Background/Objectives: Vancomycin-resistant enterococci (VRE) are significant nosocomial pathogens worldwide, potentially transmitted by food-producing animals and related products. This study investigates the epidemiological role of bovine raw milk in the transmission of VRE to humans. Methods: Bulk milk samples were screened for van gene presence using a multiplex PCR. Mastitogenic enterococci isolated from individual milk samples were tested for antimicrobial susceptibility using the broth microdilution method. Strains not susceptible to vancomycin were whole genome sequenced. Results: Overall, vanC genes were detected in 299/1026 (29.14%) bulk milk samples. Specifically, vanC1 was found in 204 samples (19.88%) and vanC2/3 in 57 samples (5.56%), with both detected simultaneously in 38 samples (3.70%). Clinically significant vanA and vanB genes were not identified. A total of 163 mastitogenic Enterococcus strains were isolated from individual milk samples. Eight different Enterococcus species were detected, with E. faecium (104/163, 63.80%) and E. faecalis (34/163, 20.86%) being the most common. Multidrug resistance was observed in 106/163 (65.03%) isolates. The most common resistance frequencies were to ciprofloxacin and erythromycin (102/163, 62.58% both), followed by quinupristin/dalfopristin (93/163, 57.06%), linezolid (65/163, 39.88%), tetracycline (58/163, 35.58%), daptomycin (46/163, 28.22%), chloramphenicol (33/163, 20.25%), ampicillin, tigecycline, and high-dosage gentamycin (8/163, 4.91% all). Resistance to teicoplanin was not observed. Two vancomycin non-susceptible strains were identified: one vanC2/3 E. casseliflavus and one vanC1 E. gallinarum. Whole genome sequencing confirmed the presence of the complete vanC gene cluster and several virulence genes in both strains. Conclusions: Our findings suggest that while raw milk is unlikely to be a source of vancomycin resistance genes of highest clinical importance (vanA or vanB), it may contribute to the spread of vanC enterococci, which are increasingly associated with human infections.

: Blue crab (Callinectes sapidus) is one of the last reported invasive species that have colonized the Italian coasts, in particular the lagoons of the northern Adriatic Sea. Their massive presence is compromising shellfish farming, causing a serious economic crisis among the valley populations. To limit the economic damage, blue crabs have been introduced as human food in the Italian markets. This study aimed to evaluate the microbiological contamination of blue crab as potential risk to human health. From July 2023 to April 2024, 72 blue crabs were collected in coastal lagoons and 5-6 nm from the coast of the Emilia Romagna Region. Microbiological analysis revealed Shewanella algae as one of the most commonly identified microorganisms (18/72, 25%). Shewanella was isolated on Chromagar Vibrio™ and TCBS after an enrichment step in Alkaline Saline Peptone Water. Colony morphology, Gram stain, and biochemical tests (oxidase, catalase) sustained Shewanella identification, later confirmed by MALDI-TOF MS (MALDI Biotyper® Sirius, Bruker Daltonics). S. algae is a Gram-negative, rod-shaped, aerobic microorganism widely distributed in seawater, freshwater, and soil. In humans, it is considered an opportunistic pathogen, especially in people with impaired immune systems. Shewanellosis has been strongly associated with seawater exposure and typically arises in hot months. Clinical manifestations include skin and soft tissue infections, otitis media, keratitis, biliary tract infections, vertebral discitis, and bloodstream infections. Notably, in the last years, S. algae infections have increased globally, even in healthy young people. The present study suggests blue crabs as a potential source of S. algae throughout the year. The increasing identification of S. algae in marine waters worldwide, as well as in marine-derived foods like blue crabs, underscores the growing importance of S. algae as an emerging pathogen and its impact on public health. Key messages: • Shewanella algae is an opportunistic pathogen, increasingly isolated in marine water and seafood. • Blue crab is an invasive species that has recently conolonised the Italian coasts.

Invasive mosquitoes are vectors of important human and animal pathogens and a serious threat to public health. Aedes albopictus (Skuse) (Diptera: Culicidae) and Culex quinquefasciatus (Say) (Diptera: Culicidae) are good examples because of their wide occurrence, host range and vector competence. An understanding of the responsiveness of mosquitoes to olfactory stimuli is essential for implementing effective surveillance and developing repellents. The present study evaluated the behavioural responses of A. albopictus and C. quinquefasciatus to CO₂ and human skin odour in an olfactometer. In addition, CO₂ synergistic effect was assessed in association with human skin odour. Mosquitoes of different ages (3-5 and 10-15 day old) were included in the study in order to determine changes in responsiveness to attractants during an insects' lifetime. The highest numbers of mosquitoes captured associated with CO₂ were (A. albopictus, 48/77, 62.34%; C. quinquefasciatus, 117/126, 92.86%) and hand odour (A. albopictus, 211/232, 90.95%; C. quinquefasciatus, 320/374, 85.56%) in the "CO₂ vs blank" and "hand vs blank" treatments. Skin odour was the most attractive for both species (A. albopictus, 279/309, 90.29%; C. quinquefasciatus, 292/306, 95.42%) in "CO₂ vs hand" experiment. The highest mosquito responsiveness was recorded in the "CO₂ + hand vs hand" bioassay (A. albopictus, 174/265, 65.66%; C. quinquefasciatus, 231/425, 54.35%). Similar trends were recorded for 10-15 and 3-5 day old mosquitoes of both species in all the experiments. In addition, a linear mixed model was used to evaluate the interactions between species, age and attractants. Human skin odour and CO₂ were effective attractants for both A. albopictus and C. quinquefasciatus (attractant-species interaction, p-value < 0.05). CO₂ synergistic effect was recorded for both species (species-attractant interaction, p-value < 0.05) even when CO₂ was not directly combined with skin odour (p-value < 0.05). The interaction between attractant and age revealed (p-value < 0.05) that in both species, 10-15 day old mosquitoes were more responsive to CO₂ and human skin odour, than younger (3-5 days) adults. The species-age interaction (p-value < 0.05) showed that 3-5 and 10-15 day old C. quinquefasciatus were more receptive to CO₂ and skin odour, especially when used in combination, than A. albopictus.

Trichinella spp. are zoonotic nematodes with a global distribution, primarily maintained through wildlife reservoirs which complicates eradication efforts. In Europe, four species—T. spiralis, T. britovi, T. nativa, and T. pseudospiralis—circulate in wild carnivores and omnivores, with T. britovi and T. spiralis being the most prevalent in wild and domestic hosts. This study aimed to assess the presence and species distribution of Trichinella spp. in wild mammals. This study presents the results of five years of wildlife surveillance carried out within the long-standing monitoring program implemented in the Emilia-Romagna region, northern Italy, which has been in place since 2006. Between 2020 and 2024, a total of 104,338 wild mammals, including red foxes, wolves, and wild boar, were tested using the magnetic stirrer digestion method in accordance with EU regulations. A total of 12 animals (0.011%) were found to be infected with Trichinella larvae. Molecular identification of positive samples, performed at the European Union Reference Laboratory for Parasites, showed the presence of T. britovi in 11 animals, while one isolate remained unidentified. T. britovi was detected primarily in wolves and red foxes. No infections were identified in wild boar during the study period. Overall, the low prevalence observed confirms that Trichinella spp. infections in wildlife in Emilia-Romagna are sporadic but persistently present. Only T. britovi was detected, supporting its role as the predominant species in this area. These findings highlight the importance of long-term surveillance for early detection and risk assessment within a One Health framework.

Salmonella is diffused worldwide, and Salmonella enterica subsp. enterica is spread worldwide with many serovars associated with the infection of domestic bovines. The most spread are S. Dublin, S. Typhimurium and S. Infantis. S. Napoli is, however very rarely reported in domestic ruminants. Here, we report an outbreak of S. Napoli on a dairy cow farm in Northern Italy (Piacenza). A total of 18 S. Napoli isolates were recovered from aborted fetuses, feces, tissues and environmental samples. Whole genome sequencing suggested that all isolates belonged to the same cluster. After the application of stringent biocontainment and biosecurity measures, no further cases were reported. However, four months after the first case, the serovar was still isolated in environmental samples, underlying the importance of adopting the correct biosecurity and biocontainment measures in order to prevent the circulation and transmission of Salmonella within the farm.