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.

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.

Spatial-temporal Trends and Factors Associated with the Bluetongue Virus Seropositivity in Large Game Hunting Areas from Southern Spain.

An epidemiological study was carried out to determine the spatial-temporal trends and risk factors potentially involved in the seropositivity to bluetongue virus (BTV) in hunting areas with presence of red deer (Cervus elaphus). A total of 60 of 98 (61.2%; CI95% : 51.6-70.9) hunting areas sampled presented at least one seropositive red deer. Antibodies against BTV were detected in juvenile animals during the hunting seasons 2007/2008 to 2013/2014 in 15 of 98 (15.3%) hunting areas, which indicates an uninterrupted circulation of BTV in this period. A multivariate logistic regression model showed that the red deer density at hunting area level (>22 individuals/km(2) ), the annual abundance of Culicoides imicola (>1.4 mosquitoes/sampling) and the goat density at municipality level (>24.1 individuals/km(2) ) were factors significantly associated with BTV seropositivity in hunting areas. Control measures against BTV in the studied area include vaccination programmes in wild and domestic ruminants, movement control in areas with high densities and abundance of red deer and C. imicola, respectively. Considering the potential risk of BTV re-emergence, red deer should be included in the BT surveillance programmes in regions where these species share habitats with livestock.

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.

Seroprevalence of Toxoplasma gondii in equids from Southern Spain.

Antibodies to Toxoplasma gondii were determined in serum samples from 616 equids (454 horses, 80 mules and 82 donkeys) in a cross-sectional study of 420 herds in Andalusia (Southern Spain), the region with the highest number of equids in Spain. Antibodies to T. gondii were found in 10.8% horses, 15.0% mules and 25.6% donkeys by using the modified agglutination test (MAT) at a cut-off of 1:25. Herd seroprevalence for horses, mules and donkeys was 14.7% (48/327), 23.9% (11/46) and 34.0% (16/47), respectively, and 75 herds (17.8%) had at least one seropositive animal. Significant differences in T. gondii seroprevalence were observed among species, with donkeys having the highest seroprevalence and horses the lowest (P=0.04). Seroprevalence was significantly higher in herds with presence of domestic ruminants. This study is the first report of the presence of T. gondii antibodies in equine species in Spain and the first reporting T. gondii infection in donkeys in Europe. The presence of antibodies is indication of contact with the parasite and therefore, consumption of equine meat could be a potential source of human infection in Spain.

Monitoring of the West Nile virus epidemic in Spain between 2010 and 2011.

West Nile virus (WNV) is a mosquito-transmitted flavivirus recognized as an emerging and re-emerging pathogen in different countries. This study describes the monitoring of the first WNV epidemic in Spain between 2010 and 2011. Between September and December 2010, 36 outbreaks of WNV in horses were reported in three different provinces of Andalusia (southern Spain), with no apparent spread outside this area. The temporal distribution and the clinical signs observed during the WNV epidemic in Spain were, in general, similar to those reported in Europe and in the Mediterranean Basin. Morbidity, mortality and fatality rate in the affected herds were 4.6, 1.4 and 35.3%, respectively. Thirty-six of 75 (47.4%) suspected herds investigated presented at least one IgM seropositive animal. The individual seroprevalence in unvaccinated animals from the infected holdings was 51.7%. RNA WNV lineage 1 virus was confirmed from blood and cerebrospinal fluid samples in a lethally infected horse. The entomological survey showed that the most abundant mosquito species detected in the affected area was Culex pipiens. A cross-sectional study was carried out in non-suspected herds between April 2010 and February 2011 in the affected area. The individual seroprevalence was 11.0%, and six of the 38 herds sampled (15.8%) presented at least one seropositive animal. The results showed active WNV circulation several months before the first outbreak was reported in horses. The seropositivity found in municipalities where clinical cases were not reported indicates a higher geographical dissemination of the virus. Significantly higher seroprevalences were detected in areas close to Morocco. Furthermore, 90 wild ruminants were tested for the presence of antibodies against WNV, but the results were all negative.