Spatial and temporal phylogeny of border disease virus in pyrenean chamois (Rupicapra p. Pyrenaica)

Border disease virus (BDV) affects a wide range of ruminants worldwide, mainly domestic sheep and goat. Since 2001 several outbreaks of disease associated to BDV infection have been described in Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in Spain, France and Andorra. In order to reconstruct the most probable places of origin and pathways of dispersion of BDV among Pyrenean chamois, a phylogenetic analysis of 95 BDV 5'untranslated sequences has been performed on chamois and domestic ungulates, including novel sequences and retrieved from public databases, using a Bayesian Markov Chain Monte Carlo method. Discrete and continuous space phylogeography have been applied on chamois sequences dataset, using centroid positions and latitude and longitude coordinates of the animals, respectively. The estimated mean evolutionary rate of BDV sequences was 2.9x10(-3) subs/site/year (95% HPD: 1.5-4.6x10(-3)). All the Pyrenean chamois isolates clustered in a unique highly significant clade, that originated from BDV-4a ovine clade. The introduction from sheep (dated back to the early 90s) generated a founder effect on the chamois population and the most probable place of origin of Pyrenean chamois BDV was estimated at coordinates 42.42 N and 1.9 E. The pathways of virus dispersion showed two main routes: the first started on the early 90s of the past century with a westward direction and the second arise in Central Pyrenees. The virus spread westward for more than 125 km and southward for about 50km and the estimated epidemic diffusion rate was about 13.1 km/year (95% HPD 5.2-21.4 km/year). The strong spatial structure, with strains from a single locality segregating together in homogeneous groups, and the significant pathways of viral dispersion among the areas, allowed to reconstruct both events of infection in a single area and of migrations, occurring between neighboring areas

Two different epidemiological scenarios of border disease in the populations of Pyrenean chamois (Rupicapra p. pyrenaica) after the first disease outbreaks

Since 2001 several outbreaks of a new disease associated with Border disease virus (BDV) infection have caused important declines in Pyrenean chamois (Rupicapra pyrenaica) populations in the Pyrenees. The goal of this study was to analyze the post-outbreak BDV epidemiology in the first two areas affected by disease with the aim to establish if the infection has become endemic. We also investigated if BDV infected wild and domestic ruminants sharing habitat with chamois. Unexpectedly, we found different epidemiological scenarios in each population. Since the disease outbreaks, some chamois populations recuperated quickly, while others did not recover as expected. In chamois from the first areas, prevalence was high (73.47%) and constant throughout the whole study period and did not differ between chamois born before and after the BDV outbreak; in all, BDV was detected by RT-PCR in six chamois. In the other areas, prevalence was lower (52.79%) and decreased during the study period; as well, prevalence was significantly lower in chamois born after the disease outbreak. No BDV were detected in this population. A comparative virus neutralisation test performed with four BDV strains and one Bovine viral diarrhoea virus (BVDV) strain showed that all the chamois had BDV-specific antibodies. Pestivirus antibodies were detected in all the rest of analyzed species, with low prevalence values in wild ruminants and moderate values in domestic ruminants. No viruses were detected in these species. These results confirm the hypothesis that outbreaks of BDV infection only affect the Pyrenean chamois, although other wild ruminants can occasionally be infected. In conclusion, two different scenarios have appeared since the first border disease outbreaks in Pyrenean chamois: on the one hand frequent BDV circulation with possible negative impact on population dynamics in some areas and on the other, lack of virus circulation and quick recovery of the chamois population.The PhD studies of LFS are funded by a University Teaching Grant (FPU: Formación de Profesorado Universitario) from the Spanish Ministry of Education (Ministerio de Educación) and ES by the Beatriu de Pinós programme (BP-DGR 2011) of the Catalan Science and Technology System. This research was supported by grant CGL2006-11518/BOS and CGL2009-09071/BOS from the Spanish government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Enhanced fatty acid oxidation through metformin and baicalin as therapy for COVID-19 and associated inflammatory states in lung and kidney

Progressive respiratory failure is the primary cause of death in the coronavirus disease 2019 (COVID-19) pandemic. It is the final outcome of the acute respiratory distress syndrome (ARDS), characterized by an initial exacerbated inflammatory response, metabolic derangement and ultimate tissue scarring. A positive balance of cellular energy may result crucial for the recovery of clinical COVID-19. Hence, we asked if two key pathways involved in cellular energy generation, AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) signaling and fatty acid oxidation (FAO) could be beneficial. We tested the drugs metformin (AMPK activator) and baicalin (CPT1A activator) in different experimental models mimicking COVID-19 associated inflammation in lung and kidney. We also studied two different cohorts of COVID-19 patients that had been previously treated with metformin. These drugs ameliorated lung damage in an ARDS animal model, while activation of AMPK/ACC signaling increased mitochondrial function and decreased TGF-β-induced fibrosis, apoptosis and inflammation markers in lung epithelial cells. Similar results were observed with two indole derivatives, IND6 and IND8 with AMPK activating capacity. Consistently, a reduced time of hospitalization and need of intensive care was observed in COVID-19 patients previously exposed to metformin. Baicalin also mitigated the activation of pro-inflammatory bone marrow-derived macrophages (BMDMs) and reduced kidney fibrosis in two animal models of kidney injury, another key target of COVID-19. In human epithelial lung and kidney cells, both drugs improved mitochondrial function and prevented TGF-β-induced renal epithelial cell dedifferentiation. Our results support that favoring cellular energy production through enhanced FAO may prove useful in the prevention of COVID-19-induced lung and renal damage.This work was supported by grants PID2019-104233RB-I00 and PID2022-136703OB-I00 funded by MCIN /AEI/10.13039/501100011033 / FEDER, Spanish National Research Council CSIC–COV19-096/PIE202020E160, Comunidad de Madrid “NOVELREN” B2017/BMD-3751 and Fundación Renal “Íñigo Alvarez de Toledo”, all from Spain. The CBMSO receives institutional support from Fundación “Ramón Areces”. VM and EA hold a Juan de la Cierva-Formación fellowship from the Spanish Ministry of Science and Innovation (FJC2020-043505-I and FJC2021-046602-I, respectively). VM was supported by a Postdoctoral Long-Term Fellowship from the Federation of European Biochemical Societies (FEBS). CR and BS are recipients of a pre-doctoral fellowship of the FPI Program (BES-2016-076735 and PRE-2020-093831, respectively) from the Spanish Research State Agenc

Epidemiology of pestivirus infection in pyrenean Chamois (rupicapra pyrenaica pyrenaica) and other wild and domestic ruminants

Des de l’any 2001, diferents brots d’una malaltia associada a la infecció per un pestivirus, concretament un virus de la malaltia de la frontera de genotip 4 (BDV-4, de l’anglès border disease virus), han causat reduccions importants en el nombre d’efectius d’isard (Rupicapra pyrenaica pyrenaica). L’objectiu principal d’aquesta tesi doctoral va consistir en estudiar l’epidemiologia de la infecció per BDV a l’isard i a altres remugants salvatges i domèstics, ja que els pestivirus no són hoste-específics estrictes. La principal zona d’estudi va consistir en els Pirineus Catalans i Andorrans, però també es van estudiar diferents zones dels Ports de Tortosa i Beseit i Sierra Nevada, a Espanya, i dels Alps italians i suïssos. Les espècies estudiades van ser l’isard, el mufló europeu (Ovis aries), el cérvol (Cervus elaphus), el cabirol (Capreolus capreolus), la daina (Dama dama), l’isard alpí (Rupicapra rupicapra), l’íbex (Capra ibex) i la cabra salvatge (Capra pyrenaica), així com l’oví, caprí i vaquí que pasturen en els prats d’alta muntanya. Es va determinar la presència d’anticossos enfront a pestivirus en sèrum mitjançant una tècnica d’ELISA. En els sèrums positius es va dur a terme un test de seroneutralització vírica comparada amb una soca del virus de la diarrea vírica bovina (BVDV, de l’anglès bovine viral diaorrhea virus) i diferents soques de BDV locals i de referència. La detecció vírica es va realitzar mitjançant una tècnica de reacció en cadena de la polimerasa inversa i/o un test ELISA. Per últim, es va seqüenciar la regió no codificant 5’ dels virus detectats i es van aïllar. L’estudi de poblacions d’isard prèviament afectades per brots de malaltia va indicar que la infecció per BDV ha esdevingut endèmica en determinades zones i que possiblement aquest fet està relacionat amb la manca de recuperació de certes poblacions. Tanmateix, en altres zones, el virus aparentment ha deixat de circular i la recuperació de la població d’isards després dels brots és bona. L’estudi a la reserva nacional de caça de Freser-Setcases, va establir que el BDV-4 és mantingut a la població d’isards sense causar cap efecte negatiu a la dinàmica poblacional. És possible que en aquesta població hi circulin soques menys virulentes de BDV-4, causant una elevada seroprevalença i evitant la dispersió d’altres soques més virulentes, i l’aparició d’un brot de malaltia. En totes les poblacions els isards seropositius van presentar títols més alts d’anticossos contra les soques BDV, i la majoria contra les BDV-4, concretament. En tots els remugants salvatges que comparteixen hàbitat amb l’isard es va detectar una seroprevalença baixa. La seroprevalenca en oví i caprí va variar depenent de l’origen geogràfic, i al vaquí sempre va ser elevada. L’oví va presentar títols d’anticossos més alts enfront a les soques BDV-4, el vaquí a BVDV i les cabres enfront a BDV-4 i BVDV. Als estudis realitzats al sud-oest dels Alps italians es va detectar una elevada seroprevalença a l’isard alpí. Aquest fet suggereix que la infecció per pestivirus és freqüent a les ambdues espècies del gènere Rupicapra. Per altra banda, als íbex i a les cabres salvatges de totes les zones es va detectar una seroprevalença molt baixa, i la majoria dels individus positius van mostrar anticossos enfront a BDV. A Suïssa, tots els cérvols analitzats i un isard alpí van mostrar anticossos enfront a BVDV, mentre que la resta d’isards van mostrar títols més alts enfront a BDV. Aquest resultat indica que els remugants salvatges de Suïssa es poden infectar amb les dos espècies de pestivirus.Since 2001 several outbreaks of a new disease associated to Border Disease Virus (genus Pestivirus) of genotype 4 infection (BDV-4) have caused important declines in the Pyrenean chamois (Rupicapra pyrenaica pyrenaica) populations. The main goal of the present research work was to study the epidemiology of BDV infection in Pyrenean chamois and other wild and domestic ruminants, since pestiviruses are not strictly host-specific. The main study areas consisted of the Catalan and Andorran Pyrenees, but also different areas from the Ports de Tortosa i Beseit and Sierra Nevada Spanish mountains and from the Italian and Swiss Alps. The studied species were the Pyrenean chamois, European mouflon (Ovis aries), red (Cervus elaphus), roe (Capreolus capreolus), and fallow deer (Dama dama), Alpine chamois (Rupicapra rupicapra), Alpine (Capra ibex) and Iberian ibex (Capra pyrenaica), and sheep, goats and cattle that share the habitat with chamois. Sera were tested for the presence of antibodies against pestivirus with a commercial blocking ELISA assay. Comparative virus neutralization tests were performed in positive sera by using a bovine viral diarrhoea virus (BVDV) strain and reference and local BDV strains. Viral detection in wild ruminants was performed by reverse transcription-polymerase chain reaction (RT-PCR) using the panpestivirus primers 324 and 326 and/or with a commercial sandwich ELISA assay. As last, the sequence analyses of the 5’UTR region from positive samples and virus isolation were performed. The study of pestivirus epidemiology in Pyrenean chamois populations previously affected by BDV-infection outbreaks revealed that in some populations the disease has become endemic and BDV circulates frequently in the chamois population, possibly having a negative impact on host population dynamics. Contrarily, in some populations BDV does not seem to circulate after the disease outbreak. The study of a Pyrenean chamois population from the Eastern Pyrenees unaffected by disease outbreaks revealed that BDV-4 is self-maintained in this population, apparently without causing negative population dynamic effects. The circulation of weakly virulent strains could have been the cause of the high seroprevalence detected, which could hinder the spread of more virulent strains within this chamois population and the appearance of disease outbreaks. The seropositive Pyrenean chamois had BDV antibodies in all the studied populations, and most of them had higher titres against the BDV-4 strains. All the wild ruminants sharing habitat with Pyrenean chamois showed a low seroprevalence. The seroprevalence in sheep and goats, varied depending on the geographical origin, while in cattle it was constantly high. Sheep had higher antibody titres against the BDV-4 strains, cattle against BVDV, while goats showed higher titres against both BVDV and BDV strains. The study of pestivirus epidemiology in the south-western Italian Alps revealed a high seroprevalence in Alpine chamois, suggesting that members of the genus Rupicapra are likely to be infected with pestiviruses. Otherwise, Alpine and Iberian Ibex from all zones had a low seroprevalence, and most of the positive ibex had BDV antibodies. In Switzerland, the analyzed red deer and one Alpine chamois, had antibodies to BVDV, and the rest of chamois had antibodies to BDV. This results shows that the wild ruminants from this country can be infected with both pestivirus species, BVDV and BDV

Epidemiology of pestivirus infection in Pyrenean chamois (rupicapra pyrenaica pyrenaica) and other wild and domestic ruminants

Des de l'any 2001, diferents brots d'una malaltia associada a la infecció per un pestivirus, concretament un virus de la malaltia de la frontera de genotip 4 (BDV-4, de l'anglès border disease virus), han causat reduccions importants en el nombre d'efectius d'isard (Rupicapra pyrenaica pyrenaica). L'objectiu principal d'aquesta tesi doctoral va consistir en estudiar l'epidemiologia de la infecció per BDV a l'isard i a altres remugants salvatges i domèstics, ja que els pestivirus no són hoste-específics estrictes. La principal zona d'estudi va consistir en els Pirineus Catalans i Andorrans, però també es van estudiar diferents zones dels Ports de Tortosa i Beseit i Sierra Nevada, a Espanya, i dels Alps italians i suïssos. Les espècies estudiades van ser l'isard, el mufló europeu (Ovis aries), el cérvol (Cervus elaphus), el cabirol (Capreolus capreolus), la daina (Dama dama), l'isard alpí (Rupicapra rupicapra), l'íbex (Capra ibex) i la cabra salvatge (Capra pyrenaica), així com l'oví, caprí i vaquí que pasturen en els prats d'alta muntanya. Es va determinar la presència d'anticossos enfront a pestivirus en sèrum mitjançant una tècnica d'ELISA. En els sèrums positius es va dur a terme un test de seroneutralització vírica comparada amb una soca del virus de la diarrea vírica bovina (BVDV, de l'anglès bovine viral diaorrhea virus) i diferents soques de BDV locals i de referència. La detecció vírica es va realitzar mitjançant una tècnica de reacció en cadena de la polimerasa inversa i/o un test ELISA. Per últim, es va seqüenciar la regió no codificant 5' dels virus detectats i es van aïllar. L'estudi de poblacions d'isard prèviament afectades per brots de malaltia va indicar que la infecció per BDV ha esdevingut endèmica en determinades zones i que possiblement aquest fet està relacionat amb la manca de recuperació de certes poblacions. Tanmateix, en altres zones, el virus aparentment ha deixat de circular i la recuperació de la població d'isards després dels brots és bona. L'estudi a la reserva nacional de caça de Freser-Setcases, va establir que el BDV-4 és mantingut a la població d'isards sense causar cap efecte negatiu a la dinàmica poblacional. És possible que en aquesta població hi circulin soques menys virulentes de BDV-4, causant una elevada seroprevalença i evitant la dispersió d'altres soques més virulentes, i l'aparició d'un brot de malaltia. En totes les poblacions els isards seropositius van presentar títols més alts d'anticossos contra les soques BDV, i la majoria contra les BDV-4, concretament. En tots els remugants salvatges que comparteixen hàbitat amb l'isard es va detectar una seroprevalença baixa. La seroprevalenca en oví i caprí va variar depenent de l'origen geogràfic, i al vaquí sempre va ser elevada. L'oví va presentar títols d'anticossos més alts enfront a les soques BDV-4, el vaquí a BVDV i les cabres enfront a BDV-4 i BVDV. Als estudis realitzats al sud-oest dels Alps italians es va detectar una elevada seroprevalença a l'isard alpí. Aquest fet suggereix que la infecció per pestivirus és freqüent a les ambdues espècies del gènere Rupicapra. Per altra banda, als íbex i a les cabres salvatges de totes les zones es va detectar una seroprevalença molt baixa, i la majoria dels individus positius van mostrar anticossos enfront a BDV. A Suïssa, tots els cérvols analitzats i un isard alpí van mostrar anticossos enfront a BVDV, mentre que la resta d'isards van mostrar títols més alts enfront a BDV. Aquest resultat indica que els remugants salvatges de Suïssa es poden infectar amb les dos espècies de pestivirus.Since 2001 several outbreaks of a new disease associated to Border Disease Virus (genus Pestivirus) of genotype 4 infection (BDV-4) have caused important declines in the Pyrenean chamois (Rupicapra pyrenaica pyrenaica) populations. The main goal of the present research work was to study the epidemiology of BDV infection in Pyrenean chamois and other wild and domestic ruminants, since pestiviruses are not strictly host-specific. The main study areas consisted of the Catalan and Andorran Pyrenees, but also different areas from the Ports de Tortosa i Beseit and Sierra Nevada Spanish mountains and from the Italian and Swiss Alps. The studied species were the Pyrenean chamois, European mouflon (Ovis aries), red (Cervus elaphus), roe (Capreolus capreolus), and fallow deer (Dama dama), Alpine chamois (Rupicapra rupicapra), Alpine (Capra ibex) and Iberian ibex (Capra pyrenaica), and sheep, goats and cattle that share the habitat with chamois. Sera were tested for the presence of antibodies against pestivirus with a commercial blocking ELISA assay. Comparative virus neutralization tests were performed in positive sera by using a bovine viral diarrhoea virus (BVDV) strain and reference and local BDV strains. Viral detection in wild ruminants was performed by reverse transcription-polymerase chain reaction (RT-PCR) using the panpestivirus primers 324 and 326 and/or with a commercial sandwich ELISA assay. As last, the sequence analyses of the 5'UTR region from positive samples and virus isolation were performed. The study of pestivirus epidemiology in Pyrenean chamois populations previously affected by BDV-infection outbreaks revealed that in some populations the disease has become endemic and BDV circulates frequently in the chamois population, possibly having a negative impact on host population dynamics. Contrarily, in some populations BDV does not seem to circulate after the disease outbreak. The study of a Pyrenean chamois population from the Eastern Pyrenees unaffected by disease outbreaks revealed that BDV-4 is self-maintained in this population, apparently without causing negative population dynamic effects. The circulation of weakly virulent strains could have been the cause of the high seroprevalence detected, which could hinder the spread of more virulent strains within this chamois population and the appearance of disease outbreaks. The seropositive Pyrenean chamois had BDV antibodies in all the studied populations, and most of them had higher titres against the BDV-4 strains. All the wild ruminants sharing habitat with Pyrenean chamois showed a low seroprevalence. The seroprevalence in sheep and goats, varied depending on the geographical origin, while in cattle it was constantly high. Sheep had higher antibody titres against the BDV-4 strains, cattle against BVDV, while goats showed higher titres against both BVDV and BDV strains. The study of pestivirus epidemiology in the south-western Italian Alps revealed a high seroprevalence in Alpine chamois, suggesting that members of the genus Rupicapra are likely to be infected with pestiviruses. Otherwise, Alpine and Iberian Ibex from all zones had a low seroprevalence, and most of the positive ibex had BDV antibodies. In Switzerland, the analyzed red deer and one Alpine chamois, had antibodies to BVDV, and the rest of chamois had antibodies to BDV. This results shows that the wild ruminants from this country can be infected with both pestivirus species, BVDV and BDV

Two different epidemiological scenarios of border disease in the populations of Pyrenean chamois (Rupicapra p. pyrenaica) after the first disease outbreaks.

Since 2001 several outbreaks of a new disease associated with Border disease virus (BDV) infection have caused important declines in Pyrenean chamois (Rupicapra pyrenaica) populations in the Pyrenees. The goal of this study was to analyze the post-outbreak BDV epidemiology in the first two areas affected by disease with the aim to establish if the infection has become endemic. We also investigated if BDV infected wild and domestic ruminants sharing habitat with chamois. Unexpectedly, we found different epidemiological scenarios in each population. Since the disease outbreaks, some chamois populations recuperated quickly, while others did not recover as expected. In chamois from the first areas, prevalence was high (73.47%) and constant throughout the whole study period and did not differ between chamois born before and after the BDV outbreak; in all, BDV was detected by RT-PCR in six chamois. In the other areas, prevalence was lower (52.79%) and decreased during the study period; as well, prevalence was significantly lower in chamois born after the disease outbreak. No BDV were detected in this population. A comparative virus neutralisation test performed with four BDV strains and one Bovine viral diarrhoea virus (BVDV) strain showed that all the chamois had BDV-specific antibodies. Pestivirus antibodies were detected in all the rest of analyzed species, with low prevalence values in wild ruminants and moderate values in domestic ruminants. No viruses were detected in these species. These results confirm the hypothesis that outbreaks of BDV infection only affect the Pyrenean chamois, although other wild ruminants can occasionally be infected. In conclusion, two different scenarios have appeared since the first border disease outbreaks in Pyrenean chamois: on the one hand frequent BDV circulation with possible negative impact on population dynamics in some areas and on the other, lack of virus circulation and quick recovery of the chamois population

Two different epidemiological scenarios of border disease in the populations of Pyrenean chamois (Rupicapra p. pyrenaica) after the first disease outbreaks

Since 2001 several outbreaks of a new disease associated with Border disease virus (BDV) infection have caused important declines in Pyrenean chamois (Rupicapra pyrenaica) populations in the Pyrenees. The goal of this study was to analyze the post-outbreak BDV epidemiology in the first two areas affected by disease with the aim to establish if the infection has become endemic. We also investigated if BDV infected wild and domestic ruminants sharing habitat with chamois. Unexpectedly, we found different epidemiological scenarios in each population. Since the disease outbreaks, some chamois populations recuperated quickly, while others did not recover as expected. In chamois from the first areas, prevalence was high (73.47%) and constant throughout the whole study period and did not differ between chamois born before and after the BDV outbreak; in all, BDV was detected by RT-PCR in six chamois. In the other areas, prevalence was lower (52.79%) and decreased during the study period; as well, prevalence was significantly lower in chamois born after the disease outbreak. No BDV were detected in this population. A comparative virus neutralisation test performed with four BDV strains and one Bovine viral diarrhoea virus (BVDV) strain showed that all the chamois had BDV-specific antibodies. Pestivirus antibodies were detected in all the rest of analyzed species, with low prevalence values in wild ruminants and moderate values in domestic ruminants. No viruses were detected in these species. These results confirm the hypothesis that outbreaks of BDV infection only affect the Pyrenean chamois, although other wild ruminants can occasionally be infected. In conclusion, two different scenarios have appeared since the first border disease outbreaks in Pyrenean chamois: on the one hand frequent BDV circulation with possible negative impact on population dynamics in some areas and on the other, lack of virus circulation and quick recovery of the chamois population

Spatial and Temporal Phylogeny of Border Disease Virus in Pyrenean Chamois (Rupicapra p. pyrenaica)

Border disease virus (BDV) affects a wide range of ruminants worldwide, mainly domestic sheep and goat. Since 2001 several outbreaks of disease associated to BDV infection have been described in Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in Spain, France and Andorra. In order to reconstruct the most probable places of origin and pathways of dispersion of BDV among Pyrenean chamois, a phylogenetic analysis of 95 BDV 5'untranslated sequences has been performed on chamois and domestic ungulates, including novel sequences and retrieved from public databases, using a Bayesian Markov Chain Monte Carlo method. Discrete and continuous space phylogeography have been applied on chamois sequences dataset, using centroid positions and latitude and longitude coordinates of the animals, respectively. The estimated mean evolutionary rate of BDV sequences was 2.9×10−3 subs/site/year (95% HPD: 1.5-4.6×10−3). All the Pyrenean chamois isolates clustered in a unique highly significant clade, that originated from BDV-4a ovine clade. The introduction from sheep (dated back to the early 90s) generated a founder effect on the chamois population and the most probable place of origin of Pyrenean chamois BDV was estimated at coordinates 42.42 N and 1.9 E. The pathways of virus dispersion showed two main routes: the first started on the early 90s of the past century with a westward direction and the second arise in Central Pyrenees. The virus spread westward for more than 125 km and southward for about 50km and the estimated epidemic diffusion rate was about 13.1 km/year (95% HPD 5.2-21.4 km/year). The strong spatial structure, with strains from a single locality segregating together in homogeneous groups, and the significant pathways of viral dispersion among the areas, allowed to reconstruct both events of infection in a single area and of migrations, occurring between neighboring areas

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 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