Leptospirosis is a zoonosis of public health concern. Its prevalence in stray animals in the South of Italy is unknown. This study aimed to investigate Leptospira spp. prevalence in 1009 stray animals. Out of them, 749 were alive animals, including 358 dogs (316 from Palermo and 42 from Ragusa) and 391 cats (359 from Palermo and 32 from Ragusa), and 260 were corpses (216 dogs and 44 cats) randomly collected in Sicily. Dogs and cats underwent a serological screening by Microscopic Agglutination Test and a molecular investigation by Real-Time PCR targeting lipL32. Corpses were subjected to Real-Time PCR. Serological analyses showed a prevalence of 1.12% (4/358) for dogs and 0.26% (1/391) for cats, with the only positive cat coming from Palermo. Serogroup Icterohaemorrhagiae serovar Icterohaemorrhagiae or Copenhageni, followed by Canicola and Bratislava, were the most spread among dogs, while the serological positive cat reacted with Hardjo serogroup. Two urine (2/32, 6.25%) and one blood (1/391, 0.26%) samples of cats, all from Ragusa, were positive at Real-Time PCR for pathogenic Leptospira spp. Sequencing analyses showed the presence of L. interrogans serogroup Icterohaemorrhagiae serovar Icterohaemorrhagiae or Copenhageni in one of the positive urine samples and in the positive blood sample. Analyses on corpses showed a prevalence of 1.85% (4/216) in Sicilian dog kidney samples, while all corpses of cats resulted in negative. Genotyping analysis showed a genetic relatedness between cat and human isolates. Results show Leptospira spp. circulation among Sicilian stray animals. The genetic relatedness between cat and human isolates suggests a possible common infection source.

Leptospirosis is a re-emerging zoonosis of worldwide significance; a wide spectrum of wild and domestic animal species act as natural or accidental hosts. Swine can act as maintenance or accidental hosts of pathogenic Leptospira spp. This study aimed at investigation of Leptospira spp. prevalence and diversity in slaughtered pigs in southern Italy (Sicily). In total, 55 samples of kidneys and blood were collected. Microscopic agglutination test and real-time PCR were performed to detect pathogenic and intermediately pathogenic Leptospira. Partial rpoB gene sequencing and multi-locus sequence typing (MLST) were performed to characterize Leptospira species. The analysis showed a seropositivity rate of 16.4%, with Australis representing the most frequently identified serogroup (63.64%); Pomona and Sejroe were detected with a prevalence of 27.27% and 9.09%, respectively. Pathogenic Leptospiral DNA was detected in 2 kidney samples (3.64%). Leptospira were identified through MLST as L. borgpetersenii serovar Tarassovi (serogroup Tarassovi). Obtained data confirmed the presence of Leptospira infection among pigs in southern Italy, suggesting that management of these animals may be considered an occupational risk for humans.

Q fever is a widespread zoonotic disease caused by the obligate intracellular bacterium Coxiella burnetii, primarily transmitted through the inhalation of contaminated aerosols. This study aimed to detect C. burnetii in two Sicilian sheep flocks, with no better defined reproductive disorders reported by the farmers. Blood, individual and bulk milk, ticks, and conjunctival swabs were collected from both flocks (A and B). Real-time and traditional PCRs were carried out to detect C. burnetii DNA and anti-C. burnetii antibodies were searched using an ELISA. In terms of Farm A, C. burnetii DNA was detected in 7.1% of blood samples, 20% of individual milk samples, bulk milk, 66.6% of conjunctival swabs, and in all the examined tick pools. Anti-C. burnetii antibodies were found in 77.0% of sera, 92.5% of individual milk samples, and bulk milk. In terms of Farm B, C. burnetii DNA was detected in 3.8% of blood samples, 39.4% of individual milk samples, bulk milk, 100% of conjunctival swabs, and in all tick pools; anti-C. burnetii antibodies were present in 53.6% of sera, 73.2% of milk samples, and in bulk milk. Our results highlight the high diffusion of C. burnetii in the two outbreaks, with widespread pathogen circulation, significant shedding in dairy products, and high environmental contamination, highlighting the need for enhanced surveillance and control measures in dairy sheep farms.

Bats, as members of the order Chiroptera, are vital to ecosystems and serve as reservoirs for numerous microorganisms, some of which can cause zoonotic diseases. Human interactions with bats are increasing due to habitat alterations, making it essential to understand their microbiota, particularly potential pathogens. This study aimed to evaluate the excretion of zoonotic bacteria and protozoa in insectivorous bats from four caves in the provinces of Ragusa, Catania, and Syracuse (Sicily, Southern Italy) using molecular biology tests for zoonotic agents, including Bartonella henselae, Borrelia, Coxiella burnetii, Leptospira, Chlamydia, Rickettsia, Anaplasma, and Piroplasmids. From December 2020 to April 2023, urine, fecal swabs, ocular conjunctival swabs, and oropharyngeal swabs were collected from 149 bats of six species, along with guano samples from the caves. Bartonella henselae DNA was detected in 3 of the 149 tested bats, one ocular conjunctival swab and two oropharyngeal swabs. Chlamydia spp. DNA was detected in a sample of guano, in feces, ocular conjunctival and oropharyngeal swabs of a bat, and in four urine samples. Piroplasmid DNA was detected in 10 of 149 fecal swabs and in 5 of 16 bat ectoparasites. No samples were positive for Leptospira spp., Borrelia spp., Coxiella burnetii, Rickettsia spp., or Anaplasma spp. These findings underscore the importance of multiple sample types in assessing bats as reservoirs for zoonotic pathogens, particularly highlighting their role in transmitting pathogens through various body habitats, including saliva, urine, and ocular excretions. This study highlights the relevance of monitoring bat populations and studying their microbiota to enhance protections for both human and animal health.

Leptospirosis is a worldwide widespread zoonosis caused by Leptospira genus. We report an acute leptospirosis case in a puppy housed at a municipal kennel and the subsequent diagnostic investigations carried out on all dogs housed in the kennel. Laboratory investigation included mainly a microagglutination test, real-time PCR, and multi-locus sequence typing (MLST) for Leptospira genus. Other agents of infection were excluded. The puppy resulted positive for Leptospira interrogans Icterohaemorrhagiae both with serological and molecular assays. All of the other 66 dogs in the kennel underwent clinical and laboratory investigations twice, 15 days apart. No other dog showed leptospirosis clinical signs. At the first sampling, eight dogs (12%) showed antibodies against Leptospira interrogans serogroup Icterohaemorragiae serovar Copenhageni. Real-time PCR on urine samples of seropositive dogs detected Leptospira spp. DNA in one sample, then identified as Leptospira interrogans serogroup Icterohaemorragiae by MLST. Fifteen days after, four of the previous seropositive dogs still showed antibodies against Leptospira spp. All urine samples collected from seropositive dogs were negative at real-time PCR. The study allowed the early confirmation of a Leptospirosis case and the identification of at least one asymptomatic carrier of pathogenic Leptospira spp. The prompt activation of all appropriate management measures allowed limiting and extinguishing the infection.

Ticks are a major concern for both animal and human health, as they are primary vectors of infectious pathogens. This study focused on ticks found in a nature reserve in southern Italy, highly frequented for recreational activities and inhabited by wild boars. Using molecular techniques, 214 ticks, including questing ticks and those removed from wild boars, were examined for tick-borne pathogens (TBPs), with a focus on zoonotic pathogens. Six tick species were identified: Hyalomma lusitanicum, Rhipicephalus pusillus, Rh. sanguineus s.l., Rh. bursa, Rh. turanicus, and Dermacentor marginatus, several of which are known vectors of zoonotic pathogens. Overall, 14% of ticks were positive for TBPs, mainly bacteria of Rickettsia genus. Molecular analyses detected Rickettsia slovaca, R. massiliae, Candidatus R. shennongii, R. conorii, R. felis, and R. barbariae. Additionally, single detections of Coxiella burnetii and Theileria annulata were recorded. Phylogenetic analyses were conducted on Rickettsia sequences. The range of ticks and TBPs present in this area highlights potential public health concerns. New associations between tick species and TBPs were documented, though vector roles need further investigation. The study highlights the importance of monitoring tick populations in both rural and urban environments to protect public health and prevent tick-borne disease spreading.