Parenteral Vaccination with Heat-Inactivated Mycobacterium Bovis Reduces the Prevalence of Tuberculosis-Compatible Lesions in Farmed Wild Boar.

In 2012, a wild boar (Sus scrofa) tuberculosis (TB) control programme was set up in a wild boar farm by means of intramuscular (IM) vaccination with a heat-inactivated Mycobacterium bovis vaccine (IV). The goal was to assess safety and efficacy of the parenterally administered IV in a large farm setting with natural M. bovis circulation. Based on preceding results under laboratory conditions, we hypothesized that vaccinated piglets would show smaller scores of TB-compatible lesions (TBCL) than unvaccinated controls. After vaccination, no adverse reactions were detected by visual inspection or at post-mortem examination (n = 668 and 97, respectively). Post-mortem data on TBCL were available for 97 vaccinated wild boar and 182 controls. The observed TBCL prevalence was 4.1% (95% CI = 0.2-8%) and 12.1% (95% CI = 7.1-17.1%) for vaccinated and control wild boar, respectively (P < 0.05). Among those animals with TBCL, no difference in the mean lesion score was found (P > 0.05). The results show that IV administered intramuscularly to wild boar piglets is safe and protects vaccinated individuals (66% reduction in TBCL prevalence) against natural challenge in a low-prevalence setting. In a context of increasing TB prevalence in wild boar in Mediterranean habitats, vaccination achieved a progressive though slow decline in lesion prevalence since the onset of the vaccination scheme. Hence, vaccination might contribute, along with other tools, to TB control in wild boar and in pigs.

Testing Eurasian wild boar piglets for serum antibodies against Mycobacterium bovis.

Animal tuberculosis (TB) caused by infection with Mycobacterium bovis and closely related members of the M. tuberculosis complex (MTC), is often reported in the Eurasian wild boar (Sus scrofa). Tests detecting antibodies against MTC antigens are valuable tools for TB monitoring and control in suids. However, only limited knowledge exists on serology test performance in 2-6 month-old piglets. In this age-class, recent infections might cause lower antibody levels and lower test sensitivity. We examined 126 wild boar piglets from a TB-endemic site using 6 antibody detection tests in order to assess test performance. Bacterial culture (n=53) yielded a M. bovis infection prevalence of 33.9%, while serum antibody prevalence estimated by different tests ranged from 19% to 38%, reaching sensitivities between 15.4% and 46.2% for plate ELISAs and between 61.5% and 69.2% for rapid immunochromatographic tests based on dual path platform (DPP) technology. The Cohen kappa coefficient of agreement between DPP WTB (Wildlife TB) assay and culture results was moderate (0.45) and all other serological tests used had poor to fair agreements. This survey revealed the ability of several tests for detecting serum antibodies against the MTC antigens in 2-6 month-old naturally infected wild boar piglets. The best performance was demonstrated for DPP tests. The results confirmed our initial hypothesis of a lower sensitivity of serology for detecting M. bovis-infected piglets, as compared to older wild boar. Certain tests, notably the rapid animal-side tests, can contribute to TB control strategies by enabling the setup of test and cull schemes or improving pre-movement testing. However, sub-optimal test performance in piglets as compared to that in older wild boar should be taken into account.

Temporal trend of tuberculosis in wild ungulates from Mediterranean Spain.

Tuberculosis (TB), a chronic disease caused by infection with the Mycobacterium tuberculosis complex, is endemic in wild boar (Sus scrofa) and red deer (Cervus elaphus) in south-central Spain. Understanding the temporal dynamics of this chronic infection requires long time series data collection over large areas. The aim of this paper was to identify the determinants of TB prevalence and severity in both species in Ciudad Real province, Spain, from 2000 to 2012. Study variables included management, population dynamics, and a range of geographical and climatological factors. The prevalence of TB in wild boar increased from 50% to 63% since the study commenced. This may be due to an increased hunting bag (a proxy for population abundance), which was correlated with TB infection rates. Low rainfall (a stochastic factor) was associated with higher individual risk of TB presence and progression, resulting in an increased proportion of severe cases of wild boar TB in dry years. This was probably a result of increased food restriction leading to a higher susceptibility to TB. In contrast, red deer TB showed an apparent stable trend, which may be a consequence of the species' higher and stable population size. Hunting management, characterized by fencing, was associated with a higher risk of TB in both wild boar and red deer, suggesting that intensive hunting management may have contributed to exacerbated TB figures. This difference was more marked in red deer than in wild boar, probably because fencing imposes less restriction on movement, population mixing and TB spread to wild boar than to deer. Our findings on TB dynamics are fundamental for assessing the impact of future disease-control actions (e.g. field vaccination). Moreover, such control plans must operate in the long term and cover large areas.

Effects of culling Eurasian wild boar on the prevalence of Mycobacterium bovis and Aujeszky's disease virus.

Worldwide, failure to eradicate a disease in livestock has sometimes been related to wildlife reservoirs of infection. We describe the effects of Eurasian wild boar (Sus scrofa) abundance reduction through increased culling on the prevalence of two chronic infectious diseases, tuberculosis (TB) and Aujeszky's disease (AD), in a region of South-central Spain (SCS). The two infections studied responded differently to an approximately 50% reduction of wild boar abundance. Wild boar TB prevalence remained stable in control sites, whereas it decreased by 21-48% in treatment sites. In one treatment site, the annual wild boar abundance was positively correlated with the annual percentage of skin test reactor cattle. In another treatment site, red deer (Cervus elaphus) M. bovis infection prevalence decreased after culling wild boar. No significant effect of wild boar culling on wild boar ADV seroprevalence was found. The reduction in wild boar TB was achieved despite no alternative M. bovis host being included in the culling strategy. We advocate that culling could become a part of integrated control strategies including habitat and game management changes and vaccination, contributing to increase their success likelihood, or reducing the total expenses.

Spatio-temporal trends and risk factors for Trichinella species infection in wild boar (Sus scrofa) populations of central Spain: a long-term study.

In south-central Spain, the harvest of Eurasian wild boar (Sus scrofa) has increased significantly during recent decades in association with more intensive management actions to increase hunting yields and with consequent effects on the health status of the wild boar populations. We investigated the spatio-temporal trends and the risk factors related to the prevalence of Trichinella spp. in wild boar in order to obtain the annual probability of occurrence for these parasites in the Ciudad Real province of south-central Spain. Based on muscle samples collected during the hunting seasons from 1998/1999 to 2009/2010, the mean prevalence for Trichinella spp. in 95,070 wild boar was 0.2% (95% confidence interval 0.17-0.23). A subsample of 1,432 wild boar was also tested by ELISA. No correlation was observed between the prevalence of infection detected by serology and by the artificial digestion of muscle. The presence of Trichinella infections in wild boar showed a decreasing trend during the study period and was negatively related with fenced wild boar populations. The predicted 'favourability' for Trichinella infections disappeared almost completely after the 2006/2007 hunting season. Risk maps based on biogeographical tools showed, however, that most hunting estates presented favourable risk factors for these parasites during at least one of the hunting seasons studied.

Monitoring of African swine fever in the wild boar population of the most recent endemic area of Spain.

Wild boars are natural hosts for African swine fever (ASF). The ASF virus (ASFV) can persist for long periods in the environment, such as in ticks and contaminated products, which may be sources of infection for wild boar populations. African swine fever was eradicated in domestic pig populations in Spain in 1995, after 35 years of significant effort. To determine whether ASFV can persist in wild boar hosts after it has been eradicated from domestic pigs and to study the role of wild boar in helping ASFV persist in the environment, we checked for the presence of ASFV in wild boars in Doñana National Park, one of the largest natural habitats of wild boar in Spain and one of the last areas where ASF was endemic prior its eradication. Samples from 158 animals collected between 2006 and 2010 were analysed using serological and nucleic acid-based diagnostic techniques recommended by the World Organization for Animal Health (OIE). None of the samples was found to be positive. These results confirm the absence of disease in wildlife in what was once one of the areas most affected by ASF in Spain, and they suggest that wild boars play a limited role in ASFV persistence. These results confirm that ASFV cannot persist in isolated wild boar populations for long periods of time without the interaction of other factors such as re-infection by contact with domestic pigs or by feeding on contaminated swill.

First report of Troglotrema acutum (Digenea, Troglotrematidae) in the Eurasian badger Meles meles in the Iberian Peninsula and presumptive lesions caused in the host.

A total of 109 badger Meles meles skulls from Catalonia (north-eastern Iberian Peninsula) were studied for helminths. The tremadode Troglotrema acutum is reported here for the first time in the Eurasian badger in the Iberian Peninsula and southern Europe. Three methodologies were used to detect this trematode: an examination for surface lesions, axial computed tomography and fresh skull dissection. The damage caused in the affected skulls is described, along with details regarding the use of computed tomography to detect hyperostosis, leakage in the sinus structure and bone surface erosion in the affected skulls.

Performance of immunochromatographic and ELISA tests for detecting fallow deer infected with Mycobacterium bovis.

Fallow deer (Dama dama) are widely distributed as natural or naturalised populations, as well as in game parks and deer farms. We used 157 fallow deer sampled in populations considered to be Mycobacterium tuberculosis complex (MTC) free and 73 Mycobacterium bovis-infected fallow deer confirmed postmortem by culture to evaluate the diagnostic performance of two tests for the detection of anti-mycobacterial antibodies: the dual path platform (DPP) VetTB assay and the bovine purified protein derivative (bPPD) ELISA. We also compared their sensitivity with that of the skin test, analyzed the effect of haemolysis degree on the antibody detection and described the relationship between the test readings and presence/absence of gross tuberculosis (TB) compatible lesions. Sensitivity of bPPD ELISA was 51% at a specificity of 96%. Depending on the cut-off value selected, the sensitivity of DPP VetTB ranged from 62 to 71%, while its specificity was 88-95%. In the subgroup of M. bovis-infected deer for which the skin test data were available (33 of 73); this method detected 76% of culture-positive animals, although the specificity of the intradermal test was not determined in this study. When the DPP VetTB and skin test data were combined, the resulting sensitivity obtained in this sub-group of M. bovis-infected deer increased to 97%. Gross pathology identified TB compatible lesions (TBL) in 89% culture-confirmed fallow deer. The infected animals with visible lesions had significantly higher readings in the DPP VetTB, but not in the bPPD ELISA. Only high levels of haemolysis decreased antibody test sensitivity and this effect was more evident for the bPPD ELISA. The results allowed inferring a number of management recommendations for rapid detection of MTC infection in live fallow deer and in surveys on hunter-harvested cervids.

Seroprevalence evolution of selected pathogens in Iberian wild boar.

A total of 1279 Eurasian wild boar (Sus scrofa) sera were collected from 2000 to 2011 in the Iberian Peninsula to reveal time changes in serum antibody prevalences against selected infectious agents (porcine circovirus type 2, PCV2; porcine reproductive and respiratory syndrome virus, PRRSV; hepatitis E virus, HEV; and Erysipelothrix rhusiopathiae) and to identify putative individual or population factors driving such changes. Overall seroprevalences were 48%, 26%, 2% and 15% for PCV2, HEV, PRRSV and E. rhusiopathiae, respectively. The global observed prevalence of antibodies against PCV2 and HEV remained stable during the study period, while the global mean antibody seroprevalence against E. rhusiopathiae declined. The mean increment in prevalence was always lower for open than for fenced sites. This study evidenced for the first time that wild boar from the Iberian Peninsula have widespread contact with E. rhusiopathiae, and confirmed high prevalences of antibodies against PCV2 and HEV. Maintained high prevalences of transmissible agents in wild boar suggest that epidemiological drivers such as aggregation and high density are still acting. This will most probably also affect the transmission rates of other disease agents and should be taken into account regarding disease control at the wildlife-livestock interface.

No evidence that wild red deer (Cervus elaphus) on the Iberian Peninsula are a reservoir of Mycobacterium avium subspecies paratuberculosis infection.

The potential role of red deer (Cervus elaphus) as a reservoir of Mycobacterium avium subspecies paratuberculosis (MAP) infection is largely unknown. A total of 332 wild red deer were investigated using post-mortem examination, bacteriology and serology. Only three animals (1.12%) were found to have lesions on histopathological examination and no MAP bacteria were recovered on culture. The results suggest it is unlikely that wild red deer make a significant contribution to the maintenance of MAP infection in the region. The cross-reactivity of the ELISAs used indicates this diagnostic modality is ineffective in the detection of MAP infection in this species. The implications of these results for the control of this important pathogen in both livestock and wildlife are discussed.

Serological, pathological and polymerase chain reaction studies on Mycoplasma hyopneumoniae infection in the wild boar.

There are evidences that wild boar and domestic pig populations share the vulnerability to certain pathogens and, in consequence, the risk of pathogen transmission. Based on serological evidences, it is known that Mycoplasma hyopneumoniae (Mhyo) is able to infect the wild boar. However, if this infection causes lesions and disease in this species is still unknown. The objectives of the present study were to assess the seroprevalence to Mhyo, as well as the rates of detection of Mhyo (by nested polymerase chain reaction, nPCR) in upper and lower respiratory tract of the wild boar. Moreover, presence of enzootic pneumonia (EP)-like gross and microscopic lesions in these animals was also assessed. Antibodies against Mhyo were detected in 92 out of 428 (21%) serum samples tested. Moreover, Mhyo DNA was detected by nPCR in 17 out of 85 (20%) nasal swabs and in 12 out of 156 (8%) lung samples. No gross EP-like lesions were observed in any of the studied pigs. Besides, presence of EP-like microscopic lung lesions was observed in 18 out of 63 (29%) animals. Mhyo DNA was detected at lung level in 2 out of these 18 (11%) animals. Presence of EP-like microscopic lesions was significantly related (p<0.05) with Mhyo detection at nasal swab swab but not (p>0.05) with its detection in lung samples; such lesions were also linked (p<0.05) with presence of Metastrongylus spp.-like parasite structures. Results of the present study confirm that Mhyo is able to infect and might be able to cause EP-like microscopic lesions in wild boar but, apparently, with a subclinical impact.