Manuel Pizarro

Discovery of an Ebolavirus-Like Filovirus in Europe

Filoviruses, amongst the most lethal of primate pathogens, have only been reported as natural infections in sub-Saharan Africa and the Philippines. Infections of bats with the ebolaviruses and marburgviruses do not appear to be associated with disease. Here we report identification in dead insectivorous bats of a genetically distinct filovirus, provisionally named Lloviu virus, after the site of detection, Cueva del Lloviu, in Spain.

Expression of αv, α4, α5 and β3 integrin subunits, fibronectin and vitronectin in goat peri-implantation

Abstract Integrins are glycoprotein heterodimers located in the cell membranes that stimulate intercellular adhesion and act as extracellular matrix (ECM) protein receptors. Although integrins have been detected in the implantation sites of various species, little is known about their participation in ruminant non-invasive placentation. The objective of this study was the detection of α v , α 4 , α 5 , β 1 and β 3 integrin subunits and of two of their ligands, fibronectin and vitronectin, to determine their participation in the caprine peri-implantation process. On Day 21 post-coitum (pc), endometrial epithelium and trophoblastic cells showed an intense α v and β 3 integrin subunits expression and moderate staining for α 4 and α 5 . On Day 23 pc, integrin expression decreased noticeably and only a weak staining of α 4 and β 3 integrin subunits were observed. No β 1 integrin subunit expression was detected on either of the days studied. Fibronectin (FN) expression in trophectodermic and endometrial epithelium was weak or moderate on the days studied while vitronectin (VN) expression in the same tissues was moderate or strong on Day 21 pc but decreased on Day 23 pc. These results suggest that α v β 3 integrin, α 4 and α 5 subunits, VN and FN are expressed in caprine endometrium and blastocyst and may play a role in the cascade of the implantation process.

Outbreak of avian tuberculosis in 48-week-old commercial layer hen flock.

SUMMARY. This report describes an outbreak of tuberculosis in a flock of 38,500 48-wk-old layer hens. Clinical characteristics of the process included a gradual drop in egg production and feed intake, as well as an increased mortality rate. Two well-defined clinical features were observed. On the one hand, a number of birds displayed good body condition and continued to lay but presented granulomatous nodular lesions, particularly in the infraorbital sinus, liver, and intestine. In contrast, other hens were emaciated and presented granulomatous lesions of various sizes throughout the internal organs. The lesions observed in the various organs of all the affected birds consisted of granulomas containing acid/alcohol-fast bacilli. The presence of Mycobacterium avium was confirmed through polymerase chain reaction techniques. This report describes the epidemiologic and histopathologic characteristics of the outbreak.

Characterization in vitro and in vivo of a pandemic H1N1 influenza virus from a fatal case.

Pandemic 2009 H1N1 (pH1N1) influenza viruses caused mild symptoms in most infected patients. However, a greater rate of severe disease was observed in healthy young adults and children without co-morbid conditions. Here we tested whether influenza strains displaying differential virulence could be present among circulating pH1N1 viruses. The biological properties and the genotype of viruses isolated from a patient showing mild disease (M) or from a fatal case (F), both without known co-morbid conditions were compared in vitro and in vivo. The F virus presented faster growth kinetics and stronger induction of cytokines than M virus in human alveolar lung epithelial cells. In the murine model in vivo, the F virus showed a stronger morbidity and mortality than M virus. Remarkably, a higher proportion of mice presenting infectious virus in the hearts, was found in F virus-infected animals. Altogether, the data indicate that strains of pH1N1 virus with enhanced pathogenicity circulated during the 2009 pandemic. In addition, examination of chemokine receptor 5 (CCR5) genotype, recently reported as involved in severe influenza virus disease, revealed that the F virus-infected patient was homozygous for the deleted form of CCR5 receptor (CCR5Δ32).

Caprine testicular hypoplasia associated with sexual reversion decreases the expression of insulin-like growth factor II (IGF-II) mRNA in testes.

The genetic expression of insulin-like growth factor II (IGF-II) mRNA was studied in healthy adult testes and in hypoplastic testes of polled Murciano-Granadina goats by means of in situ hybridization. A positive reaction in spermatogonia, pachytene spermatocytes and a few peritubular myoid cells was observed using the ovine antisense oligonucleotide in healthy testes. The hypoplastic testes displayed a loss of germinal epithelium and a slight thickening of the basement membranes. A limited number of immature germinal cells displayed a lesser hybridization reaction, while the expression of IGF-II mRNA observed in the peritubular myoid cells was similar to that seen in healthy testes. In hypoplastic testes, IGF-II mRNA expression within germinal cells decreased with increasing hypoplasia within the seminiferous epithelium and there was no hybridization within the tubules in cases of severely disrupted spermatogenesis. These results suggest that testicular hypoplasia is associated with changes in the expression of IGF-II mRNA.

Ulcerative Enteritis (Quail Disease) in Lories

Abstract Ulcerative enteritis is found in a wide range of avian hosts but has not been described in psittacine birds. This case report describes ulcerative enteritis in four lories (two Trichoglossus sp. and two Eos sp.) that were found dead without any previous sign of disease. Macroscopically, all four birds showed good body condition. The only remarkable finding was a moderate dilatation of the small intestine with the presence of multiple yellow foci. Histologically, multiple ulcers extended into the submucosa and were filled with necrotic debris; bacteria and fibrin were observed in the intestinal mucosa. The liver and spleen exhibited a multifocal fibrinoid necrosis associated with a very moderate inflammatory reaction. Microbiological isolation revealed colonies of Clostridium colinum and Clostridium perfringens in the intestinal tract of the investigated birds.

Filariasis (Pelecitus sp.) in the Cervical Subcutaneous Tissue of a Pigeon with Trichomoniasis

Filariasis due to Pelecitus was found in the subcutaneous tissue of the neck of a domestic pigeon from Spain that died from trichomoniasis. Macroscopically, filariae were observed in the congested and hemorrhagic cervical connective tissue. Microscopically, a particular distribution of parasites with no inflammatory infiltrates was observed. Adult nematodes were located in peritracheal connective tissue and in the deep zone of the subcutaneous tissue. Microfilariae were mainly located in the superficial zone of the subcutaneous tissue. The only other changes observed were a moderate catarrhal tracheitis and congestion of the viscera.

Skin and abdominal fascia melanization in broiler chickens

Abstract Increased trimming and consequent carcass downgrade was experienced in a broiler flock. Condemned birds had gray to black pigmentation of the subcutaneous tissue and fat of the abdomen, and bluish pigmentation of the shanks. Histopathology revealed accumulation of melanin in these tissues. No significant bacteria were isolated. Accumulation of melanin in the skin and fascia is because of the cumulative and interactive effects of several genes. Additionally, pigmentation of abdominal fat has a high correlation with accumulation of melanin in the skin of the shanks. Although melanization of the skin and abdominal fascia is not harmful to people, it may cause severe economic losses to the producer.

Changes in eggshell thickness and ultrastructure in the Bearded Vulture (Gypaetus barbatus) Pyrenean population: A long-term analysis

The Spanish Bearded Vulture (Gypaetus barbatus) population has suffered from negative trends in a number of reproductive parameters that could jeopardize its long-term viability. From 1989 to 2012, 27 entire eggs and 63 eggshell fragments were collected from nests after breeding failure and/or fledging. Longer-term changes in eggshell thickness were made by examining 69 eggs collected in Spain from 1858 to 1911, and now held in European museums. Low levels of contamination with organochlorine pesticides and polychlorinated biphenyls were found in whole eggs and in conjunction with the high fertility rates observed in the field (66.7%) do not indicate a population suffering from the effects of organochlorine contamination. However, a decrease in the Ratcliffe Index and eggshell thickness were observed in eggs collected since 2001, increasingly so in samples post-2004, indicating an abrupt loss of egg quality. We found no significant relationship between organochlorine residues and eggshell in any of the variables measured. In contrast, we found a positive relationship between food availability and the Ratcliffe Index, eggshell thickness and eggshell surface area. A density-dependent explanation of reduced egg quality could arise from ecological constraints as the decrease of food resources. The impact of sudden changes in food availability due to sanitary regulations between 2006 and 2011 could be related with the loss of Bearded Vulture egg quality.

Capillaria contorta parasitism in red-legged partridge under farm conditions in Spain: histopathology of the upper digestive system

Capillaria contorta is a parasitic nematode of the oesophagus and crop of a number of domestic and wild birds. This study deals with three cases of capillariasis in the red-legged partridge. Infected individuals in two of these cases presented a history of weight loss, weakness, feather alterations and death, while morbidity and mortality were limited. In the third case, deaths occurred only after the birds were freed for repopulation purposes. Congestion of the crop was seen at necropsy. Histological examination revealed the presence of parasites and their eggs in the epithelium of the oesophagus and crop of every partridge. The worms displayed a characteristic distribution in all three cases: adults were found in the basal portion of the epithelium while the eggs were located in the middle. In one bird, parasites were observed in the anterior segment of the proventriculus. The fundamental epithelial lesion consisted of spongiosis of the oesophagus and crop. In the individual with proventricular parasitism, epidermoid metaplasia was also seen. The inflammatory reaction observed in the lamina propria was discrete and diffuse, except in those cases in which epithelial necrosis and bacterial colonies were detected.