Juan Francisco García Marín

Tuberculosis due to Mycobacterium bovis and Mycobacterium caprae in sheep

Tuberculosis was diagnosed in three flocks of sheep in Galicia, Spain, in 2009 and 2010. Two flocks were infected with Mycobacterium bovis and one flock was infected with Mycobacterium caprae. Infection was confirmed by the comparative intradermal tuberculin test, bacteriology, molecular analysis and histopathology. Sheep have the potential to act as a reservoir for tuberculosis.

Presence of focal and multifocal paratuberculosis lesions in mesenteric lymph nodes and the ileocaecal valve of cattle positive to the tuberculin skin test.

The tuberculin skin test is commonly used in tuberculosis (TB) eradication programmes in cattle (Monaghan et al., 1994). The detection of false-positive reactors is important particularly in areas where there is a relatively low incidence of the disease, and can be attributed, at least in part, to immunological cross-reactivity with other mycobacteria infections. Of particular concern is Mycobacterium avium subspecies paratuberculosis (Map) infection, which is characterized by a chronic granulomatous enteritis in ruminants. The prevalence ofMap seems to be increasing in Spain where it now accounts for approximately 30.4% of positive reactors in cattle (Juste et al., 2000). Paratuberculosis immunization is also know to interfere with the diagnosis of bovine TB when single skin test is performed (Kohler et al., 2001). In order to differentiate between both mycobacterial diseases, a comparative skin test has to be carried out and the response to avian purified protein derivative (PPD) is greater than to bovine PPD in paratuberculosis infected cattle (Chiodini et al., 1984). The aim of the present study was to describe the lesions associated with paratuberculosis in adult cattle positive to the single tuberculin skin test, and to compare these with cattle negative to the tuberculin test. During the 11-month period of study, from February to December 1999, a total of 134 cattle, 4–9 years of age, was examined. Ninety-one (group 1) were positive tuberculin reactors to the single skin test, selected from 49 herds. Tuberculin skin test was performed by the in-

Endothelialization of Nonapposed Stent Struts Located over the Origin of a Side Branch: Results with Different Carbofilm‐Coated Stents

OBJECTIVE To evaluate the degree of endothelialization of the nonapposed struts located at the ostia of side branches. BACKGROUND Endothelialization of coronary stents has got considerable relevance because of the phenomenon of late thrombosis. Bifurcation location and incomplete stent apposition have been linked to this complication. METHODS Domestic pigs (n = 11; weight: 25 +/- 3 kg) were anesthetized and had one stent per coronary artery implanted: one stainless steel (Tecnic), one cobalt-chromium (Chrono), and one tacrolimus-eluting stent (Janus), all of them being Carbofilm-coated (Sorin). One, three, or seven days postprocedure, the pigs were sacrificed, the hearts explanted, and longitudinal sections examined by surface electron microscopy to quantify the percentage of the strut endothelialized over the branches and in the total surface. RESULTS Forty-four side branches (25 stents) that had stent struts over their origin were evaluated. Different patterns of endothelialization were observed, from the total absence to the complete endothelialization. There were no significant differences in relation to type of stent or to the artery treated. The predictors of higher percentage of endothelialization were the ratio of metal to branch diameter (P = 0.04) and better endothelialization in the rest of the stent (P = 0.0002), only this parameter maintaining significant correlation (P = 0.03) in multivariate analysis. CONCLUSIONS Carbofilm-coated stent struts located over the origin of side branches follow the pattern of endothelialization for the rest of the stent, even in the case of tacrolimus-eluting stent.

Louping ill in goats, Spain, 2011.

Although louping ill affects mainly sheep, a 2011 outbreak in northern Spain occurred among goats. Histopathologic lesions and molecular genetics identified a new strain of louping ill virus, 94% identical to the strain from Britain. Surveillance is needed to minimize risk to domestic and wildlife species and humans.

Impact of maedi-visna in intensively managed dairy sheep.

Maedi-visna (MV) is a slow lentiviral disease of sheep that has a significant economic impact in many sheep-producing regions although there remains a paucity of data relating to actual production losses resulting from this disease. The objective of this study was to evaluate direct losses, through death or culling, from two dairy sheep flocks with high seroprevalences of infection over a 2 year period. Maedi-visna was found, either alone or in combination with other diseases, to be the most common disease diagnosed in these sheep, and the major cause of direct animal losses in the two flocks. Moderate to severe lesions associated with MV were found in 52% and 80% of the sheep, respectively, affecting the lungs, brain and/or mammary glands. Despite the similarity of the two flocks under study in terms of breed, number of animals, geographical proximity, and inter-change of rams, a striking difference was observed regarding the clinical presentation of the disease: in one flock the respiratory form was dominant while in the other 70% of animals died or were culled because of neurological signs.

Clostridium sordellii in a Brown Bear (Ursus arctos) from Spain

Abstract Clostridium sordellii is found in the environment and occasionally in animal (including human) intestines and may cause myonecrosis and large outbreaks of enterotoxemia. A few cases of fatal clostridial infection in bears (Ursus spp.) have been described worldwide but none attributed to C. sordellii. We describe a fatal case of septicemia caused by C. sordellii in an illegally trapped brown bear (Ursus arctos). At necropsy, acute gangrenous myositis was the primary lesion. Serohemorrhagic edema was observed in the abdominal cavity, thorax, pericardium, and skeletal muscle, mostly affecting femoral, humeral, and scapular muscles. Hemorrhage was observed in the heart, skeletal muscles, stomach, and intestine. Liver, spleen, and kidney appeared with loss of consistency, hemorrhages, and edema. Microscopically, primary lesions were in skeletal muscle, stomach, and small intestine, with gram-positive, clostridial-like bacilli. Biochemical and molecular tests identified C. sordellii in cultures from liver, muscle, and intestine. Sequences showed a homology of >99% with the 16S rRNA gene sequence of C. sordellii. The severity of effects of the C. sordellii infection reveal the importance of this pathogen as a wildlife health risk with conservation concerns, as well as the need to consider possible infection with this pathogen in management actions involving immobilization, stress, or severe muscular activity of wild brown bears.

Identification and characterization of a novel tick-borne flavivirus subtype in goats (Capra hircus) in Spain

In 2011, a neurological disease was reported in a herd of goats (Capra hircus) in Asturias, Spain. Initial sequencing identified the causative agent as louping ill virus (LIV). Subsequently, with the application of whole genome sequencing and phylogenetic analysis, empirical data demonstrates that the LIV-like virus detected is significantly divergent from LIV and Spanish sheep encephalitis virus (SSEV). This virus encoded an amino acid sequence motif at the site of a previously identified marker for differentiating tick-borne flaviviruses that was shared with a virus previously isolated in Ireland in 1968. The significance of these observations reflects the diversity of tick-borne flaviviruses in Europe. These data also contribute to our knowledge of the evolution of tick-borne flaviviruses and could reflect the movement of viruses throughout Europe. Based on these observations, the proposed name for this virus is Spanish goat encephalitis virus (SGEV), to distinguish it from SSEV.

Perivascular inflammatory cells in ovine Visna/maedi encephalitis and their possible role in virus infection and lesion progression

We examined the distribution in the perivascular spaces of Visna/maedi antigen, T cells (CD3+, CD4+ and CD8+), B cells and macrophages by immunohistochemistry in 22 natural cases of Visna/maedi encephalitis. Sheep showed lymphocytic or histiocytic lesions. In mild lymphocytic lesions, the viral antigen was detected in perivascular cuffs where CD8+ T cells predominated, but in severe lymphocytic lesions, sparse antigen was identified, and CD8+/CD4+ T cells appeared in a similar proportion in multilayer perivascular sleeves. In histiocytic lesions, vessels were surrounded by macrophages with abundant viral antigen, with CD8+/CD4+ T cells and B cells in the periphery. These results could reflect different stages of virus neuroinvasion and clarify the neuropathogenesis of Visna/maedi encephalitis.

Canine adenovirus type 1 (CAdV-1) in free-ranging European brown bear (Ursus arctos arctos): A threat for Cantabrian population?

Canine adenovirus type 1 (CAdV-1) is responsible for infectious canine hepatitis. The disease has been described in captive American black bear (Ursus americanus) and European brown bear (Ursus arctos arctos), with just one recently reported case in a cub of a free-ranging brown bear (Ursus arctos horribilis) from Alaska. The aim of this work is to summarize findings related to presence and associated mortality of CAdV-1 in 21 free-ranging Cantabrian brown bears (Ursus arctos arctos) submitted to necropsy in Asturias and Castilla y León (northwestern Spain) from 1998 to 2018. On the basis of the anatomopathological findings and laboratory results three free-ranging brown bears died due to infectious canine hepatitis, which is to our knowledge the first description of death due to this disease in free-ranging bears in Europe. Gross lesions consisted of petechial haemorrhages and congestion in different internal organs, haemorrhagic fluid in internal cavities, friable and yellowish liver and thickening of gall bladder. Microscopic lesions were observed mainly in liver, kidney and brain and consisted of multifocal necrosis of cells with presence of basophilic intranuclear inclusions. Immunohistochemical (IHC) and real-time polymerase chain reaction (qPCR) techniques were used to assess the presence of CAdV-1 in paraffin-embedded liver samples. Viral antigens were detected by IHC labelling within hepatocytes and Küppfer cells in the three animals. The presence of viral DNA was confirmed by qPCR in one of them. In order to evaluate the circulation of CAdV-1 in brown bears, a retrospective study was performed using both IHC and qPCR techniques in 11 and 12 additional brown bears, respectively. An extra brown bear was found positive by IHC. This study shows that CAdV-1 surveillance of brown bears and sympatric carnivores should be considered as major concern for the monitoring the population evolution throughout time in this endangered species.

Fate of the true-vacuum bubbles

We investigate the bounce solutions in vacuum decay problems. We show that it is possible to have a stable false vacuum in a potential that is unbounded from below.