Mycobacterium bovis populations in countries with persistent bovine tuberculosis usually show a prevalent spoligotype with a wide geographical distribution. This study applied mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) typing to a random panel of 115 M. bovis isolates that are representative of the most frequent spoligotype in the Iberian Peninsula, SB0121. VNTR typing targeted nine loci: ETR-A (alias VNTR2165), ETR-B (VNTR2461), ETR-D (MIRU4, VNTR580), ETR-E (MIRU31, VNTR3192), MIRU26 (VNTR2996), QUB11a (VNTR2163a), QUB11b (VNTR2163b), QUB26 (VNTR4052), and QUB3232 (VNTR3232). We found a high degree of diversity among the studied isolates (discriminatory index [D] = 0.9856), which were split into 65 different MIRU-VNTR types. An alternative short-format MIRU-VNTR typing targeting only the four loci with the highest variability values was found to offer an equivalent discriminatory index. Minimum spanning trees using the MIRU-VNTR data showed the hypothetical evolution of an apparent clonal group. MIRU-VNTR analysis was also applied to the isolates of 176 animals from 15 farms infected by M. bovis SB0121; in 10 farms, the analysis revealed the coexistence of two to five different MIRU types differing in one to six loci, which highlights the frequency of undetected heterogeneity.

Wildlife vaccination is increasingly being considered as an option for tuberculosis control. We combined data from laboratory trials and an ongoing field trial to assess the risk of an oralMycobacterium bovisBCG vaccine and a prototype heat-inactivatedMycobacterium bovispreparation for Eurasian wild boar (Sus scrofa). We studied adverse reactions, BCG survival, BCG excretion, and bait uptake by nontarget species. No adverse reactions were observed after administration of BCG (n= 27) or inactivatedM. bovis(n= 21). BCG was not found at necropsy (175 to 300 days postvaccination [n= 27]). No BCG excretion was detected in fecal samples (n= 162) or in urine or nasal, oral, or fecal swab samples at 258 days postvaccination (n= 29). In the field, we found no evidence of loss of BCG viability in baits collected after 36 h (temperature range, 11°C to 41°C). Camera trapping showed that wild boar (39%) and birds (56%) were the most frequent visitors to bait stations (selective feeders). Wild boar activity patterns were nocturnal, while diurnal activities were recorded for all bird species. We found large proportions of chewed capsules (29%) (likely ingestion of the vaccine) and lost baits (39%) (presumably consumed), and the proportion of chewed capsules showed a positive correlation with the presence of wild boar. Both results suggest proper bait consumption (68%). These results indicate that BCG vaccination in wild boar is safe and that, while bait consumption by other species is possible, this can be minimized by using selective cages and strict timing of bait deployment.

The presence of AMR bacteria in the human–animal–environmental interface is a clear example of the One Health medicine. Several studies evidence the presence of resistant bacteria in wildlife, which can be used as a good indicator of anthropization level on the ecosystem. The fast increase in AMR in the environment in the last decade has been led by several factors as globalization and migration. Migratory birds can travel hundreds of kilometers and disseminate pathogens and AMR through different regions or even continents. The aim of this study was to compare the level of AMR in three migratory bird species: Ciconia ciconia, Larus fuscus and Chroicocephalus ridibundus. For this purpose, commensal Escherichia coli has been considered a useful indicator for AMR studies. After E. coli isolation from individual cloacal swabs, antimicrobial susceptibility tests were performed by the disk-diffusion method, including 17 different antibiotics. A total of 63.2% of gulls had resistant strains, in contrast to 31.6% of white storks. Out of all the resistant strains, 38.9% were considered multi-drug resistant (50% of white storks and 30% of seagulls). The antibiotic classes with the highest rate of AMR were betalactamics, quinolones and tetracyclines, the most commonly used antibiotic in human and veterinary medicine in Spain.