Small ruminant lentiviruses (SRLVs) are retroviruses infecting small ruminants, characterized by a high genetic variability. SRLVs infection may result in a variety of clinical signs in goats and sheep and is not species-specific. SRLVs are worldwide distributed with four distinct genotypes and numerous sub-genotypes described so far. Full understanding of SRLVs epidemiology is still a matter of research and genetic characterization remains a key tool for unrevealing the complexity of such infections. A previous study, exploring the genetic landscape of SRLV in Italy, reported considerable genetic heterogeneity across geographic regions and suggested the presence of new sub-genotypes. In light of these previous results, our study provides an update on the genetic characterization of SRLVs based on phylogenetic analysis of SRLVs positive samples collected during passive surveillance in 16 out of 20 Italian regions between 2019 and 2024. Our findings revealed for the first time the broad genetic diversity of SRLVs circulating across Italy with apparently restricted circulation of some sub-genotypes to specific areas/regions. The observed genetic clusters are region-specific and do not align with the currently recognized sub-genotype nomenclature, suggesting that the existing classification system may necessitate an update to describe the current SRLVs diversity more accurately. Continuous monitoring through full gene and genome sequencing, coupled with an updated and expanded genetic classification framework, is essential for improving our understanding of this complex and evolving viral infection in small ruminants.

The swine production chain represents a critical point for the spread of antimicrobial resistance, with slaughterhouses acting as potential hotspots for cross-contamination. The veterinary use of phenicols may promote resistance to oxazolidinones, a last-resort class of antibiotics. This study investigated the presence of oxazolidinone-resistant enterococci in a pig slaughterhouse in central Italy, analyzing 376 samples, 25.8% of which tested positive for Enterococcus spp. resistant to florfenicol. Environmental samples collected at the end of the day of slaughter showed higher contamination levels compared to those collected beforehand. PCR analysis of 42 isolates of Enterococcus faecalis and Enterococcus faecium revealed a high prevalence of the optrA gene, followed by poxtA and cfr(D). The data indicated high resistance rates to tetracycline and erythromycin, consistent with existing literature. Linezolid resistance was observed in 73.8% of isolates, while no vancomycin resistance was detected. Whole Genome Sequencing (WGS) analysis revealed diverse sequence types, complex resistomes, multiple virulence factors, and considerable variability in the genetic elements carrying cfr/cfr(D), optrA, and poxtA. These determinants were often plasmid-borne and co-localized with other antibiotic and heavy metal resistance genes, which could contribute to their persistence and dissemination through co-selection mechanisms. Overall, the findings highlight the widespread occurrence of oxazolidinone-resistant enterococci in the pig production chain and their potential circulation across different production stages. An integrated approach encompassing prudent antibiotic use in livestock, good hygiene practices at the slaughterhouses, and increased awareness of the problem will be essential to mitigate the spread of antimicrobial resistance along the food production chain.

Fishery and aquaculture products increasingly represent an important food source for the world population. The intensification of aquaculture guided an increase in the use of antimicrobials in this sector as well. The aquatic environment represents a link among humans, animals and environmental antibiotic resistance. The rising problem of antibiotic resistance leads to the need for the identification of indicator bacteria and the development of monitoring methods, which are poorly standardized for the aquatic environment. Aeromonas spp., a ubiquitous bacterium in aquatic environments, is among the possible candidates as bacterial indicator. The goals of this study were to: (i) evaluate the antibiotic-resistance profile of different antimicrobial classes in Aeromonas spp. collected from seafood sold in superstores of central Italy; (ii) propose possible epidemiological cut-offs for the genus Aeromonas and evaluate its effectiveness as a possible bacterial indicator for monitoring antibiotic resistance in seafood. The results obtained revealed a high presence of Aeromonas spp. in seafood categories, highlighting its candidature as a bacterial indicator. The presence of isolates resistant to some of the tested antibiotics has been described, even if in rare occasion. This data raises concerns about the food safety of seafood for consumption, which may represent a risk for public health and consumers. In this study, we described the epidemiological cut-offs which agree with the literature, representing a contribution to the definition of reliable epidemiological cut-offs for the genus Aeromonas spp.

This study assessed Escherichia coli as an indicator of antimicrobial resistance (AMR) in retail seafood, integrating microbiological and genomic analyses within a One Health perspective. Overall, E. coli was cultured from 55 (13%) of 422 raw samples from seven seafood categories commonly consumed in Italy. E. coli was detected across multiple seafood types, but particularly in bivalves (23%) and crustaceans (20%). Because these species can accumulate or acquire faecal contaminants from the aquatic environment or along the post-harvest chain, they represent relevant matrices for AMR monitoring. In parallel, we investigated the presence of Extended-β-Lactamase ESBL-, AmpC- and carbapenemase-producing Enterobacteriaceae. Cefotaxime-resistant E. coli isolates were identified at low but relevant frequencies, especially in bivalves, and were characterised phenotypically and by whole-genome sequencing. Most isolates carried clinically important ESBL or AmpC genes, more frequently blaCTX-M-15, often on contigs predicted as plasmid-associated, indicating their potential for horizontal spread. An ESBL-producing Atlantibacter hermannii harbouring blaCTX-M-55 was also recovered from frozen shrimps. Additional AmpC-producing Enterobacteriaceae, including Citrobacter and Enterobacter species, were detected in a subset of samples.Overall, our findings support the suitability of E. coli as an AMR indicator organism for retail bivalves and crustaceans. The detection of ESBL-producing strains, carrying important plasmid-borne resistance determinants, highlights the need to strengthen AMR surveillance in these seafood commodities.

Alternaria toxins (ATs) are a group of toxins produced by Alternaria fungi that frequently contaminate tomatoes and tomato products. Recently, the European Food Safety Authority evaluated ATs for their genotoxic and carcinogenic properties. Alternaria infestation is often controlled using ad hoc treatment strategies (fungicides). In this study, two analytical methods were developed, validated and applied for the determination of five ATs and 195 pesticides in tomato products collected from the Italian market. Two distinct groups, organic (n = 20) and conventional (n = 20) Italian tomato sauces, were characterized in depth. Tenuazonic acid, alternariol and alternariol monomethyl ether were found up to 517, 27 and 7.1 µg/kg, respectively, while pesticides were detected between 0.0026 and 0.0421 mg/kg in conventional products, and, interestingly, up to 0.0130 mg/kg in organic products. No correlation emerged between the detected levels of ATs and pesticides and the type of tomato cultivation, but the probability of pesticide contamination in conventional products was eight times higher than in organics. Some considerations about exposure assessment and risk characterization for ATs were also proposed in the overall population and in more sensitive and/or exposed subgroups, underlining the need for new focused toxicological and monitoring studies to establish reliable reference values. Moreover, these data highlight that fungicide treatments may not protect tomatoes from ATs contamination, although it may remove fungi infestation. As organic product consumption is increasing, it is important to lay down dedicated regulations for maximum permitted levels to ensure the food safety of these products that are often perceived by consumers as a healthier and environmentally friendlier choice.

Background: Brucella canis is a zoonotic pathogen associated with reproductive disorders in dogs and represents an emerging public health concern. Dogs are the only known source of infection for humans, and transmission is often associated with close contact, particularly in occupational settings. Reports of canine and human infections in Europe are increasing, underscoring the need for integrated surveillance to assess the risk of introduction and spread. Objectives: This study aimed to investigate the possible circulation of B. canis in different subgroups of dogs from Central Italy, representing diverse risk contexts (stray, breeding, blood donor, refugee-associated, and previously outbreak-linked dogs), and to generate sentinel data to inform further risk-based surveillance and zoonotic risk assessment. Methods: A comprehensive serological, molecular, and bacteriological survey was conducted on 128 dogs sampled in the Umbria region, covering animals from different backgrounds and risk contexts. Blood samples were tested using bacterial culture, real-time PCR, serum agglutination test, complement fixation test, and/or indirect immunofluorescence antibody test. Results: All tested dogs were negative for B. canis. The upper 95% confidence limit for prevalence was 3.5%, suggesting that widespread circulation is unlikely, although a low/moderate prevalence in specific groups cannot be excluded. Conclusions: Although no cases of B. canis were detected, the results provide sentinel information and highlight the need for continued risk-based surveillance, particularly in low-prevalence areas to prevent introduction of the infection and to enable early detection in case of occurrence. As dogs are the only known source of human infection, veterinary monitoring plays a pivotal role in mitigating zoonotic risks and supporting One Health strategies for evidence-based control.

Seafood is a crucial global food source, but its role as a reservoir for antibiotic-resistant pathogens is concerning. This study focuses on the prevalence of antibiotic resistance in retail seafood, using Enterococcus spp. as an indicator. Seven categories of raw seafood were collected from retail markets in Central Italy. Out of 422 samples, 288 tested positive for Enterococcus spp., resulting in a prevalence rate of 68.25% (CI95%: 63.66%-72.51%). The most common species were E. faecalis (48%) and E. faecium (17.4%). Enterococcus spp. was most frequently found in cephalopods, salmon, bivalves, and crustaceans than in sea bass and bream. The odds of Enterococcus spp. recovery were higher in frozen than in fresh samples, while there was no difference between farm-raised and wild-caught seafood. A subset of 102 isolates was selected for antibiotic resistance testing, showing the highest resistance rates to quinupristin/dalfopristin (52.94%) and tetracycline (27.45%). Over 70% of isolates were multidrug-resistant. Additionally, strains resistant to vancomycin and oxazolidinones, two last-resort antimicrobials, were detected. Vancomycin resistance was observed in E. casseliflavus and E. gallinarum, which are naturally resistant due to the vanC chromosomal cluster. The optrA gene, responsible for resistance to oxazolidinones and phenicols, was found in E. thailandicus and E. faecium, where it was located on to a transferable plasmid. In conclusion, this study confirms the widespread presence of Enterococcus spp. in retail seafood. The detection of multi-resistant isolates and resistance to last-resort antimicrobials highlights significant human health risks, stressing the importance of including Enterococcus spp. in seafood antibiotic resistance surveillance.

Canine prostatic serum esterase (CPSE) is considered a useful tool to identify prostate disorders in dogs, with increasing interest in ultrasound (US)-based sonoelastography to non-invasively detect prostate disorders. Since no report is available about a possible correlation between these diagnostic tools, we aimed to investigate a possible correlation between strain elastography (SE) and 2D-shear wave elastography (SWE) and CPSE. Twenty-one dogs were included and, on each animal, CPSE was evaluated followed by a complete US examination and SE and 2D-SWE application. Healthy dogs were identified based on the CPSE results. All the dogs included were characterized by normal CPSE values (<52.3ng/mL) and normal US prostate appearance. The prostate was characterized by intermediate stiffness with SE (pattern III - 84.7% for the left lobe and 79.27% for the right lobe) and softer than the abdominal wall (SR 0.6 for the left lobe and 0.56 for the right lobe), with low values for both m/s and kilopascals (kPa) for 2D-SWE, pointing that the healthy tissue is not hard. 2D-SWE results were, respectively, 13.515.55kPa and 2.31 +/- 0.42m/s for the left lobe and 18.05 +/- 6.47kPa and 2.39 +/- 0.43m/s for the right lobe. The significant difference between the right and left measurements expressed with kPa, not evidenced with m/s, can be considered indicative of m/s as the most reliable measurement to be considered regarding the prostate parenchyma. Even though no linear correlation was detected between CPSE and elastography values, these preliminary results evidence that the healthy prostates were characterized by a similar elastographic pattern, thus pointing that these techniques can be potentially useful to be applied in case of prostatic disorders to improve the accuracy of the final diagnosis in a non-invasive way.

Companion dogs may be valuable sentinels to better understand the environmental determinants of morbidity and mortality in humans. This study aimed to assess the dog population and mortality in Umbria Region. The source of data was the local Canine Registry. Attribute-specific crude mortality rates by sex, age, and breed were produced on a five-year basis (2014–2018). The human ICD-10 was employed to code the causes of deaths. Over 2014–2018, an annual average population of 226,875 specimens and a total of 46,743 deaths were estimated. Mortality rate was higher in young males than in young females. A specific cause of death was reported for 5,209 dogs; the 62.8 per cent (95

Poultry is the most likely source of livestock-associated Extended Spectrum Beta-Lactamase (ESBL) and plasmid-mediated AmpC (pAmpC)-producing E. coli (EC) for humans. We tested the hypothesis that farming methods have an impact on the load of ESBL/pAmpC-EC in the gut of broilers at slaughter. Isolates (n = 156) of antibiotic-free (AF), organic (O), and conventional (C) animals were characterized for antibiotic susceptibility and antibiotic resistance genes. Thirteen isolates were whole-genome sequenced. The average loads of ESBL/pAmpC-EC in cecal contents were 4.17 Log CFU/g for AF; 2.85 Log CFU/g for O; and 3.88 Log CFU/g for C type (p < 0.001). ESBL/pAmpC-EC isolates showed resistance to antibiotic classes historically used in poultry, including penicillins, tetracyclines, quinolones, and sulfonamides. Isolates from O and AF farms harbored a lower proportion of resistance to antibiotics than isolates from C farms. Among the determinants for ESBL/pAmpC, CTX-M-1 prevailed (42.7%), followed by TEM-type (29%) and SHV (19.8%). Avian pathogenic E. coli (APEC), belonging to ST117 and ST349, were identified in the collection. These data confirm the possible role of a broiler as an ESBL/AmpC EC and APEC reservoir for humans. Overall, our study suggests that antibiotic-free and organic production may contribute to a reduced exposure to ESBL/AmpC EC for the consumer.

Among the fungi responsible for deep mycosis, the genus Aspergillus plays a predominant role both in human and veterinary medicine. From a "One Health" perspective, infections by Aspergillus spp. often represent a public health problem linked to specific occupational categories that could have a greater risk of inhaling spores and developing any respiratory disease. This preliminary investigation allowed to acquire information about the spread of Aspergillus spp. in avian livestock of the Umbria region (Central Italy), their sensitivity to antifungals, and the presence of mycotoxins in the considered farms. Environmental, feed, animal, and human samples were collected for mycological investigations; chemical analyses were also performed in feed samples. Moreover, prevalence estimated of the fungal isolates were provided for each individual farm sampled. Direct fungal identification was possible in 298 out of the 559 total samples; 162 of the samples were positive for Aspergillus spp. Mycotoxins were detected in 5 out of the 21 feed samples collected. All the aspergilli tested for antifungal susceptibility were resistant to fluconazole. The results obtained show how much the genus Aspergillus is widespread in the investigated farms; therefore, the poultry livestock represents a favorable environment for the maintenance and spread of fungal spores and their potential transmission to animals and humans.

Although its importance, little information is available on antibiotic-resistance in cow-calf beef farms. This study aimed to determine prevalence and risk factors for antibiotic-resistant organisms in this livestock system. Fifty-four farms from Central Italy were included to assess the presence of antibiotic-resistant indicator Escherichia coli and of ESBL and/or AmpC-producing E. coli (ESBL/AmpC-EC) in calves. Antimicrobial usage (AMU) was recorded, and farm-related variables were collected through questionnaires. Potential risk factors were tested using a mixed-effects logistic regression model. The presence of resistant-E. coli was recorded in 75.9% of farms (95% confidence interval [CI]: 62.4-86.5) with resistance to tetracyclines, sulfonamides, penicillins, and fluoroquinolones as the most frequent. The prevalence of farms positive for ESBL/AmpC-EC was 35.2% (95% CI: 22.7-49.4). AMU on the farms originating a resistant-E. coli was higher than that on the farms originating a susceptible-E. coli. The same difference was found for the consumption of beta-lactams (beta-DCD/year) and AMU via the parenteral route, which resulted also associated with the presence of ESBL/AmpC-EC. Farms with higher beta-DCD/year had an increased risk of being positive for resistant-E. coli, whereas farms with higher overall AMU had an increased risk for ESBL/AmpC-EC presence. Among farm-related factors, only farm size was associated with the presence of ESBL/AmpC-EC (odds ratio: 5.8, 95% CI: 1.3-26.3). Our findings highlight a reduction of the risk of ESBL/AmpC-EC in small cow-calf farms, and a strong association between AMU and antibiotic-resistance. Antibiotic stewardship programs are needed to improve the health status of cow-calf farms and ensure their long-term sustainability.

African swine fever (ASF) is one of the most important swine diseases, both for its wide geographical spread and for the relevant impact that it has on pig industry; it is included in the list of notifiable diseases of the World Organization for Animal Health (OIE), together with all the other diseases that represent a danger to animal and public health. The constant spread of African Swine Fever virus (ASFV), together with its ability to adapt to different environments, its high resistance in the environment and biological products and the lack of a vaccine, make it difficult to eradicate. The disease was confined to Africa until the end of the 1950s, until the virus belonging to genotype I appeared in Portugal in 1957. In 2007, the ASFV (genotype II) reached the Caucasus region and entered the European Union (EU) in 2014, reaching progressively several Member States such as: Bulgaria, Slovakia, Estonia, Hungary, Latvia, Lithuania, Poland, Romania, and Germany. The infection also appeared in the Czech Republic and Belgium, but it was successfully eradicated. The knowledge about the consequences of this infection evidences many multidisciplinary aspects: the disease has consequences in many areas, from the socio-economic one to the medical one, passing through the psychological and cultural sphere. The aim of this paper is to propose a One Health approach for ASF because of its strict linkage between animal and human health as well, although it is not transmissible to humans.

The emergence of Clostridioides difficile as the main agent of antibiotic-associated diarrhoea has raised concerns about its potential zoonotic role in different animal species. The use of antimicrobials is a major risk factor for C. difficile infection. Here, we provide data on C. difficile infection in dairy and beef calves in Umbria, a region in central Italy. This cross-sectional study focuses on prevalence, risk factors, ribotypes, toxinotypes and antimicrobial resistance profiles of circulating ribotypes. A prevalence of 19.8% (CI95%, 12–27.6%) positive farms was estimated, and the prescription of penicillins on the farms was associated with C. difficile detection (OR = 5.58). Eleven different ribotypes were found, including the ST11 sublineages RT-126 and -078, which are also commonly reported in humans. Thirteen isolates out of 17 showed resistance to at least one of clindamycin, moxifloxacin, linezolid and vancomycin. Among them, multiple-drug resistance was observed in two isolates, belonging to RT-126. Furthermore, RT-126 isolates were positive for tetracycline resistance determinants, confirming that tetracycline resistance is widespread among ST11 isolates from cattle. The administration of penicillins increased the risk of C. difficile in calves: this, together with the recovery of multi-resistant strains, strongly suggests the need for minimising antibiotic misuse on cattle farms.

The objective of this research is, once the maximum time for which a critical activity / service can remain suspended in an emergency is known, to determine the resources for the recovery of each activity / service and define the priorities for recovery. This activity could reduce the vulnerability and therefore increase the resilience of companies in the agro-food sector, inserted in an area exposed to natural risks, to protect food safety and production. A pilot study (Current Research Project 16/2012 - Pilot study on business continuity planning in the agri-food field, in natural hazards exposed area) it was developed to apply Business Continuity Management (BCM) procedures for companies in the agro-food sector. The study involved two Regions of central Italy exposed to different natural risks (seismic, hydraulic and hydrogeological), characterized by small / medium-sized enterprises scattered throughout the territory of Umbria and Marche Regions. A study of the Territory was carried out to identify the threats that persist and to define possible scenarios at which to develop strategies and business continuity plans. The work developed through several consequent phases: Identification of all threats to business continuity; Definition of the threat scenarios on which to develop business continuity strategies and plans; Assessment of the organization's vulnerability for each type of threat considered and its exposure; Verification of the activities implemented to mitigate the damage suffered.On two pilot supply chains with different characteristics (dairies attached to farms and cured meat factories) studies and evaluations were carried out, breaking down and detailing the entire process into a well-defined number of processes. The supply chain management strategies and practices are the key factors in creating and distributing value along the steps of the same. Each of these subjects and steps was considered as a single process (business unit), so two types of risk assessment were applied in the selected companies: General assessment to identify specific exposures that may require protective measures; Assessment by business unit: to identify those functions that are most likely to disrupt the business and identify those resources on which the business strictly depends. Through a questionnaire administered to local producers, timely information was obtained for the evaluation of critical processes for operational and business continuity in the event of a disaster. The objective of this project is to evaluate whether the ready availability of data on the Operational Continuity (CO) of a company could shorten the Recovery Time Objective (RTO), that is the time necessary for the full recovery of the operation of a system or a organizational process and assess whether the assumed time corresponds to the maximum duration, foreseen or tolerated, of the down time (time of out of service) that has occurred. We have attributed primary importance to the quantification of the RTO value, known and verified, bearing in mind that a long down time damages the possibility of restoring the service much more than a short one. The greatest damage results from the lack of knowledge of the expected time to restore the damaged services. In general, the shorter the RTO, the more expensive the recovery strategy is, so we also want to evaluate whether the applied strategy could reduce not only the time but also the costs necessary for recovery. In order to make data on company CO readily available, the Emergenza 2.0 IT platform was optimized by implementing it with the information obtained from the questionnaires. In practice, the information necessary to assess their vulnerability or ability to survive a catastrophe was associated with the veterinary facilities

Clostridium (C.) perfringens is the causative agent of several diseases and enteric infections in animals and humans. The pathogenicity of the bacterium is largely mediated by the production of a wide range of toxins. Individual C. perfringens strains produce only subsets of this toxin repertoire, which permits the classification in seven toxinotypes (A–G). In addition, a variety of minor toxins further characterizes the single strains. The aim of this work was to evaluate, using Polymerase Chain Reaction (PCR) assays, the diversity of 632 C. perfringens strains isolated in Italy over 15 years. The genotyped strains were analyzed to determine the presence of major and minor toxins (cpe, consensus, and atypical cpb2), their geographical origins, and the source of isolation (animal species or food). Our study shows that toxinotype A had the greatest representation (93%) and correlated mainly with consensus cpb2 in a variety of animal species, as well as with atypical cpb2 in the five food samples. Type D, associated with cpe and atypical cpb2 minor toxins, was identified in 3% of the cases, and type F was identified in 2.5%. Seven type C isolates (1.1%) were detected in cattle, whereas the only type B atypical cpb2 isolated in Italy was detected in a goat, and one type E cpe+atypical cpb2 was detected in a sheep. Type G was not detected.

The study was aimed at assessing heat load-related risk of clinical mastitis (CM) in dairy cows. Records of CM for the years 2014 and 2015 were obtained from a large conventional dairy farm milking about 1,200 Holstein cows in central Italy. A case of CM was defined by the presence of clinical signs and veterinary confirmation. Quarter milk samples were collected and bacteriological investigated for each CM. Etiological agents were identified and classified as environmental or contagious pathogens. Hourly weather data from the nearest weather station were used to calculate heat load index (HLI). Upper and lower thresholds of HLI, at which the animal accumulates or dissipates heat, were settled and used to measure heat load balance through the accumulated heat load (AHL) model. Zero and positive values of AHL indicate periods of thermo-neutral and heat accumulation, respectively. Each case of CM was associated with HLI-AHL values recorded 5 d before the event. The risk of CM was evaluated using a case-crossover design. A conditional logistic regression model was used to calculate the odds ratio and 95% confidence intervals of CM recorded in thermo-neutral (AHL = 0) or heat load (AHL > 0) days, pooled or stratified for pathogen type (environmental or contagious). Classes of AHL as low (<6.5), medium (6.6-34.9), and high (>35) were included in the model. Other variables included in the model were milk yield as liters (<20, 20-30, and >30), days in milk (<60, 60-150, and >150), and parity (1, 2-3, and >3). A total of 1,086 CM cases were identified from 677 cows. Escherichia coli, Streptococcus spp., and Streptococcus uberis were the environmental pathogens isolated with the highest frequency; Staphylococcus aureus prevailed within contagious species. The analysis of pooled data indicated a significant effect of heat load on the occurrence of CM in the contagious pathogen stratum. Higher milk yield, middle and late stage of lactation, and older parity increased the risk of CM under heat load conditions. However, the association between pathogen type and these factors was not clear because the model provided significant odds ratios within all pathogen categories. The present study provided the first evidence of an association between HLI and CM in dairy cattle and suggested the ability of the AHL model to assess the risk of mastitis associated with heat load.