First report of a primitive neuroectodermal tumor of the bladder in a newborn.

Primitive neuroectodermal tumor (PNET) is part of the Ewing sarcoma family of tumors. The present case reports a primitive neuroectodermal tumor (PNET) of rare location in the bladder in a newborn. It was evaluated with prenatal ultrasound and postnatal tomography that revealed a mass in the posterior wall of the bladder. The patient underwent partial cystectomy with subsequent analysis of the surgical piece removed, the histopathological study indicated a tumor of mesenchymal origin, and immunohistochemical staining confirmed the diagnosis of PNET of the bladder. Satisfactory result and short-term follow-up.

Epithelial-Mesenchymal Transition in a Case of Metastatic Thyroid Carcinoma in a Brown Bear (Ursus arctos).

A 20-year-old male brown bear (Ursus arctos) with a 20 × 25 cm necrotic mass adjacent to the trachea was diagnosed as having an anaplastic thyroid carcinoma. Metastases were observed in the lungs and one adrenal gland and, histologically, these had anaplastic and follicular carcinoma patterns, respectively. E-cadherin labelling was observed in the adrenal mass only, while N-cadherin immunolabelling was detected in the thyroid gland and lung masses. Thyroid-specific markers (thyroid transcription factor-1, thyroglobulin) were expressed in the adrenal gland metastasis. This case illustrates an example of a primary epithelial-mesenchymal transition (EMT) enabling metastasis to distant organ sites, followed by a mesenchymal-epithelial transition within the adrenal gland microenvironment, allowing invasion and reacquisition of thyroid epithelial cell features. EMTs help to understand the phenomenon of carcinoma cell plasticity in enabling colonization and growth of metastases.

From the bluetongue vaccination campaigns in sheep to overimmunization and ovine ASIA syndrome.

The use of vaccines has proven to be very effective in controlling and eradicating infectious diseases, both in veterinary and human medicine; however, vaccines can be also the source of an array of problems caused by procedures such as overimmunization. Bluetongue, an orbiviral disease that affects ruminants, is best controlled by the use of inactivated vaccines. During the last years of the past decade, these vaccines were applied all over Europe to control the spreading of the disease, a goal that was accomplished; however, at the same time, several adverse effects related to the vaccination were reported. Especially in sheep, this vaccination campaign brought out a new cachectic and neurologic disease with harmful consequences for the ovine industry. This disease is now recognized as the ovine version of the autoimmune/inflammatory syndrome induced by adjuvants (ASIA syndrome) and poses an immense challenge in veterinary medicine, immunology, and vaccinology.

Small Ruminant Lentiviruses in Sheep: Pathology and Tropism of 2 Strains Using the Bone Marrow Route.

The objective of this work was to comparatively study the tissue tropism and the associated pathology of 2 autochthonous small ruminant lentivirus (SRLV) field strains using an experimental infection in sheep through the bone marrow. Fifteen male, SRLV-free lambs of the Rasa Aragonesa breed were inoculated with strain 697 (nervous tissue origin, animals A1-A6), with strain 496 (articular origin, animals B1-B6), or with uninfected culture medium (C1-C3). Clinical, serologic, and polymerase chain reaction (PCR) evaluations were performed periodically. Two lambs from each infected group and a control animal were euthanized at 134, 273, and 319 days postinfection. Tissues were analyzed by gross and histopathologic evaluation; immunohistochemistry for CD3, CD4, CD8, CD68, and FoxP3 cell markers; lung morphometric evaluation; and tissue proviral quantification by PCR. All infected animals became positive either by enzyme-linked immunosorbent assay and/or PCR, with group B lambs showing the highest serologic values and more consistently positive PCR reactions. Group A lambs showed representative lung lesions but only mild histopathologic changes in the central nervous system (CNS) or in carpal joints. Contrarily, group B lambs demonstrated intense carpal arthritis and interstitial pneumonia but an absence of lesions in the CNS. Proviral copies in tissues were detected only in group B lambs. Experimental infection with these SRLV strains indicates that strain 496 is more virulent than strain 697 and more prone to induce arthritis, whereas strain 697 is more likely to reproduce encephalitis in Rasa Aragonesa lambs. Host factors as well as viral factors are responsible for the final clinicopathologic picture during SRLV infections.

Small ruminant lentivirus infections and diseases.

Small ruminant lentiviruses include viruses with diverse genotypes that frequently cross the species barrier between sheep and goats and that display a great genetic variability. These characteristics stress the need to consider the whole host range and to perform local surveillance of the viruses to opt for optimum diagnostic tests, in order to establish control programmes. In the absence of effective vaccines, a comprehensive knowledge of the epidemiology of these infections is of major importance to limit their spread. This article intends to cover these aspects and to summarise information related to characteristics of the viruses, pathogenesis of the infection and description of the various syndromes produced, as well as the diagnostic tools available, the mechanisms involved in transmission of the pathogens and, finally, the control strategies that have been designed until now, with remarks on the drawbacks and the advantages of each one. We conclude that there are many variables influencing the expected cost and benefits of control programs that must be evaluated, in order to put into practice measures that might lead to control of these infections.

Small ruminant lentivirus-induced arthritis: clinicopathologic findings in sheep infected by a highly replicative SRLV B2 genotype.

We describe the clinicopathologic features of an arthritis outbreak in sheep induced by small ruminant lentivirus (SRLV), linked to the presence of a new SRLV isolate phylogenetically assigned to caprine arthritis encephalitis virus-like subgroup B2. Thirteen SRLV seropositive Rasa Aragonesa adult ewes were selected from 5 SRLV highly infected flocks (mean seroprevalence, 90.7%) for presenting uni- or bilateral chronic arthritis in the carpal joint. A complete study was performed, including symptomatology, histopathology, immunocytochemistry, immunohistochemistry, in situ hybridization, and microbiology. The carpus was the joint almost exclusively affected, with 10 sheep (76%) showing a moderate increase in carpal joint size (diameter range, 18-20 cm; normal range, 15-16 cm) without signs of locomotion problems and with 3 ewes (23%) showing severe inflammation with marked increase in diameter (21-24 cm), pain at palpation, and abnormal standing position. Grossly, chronic proliferative arthritis was observed in affected joints characterized by an increased thickness of the synovial capsule and synovial membrane proliferation. Microscopically, synovial membrane inflammation and proliferation and hyperplasia of synoviocytes were observed. More positive cases of SLRV infection were detected by immunocytochemistry of articular fluid than of bronchoalveolar lavage fluid. Immunohistochemistry and in situ hybridization also detected positive cells in the subsynovial connective tissue, lung, mediastinal lymph node, mammary gland, and mammary lymph node. All animals were negative for the presence of Mycoplasma or other bacteria in the articular space. The present outbreak likely represents an adaptation of a caprine virus to sheep. Our results underline the importance of the arthritis induced by SRLV in sheep, a clinical form that might be underestimated.

Successful Visna/maedi control in a highly infected ovine dairy flock using serologic segregation and management strategies.

A control system for Visna/maedi virus (VMV) infection based on serologic segregation and management strategies was applied in an infected Spanish dairy Manchega breed sheep flock (n=670) that was affected by a severe respiratory process associated to VMV. The control started in 2004 and consisted on the serological study of animals, segregation in two different flocks (seropositive and seronegative), separate management of flocks, selection of young female lambs for replacement only from seronegative ewes offspring, immediate removal of seropositive animals detected in the seronegative flock and a management tending toward the reduction and final culling of the seropositive flock. The serological control was repeated yearly or twice a year, approximately. Initial VMV seroprevalence of the undivided flock was 66.4% (January 2004) that descended to 47.3%, 12.8%, 2.2% and 0.2% between July 2004 and May 2006. Residual seroprevalence fluctuated slightly thereafter with a peak of 2.2% in April 2008. After segregation, number of animals in the seronegative flock was 378 that descended to 323 in October 2005. Since then, this number has increased steadily reaching 650 sheep in December 2011. The seropositive flock was progressively reduced by culling until its total disappearance in June 2010. This work presents the detailed results obtained in the control strategy against VMV in a single dairy sheep flock by implementing a segregation system based on serologic testing. The system is highly successful, as it reduces to residual levels VMV infection in about two years without the need of culling a high number of animals, as required by other methods. Moreover, the original size flock was been recovered within 8 years and has led to a subjective improvement of animal health and welfare in the flock. The residual seroprevalence could be eliminated at this stage by applying more sensitive molecular or other serological techniques to reach eradication.

An insight into a combination of ELISA strategies to diagnose small ruminant lentivirus infections.

A single broadly reactive standard ELISA is commonly applied to control small ruminant lentivirus (SRLV) spread, but type specific ELISA strategies are gaining interest in areas with highly prevalent and heterogeneous SRLV infections. Short (15-residue) synthetic peptides (n=60) were designed in this study using deduced amino acid sequence profiles of SRLV circulating in sheep from North Central Spain and SRLV described previously. The corresponding ELISAs and two standard ELISAs were employed to analyze sera from sheep flocks either controlled or infected with different SRLV genotypes. Two outbreaks, showing SRLV-induced arthritis (genotype B2) and encephalitis (genotype A), were represented among the infected flocks. The ELISA results revealed that none of the assays detected all the infected animals in the global population analyzed, the assay performance varying according to the genetic type of the strain circulating in the area and the test antigen. Five of the six highly reactive (57-62%) single peptide ELISAs were further assessed, revealing that the ELISA based on peptide 98M (type A ENV-SU5, consensus from the neurological outbreak) detected positives in the majority of the type-A specific sera tested (Se: 86%; Sp: 98%) and not in the arthritic type B outbreak. ENV-TM ELISAs based on peptides 126M1 (Se: 82%; Sp: 95%) and 126M2 0,65 0.77 (Se: 68%; Sp: 88%) detected preferentially caprine arthritis encephalitis (CAEV, type B) and visna/maedi (VMV, type A) virus infections respectively, which may help to perform a preliminary CAEV vs. VMV-like typing of the flock. The use of particular peptide ELISAs and standard tests individually or combined may be useful in the different areas under study, to determine disease progression, diagnose/type infection and prevent its spread.

Ovine TRIM5α can restrict visna/maedi virus.

The restrictive properties of tripartite motif-containing 5 alpha (TRIM5α) from small ruminant species have not been explored. Here, we identify highly similar TRIM5α sequences in sheep and goats. Cells transduced with ovine TRIM5α effectively restricted the lentivirus visna/maedi virus DNA synthesis. Proteasome inhibition in cells transduced with ovine TRIM5α restored restricted viral DNA synthesis, suggesting a conserved mechanism of restriction. Identification of TRIM5α active molecular species may open new prophylactic strategies against lentiviral infections.

Visna/Maedi virus genetic characterization and serological diagnosis of infection in sheep from a neurological outbreak.

An extensive outbreak characterized by the appearance of neurological symptoms in small ruminant lentivirus (SRLV) infected sheep has been identified in Spain, but the genetic characteristics of the strain involved and differential diagnostic tools for this outbreak remain unexplored. In this work, 23 Visna-affected naturally infected animals from the outbreak, 11 arthritic animals (both groups presenting anti-Visna/Maedi virus serum antibodies), and 100 seronegative animals were used. Eight of the Visna-affected animals were further studied post-mortem by immunohistochemistry. All had lesions in spinal cord, being the most affected part of the central nervous system in six of them. A representative strain of the outbreak was isolated. Together with other proviral sequences from the outbreak the virus was assigned to genotype A2/A3. In vitro culture of the isolate revealed that viral production was slow/low in fibroblast-like cells but it was high in blood monocyte-derived macrophages. The long terminal repeat (LTR) of the viral genome of this isolate lacked an U3-duplication, but its promoter activity in fibroblast-like cells was normal compared to other strains. Thus, viral production could not be inferred from the LTR promoter activity in this isolate. Analysis of the viral immunodominant epitopes among SRLV sequences of the outbreak and other known sequences allowed the design of a synthetic SU peptide ELISA that detected the Visna affected animals, representing a tool of epidemiological interest to control viral spread of this highly pathogenic strain.

Novel group A rotavirus G8 P[1] as primary cause of an ovine diarrheic syndrome outbreak in weaned lambs.

Rotavirus is a worldwide major cause of diarrhea outbreaks in neonatal ruminants. An outbreak of ovine diarrheic syndrome (ODS) in 50-75 days-old lambs (weaned lambs) is described. Fecal immunochromatography and intestinal immunohistochemistry for rotavirus group A were performed. In addition, semi-nested multiplex RT-PCR for G and P rotavirus genotyping in combination with sequencing were performed, to support the diagnosis and identify the viral strain. A novel ovine rotavirus group A G8 P[1] strain was determined as the main cause of the ODS observed, whereas other pathogens were ruled out.

Visna/maedi virus serology in sheep: survey, risk factors and implementation of a successful control programme in Aragón (Spain).

A serological survey of Visna/maedi virus (VMV) infection involving 274,048 sheep from 554 flocks was undertaken during 2002-2007 in Aragón, North-East Spain. One hundred and two of these flocks enrolled in a VMV control programme to reduce seroprevalence by selecting replacement lambs from seronegative dams and gradual culling of seropositive sheep. Twenty-five flocks were also visited to collect flock management and housing data. All study flocks had seropositive animals and 52.8% of animals tested were seropositive. Among flocks that joined the control programme 66 adopted the proposed measures and reduced seroprevalence significantly by between 26.1% and 76.9% whereas the remaining 36 flocks did not apply the measures and seroprevalence significantly increased. Seroprevalence increased with flock size and the number of days the sheep were housed, and decreased with increasing weaning age and shed open area, suggesting a reduced risk of VMV infection in sheep associated with better ventilation. At the end of the period, 24 flocks were certified as VMV-controlled with a seroprevalence <5%, and seven as VMV-free with 0% seroprevalence. These are the first officially recognised VMV-free flocks in Spain and represent a nucleus of VMV-free replacement animals for other flocks. Moreover, they are evidence of the possibility of eliminating VMV infection without resorting to whole-flock segregation or culling of seropositive sheep.

Effects of housing on the incidence of visna/maedi virus infection in sheep flocks.

The incidence of seroconversion to visna/maedi virus (VMV) infection and its relationship with management and sheep building structure was investigated in 15 dairy sheep flocks in Spain during 3-7years. Incidence rates were 0.09 per sheep-year at risk in semi-intensive Latxa flocks and 0.44 per sheep-year at risk in intensive Assaf flocks and was greatest for the one year old Assaf replacement flock. Separate multivariable models developed for replacement and adult flocks indicated that in both cases seroconversion was strongly associated to direct contact exposure to infected sheep and to being born to a seropositive dam. The latter effect was independent of the mode of rearing preweaning and the risk of seroconversion was similar for sheep fed colostrum and milk from a seropositive or a seronegative dam. These results are further evidence of the efficiency of horizontal VMV transmission by close contact between sheep and also suggest a inheritable component of susceptibility and resistance to infection. In contrast, indirect aerogenous contact with seropositive sheep was not associated with seroconversion as evidenced in replacement sheep housed in separate pens in the same building as adult infected sheep for one year. Consequently, VMV may not be efficiently airborne over short distances and this is important for control of infection. Moreover, there was no relationship between seroconversion and shed open areas. The latter could be related to having examined few flocks in which high infection prevalence dominated the transmission process while ventilation, may depend on a variety of unrecorded factors whose relationship to infection needs to be further investigated.

Educación nutricional en el control de la obesidad mediante intervenciones no tradicionales / Controlling obesity through nutrition education using unconventional projects

Introducción: la radio ha demostrado ser un apoyo importante en la generación de proyectos con base comunitaria persiguen la búsqueda y aplicación de mecanismos viábles de participación y autogestión. El presente trabajo pretende evaluar la efectividad de la intervención en la comunidad con impacto en las creencias, opiniones y hábitos alimentarios de las personas adultas utilizando la radio como elemento central de la metodología ECCA. Material y método: Se utilizói un diseño evaluativo cuasiexperimental pretest-postest con un solo grupo según clasificación de Campbell. La población (universo) estuvo conformada por todos sujetos (personas de más de 18 años) que se matricularon en los (2) cursos de obesidad a lo largo del 2008 (N=2.888). La muestra, no intencional y no probabilística, estuvo conformada por (n=471) sujetos distribuídos por todas las islas. Se elaboró un curso de prevención de la obesidad (en niño, en la mujer, en personas mayores, etc.), de 12 unidades. Se elaboró un cuestionario al comienzo del curso, en el momento de la matricula y un mes despues del curso. Para comparar los porcentajes correspondientes a las encuestas inicial y final se utilizó el test de McNemar y para la comparación de medias el t-test para datos emparejados (paired t-test). Resultados: La actividad física, desayuno diario, e IMC se modificaron de forma significativa; también ayudo a sentirse en forma o a controlar el peso; aumento la actividad ffsica en pareja, disminuyo el tiempo delante del televisor, mejoraron algunas creencias sobre la alimentación, mejoraran algunas hábitos en cuanto a la calidad y cantidad del desayuno antes de ir a trabajar, mejoran las comidas saludables tanto al mediodía como en la noche, mejora el equilibrio nutricional en las dietas habituales así como la disciplina en los horarios de comida en la familia. Conclusiones: La educación para la salud en el ámbito de la pnvención de la obesidad a través de la radio, siguiendo la metodología ECCA, generó cambios positivos en cuanto a los hábitos personales, alimentarios y dieteticos no solo a nivel personal sino también familiar(AU)

Background: Radio has proven to be an important support in the generation of community based projects aimed at searching and applying viable mechanisms of participation and self-management. This article attempts to evaluate the project effectiveness in the community; the impact on adult´s beliefs, opinions and eating habits, using the radio as the central element in the ECCA methodology. Method: A quasi-experimental pre-test – post-test evaluation design was used with a single group, according to Campbell´s classification. The population (universe) was made up of all subjects (people over 18 years of age) that were registered in the courses (2) on obesity throught 2008 (N=2.888). The sample, unintentional and not probabilistic, was made up of (n=471) subjects, distributed in all the islands. A twelve unit course on obesity prevention (in children, women, senior citizenx, etc.) was created. A questionnaire on beliefs, opinions and on personal and eating habits was drafted to be self-administered before the start of the course during registration and 1 month after ending the course. In order to compare the initial and final survey percentages the MxNemar test was used and the paired t-test for the averages. When the corresponding p-value was equal of inferior to 0.05 the hypothesis contrast was considered statistically significant. Results: Physical activity, daily breakfast, and BMI were modified significantly; it also contributed to improving the feeling of being fit and controlling weight; it increased the couple´s physical activity, reduced time watching TV, improved some eating related belief, improved the quality and quantity of before work breakfast, improved mid-day and night-time healthful meals, improved nutritional balance in routine diets, as well discipline in family mealtime schedules. Conclusions: Health education in the field of obesity prevention using the radio, following the ECCA methodology generated positive changes in personal nutritional and diet habits, not only at the personal but also on the family level(AU)

Hábitos alimentarios y estilos de vida en trabajadores de servicio sometidos a turnicidad del Ayuntamiento de Las Palmas de Gran Canaria / Food habits and life styles of shift workers in the Las Palmas de Gran Canaria town council

Fundamentos: El trabajo a turnos induce hábitos de vida anómalos, lo cual se asocia a mayor morbilidad en los trabajadores. En este estudio se realiza una valoración de los hábitos nutricionales durante la semana laboral según el turno de trabajo en un grupo de trabajadores del Ayuntamiento de Las Palmas de Gran Canaria. Métodos: La población de estudio estuvo compuesta por 255 peones de limpieza viaria, 28 recolectores de recogida de residuos sólidos y 18 trabajadores del servicio de limpieza; sometidos a turnos de trabajo (mañanas, tardes, noches y rotatorio), que representan el 87% del colectivo municipal en estudio. El número de encuestas estudiadas ha sido 262. Se determinó la ingesta dietética mediante cuestionario de frecuencia de consumo de alimentos autoadministrado con ochenta y dos alimentos, junto a una valoración del turno de trabajo, estilos de vida, actividad física, hábitos de consumo de las comidas principales, actividad física, hábitos tabáquicos, formas de vivir, categoría profesional, edad y sexo. Los datos fueron analizados mediante el paquete estadístico SPSS Base v. 15.0 para entorno Windows. Se calculó el consumo medio de alimentos (gr. /día) según turnicidad. La proporción de la distribución de las variables categóricas se realizó mediante la prueba de Chi-cuadrado. Resultados: Se observa que sólo un 34% desayunan en casa al levantarse, y un 76% hacen una comida a media mañana. Los postres en la comida son fruta (50%) y pasteles o lácteos (50%), y en la cena son mayoritariamente lácteos y pasteles (70%) y en menor proporción fruta (30%). Existen pocas diferencias en función del turno, a excepción del hábito de comer entre horas o snacks. Un 30% de la población cena un vaso de leche con galletas. Conclusiones: El hábito de tomar determinados alimentos tiene relación con el turno laboral. Es necesario mejorar localidad nutricional de la totalidad del colectivo estudiado mejorando el desayuno y la cena, y disminuyendo las comidas entre horas y los picoteos. Educar tanto al colectivo en estudio como a todos los trabajadores sometidos a turnicidad, mediante estudios previos al comenzar los cambios de turnos y con Guías adaptadas a cada turno de trabajo es una necesidad a corto plazo(AU)

Background. Shift work induces anomalous life styles, associated to increased morbidity in the workers. In this study and assessment of nutritional habits is made (breakfast, meals, outer suppers and exits) during the labour week according to the work turn in a group of workers of Las Palmas de Gran Canaria Town Council. Methods: Descriptive observational study cross-sectional character. The study population was composed by 255 shiftworkers of road cleaning, 28 workers collecting of solid residues and 18 workers of the service of cleaning; put under work turns (mornings, afternoons, nights and rotator), that represent 87% of the municipal group in study. The number of studied surveys has been 262. Dietary intakes was assessed by a self registered food frequency questionnaire with eighty and two foods, next to a valuation of the work turn (shift workers), styles of life, physical activity, habits of consumption of the main meals, physical activity, tobacco habits, forms to live, professional category, age and sex. The data were analyzed by means of statistical package SPSS Base v.15.0 for surroundings Windows. The food mean consumption calculated (gr./day) according to shift work. The proportion of the distribution of the categorical variables was made by means of the test of Chi-square. Results: In this group it is observed that only a 34% have breakfast in house when rising, and a 76% make a food to mid-morning. The desserts in the food are milky fruit (50%) and pies (70%) and in smaller proportion fruit (30%). Few differences based on the turn exist, with the exception of the habit to eat between hours or snacks. A 30% of the population have supper a milk glass with cakes. Conclusions: the habit to take certain foods has relation with the labour turn. It is necessary to improve nutritional quality of the totality of the studied group being improved the breakfast and the supper, and diminishing the meals between hours and the snacks. To as much educate to the group in study like a all the submissive shift workers, by means of previous studies when beginning the changes of turns and with Guides adapted to each turn of work is a necessity in the short term(AU)

Use of B7 costimulatory molecules as adjuvants in a prime-boost vaccination against Visna/Maedi ovine lentivirus.

RNA transcripts of the B7 family molecule (CD80) are diminished in blood leukocytes from animals clinically affected with Visna/Maedi virus (VMV) infection. This work investigates whether the use of B7 genes enhances immune responses and protection in immunization-challenge approaches. Sheep were primed by particle-mediated epidermal bombardment with VMV gag and env gene recombinant plasmids together with plasmids encoding both CD80 and CD86 or CD80 alone, boosted with gag and env gene recombinant modified vaccinia Ankara virus and challenged intratracheally with VMV. Immunization in the presence of one or both of the B7 genes resulted in CD4+ T cell activation and antibody production (before and after challenge, respectively), but only immunization with CD80 and CD86 genes together, and not CD80 alone, resulted in a reduced number of infected animals and increased early transient cytotoxic T lymphocytes (CTL) responses. Post-mortem analysis showed an immune activation of lymphoid tissue in challenge-target organs in those animals that had received B7 genes compared to unvaccinated animals. Thus, the inclusion of B7 genes helped to enhance early cellular responses and protection (diminished proportion of infected animals) against VMV infection.

Phylogenetic analysis of SRLV sequences from an arthritic sheep outbreak demonstrates the introduction of CAEV-like viruses among Spanish sheep.

Small ruminant lentiviruses (SRLVs) cause different clinical forms of disease in sheep and goats. So far in Spain, Maedi visna virus-like (MVV-like) sequences have been found in both species, and the arthritic SRLV disease has never been found in sheep until a recent outbreak. Knowing that arthritis is common in goats, it was of interest to determine if the genetic type of the virus involved in the sheep arthritis outbreak was caprine arthritis encephalitis virus-like (CAEV-like) rather than MVV-like. Alignment and phylogenetic analyses on nucleotide and deduced amino acid sequences from SRLV of this outbreak, allowed a B2 genetic subgroup assignment of these SRLV, compatible with a correspondence between the virus genetic type and the disease form. Furthermore, an isolate was obtained from the arthritic outbreak, its full genome was CAEV-like but the pol integrase region was MVV-like. Although its LTR lacked a U3 repeat sequence and had a deletion in the R region, which has been proposed to reduce viral replication rate, its phenotype in sheep skin fibroblast cultures was rapid/high, thus it appeared to have adapted to sheep cells. This outbreak study represents the first report on CAEV-like genetic findings and complete genome analysis among Spanish small ruminants.

AA amyloidosis induced in sheep principally affects the gastrointestinal tract.

AA amyloidosis was initiated experimentally in adult sheep by induction of gangrenous pneumonia, an inflammatory process known to be associated with amyloid formation. A vegetable fragment contaminated with rumen content was instilled into the lungs of 4 experimental animals. A fifth animal was not inoculated and served as control. The animals were examined daily and blood and urine were sampled biweekly post-inoculation. One sheep was killed 18 days post-inoculation (dpi), another 49dpi, and the remaining two (as well as the control animal) 63dpi. Respiratory signs, diarrhoea and/or soft, unformed stool were observed in all inoculated sheep. All experimental animals developed gangrenous pneumonia with hypoalbuminaemia and hypergammaglobulinaemia, and elevated urinary protein, creatinine, gamma glutamyl transferase and ss-glucuronidase. Amyloid deposition was most pronounced in the gastrointestinal tract and was evident from 18dpi. Amyloid was present from the tongue to the rectum, but was most prominent in the duodenum where the deposits disrupted the normal mucosal architecture. Other body organs had only mild amyloid deposition. Immunohistochemistry confirmed that the deposits were AA amyloid. These findings suggest that the gastrointestinal tract is the main target organ for AA amyloid deposition in sheep. The observations in this experimental model must now be confirmed in animals with spontaneously arising AA amyloidosis.

Systemic DNA immunization against ovine lentivirus using particle-mediated epidermal delivery and modified vaccinia Ankara encoding the gag and/or env genes.

To determine whether systemic immunization with plasmid DNA and virus vector against visna/maedi virus (VMV) would induce protective immune responses, sheep were immunized with VMV gag and/or env sequences using particle-mediated epidermal bombardment and injection of recombinant modified vaccinia Ankara. The results showed that immunization induced both humoral and cell-mediated responses prior to and after virus challenge. The vaccination protocol did not prevent infection, but immunization with the gag gene or a combination of gag and env genes resulted in significantly reduced provirus loads in blood and mediastinal lymph node, respectively. Provirus loads in lung and draining lymph node were unaffected, but p25 expression was undetectable in lungs of animals immunized with a combination of gag and env genes. Analysis of target tissues for lesions at post-mortem showed that immunization with the env gene caused a significant increase in lesion score, while the gag gene or a combination of gag and env genes had no effect. Inclusion of the ovine interferon-gamma gene in the initial priming mixture had minimal effect on immune responses, provirus load, or lesion development, although it resulted in a decreased p25 expression in the lung. The results thus show that systemic immunization with gag or a combination of gag and env genes reduces provirus load in blood and lymphoid tissue, respectively whereas env immunization has no effect on provirus load but increased lesion development.