Caracterización molecular de Trypanosoma vivax en ovinos naturalmente infectados en dos hatos de los municipios San Fernando y Biruaca estado Apure Venezuela / Molecular characterization of Trypanosoma vivax in naturally-infected sheep from two farms at San Fernando and Biruaca municipalities Apure state Venezuela

Este estudio tuvo como objetivo efectuar una caracterización molecular de Trypanosoma vivax en ovinos de dos hatos en los cuales estos rumiantes, conjuntamente con vacunos y búfalos de agua, comparten la misma área agroecológica, estableciendo el potencial papel de los ovinos como fuente de infección de tripanosomosis por T. vivax para los grandes rumiantes. La técnica de microcentrifugación capilar (TMC) fue usada para establecer el porcentaje de infecciones activas por tripanosomas existente en los animales evaluados. Se empleó un ensayo de reacción en cadena de la polimerasa (PCR) para confirmar la identificación de especie, mientras que un ensayo de PCR-RFLP (polimorfismo en la longitud de los fragmentos de restricción) permitió evaluar la variabilidad intraespecífica entre los aislados de T. vivax detectados en ovinos vs. aquellos provenientes de bóvidos (vacunos y búfalos de agua), colectados en la misma área de producción. De las 320 muestras de sangre de ovinos colectadas, la TMC detectó positividad en 11 (4,35 por ciento), lo que es de gran relevancia epidemiológica debido a la baja sensibilidad de esta metodología. Los resultados de PCR permitieron caracterizar a T. vivax como la especie presente en todas las infecciones activas detectadas. Todos los animales infectados mostraron un valor de hematocrito inferior (P<0,05) al registrado en animales no infectados (22,435 vs. 31,450). El ensayo de PCR-RFLP permitió observar la existencia de perfiles de restricción similares entre los aislados de T. vivax evaluados, sugiriéndose la ausencia de variación intraespecífica para el marcador molecular en estudio, independientemente del origen de hospedador del que provino la muestra (ovinos, vacunos, búfalos de agua). Estos resultados permiten sugerir que los aislados de T. vivax que infectan ovinos, vacunos y búfalos de agua en el área de estudio pudiesen estar estrechamente relacionados desde un punto de vista genético y, consecuentemente...

This study was made to achieve a molecular characterization of Trypanosoma vivax in two Venezuelan farms where both small ruminants (mainly ovines) and bovines (cattle and water buffaloes) share the same agroecological area. In addition, it was made to assess the role of sheep as source of T. vivax infection for cattle and buffalo herds. The microhematocrit centrifugation technique (MHC) was used to establish the percentage of current trypanosome infection. A PCR-based assay was used to confirm the species identification while a PCR-RFLP assay was used for studying intra-specific variation among T. vivax from sheep vs. those from other livestock from the same area. From 320 sheep blood samples, MHC detected 11 (4,35 percent) which is of remarkable epidemiological significance due to the low sensitivity of this method. Based on PCR results, T. vivax was characterized as the only species responsible for all sheep infections. All infected animals showed a lower packed cell volume value (P<0,05) when compared with the non-infected (22,435 vs. 31,450). The PCR-RFLP technique revealed similar profiles among T. vivax isolates suggesting a non intra-specific variation within the molecular marker amplified regardless the host (sheep water buffaloes or cattle). Thus, it was suggested that T. vivax infecting sheep, cattle, and buffaloes in the study area could be genetically closely related. These findings show that sheep may play an important role in the epidemiology of livestock trypanosomiasis in this area and they might be incorporated into therapeutic and preventive programs against livestock trypanosomiasis.

Trypanosoma vivax nos tecidos testicular e epididimário de ovinos experimentalmente infectados / Trypanosoma vivax in testicular and epidydimal tissues of experimentally infected sheep

Quatro ovinos machos, com cerca de 12 meses de idade (Ovinos 1-4), foram infectados por via intravenosa com aproximadamente 1,25x10(5) tripomastigotas de Trypanosoma vivax, outros quatro ovinos (Ovinos 5-8) destinaram-se ao grupo controle. Após a infecção, exames clínicos visando avaliar temperatura retal, freqüências cardíaca e respiratória e parasitemia foram realizados diariamente por 30 dias, tempo estabelecido para o término do experimento. A avaliação do hematócrito foi realizada a cada cinco dias. Ao final do período experimental, os animais foram castrados e os testículos e epidídimos submetidos ao exame anatomopatológico. Amostras destes órgãos dos Ovinos 1, 4 e 5 foram tomadas para a realização da reação em cadeia da polimerase (PCR). Os parâmetros clínicos (hipertermia, aumento das freqüências cardíaca e respiratória, aumento de volume dos linfonodos e palidez das mucosas) mantiveram-se para o grupo infectado acima dos valores mostrados pelo grupo controle durante todo o período experimental. A parasitemia foi observada a partir do 3º dia pós-infecção (dpi) com picos nos 6-10os dpi e nos 15-18os dpi. Os Ovinos 1 e 4 apresentaram, a partir do 25º dpi, anemia acentuada. Macroscopicamente, todos os testículos dos animais do grupo infectado apresentaram-se flácidos e com coloração pálida. Microscopicamente, observaram-se degeneração testicular moderada a acentuada, epididimite multifocal e hiperplasia do epitélio epididimário. A análise por PCR de T. vivax nos tecidos testicular e epididimário resultou em 100% de positividade para ovinos infectados experimentalmente. As lesões epididimárias e testiculares associadas à presença do parasita nesses órgãos, detectada por PCR, sugerem a participação do parasita no mecanismo etiopatogênico de danos reprodutivos.(AU)

Four adult sheep (number 1, 2, 3 and 4), all males, were inoculated intravenously with 1ml of blood containing 1.25x10(5) trypomastigotes of Trypanosoma vivax, and Sheep 5, 6, 7 and 8 were used as control. After infection, clinical exams considering rectal temperature, respiratory and cardiac frequencies, and parasitaemia were recorded daily for a 30-day experiment period. Blood samples were obtained for 5-day intervals to hematocrit analysis. At the end of the experimental period, the sheep were orquiectomized. Testes and epididymides from these animals were studied anatomopathologically. Samples from these tissues of Sheep 1, 4 and 5 were taken to polymerase chain reaction (PCR). Clinical parameters remained for the infected group above the values observed in the control group during the experimental period. Parasitaemia was observed on day 3 post-infection, and the highest values occurred between day 6 and 10, and day 15 and 18 post-infection. Sheep 1 and 4 showed severe anemia on day 25 post-infection. All sheep of the infected group showed flabby and palid testes. Histologically, moderate to severe testicular degeneration, multifocal epididymitis and hyperplasia of epididymal epithelium were observed. The result of T. vivax PCR analysis in the testes and epididymal tissues was positive in 100% of the samples of the experimentally infected sheep. Epididymal and testicular lesions associated with the presence of the parasite in these tissues, shown by PCR, suggest the participation of T. vivax in the pathophysiological mechanism of reproductive damage.(AU)

Uso de proteínas como alternativa diagnóstica para discriminar infecciones entre Trypanosoma vivax y Trypanosoma evansi / Use of proteins as diagnostic tool to discriminate between Trypanosoma vivax and Trypanosoma evansi infections

Tripomastigotes sanguícolas de Trypanosoma vivax y Trypanosoma evansi obtenidos de infecciones experimentales en ovejas y ratones respectivamente, fueron utilizados para purificar y caracterizar proteínas citosólicas mediante el método de partición con Tritón X-114. Los resultados revelan diferencias en los patrones proteicos entre las dos especies. Asimismo, las reacciones antigénicas mediante Western blot utilizando suero de animales naturalmente infectados, permitió discriminar las infecciones entre ambos parásitos. Se sugiere la utilización de esta metodología como una prueba diagnóstica confiable.

The parasitological and serological prevalence of tsetse-transmitted bovine trypanosomosis in the Eastern Caprivi (Caprivi District, Namibia).

Between August 1995 and June 1997 a survey to determine the distribution of tsetse-transmitted trypanosomosis was conducted in the Eastern Caprivi (Caprivi District, Namibia). A total of 1,481 adult cattle was examined at 33 sampling sites. Direct parasitological diagnostic tests were used and eluted blood spots were screened for the presence of anti-trypanosomal antibodies. Tsetse-transmitted trypanosomal infections were detected in 66 animals (4.5%) from 14 different locations. The parasitological and serological prevalence of trypanosomosis was highest in the Mamili area. Trypanosomosis was virtually absent in the Linyanti/Chobe area and the target barrier along the Kwando River had significantly reduced the prevalence of trypanosomosis in cattle grazing to the east of it. This suggests that anti-trypanosomal antibody prevalence data can be used to evaluate and monitor the effectiveness of tsetse control measures. Survey results suggest that in the Katima Mulilo area, trypanosomal infections were being acquired when cattle grazed along the Zambezi River. Moreover, survey results indicate that tsetse have not been able to establish themselves in the Katima Mulilo area. The parasitological prevalence in a herd and the respective prevalence of anti-trypanosomal antibodies was significantly correlated to the percentage of anaemic animals in that herd. Furthermore, the parasitological prevalence in a herd was positively correlated with the prevalence of anti-trypanosomal antibodies of that herd. It is concluded that the prevalence of anti-trypanosomal antibodies in a herd can be used as an additional indicator of the extent of infection in that particular herd.

On the interpretation of age-prevalence curves for trypanosome infections of tsetse flies.

Epidemiological models are used to analyse 8 published data sets reporting age-prevalence curves for trypanosome infections of the tsetse fly Glossina pallidipes. A model assuming a fixed maturation period and a rate of infection which is independent of fly age is adequate for Trypanosoma vivax-type infections, explaining 98% of observed variance in prevalence by site and age, allowing that the rate of infection may be site dependent. This model is not adequate for T. congolense-type infections and the fit can be improved by allowing (i) the rates of infection to decline with age (although non-teneral flies remain susceptible), (ii) a fraction of resistant flies, which may vary between sites, (iii) increased mortality of infected flies and (iv) variation in the maturation period. Models with these features can explain up to 97% of observed variance. Parameter estimates from published experimental data suggest that all may contribute in practice but that (i) and/or (ii) are likely to be the most important.