We investigated the occurrence of organochlorine pollutants (OCs) in the muscle of brown trout and evaluated their potential modulation of parasite infection. The toxicological risk for consumer health was assessed, too. Trout were collected from the Sila National Park (Calabria region, South of Italy). The highest concentrations emerged for the sum of the 6 non-dioxin-like (ndl) indicator polychlorinated biphenyls (Σ6ndl-PCBs), followed by the 1,1,1-trichloro-2,2-di(4-chlorophenyl)-ethane (DDT), dioxin-like PCBs, hexachlorobenzene (HCB), and dieldrin. Measured on lipid weight (LW), the mean value of Σ6ndl-PCBs amounted to 201.9 ng g⁻¹, that of ΣDDTs (the sum of DDT-related compounds) to 100.2 ng g⁻¹, with the major contribution of the DDT-metabolite p,p’-DDE which was detected in all sample units (97.6 ng g⁻¹ on average). Among dioxin-like congeners, PCB 118 showed the highest mean concentration (21.96 ng g⁻¹ LW) and was detected in all sample units. Regression analysis of intestinal parasites on OC concentration was performed, controlling for two potential confounding factors, namely sex and sexual stage. The results evidenced the existence of interactions between the dual stressors in the host-parasite system in the wild. A negative and statistically significant correlation was estimated, suggesting that OCs may decrease parasite infection degree. Regarding the toxicological risk evaluation, OC concentrations were consistently below the current European Maximum Residue Limits.

This study, for the first time in Italy, analyses by WGS a Streptococcus agalactiae strain isolated from a non-pregnant adult affected by Meningitis and without common risk factors. The S. agalactiae strain was classified as a serotype II (SS2), sequence type ST569. Molecular characterization evidenced the presence of resistance genes to tetracycline and macrolide (tet(M) and mre(A)) and several virulence genes coding for adhesion and immune evasion factors (bca, cps family, neu family, scpB, gbs family, pil family and hylB), toxins (cfa/cfb, cyl family), pro-inflammatory factors (lepA), and two homologous genes that contributed to bacterial escape from the host immune system (lmb, luxS). SNP analysis showed 18 different alleles, with 9 missense SNP mutations related to genes involved in cellular metabolism (dhaS, ftsE, ligA, nrdD and secA), virulence (bgrR and galE) and antimicrobial resistance (glpK and mutL). SNPs in glpK and mutL genes might reduce susceptibility to drugs. The SNP analysis highlighted the presence of mutations conferring pathogenicity to the strain. The evidence in this study could explain the development of Meningitis in a healthy patient. This case highlights the importance of using molecular methods to characterize the complete genome of a bacterial species that could seriously affect human health.

Equine Infectious Anemia Virus (EIAV), a lentivirus marked by considerable genetic variability, poses significant diagnostic challenges. Existing diagnostic tools encompass the Agar Gel Immunodiffusion Assay (AGID), enzyme-linked immunosorbent assay (ELISA), and Western blotting (WB). ELISA and AGID mainly utilize the p26 capsid protein, often sourced from the Wyoming reference strain. To broaden the range of viral proteins and strains employed in these immunoassays, we previously developed a novel p26/double-strain gp45 indirect ELISA. In this study, we evaluated the performance of this ELISA in comparison to two commercial EIAV ELISAs using Cohen’s Kappa test and Bayesian Latent Class Analysis (BLCA), a statistical method that estimates test performance without requiring a perfect reference standard. A comparison with the official classification of the sera by the Italian Veterinary Service was also performed. A total of 372 serum samples, including 96 that were positives by all three tests, were analyzed. Results from both Cohen’s Kappa test and BLCA, alongside comparison with official classifications, affirm the diagnostic reliability of the two commercial ELISAs and suggest that the novel ELISA, with its enhanced antigenic diversity, could offer an accurate and reliable diagnostic option for EIAV. This novel assay enhances existing commercial ELISAs and has the potential to strengthen routine diagnostic workflows.

Feline leishmaniosis (FeL) is still considered an emerging and neglected disease. Cats, once considered accidental hosts, are now recognized as adjunctive reservoirs of the disease, especially in areas where canine (CanL) and human (HumL) leishmaniosis are widespread. Although often asymptomatic, infected cats could contribute to the transmission cycle of the parasite. Recent studies in Campania (Italy) have found a significant prevalence of feline infection, indicating the need to implement diagnostic and surveillance protocols to prevent the spread of the disease. The aim of the study was to outline the current scenario by studying the prevalence of FeL in Campania to identifying the potential zoonotic risk and in addition to validate the Immunofluorescence Antibody Test (IFAT) method for the diagnosis of leishmaniosis in cats. The study involved initially 702 cats; for each cat, a clinical record was compiled, including identification data, anamnesis, and clinical findings. Due to incomplete information, statistical analysis was performed only on a subset of 601 cats. A blood sample was collected to obtain serum/plasma specimens. When feasible, a lymph node fine-needle aspiration was performed. The observed seroprevalence rate was 32.1% (193/601), with a higher seroprevalence in outdoor cats and the presence of asymptomatic seropositive animals (28.0%;54/193), suggesting that felines may act as silent reservoirs of Leishmania infantum. An excellent result was obtained for the validation and standardization of the analytical IFAT method for the diagnosis of feline leishmaniasis; therefore, an inter-laboratory test has been carried out to establish the dilution cut-off at ≥1:80 as compatible with infection. Furthermore, a xenodiagnosis examination was conducted on a cat that was infected to more accurately evaluate the possibility of asymptomatic cats acting as carriers of the infection; however, this test resulted negative.

This study, for the first time in Italy, analyses by WGS a Streptococcus agalactiae strain isolated from a non-pregnant adult affected by meningitis and without common risk factors. The S. agalactiae strain was classified as a serotype II (SS2), sequence type ST569. Molecular characterization evidenced the presence of resistance genes to tetracycline and macrolide (tet(M) and mre(A)) and several virulence genes coding for adhesion and immune evasion factors (bca, cps family, neu family, scpB, gbs family, pil family and hylB), toxins (cfa/cfb, cyl family), pro-inflammatory factors (lepA), and two homologous genes that contributed to bacterial escape from the host immune system (lmb, luxS). SNPs analysis showed 18 different alleles, with 9 missense SNP mutations related to genes involved in cellular metabolism (dhaS, ftsE, ligA, nrdD and secA), virulence (bgrR and galE) and antimicrobial resistance (glpK and mutL). SNPs in glpK and mutL genes might reduce susceptibility to drugs. The SNPs analysis highlighted the presence of mutations conferring pathogenicity to the strain. The evidence in this study could explain the development of meningitis in a healthy patient. This case highlights the importance of using molecular methods to characterize the complete genome of a bacterial species that could seriously affect human health.

Wolves have reclaimed much of their former range in Europe, including areas with a high human footprint, yet the physiological impacts of human‐associated stressors remain poorly understood. We first examined methodological factors affecting hair cortisol concentrations (HCC) in wolves by using a ‘shave/re‐shave' design to assess hair growth rate and seasonal variation in four captive individuals. We also evaluated the effects of body region, collection method, and the relationship between faecal glucocorticoid metabolites (FGCMs) and HCC. Hair grew faster in summer, but HCC peaked in hair grown during winter, likely reflecting seasonal elevations in glucocorticoids (GCs) due to reproduction and varying metabolic demands. FGCMs showed comparable seasonal fluctuations and were significantly correlated with HCC. Additional samples collected by brushing the belly showed higher HCC than those from the back. To then assess the effects of urbanisation on HCC in free‐ranging wolves, we analysed hair samples from 254 carcasses from Italy and quantified human influence using the human footprint index (HFI). Contrary to our prediction, the HFI did not explain variation in HCC. This result may reflect the absence of individuals exposed to extreme urbanisation, uncertainty regarding whether carcass locations represented true home ranges, and the possibility that broad landscape indices such as the HFI fail to capture more immediate anthropogenic stressors. In contrast, health status and season significantly predicted HCC: wolves with chronic injuries or infectious diseases exhibited elevated HCC, and individuals that died in winter had higher HCC than those that died in other seasons. We discuss these results in the context of the wolf breeding season and dispersal period, and highlight the importance of considering seasonal demands, health status, and methodological constraints when interpreting GC patterns in wolves.