Camilla Smoglica

Mammarenaviruses are enveloped, ambisense, single-stranded RNA viruses capable of causing fatal hemorrhagic fevers and severe neurological disorders in humans. Although muroid rodents have historically been recognized as the primary reservoirs for major pathogens like Lassa virus, recent surveillance has revealed a significant expansion of their host range. This review aims to synthesize current global data regarding the epidemiology of mammarenaviruses in conventional reservoirs and the emergence of novel arenaviruses in non-traditional mammalian hosts. To achieve this, we comprehensively analyzed recent molecular and metagenomic surveillance data, evolutionary studies, and epidemiological reports published worldwide. Key discoveries include Wenzhou virus in Asian house shrews, Plateau Pika virus in plateau pikas, and an independent, geographically clustered of hedgehog-associated arenaviruses across Europe. Ultimately, this review underscores the global distribution of these pathogens and the critical need for continued, multi-host surveillance worldwide.

The Apennine wolf (Canis lupus italicus) is a distinct subspecies whose ongoing population recovery in Italy has progressively increased the demand for live capture protocols validated for scientific monitoring and conservation management. Despite the widespread use of mechanical and chemical immobilization in European wolf management, no study has to date systematically evaluated the combined use of the Fremont™ humane foot snare with a medetomidine-ketamine-acepromazine (MKA) protocol in this subspecies, nor characterized the associated cardiorespiratory, thermal, and hematobiochemical parameters under operational field conditions. Between June 2010 and July 2017, thirteen free-ranging Apennine wolves were captured in Maiella National Park (central Apennines, Italy) using the Fremont™ snare and immobilized with a standardized MKA protocol; only animals immobilized with this protocol are reported here, as three additional capture events employed different drug combinations. Cardiorespiratory parameters, body temperature, peripheral oxygen saturation, venous blood gas values, and a comprehensive hematological and serum biochemical panel were recorded during immobilization. Mean heart rate was 100 ± 15 bpm, respiratory rate 24 ± 13 breaths/min, body temperature 38.1 ± 1.3 °C, and mean SpO₂ 88 ± 11% (range: 66–97%; n = 12). No clinically significant hyperthermia requiring active intervention was recorded in the cohort as a whole. Hematological and biochemical values were broadly consistent with published reference ranges for the species, with condition-specific deviations identified in two individuals—one pregnant female and one juvenile presenting signs of transient capture-related myopathy—both of which resolved without clinical sequelae. No capture-related mortality occurred. All thirteen individuals survived the minimum post-capture monitoring period. Preliminary GPS observations in a subset of individuals (n = 3) suggest a transient reduction in movement activity in the immediate post-release period. These findings support the safety and operational feasibility of the combined Fremont™ snare–MKA protocol for the Apennine wolf, and provide baseline physiological and hematobiochemical reference data for Canis lupus italicus relevant to future capture and conservation management programmes.

The Apennine wolf (Canis lupus italicus) is a distinct subspecies whose ongoing population recovery in Italy has progressively increased the demand for live capture protocols validated for scientific monitoring and conservation management. Despite the widespread use of mechanical and chemical immobilization in European wolf management, no study has to date systematically evaluated the integrated combina-tion of a humane mechanical restraint system and a structured chemical immobiliza-tion protocol — and specifically the association of the Fremont™ humane foot snare with a medetomidine-ketamine-acepromazine (MKA) protocol, in terms of their joint physiological effects and welfare implications for this subspecies under operational field conditions. Between June 2010 and July 2017, thirteen free-ranging Apennine wolves were captured in Maiella National Park (central Apennines, Italy) using the Fremont™ snare and immobilized with a standardized MKA protocol. Cardiorespira-tory parameters, body temperature, peripheral oxygen saturation, venous blood gas values, and a comprehensive hematological and serum biochemical panel were rec-orded during immobilization. Mean heart rate was 100 ± 15 bpm, respiratory rate 24 ± 13 breaths/min, and body temperature 38.1 ± 1.0°C. No clinically significant hyper-thermia was recorded in the cohort as a whole. Hematological and biochemical values were broadly consistent with published reference ranges for the species, with condi-tion-specific deviations identified in two individuals (one pregnant female and one ju-venile presenting signs of transient capture-related myopathy), both of which resolved without clinical sequelae. No capture-related mortality occurred. All thirteen individ-uals survived the minimum post-capture monitoring period, and preliminary GPS da-ta suggest a transient reduction in movement activity in the immediate post-release period. These findings support the welfare adequacy and operational feasibility of the combined Fremont™ snare–MKA protocol for the Apennine wolf, and provide base-line physiological and hematobiochemical reference data for Canis lupus italicus rele-vant to future capture and conservation management programmes.

The Apennine wolf (Canis lupus italicus) increasingly inhabits human-modified land-scapes, where exposure to anthropogenic environments may influence pathogen cir-culation and antimicrobial resistance (AMR). Despite this relevance, no shotgun met-agenomic data are available for this subspecies during rehabilitation. A juvenile male wolf admitted to a Wildlife Rehabilitation Center (WRC) after traumatic injury and treated with multiple antibiotics was sampled at admission (T0) and after 11 months of rehabilitation (T1). Shotgun metagenomic sequencing (Illumina NovaSeq) was used to characterize fecal microbial communities, potential pathogens, and antimicrobial resistance genes (ARGs) using Kraken2 and CARD-RGI. Bacterial diversity increased from T0 to T1. Microbial composition shifted from Enterobacterales-dominated pro-files to more diverse communities. Reads associated with animal and human patho-gens were detected at both time points, together with human-associated taxa and viral reads at T1. ARGs were abundant (444 at T0; 417 at T1), mainly involving efflux pumps and β-lactamases. Genes related to Highest Priority Critically Important Anti-microbials—including mcr variants, van clusters, and oxazolidinone resistance deter-minants—were identified. Shotgun metagenomics revealed marked microbiome changes and high ARG diversity in an Apennine wolf during rehabilitation. These findings highlight wolves as potential sentinels of environmental AMR and emphasize the importance of biosecurity measures in WRCs.

Bacterial antimicrobial resistance is a significant global threat to public health, closely linked to the misuse of antimicrobials in human and veterinary medicine, aquaculture, and agriculture. The consequences of antimicrobial resistance overcome species boundaries and require a holistic approach for mitigation actions. The study of antimicrobial resistance in wildlife is thus increasingly relevant to understand the spread of antimicrobial resistance in the environment and the animal community, as well as to investigate the role of wildlife either as a carrier, reservoir, spillover, or indicator of antimicrobial resistance. The aim of this study is to describe the prevalence and type of antimicrobial resistance in bacterial isolates from wild mammals through systematic review and meta-analysis of the available literature, following the Preferred Reporting Items for Systematic reviews and Meta-Analysis (PRISMA) guidelines. Out of 5052 collected documents, 3795 were screened, and finally 139 studies on antimicrobial resistance in free-ranging wild mammals were included in the meta-analysis. The studies covered 37 countries, mostly European. The Enterobacterales Escherichia coli and Salmonella spp., as well as Campylobacter spp., were the most frequently targeted bacterial species, mainly in the Artiodactyla order and specifically in the Suidae and Cervidae families. Low to moderate prevalences of antimicrobial resistance were found in all the continents, countries, bacteria, host taxa, and antimicrobials included in the meta-analysis, even for critically important antimicrobials as defined by the World Health Organisation, with higher values in Africa and Asia, in carnivores, and in animal species with high adaptability to diverse habitats. This meta-analysis showed that antimicrobial resistance in wild mammals is widespread and variable according to taxonomy, trophic source, and geographic location. The meta-analysis highlighted methodological gaps that need to be addressed to improve the interpretation and conclusions obtained from the data. Genetic analyses on antimicrobial resistance and population ecological data should be included in future analysis to achieve a standardised methodology and overcome current limitations. To date, wildlife appears to be an environmental indicator of antimicrobial resistance and should be included in antimicrobial resistance surveillance plans not only because this sentinel role but also to monitor potential spill-back to livestock and/or humans.

Sonoelastography is a rapid and non-invasive ultrasound-based technique to assess tissue elasticity. Interest in this technique’s application in veterinary medicine is progressively increasing, with preliminary reports also available to assess canine testicular parenchyma abnormalities. Since elastographic techniques are different and it is of paramount importance to standardize procedures, this work aimed to compare two different elastographic techniques, SE and 2D SWE, on the same testes and compare the data with the current literature. Thirty healthy canine testes were included, and complete B-mode, color and power Doppler examinations were performed on each one. SE was performed qualitatively through an elastogram evaluation and semiquantitatively comparing the parenchyma with peripheral skin. 2D SWE was performed and m/s measurements were taken. Each measurement was collected in the cranial, middle and caudal portions, considering the sections above and below the mediastinum separately. The healthy testes appeared to be intermediate in stiffness with differences in the various portions for SE and expressed a mean value from 1.3 m/s to 1.4 m/s for 2D SWE, consistent with the previous literature. Considering the technical specifics of the software for the two elastography methods, both of them are valuable in assessing canine testis stiffness and are potentially clinically appliable.

Rose Bengal antigen and smooth lipopolysaccharide (s-LPS) were produced from a field strain of Brucella ceti (“homologous” antigens) and from the reference strain B. abortus S99 (“heterologous” antigens); they are currently used for the diagnosis of brucellosis in cattle, water buffaloes, sheep, goats, and pigs, as recommended in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (WOAH). “Homologous” and “heterologous” antigens were used in a rapid serum agglutination test (Rose Bengal test, RBT) and a competitive ELISA assay (c-ELISA) to test a panel of sera, blood, and other body fluids (cerebrospinal fluid, pericardial fluid, tracheal fluid, and aqueous humor) collected from 71 individuals belonging to five cetacean species (Stenella coeruleoalba; Tursiops truncatus; Grampus griseus; Globicephala melas; and Ziphius cavirostris), which were found stranded on the Italian coastline. Six animals were positive for Brucella spp. for bacterial isolation and/or PCR, and 55 animals were negative; for the remaining 10 animals, no PCR/isolation data were available. A total of 90 samples were tested. Results obtained from the two tests were compared in order to identify the most suitable antigen for the serological diagnosis of Brucella infection in cetaceans. The RBT performed with the “homologous” antigen showed the best results in comparison with the “heterologous” antigen: diagnostic sensitivity, specificity, and accuracy were 80.0%, 44.1%, and 46.9% for the “homologous” antigen and 80.0%, 17.0%, and 21.9% for the “heterologous” antigen. For the c-ELISA, “homologous” and “heterologous” s-LPS showed similar results (diagnostic sensitivity 66.7%, diagnostic specificity 97.3%, and diagnostic accuracy 95.0%). Therefore, the RBT using the “homologous” antigen and c-ELISA with “homologous” or “heterologous” s-LPS could be used in parallel for the detection of antibodies against Brucella spp. in cetaceans.

Strain elastography (SE) evaluates tissue stiffness, providing qualitative and semiquantitative evaluation, with a strain ratio (SR) > 1 indicating that the target lesion is stiffer than the reference tissue. The patellar tendon has been described as soft in healthy dogs and hard in dogs with cranial cruciate ligament rupture, but SR usefulness has not been assessed. Dogs were divided into healthy (G1) and at least 1 month after surgery (G2) groups. Thickness was recorded, and a B-mode score of 0–3 was determined based on the abnormality’s severity. SE was qualitatively performed with a score of 1–4, and EI was recorded on the whole ligament and in proximal, intermediate and distal areas; SR was collected with the fat pad and cutis/subcutis. G1 was significantly thinner, with a lower score than G2 and a more elastic pattern. EI for G1 was significantly lower than G2 for the whole tendon and the single portions, and EI for the distal portion was significantly lower than the proximal and intermediate one in G2. SR was 1 in G2 for both the fat pad and cutis/subcutis. These data suggest that after surgery, the tendon may become non-uniformly stiffer. The cutis/subcutis appears to be a more reliable reference tissue than the infrapatellar fat pad.

In this study, the antimicrobial resistance profiles of bacterial strains obtained from wild avian species recovered in wetlands of Northern Italy were described. Cloacal swabs collected from 67 aquatic birds, hunted or found dead in two private hunting grounds, were submitted to microbiological investigations and antimicrobial susceptibility testing using the Vitek 2 system, while specific PCR protocols were applied to screen for genes associated with the resistance. One hundred fifty‐seven bacterial strains were characterized. The most frequent isolates were Enterococcus faecalis (36/157; 22.9%) and Escherichia coli (23/157; 14.6%). Seventy‐seven isolates (77/157; 49%) were resulted resistant to at least one antibiotic, and eight isolates (8/157; 5%) were classified as multidrug resistant bacteria. Resistance for critically important antibiotics (linezolid, vancomycin, carbapenems, third‐generation cephalosporins, and fluoroquinolones) was also described. Salmonella spp. was obtained from a Eurasian teal (Anas crecca), and it was subsequently analyzed by whole genome sequencing, revealing the serovar Salmonella Braenderup ST22. The phylogenetic analysis, performed with all ST22 described in 2021 and 2022, placed the strain under study in a large clade associated with human salmonellosis cases. These results suggest that migratory aquatic birds may be considered as relevant carriers of critically important antibiotic resistant bacteria and zoonotic food‐borne pathogens potentially able to impact public health.

In this study, a culture-independent approach was applied to compare the microbiome composition and the abundance of the antimicrobial resistance genes (ARGs) aadA2 for aminoglycosides, tet(A), tet(B), tet(K), and tet(M) for tetracyclines, and mcr-1 for colistin in broiler litter samples collected from conventional and antibiotic-free flocks located in Central Italy. A total of 13 flocks and 26 litter samples, collected at the beginning and at the end of each rearing cycle, were submitted to 16s rRNA sequence analysis and quantitative PCR for targeted ARGs. Firmicutes resulted in the dominant phylum in both groups of flocks, and within it, the Clostridia and Bacilli classes showed a similar distribution. Conversely, in antibiotic-free flocks, a higher frequency of Actinobacteria class and Clostridiaceae, Lactobacillaceae, Corynebacteriaceae families were reported, while in the conventional group, routinely treated with antibiotics for therapeutic purposes, the Bacteroidia class and the Enterobacteriaceae and Bacillaceae families were predominant. All investigated samples were found to be positive for at least one ARG, with the mean values of aadA2 and tet(A) the highest in conventional flocks by a significant margin. The results suggest that antibiotic use can influence the frequency of resistance determinants and the microbial community in poultry flocks, even though other environmental factors should also be investigated more deeply in order to identify additional drivers of antimicrobial resistance.