BackgroundThe role of wildlife as a brucellosis reservoir for humans and domestic livestock remains to be properly established. The aim of this work was to determine the aetiology, apparent prevalence, spatial distribution and risk factors for brucellosis transmission in several Iberian wild ungulates.MethodsA multi-species indirect immunosorbent assay (iELISA) using Brucella S-LPS antigen was developed. In several regions having brucellosis in livestock, individual serum samples were taken between 1999 and 2009 from 2,579 wild bovids, 6,448 wild cervids and4,454 Eurasian wild boar (Sus scrofa), and tested to assess brucellosis apparent prevalence. Strains isolated from wild boar were characterized to identify the presence of markers shared with the strains isolated from domestic pigs.ResultsMean apparent prevalence below 0.5% was identified in chamois (Rupicapra pyrenaica), Iberian wild goat (Capra pyrenaica), and red deer (Cervus elaphus). Roe deer (Capreolus capreolus), fallow deer (Dama dama), mouflon (Ovis aries) and Barbary sheep (Ammotragus lervia) tested were seronegative. Only one red deer and one Iberian wild goat resulted positive in culture, isolating B. abortus biovar 1 and B. melitensis biovar 1, respectively. Apparent prevalence in wild boar ranged from 25% to 46% in the different regions studied, with the highest figures detected in South-Central Spain. The probability of wild boar being positive in the iELISA was also affected by age, age-by-sex interaction, sampling month, and the density of outdoor domestic pigs. A total of 104 bacterial isolates were obtained from wild boar, being all identified as B. suis biovar 2. DNA polymorphisms were similar to those found in domestic pigs.ConclusionsIn conclusion, brucellosis in wild boar is widespread in the Iberian Peninsula, thus representing an important threat for domestic pigs. By contrast, wild ruminants were not identified as a significant brucellosis reservoir for livestock.

SummaryNinety-five adult fallow deer, legally hunted in the Regional Hunting Reserve of El Sueve (Northern Spain), were subjected to a post-mortem examination for paratuberculosis, samples being taken from the proximal and distal jejunum, proximal and distal ileum, ileocaecal valve and associated lymph nodes. The lesions were divided into four categories. Focal lesions (n ¼ 19 cases) consisted of small granulomas, mainly in the jejunal and ileal lymph nodes. Multifocal lesions (n ¼ 4) consisted of well-demarcated granulomas in the intestinal lymphoid tissue and also in the intestinal lamina propria. Diffuse multibacillary lesions (n ¼ 2) were characterized by a severe granulomatous enteritis and lymphadenitis. Macrophages and numerous Langhans giant cells containing many mycobacteria were present, resulting in macroscopical changes in the normal gut morphology. These changes were found from the proximal jejunum to the ileocaecal valve, but lesions were always particularly severe in the distal jejunum. In diffuse intermediate (multibacillaryelymphocytic) lesions (n ¼ 3) the infiltrate consisted of lymphocytes, macrophages and Langhans giant cells, with small numbers of mycobacteria. Mycobacterium avium subspecies paratuberculosis was identified by a polymerase chain reaction technique. The widespread occurrence of paratuberculosis in fallow deer in this Reserve represents a potential source of infection for other susceptible species.

In Europe, badgers (Meles meles) are recognized as major tuberculosis (TB) reservoir hosts with the potential to transmit infection to associated cattle herds. Recent studies in Spain have demonstrated that vaccination with a heat-inactivated Mycobacterium bovis vaccine (HIMB) successfully protects captive wild boar and red deer against progressive disease. The aim of this study was to evaluate the efficacy of two oral vaccines against TB in a badger model: the live-attenuated M. bovis bacillus Calmette-Guérin BCG vaccine (Danish strain) and a HIMB vaccine. Twenty-four badgers were separated in three treatment groups: oral vaccinated with live BCG (10 8 CFU, n = 5), oral vaccinated with HIMB (10 7 CFU, n = 7), and unvaccinated controls (n = 12). All badgers were experimentally infected with M. bovis (10 3 CFU) by the endobronchial route targeting the right middle lung lobe. Throughout the study, clinical, immunological, pathological, and bacteriological parameters of infection were measured. Both vaccines conferred protection against experimental TB in badger, as measured by a reduction of the severity and lesion volumes. Based on these data, HIMB vaccination appears to be a promising TB oral vaccine candidate for badgers in endemic countries.

We describe the pathology, isolation and characterisation of a virus responsible for an outbreak of a systemic haemorrhagic disease causing high mortality in tadpoles of the common midwife toad Alytes obstetricans in the 'Picos de Europa' National Park in northern Spain. The virus, provisionally designated as the common midwife toad virus (CMTV), was isolated from homogenates of visceral tissue from diseased toad tadpoles following inoculation on epithelioma papulosum cyprini (EPC) cells. Molecular characterisation of the virus, including sequence analysis of the DNA polymerase and major capsid protein genes, showed that the isolated virus was a ranavirus with marked sequence identity to other members of the genus Ranavirus. A rabbit antiserum raised against purified virions was prepared and used to definitively demonstrate systemic distribution of the virus in diseased tadpoles, indicating that the isolated virus was the primary pathogen.

This report describes the isolation and characterisation of the common midwife toad virus (CMTV) from juvenile alpine newts (Mesotriton alpestris cyreni) and common midwife toad (CMT) tadpoles (Alytes obstetricans) in the Picos de Europa National Park in Northern Spain in August 2008. A comparative pathological and immunohistochemical study was carried out using anti-CMTV polyclonal serum. In the kidneys, glomeruli had the most severe histological lesions in CMT tadpoles, while both glomeruli and renal tubular epithelial cells exhibited foci of necrosis in juvenile alpine newts. Viral antigens were detected by immunohistochemical labelling mainly in the kidneys of CMT tadpoles and in ganglia of juvenile alpine newts. This is the first report of ranavirus infection in the alpine newt, the second known species to be affected by CMTV in the past 2 years.