Jorge Paniagua

Seroprevalence and risk factors associated to Mycobacterium bovis in wild artiodactyl species from southern Spain, 2006-2010.

The control of bovine tuberculosis (bTB) is at a critical point in the last stage of eradication in livestock. Wildlife species recently have emerged infected with TB in Europe, particularly ungulates in the Iberian Peninsula. Epidemiological information regarding TB in wild ungulates including affected species, prevalence, associated risk factors and appropriate diagnostic methods need to be obtained in these countries. A cross-sectional study was carried out on wild artiodactyl species, including Eurasian wild boar (Sus scrofa) red deer (Cervus elaphus), roe deer (Capraelus capraelus), fallow deer (Dama dama), Spanish ibex (Capra pyrenaica hispanica) and mouflon (Ovis musimon), in Spain to assess the seroprevalence against Mycobacterium bovis or cross-reacting members of the Mycobcaterium tuberculosis complex (MTBC), and to provide information on associated risk factors. Previously, two in-house indirect enzyme linked immunosorbent assays (bPPD-ELISA and MPB83-ELISA) were developed using known TB status sera. Positive reference sera were selected from infected animals confirmed by culture. The M. bovis isolates belonged to spoligotypes SB0121, SB0120, SB0295, SB0265 and SB0134. Two hundred and two out of 1367 (7.5%; 95% CI: 6.1–8.9) animals presented antibodies against M. bovis by both bPPD-ELISA and MPB83-ELISA. Significantly higher TB seroprevalence was observed in wild boar compared to the other species analyzed. Interestingly, seropositivity against M. bovis was not found in any out of 460 Spanish ibex analyzed. The logistic regression model for wild boar indicated that the seropositivity to M. bovis was associated with age, location and year of sampling, while the only risk factor associated with M. bovis seroprevalence in red deer and fallow deer was the age. The seroprevalence observed indicates a widespread exposure to MTBC in several wild artiodactyl species in southern Spain, which may have important implications not only for conservation but also for animal and public health.

Campylobacter infection in wild artiodactyl species from southern Spain: Occurrence, risk factors and antimicrobial susceptibility

A cross-sectional study was performed to assess the occurrence of Campylobacter species and to identify potential associated risk factors for wild artiodactyl species in southern Spain. Campylobacter species were isolated in 55 of 363 (15.2%) faecal samples. Campylobacter was identified in faeces from wild boar (49/126; 38.9%), red deer (5/179; 2.8%) and mouflon (1/13; 7.7%) but not from fallow deer (0/45). The isolated Campylobacter species were identified as C. jejuni (2 isolates; 3.6%), C. coli (11 isolates; 20.0%) and C. lanienae (37 isolates; 67.3%). Five isolates (9.1%) could not be identified at the species level. This report is the first to describe C. lanienae infection in wild ruminant species. Resistance to erythromycin (4.8%), ciprofloxacin (37.5%), tetracycline (52.9%) and streptomycin (55%) were detected. C. lanienae presented a significantly higher number of susceptible isolates to ciprofloxacin and tetracycline than C. coli. Due to the low number of positive wild ruminants, a Generalised Estimating Equations model was only carried out for wild boar. The model indicated that the risk factors associated with Campylobacter infection were the density of wild boar (>10/100ha) (OR: 3.05; CI95%: 2.2-4.3), the presence of artificial waterholes (OR: 3.67; CI95%: 1.3-10.5) and the winter season (OR: 3.30; CI95%: 1.9-5.8). Campylobacter infection is widespread in wild boar populations in southern Spain. These findings suggest that wild artiodactyls, particularly wild boar, constitute a reservoir of Campylobacter species, including resistant and multi-resistant strains, which may be of public health concern.

Absence of circulation of Pestivirus between wild and domestic ruminants in southern Spain

Ruminant pestiviruses (family Flaviviridae) affect both wild and domestic ruminants worldwide, causing reproductive disorders and severe economic losses. Wild (n=1442) and domestic (n=373) ruminants from southern Spain were tested for the presence of antibodies to pestiviruses. Seropositivity was detected by both ELISA and virus neutralisation test in 1/892 (0.1 per cent) red deer, 29/125 (23.2 per cent) cattle and 17/157 (10.8 per cent) sheep. Pestivirus-specific antibodies to bovine viral diarrhoea virus (BVDV) and border disease virus (BDV) genotypes 1, 4 and 5 were detected. Significantly higher antibody titres to BDV-4 compared with BVDV-NADL were found in one cattle farm. Results indicate that wild ruminants may act as spillover hosts from domestic ruminants, and therefore they do not represent a risk for domestic ruminants in southern Spain. The wide distribution of pestiviruses among sheep and cattle suggests that transmission between these species may occur.

Toxoplasma gondii in sympatric domestic and wild ungulates in the Mediterranean ecosystem

Toxoplasma gondii is a zoonotic protozoan of worldwide distribution. The present study provides information on risk factors affecting T. gondii infection in domestic and free-ranging wild ungulates sharing habitats in Mediterranean ecosystems in Spain. Serum samples from 482 extensively reared domestic ruminants and 2351 wild ungulates were tested for T. gondii antibodies using the modified agglutination test (MAT, cut-off 1:25). Toxoplasma gondii seroprevalence was 41.2% of 194 sheep, 18.6% of 199 cattle and 5.6% of 89 goats. The main risk factors associated with infection in livestock were the presence of cats, feeding on the ground and at stubble fields. In wild ungulates, T. gondii antibodies were detected in 10.5% of 1063 red deer, 15.6% of 294 fallow deer, 5.6% of 216 European mouflon, 5.6% of 90 Spanish ibex, 13.6% of 22 roe deer and 18.6% of 666 wild boars. The risk factors affecting T. gondii infection in wildlife were species, age and hunting season. Significantly higher seroprevalence was found in domestic ruminants, particularly in sheep, compared to the wild species tested. The present study indicates widespread exposure to T. gondii among domestic and wild ungulates in Southern Spain, with significant differences among species sharing the same ecosystem. The high seroprevalence observed in domestic ruminants, particularly in sheep, reinforces the need for farm management practices to control the risk factors associated with T. gondii infection in extensively reared livestock. Consumption of raw and undercooked food products from domestic and wildlife species may have important implications for public health.

Monitoring of Schmallenberg virus in Spanish wild artiodactyls, 2006-2015

Schmallenberg disease is an emerging disease that affects domestic and wild ruminants in Europe. An epidemiological survey was carried out to assess exposure to Schmallenberg virus (SBV) in wild artiodactyls in Spain between 2006 and 2015. A total of 1751 sera from wild artiodactyls, including 1066 red deer, 304 fallow deer, 192 mouflon, 109 wild boar, 49 roe deer and 31 Spanish ibex were tested for antibodies against SBV by ELISA and confirmed by virus neutralization test. SBV was not detected between the 2006/2007 and the 2010/2011 hunting seasons. Overall seroprevalence (including samples collected between the 2011/2012 and 2014/2015 hunting seasons) was 14.6% (160/1099; 95%CI: 12.7–16.6). Mean SBV seroprevalence was 13.3±2.6% in red deer, 23.9±4.2% in fallow deer, 16.4±6.1% in mouflon and 2.8±3.1% in wild boar. No antibodies against SBV were found in roe deer or Spanish ibex. The presence of SBV RNA was confirmed in three of 255 (1.2%) spleen samples from wild ruminants analysed by rRT-PCR. In a multivariate mixed-effects logistic regression model, the main risk factors associated with SBV seroprevalence were: species (fallow deer, red deer and mouflon), age (adults) and interactions between hunting areas of more than 1000 hectares and hunting season (2012/2013, 2013/2014 and 2014/2015). The hypothesis of endemic circulation of SBV in the last few years is supported by the detection of SBV RNA in animals sampled in 2011 and 2015, as well as antibodies detected at low level in juveniles in 2012, 2013 and 2014. The results indicate that SBV circulated in wild ruminant populations in Spain during the same period when the virus was first reported in northern Europe, and at least five months before the first case was officially reported in livestock in Spain.

Absence of Rift Valley fever virus in domestic and wild ruminants from Spain

RIFT Valley fever virus (RVFV) is a mosquito-borne virus, member of the genus Phlebovirus (family Bunyaviridae ), which affects mainly ruminant species. It may cause severe economic losses through abortions and high mortality among newborns, and through the heavy control costs and the trade restrictions imposed. Rift Valley fever (RVF) is also a zoonotic disease with potentially severe consequences for infected people, including haemorrhagic fever, meningoencephalitis, renal failure, blindness and, in some cases, death. RVFV is transmitted through the bites of various species of mosquitoes (typically the Aedes or Culex genera) and also through infected tissues such as aborted fetuses, meat and blood (EFSA 2013). RVF is considered to be endemic in sub-Saharan African countries, with sporadic major outbreaks associated with periods of heavy rainfall and flooding. In the last few years, Mauritania and Senegal have been repeatedly affected by RVF (OIE-WAHID 2015). RVFV was thought to be restricted to Africa; however, in 2000, the virus emerged in Yemen and Saudi Arabia. Currently, the risk of expansion of RVFV to North African countries is considered high, and in the event of RVFV introduction into Morocco, the risk to Spain would significantly increase (CCAES 2014). The aim of the present study was to detect RVFV circulation in Andalusia (southern Spain), the highest risk area in Spain (Sanchez-Vizcaino and others 2013), or alternatively provide …