Amplicon sizes were estimated by electrophoresis on a 1.5% agarose gel at 45 V during 2 h, using 100-bp ladder (Biotools B&M). Figure 2 presents the spoligotyping patterns, VNTR allelic profiles and typing Cyclosporin A in vitro pattern (TP) codes defined for this study. Figure 2 Spoligotyping patterns, VNTR allelic variants, and codes used to define typing patterns (TPs) in this study. 1) VNTR allelic variants for MIRU10 were always 2, for MIRU16 always 3, for MIRU23 always 4, for MIRU26 always 5, for MIRU31 always 3 and for MIRU40 always 2. 2) Isolates with TP codes A4, G1, G6, H1 and I4 as in Romero et al. (2008). Statistics Chi-square tests were used for between-pair comparisons of prevalences. To test for the effect of

host species vs site regarding the mycobacterial isolates, we used the Czechanovsky similarity index [44]. This index AZD1480 in vitro considers the list of mycobacterial

species recorded in a given host type or in a given study area. It is calculated by dividing two times the species Omipalisib cell line shared between two lists, by the total number of species of both lists, as follows: Considering the animals in which any mycobacterial infection was diagnosed, three generalized linear mixed models (GLMM, SAS 9.0 software, GLIMMIX procedure) were explored to test different explanatory variables that affect the presence of a mycobacterial type or group. The most common mycobacterial groups were: (i) M. bovis (ii) M bovis A1 and (iii) M. scrofulaceum. The presence or absence of infection in a mycobacterial group was considered as a binary variable. The model was fitted using a logit link function. The model considered social group as a random effect. The model included enough host species (wild boar, fallow deer and red deer), the study area and age (juvenile: less than 2 years, adult: older than 2 years) as categorical explanatory variables. The distance to the water (log10-trasnformed) was included as a continuous predictor. To compare the spatial associations

of infection by specific mycobacterial type and hosts, we included as explanatory continuous variable the ratio (log10-transformed) between the nearest neighbor distance from host to a different host species with the same type of mycobacteria relative to the nearest distance to a con-specific host with the same type of mycobacteria (calculated using ArcGis version 9.2, ESRI, Redlands, CA). A ratio >1 indicates that the nearest distance to a host with the same spoligotype is higher for a different host species. All the aforementioned explanatory variables we also included in the models interacting with the host species. Due to over-parameterization of the models and zero inflated data, no interactions were included in the M. bovis A1 and M. scrofulaceum models. P-value was set as ≤ 0.05. We estimated exact confidence limits for prevalence (proportions) using Sterne’s exact method. Results Mycobacteria species and molecular types We obtained a total of 154 mycobacterial isolates from DNP wildlife.