5. The importance of biosecurity measures was attributed by experts. To consider geographic risk factors, we classified pig farms based on local density of farmed pigs, and on the estimated wild boar population density. The combination of RPC's with geographical risk factors resulted into a final ranking of pig farms in terms of the risk of ASFV introduction. Furthermore, the estimation of frequency and levels of non-compliance with biosecurity measures was used to identify weak points in risk prevention at farm level. The outcome of the risk assessment was affected by choices in assigning non-compliance scores and importance to specific components of biosecurity. The method was applied in 60 commercial farms in major pig production areas in Italy. Furthermore, we applied a reduced version of our checklist in 12 non-commercial/small commercial (≤20 pigs) farms in the northern Apennines. In commercial farms, highest RPC's were obtained for biosecurity measures associated with personnel practices and farm buildings/planimetry. Intervention should be addressed to training of personnel on biosecurity and ASF, to avoid contacts with other pig herds, and to improve practices in the entrance into the farm. Sharing trucks with other farms, and loading/unloading of pigs were other weak points. Fencing was classified as insufficient in 70% of the commercial farms. Among these farms, breeding units were characterised by the lowest risk of ASFV introduction (although differences among median ranks were not statistically significant: P-value = 0.07; Kruskal–Wallis test), and increasing herd size was not significantly correlated with a higher risk (Kendall's τ = −0.13; P-value = 0.14). Density of farmed pig was greatest in the main pig production area in northern Italy. Conversely, exposure to wild boars was greatest for non-commercial/small commercial farms on the Apennines, which were also characterised by non-compliance with critical biosecurity measures.

In a previous study, a semi-quantitative risk assessment was developed to rank pig holdings in terms of likelihood of introducing African swine fever virus (ASFV) by assessing their compliance with biosecurity and exposure to geographical risk factors. The method was initially developed for confined pig holdings, but given that ASF is endemic in wild boar of several countries, we modified the approach to make it suitable for free-range farms as well. In the current study, a total of 41 outdoor pig farms were assessed in an area where exposure to wild boar was generally high (density from 2.3 to 10.3 wild boar per Km2). As expected, non-compliance with biosecurity measures was frequent in outdoor farms, and the frequency of non-compliance indicated that the absence of adequate separation of pigs from the external environment was the major weakness in the farms assessed. In 46.3% of them, there was no fence or, if present, it was not adequate to avoid contact with wild boar. However, the approach adopted proved to be suitable for identifying intervention priorities to mitigate the risk of ASFV spread in free-range pig herds and for identifying the weaknesses of individual farms, as recommended by EFSA in 2021, which suggests implementing tools to improve biosecurity by favoring higher-risk farms.

Staphylococcal infections represent a challenge in companion animals and hospitalized patients. This study aimed to assess the resistance of Staphylococcus pseudintermedius isolates, against a broad panel of antibacterials, including exclusive to human medicine. A total of 40 S. pseudintermedius were collected from clinical specimens of dogs (n = 31) and cats (n = 5). All strains were tested for 20 antibacterials, namely 14 Critical Important and eight Highly Important Antibacterials (CIA and HIA, respectively), indicative for 18 antimicrobial classes. All strains were susceptible to seven antibiotics (daptomycin, fosfomycin, fusidic acid, linezolid, quinupristin-dalfopristin, teicoplanin/vancomycin, tigecycline). The highest resistance was against penicillin (97.5% Confidence Interval [CI]: 83.8–100.0), whereas the lowest against telavancin (2.5%, CI: 0.0–16.2). Resistance versus Highest Priority CIA was observed, namely against macrolides (70.0, CI: 52.1–84.3), quinolones (62.5, CI: 44.5–78.3), 5th generation cephalosporins (7.5, CI: 1.3–21.6), and glycopeptides (2.5%, CI: 0.0–14.2). Among High Priority CIA, strains were resistant only to aminoglycosides (65.0, CI: 47.0–80.4) and ansamycins (12.5, CI: 3.8–28.1). We observed the highest resistance against veterinary medicine antibacterials, but there was also resistance against antibacterials exclusive to human medicine, namely ceftaroline (7.5, CI: 1.0–23.8) and telavancin. S. pseudintermedius zoonotic potential and its rate of acquisition of new resistance should encourage surveillance on a broad spectrum of antibacterials.

To understand the effectiveness of a probiotic mixture on intestinal morphology, mucus layer composition, and cecal microbiota diversity, 40 10-day-old Guinea fowls (Numida meleagris) were assigned to two groups: the control group (C), receiving drinking water, and the treated group (P), receiving water plus a commercial multi-strain probiotic (Slab51®, 2 × 1011 CFU/L). Birds were slaughtered after 4 months, and the intestines were collected. Samples from the duodenum, ileum, and cecum were processed for morphological and morphometric studies, and conventional glycohistochemistry. Cecal samples were also used to assess the microbiota by 16S metataxonomic approach. Group P showed significant increase in the villus height (p < 0.001 in the duodenum and p < 0.05 in the ileum and cecum), villus width (p < 0.05 in all investigated tracts), depth of crypts (p < 0.001 in the duodenum and cecum; p < 0.05 in the ileum), and goblet cells per villus (p < 0.001 in all investigated tracts) compared with group C. Cecal microbiota of the birds varied considerably and comparing the relative abundance of the main observational taxonomic units (OTUs), a positive enrichment of several beneficial taxa, such as Oscillospira, Eubacterium, Prevotella, and members of the Ruminococcaceae, was observed. The enrichment of those taxa can improve microbiota stability and resilience facing environmental stresses, enhancing its resistance against invading pathogens. Ruminococcaceae, which represent the most important taxon in both groups, and Prevotella have a key role in the gut physiology due to the production of short-chain fatty acids (SCFAs), which are a vital energy source for enterocytes, improve glucose metabolism, and exert an overall anti-inflammatory effect. Probiotic administration enriches the presence of Coprococcus, Oscillospira, and Eubacterium taxa that produce butyrate, which exerts a beneficial effect on growth performance, structure of villi, and pathogen control and has anti-inflammatory properties too. This study indicates that Slab51® supplementation positively affects the morphology and microbiota diversity of the guinea fowl intestine.

Wild and domestic animals can be usefully employed as sentinels for the surveillance of diseases with an impact on public health. In the case of tick-borne encephalitis virus (TBEV), the detection of antibodies in animals can be more effective than screening ticks for detecting TBEV foci, due to the patchy distribution of the virus. In the Piedmont region, northwestern Italy, TBEV is considered absent, but an increase in tick densities, of Ixodes ricinus in particular, has been observed, and TBEV is spreading in bordering countries, e.g., Switzerland. Therefore, we collected sera from wild ungulates during the hunting season (October–December) from 2017 to 2019 in the Susa Valley, Italian western Alps, and screened them for TBEV antibodies by a commercial competitive ELISA test. We collected 267 serum samples by endocranial venous sinuses puncture from red deer, roe deer and northern chamois carcasses. The animals were hunted in 13 different municipalities, at altitudes ranging between 750 and 2800 m a.s.l. The serological survey for TBEV yielded negative results. Borderline results for five serum samples were further confirmed as negative for TBEV by a plaque reduction neutralisation test. To date, our results indicate that TBEV is not circulating in western Piedmont. However, monitoring of TBEV should continue since TBEV and its vector are spreading in Europe. The wide-range distribution of wild ungulates and their role as feeding hosts, make them useful indicators of the health threats posed by Ixodid ticks.

A questionnaire survey was promoted under the COST Actions “Network for Evaluation of One Health-NEOH” and “European Network for Neglected Vectors and Vector-Borne Infections-EURNEGVEC”, from June 2016 to April 2017, to collect information on the existence of One Health (OH) collaboration and implementation of OH initiatives in 37 EU COST Countries. The questionnaire was to be answered by key respondents representing the three major OH components: (i). Animal Health; (ii). Human Health/Public Health; (iii). Environmental Health. A target respondent rate of nine respondents/country was aimed for, representing the following categories: (i). ministries; (ii). academia-research; (iii). private sector and NGOs, associations and scientific societies. The questionnaire, composed of 27 questions organized in six sections, was circulated to target respondents by Committee Members of the two COST actions. A total of 171 respondents from 34 countries completed the questionnaire, mainly belonging to academic and research institutions (55.5%), and to Animal Health/Animal Science fields (53.8%). Although the majority (57.9%) declared they had heard about OH, few respondents (10.7%) provided a complete definition. The “human” and “animal” elements prevailed over other key elements of OH definition (ecosystem, intersectoral, transdisciplinary, holistic, collaboration). Overall, 62.6% respondents declared to take part in OH initiatives. Antimicrobial resistance, avian influenza and environmental pollution were cited as the top three OH issues over the past 5 years. Limitations and gaps in intersectoral collaboration included communication and organizational problems resulting in poor networking, differing priorities and a lack of understanding between sectors. Regarding control and monitoring of zoonotic diseases, respondents from different sectors preferentially selected their own directorates/ministries while actually in most countries both Ministry of Health and Ministry of Agriculture are engaged. According to respondents, the level of awareness of OH amongst the general public is limited. Similarly, a dearth of opportunities of collaborations at different institutional and/or professional levels was described. Our survey provided an overview of how respondents in COST countries perceived and experienced OH and current limits to OH implementation. Identifying how initiatives are currently working and knowing the promoting and hindering factors allowed suggesting strategies to promote efficiency and effectiveness of OH implementation in the future.

The breeding of meat rabbits is an important sector in the livestock industry in Italy. The focus of this study was to describe the antibiotic resistance profile distribution among the Methicillin-sensitive Staphylococcus aureus isolated in a rabbit farm. From 400 animals of different ages and three farm workers, 96 randomly selected strains isolated from various anatomical sites and lesions were analysed. According to spa typing and the resistance profiles towards veterinary and human antibiotics, 26 pathotypes were identified. The highest resistance was observed against Tetracyclines (92.3%) and Macrolides (80.8%), while almost all were susceptible to Penicillins, according to the limited use of β-lactams on the farm. In total, 92.3% of pathotypes were multidrug resistant (MDRs). Two MDR pathotypes belonging to the t2802 spa type were isolated from both farmers and rabbits. Age categories harboured significantly different pathotypes (p = 0.019), while no association was found between pathotypes and lesions (p = 0.128) or sampling sites (p = 0.491). The antibiotic resistance was observed to increase with the time spent in the farm environment (age category). The selective pressure exerted by antibiotic use acted by giving advantage to more resistant strains rather than by lowering susceptibility to various drug categories within strains.

A study in an intensive Italian rabbit farm was carried out to assess the effect of age category and anatomical site on detection of S. aureus and to describe the diversity of spa-types within farm, including workers. On 400 rabbits of different age, 2066 samples from the ear, nose, axilla, groin, perineum and lesions were analyzed and 595 S. aureus were isolated. In total, 284 (71.0%) were colonized by S. aureus in at least one anatomical site and 35 animals (8.8%) had lesions. S. aureus prevalence was higher in adults than other age categories. Ear (29.4%) and nose (27.2%) were the most frequently colonized sites. The presence of lesions due to S. aureus was strongly associated with the colonization of at least one body site and the risk of lesions was proportionate to the number of sites colonized by S. aureus. In addition, a genotypic characterization was performed for 96 S. aureus strains randomly selected. All isolates resulted as low-virulence methicillin-sensitive-Staphylococcus aureus (LV-MSSA). Five different clonal lineages were found: spa-t2802, the most frequently detected (53.1%), also in human samples; spa-t491 (38.5%); spa-t094; t605; and spa-t2036. Strains t094, t491 and t2036 have not been isolated in Italy before.