Salmonella enterica serovar Choleraesuis is the aetiological agent of swine paratyphoid being a highly invasive zoonotic pathogen. Wild boar natural populations are experiencing a demographical expansion as well as some farms are breeding this species to release for hunting with management sometimes identical to that of domestic pigs, including supplementation, grouping, and antibiotic treatments. This situation increases the chance of contact between wild boars and livestock, and potentially induces stress, with different sanitary consequences. The present work aims to describe the clinical features of recent outbreaks caused by S. Choleraesuis in wild boar from central-western Spain, as well as the antimicrobial resistance and phylogenetic relationships of isolates involved. 28 strains of S. Choleraesuis were isolated from 28 different wild boars belonging to 10 different game states located in central western Spain and submitted to the Clinical Veterinary Hospital (CVH) of the University of Extremadura. Samples were taken from different organs and cultured according to the ISO 6579:2002 procedure. Suspicious colonies were identified by PCR and antimicrobial resistance was evaluated by disc diffusion susceptibility test and the presence of the main resistance genes as well as 18 plasmid replicons frequently found among the Enterobacteriaceae was verified by PCR. Pulsed field gel electrophoresis was applied to determine the genetic relationship between isolates. The outbreaks under study were characterized by high mortality (35%-84%) and a septicaemic presentation. S. Choleraesuis was isolated from all the wild boars analysed, and 26 of the 28 isolates presented resistance to at least one antibiotic. The predominant resistances found were against sulphonamide, streptomycin, tetracycline, and doxicicline and sul1, strA-strB, and tetA were the most prevalent resistance genes among isolates. 10 strains carried FIIA, FIB+H/1 or FIIA+H/1 plasmids. PFGE classified the isolates into four different profiles, grouped into two clusters. This results show that prevention against S. Choleraesuis must be considered in the sanitary programs of the wild boar breeders.

The aim of this study was to investigate the presence of methicillin-resistant Staphylococcus (MRS) strains in non-managed wild ungulates present in a typical Mediterranean forest in Spain. For this purpose, nasal swabs were obtained from 139 animals: 90 wild boar (Sus scrofa), 42 red deer (Cervus elaphus) and 7 fallow deer (Dama dama), which were subsequently pre-enriched in BHI+ NaCl (6.5%) (24 h/37 °C), and then seeded in Columbia blood agar (24 h/37 °C)). The presence of the mecA gene was investigated by PCR, first from the confluent and then from individual colonies. A total of 10 mecA+ colonies were obtained of which only seven showed phenotypic resistance to oxacillin/cefoxitin (methicillin resistance). All MRS strains belonged to the Staphylococcus sciuri group. Methicillin-resistant Staphylococcus aureus (MRSA) was not detected. In addition, a significant number of MRS strains showed resistance to other antimicrobials, mainly β-lactam (7/7), gentamicin (7/7), fusidic acid (6/7) and quinupristin–dalfopristin (6/7), showing an irregular correlation with their coding genes. The genetic profiles grouped the seven strains obtained according to the bacterial species but not in relation to the animal source or the geographical place of origin. The presence of SCCmec type III, common to animals and humans, has been detected in three of the strains obtained. In conclusion, the study reveals that the wild ungulates investigated play a role as potential reservoirs of multi-resistant strains of MRS. Such strains, due to their characteristics, can be easily transferred to other wild or domestic animal species and ultimately to humans through their products.

Summary The importance of wild boars as game species in Spain is well known. Their feeding habits and intrusive behaviour, together with a progressively wider spreading of populations, increases the interactions of these animals with livestock and humans. Considering that wild boars could have a potential role in the transmission of certain pathogens as salmonellae, the aims of this study were to determine the prevalence of Salmonella spp. in wild boars hunted in central‐western Spain, the occurrence of this pathogen in tonsils, mandibular lymph nodes and faeces (as markers for transmission risk), and to define the phylogenetic relationships among isolated strains, in order to investigate the circulation pathways of bacteria among tissues, animals and estates. Samples from 1,041 hunted wild boars were analysed for the presence of Salmonella spp. by bacteriological culture. Isolates were confirmed by PCR and serotyped in the Spanish national reference laboratory. The genetic relationships between strains were determined by PFGE. The results showed a 7.7% of positive animals (81 wild boars), being tonsils the organ most frequently colonised by Salmonella spp. (18.7%), followed by lymph nodes (5.1%) and faecal samples (2.9%). Serovars Enteritidis and Newport were the most frequent amongst the 34 different serovars obtained. The pulsed‐field gel electrophoresis (PGFE) analysis showed a great genetic diversity, with serovars that exhibited different pulsotypes when isolated from different estates and multiple serovars in the same estate. In conclusion, this study reveals the importance of wild boars as carriers and possible transmitters of virulent and/or antimicrobial‐resistant clones of Salmonella spp. to livestock and humans.

The restoration of Iberian lynx (Lynx pardinus) populations in Extremadura (Southwestern Spain) have been carried out since 2014. One of the measures to ensure the success of this program is to examine the effects that diseases may have on reintroduction. Since diseases may be greatly located at certain sites because of the specific ecological requirements of the pathogens and/or vectors, reintroduced individuals may present a risk of infection once released. To determine which pathogens the reintroduced individuals may encounter, we performed a molecular and sero-epidemiological survey in reintroduced and wild-born lynxes. From 2015 to 2019, 69 Iberian lynxes (40 reintroduced and 29 wild-born) were sampled and screened against 10 viral, bacterial and piroplasmid agents. In parallel, 195 sympatric carnivores from the families Canidae, Felidae, Viverridae, Herpestidae and Mustelidae were tested against current or past infections to six common canine/feline viruses. In the Iberian lynx, low contact rates of active infection were obtained for the feline leukemia provirus (FeLV: 1.5%; 1/67), feline parvovirus (FPV: 1.5%; 1/67) and Cytauxzoon sp. (6.7%; 1/15). We also confirmed the emergence of Aujeszky’s disease (suid herpesvirus-1) in this population (SuHV-1: 11.8%; 2/17). Evidence of previous exposure was detected for canine distemper virus (CDV: 5.8%; 3/52), feline coronavirus (1.9%; 1/52), FPV (7.7%; 1/13) and feline calicivirus (FCV: 5.3%; 1/19). From 25 recovered lynx carcasses, we could confirm infectious etiology involvement in the death of four individuals (SuHV-1 in two individuals, coinfection of Cytauxzoon spp. and Aeromonas veronii in one lynx and a Streptococcus canis myositis in another lynx). We confirmed the circulation of CDV, FPV, FeLV, FCV and the feline immunodeficiency virus within the sympatric carnivore community. Due to the low contact rate of infectious agents in such a small, endangered population, we recommend continuing a disease surveillance program to determine the prognostic factors of survival, understand the role that disease may play during the reintroduction and anticipate disease outbreaks that may pose a risk for the entire reintroduced population.

The Salmonellaenterica serovar Choleraesuis affects domestic pig and wild boar (WB), causing clinical salmonellosis. Iberian swine production is based on a free-range production system where WB and Iberian pig (IP) share ecosystems. This study focuses on the negative impact on the pork industry of infections due to this serotype, its role in the spread of antibiotic resistance, and its zoonotic potential. Antibiotic resistance (AR) and genetic relationships were analyzed among 20 strains of S. Choleraesuis isolated from diseased WB and IP sampled in the southwest region of the Iberian Peninsula. AR was studied using the Kirby–Bauer method with the exception of colistin resistance, which was measured using the broth microdilution reference method. Resistance and Class 1 integrase genes were measured using PCR, and the genetic relationship between isolates and plasmid content by pulsed field gel electrophoresis. The results show a higher incidence of AR in isolates from IP. Phylogenetic analysis revealed seven profiles with two groups containing isolates from IP and WB, which indicates circulation of the same clone between species. Most pulsotypes presented with one plasmid of the same size, indicating vertical transmission. AR determinants blaTEM and tetA were routinely found in IP and WB, respectively. One isolate from IP expressed colistin resistance and presented the mcr-1 gene carried by a plasmid. This study suggests that S. Choleraesuis circulates between WB and IP living in proximity, and also that the mobilization of AR genes by plasmids is low. Furthermore, the detection of plasmid-mediated colistin resistance in bacteria from IP is alarming and should be monitored.

Haemophilus parasuis is a recognized pathogen in domestic pigs; the pathogen has been also isolated from healthy wild boar (Sus scrofa). In the current report, a case of fatal H. parasuis infection in a wild boar piglet from central Spain is described. The affected animal presented severe pneumonic lesions, inflammation in tarsal joints with presence of fibrinous deposits, and epidural hemorrhage in the atlanto-occipital joint. Pure growth of H. parasuis was obtained from lungs and tarsal joints. The current case illustrates the susceptibility of wild boar to this agent. The gross pathology results were similar to that described in domestic pigs, but there were no fibrinous deposits on serosal surfaces.

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The aim of this study was to characterize methicillin-resistant coagulase-negative staphylococci (MRCoNS) isolates from the healthy staff of a university veterinary hospital in order to assess their importance as a reservoir of antimicrobial resistance and to determine their population structure and evolution. The study duration was over two years (2020–2021), 94 individuals were analyzed in duplicate, and 78 strains were obtained. The overall prevalence of methicillin-resistant strains detected throughout the study was 61.7%, with point prevalence values of 53.2% in 2020 and 31.5% in 2021. A total of 19.1% of the individuals analyzed were carriers throughout the study. The most frequently identified MRCoNs were Staphylococcus epidermidis (92.3%) and S. warneri (3.8%). A total of 75.6% of the isolates obtained showed the development of multi-resistance, preferentially against erythromycin, gentamicin, and tetracycline, and to a lesser extent against fusidic acid, norfloxacin, and clindamycin; these antimicrobials are frequently used in the veterinary field. Although most of the S. epidermidis isolates obtained showed wide genetic variability and low dispersion, which are characteristic of community-associated isolates, a small number of strains spread between individuals in close physical proximity and were maintained over time, forming stable clones. These clones generally maintained the same type of staphylococcal cassette chromosome mec (SCCmec) and had a similar antimicrobial resistance pattern.