Individual antibody titres from the analyzed goats from VAPS against four pestivirus strains.

<p>Antibody titres of 35 goats from VAPS against one <i>Bovine viral diarrhoea virus</i> (BVDV) strain of type 1 (BVDV-NADL), two <i>Border disease virus</i> (BDV) strains of type 4 (BDV-Esp97 and -Aran-1) and one BDV strain of type 1 (BDV-137/4). Neutralizing antibody titres are expressed as the reciprocal of the highest dilution that neutralized 100 tissue culture infective doses (100 TCID₅₀) in all cultures.</p

Prevalence of pestivirus antibodies in the analyzed species.

<p>Prevalence and 95% confidence interval (exact 95% binomial confidence intervals) of the pestivirus NS3 antibodies in all the analyzed species. In the Pyrenean chamois prevalence is classified according to hunting season.</p><p>CI = Confidence interval.</p><p>n = number of tested animals.</p><p>ND = not done.</p><p>VAPS = Val d’Aran and Pallars Sobirà regions.</p><p>CAUBS = Cerdanya, Alt Urgell and Berguedà regions.</p

Individual antibody titres from the analyzed red, fallow and roe deer and mouflon against five pestivirus strains.

<p>Individual antibody titres of positive red, fallow and roe deer and mouflon against one <i>Bovine viral diarrhoea virus</i> (BVDV) strain of type 1 (BVDV-NADL), three <i>Border disease virus</i> (BDV) strains of type 4 (BDV-Esp97, -Aran-1 and -Cadí-6) and one BDV strain of type 1 (BDV-137/4). Neutralizing antibody titres are expressed as the reciprocal of the highest dilution that neutralized 100 tissue culture infective doses (100 TCID₅₀) in all cultures.</p>a<p>Sera from VAPS were tested against BDV-4 strain Aran, while sera from CAUBS were tested against BDV-4 strain Cadí-6.</p><p>VAPS = Val d’Aran and Pallars Sobirà regions.</p><p>CAUBS = Cerdanya, Alt Urgell and Berguedà regions.</p

Significant differences between titres against different strains in sheep, goat, cattle and Pyrenean chamois in the two study areas.

<p>Significant differences between pairwise comparisons in titres against five pestivirus strains (one <i>Bovine viral diarrhoea virus</i> (BVDV) strain of type 1 (BVDV-NADL), three <i>Border disease virus</i> (BDV) strains of type 4 (BDV-Esp97, -Aran-1 and -Cadí-6) and one BDV strain of type 1 (BDV-137/4)) in sheep, goat, cattle and Pyrenean chamois in the two study areas.</p><p>S = Sheep.</p><p>G = Goat.</p><p>Ca = Cattle.</p><p>Ch = Chamois.</p>a<p>Sera from VAPS were tested against BDV-4-Aran-1, while sera from CAUBS were tested against BDV-4-Cadí-6.</p

Distribution of wild ruminant samples by year and species.

<p>n = number of tested animals.</p><p>VAPS = Val d’Aran and Pallars Sobirà regions.</p><p>CAUBS = Cerdanya, Alt Urgell and Berguedà regions.</p

Unrooted neighbour-joining phylogenetic tree based on the 5′UTR sequence among pestiviruses.

<p>The strains detected in six chamois (ARAN-9, -10, -11.-12, -13, and -14) showed genetic identity and subsequently the analysis was performed only with one sequence (ARAN-9). The detected strains cluster with other chamois viruses isolated in the bordering NHR of Alt Pallars-Aran (ARAN-1, and -2). The numbers on the branches indicate the bootstrap values (in percentage; 1,000 replicates). Sequences of strains were taken from Gen Bank with following accession numbers: ARAN-9 (HE818617), ARAN-10 (HE818618), Aran-11 (HE818619), ARAN-12 (HE818620), ARAN-13 (HE818621), ARAN-14 (HE818622), ARAN-1 (AM765800), ARAN-2 (AM765801), Chamois-Spain02 (AY641529), FRESER 2 (FN691777), ORLUR36 (DQ898294), CERDANYA-2 (AM905931), CERDANYA-1 (AM905930), CADI-6 (AM905923), CADI-1 (AM905918), BU1-C3 (DQ361068), Espana-97 -1 (FR714860), AV (EF693984), 137/4 (U65052), Moredun (U65023), CASIMIR (AB122085), BVDV NADL (M31182). The sequence of strain FRESER-1 is not deposited in the GenBank. The scale bar represents 0.05 substitutions per nucleotide site.</p

Accession numbers, references, localities and collection years of domestic ungulates BDV sequences included in the data set.

<p>Accession numbers, references, localities and collection years of domestic ungulates BDV sequences included in the data set.</p

The inferred spatiotemporal dynamics of BDV in Pyrenean chamois.

<p>The figure summarize the most significant migration links in the interested Pyrenean area. The putative root of BDV strains is highlighted with a orange circle. More detailed results are reported in supplementary panels (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0168232#pone.0168232.s002" target="_blank">S2 Fig</a>) and at <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0168232#pone.0168232.s003" target="_blank">S1 Video</a>.</p

Time of the most common ancestor estimates of Pyrenean chamois BDV, credibility interval (95% HPD) of the main clades observed in the MCC tree, with the corresponding most probable locations, and state posterior probability.

<p>Time of the most common ancestor estimates of Pyrenean chamois BDV, credibility interval (95% HPD) of the main clades observed in the MCC tree, with the corresponding most probable locations, and state posterior probability.</p

The maximum clade credibility tree of BDV 5’ UTR from Pyrenean chamois.

<p>The branches are coloured on the basis of the most probable location of the descendent nodes (Alt Pallars = APA, Alta Ribagorça = ARI, Andorra = AND, Aran = ARA, Cadí = CAD, Cerdanya-Alt Urgell = CER, Freser-Setcases = FRE). The numbers on the internal nodes indicate significant posterior probabilities (pp>0.8), and the scale at the bottom of the tree represents the number of years before the last sampling time (2011). The main geographical clades corresponding to Western, Eastern, and Central Pyrenean areas have been highlighted.</p