TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS
Procedures were evaluated for the rapid and efficient purification of oocysts and esporocysts of Sarcocystis aucheniae (Sa) from the small intestine of experimentally infected dogs with 400 Sa macrocysts obtained from meat of naturally infected alpacas. The mucosa of the small intestine was removed. Samples were washed by centrifugation with PBS, homogenized and centrifuged. Large quantities of oocysts attached to the intestinal microvilli were obtained. In the first procedure, an ezimatic treatment with 5% tripsin was used resulting in 50% oocyst separation. In the second procedure samples were treated with 2.6% sodium hypoclorite solution achieving 80% separation. In the third procedure, samples were centrifuged with a discontinuous density gradient of potassium bromide rendering 99% oocyst separation. The later procedure allowed obtaining relevant quantities of purified Sa oocysts and without loss of viability.
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Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria
2013
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oai:ojs.csi.unmsm:article-25902020-03-31T00:44:43Z TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS TÉCNICAS DE AISLAMIENTO Y PURIFICACIÓN DE OOQUISTES DE Sarcocystis aucheniae A PARTIR DE INTESTINO DELGADO DE PERROS EXPERIMENTALMENTE INFECTADOS Zacarías S., Fiorella Sam T., Rosa Ramos D., Daphne Lucas A., Orlando Lucas L., Juan Ooquistes Sarcocystis bromuro de potasio purificación oocysts Sarcocystis potassium bromide purification Procedures were evaluated for the rapid and efficient purification of oocysts and esporocysts of Sarcocystis aucheniae (Sa) from the small intestine of experimentally infected dogs with 400 Sa macrocysts obtained from meat of naturally infected alpacas. The mucosa of the small intestine was removed. Samples were washed by centrifugation with PBS, homogenized and centrifuged. Large quantities of oocysts attached to the intestinal microvilli were obtained. In the first procedure, an ezimatic treatment with 5% tripsin was used resulting in 50% oocyst separation. In the second procedure samples were treated with 2.6% sodium hypoclorite solution achieving 80% separation. In the third procedure, samples were centrifuged with a discontinuous density gradient of potassium bromide rendering 99% oocyst separation. The later procedure allowed obtaining relevant quantities of purified Sa oocysts and without loss of viability. Se evalúan procedimientos para la rápida y eficiente purificación de ooquistes y eporoquistes de Sarcocystis aucheniae (Sa), a partir del intestino delgado de perros experimentalmente infectados vía oral con 400 macroquistes de Sa provenientes de carne de alpacas naturalmente infectadas. Se recolectó la mucosa del intestino delgado. Las muestras fueron lavadas por centrifugación con buffer salino fosfato y homogenizadas y centrifugadas. Se obtuvo grandes cantidades de ooquistes adheridos a las microvellosidades intestinales. En el primer procedimiento, los ooquistes recibieron un tratamiento enzimático con tripsina al 5% resultando 50% de separación. En el segundo fueron tratados con una solución de hipoclorito de sodio al 2.6% lográndose una separación parcial (80%). En el tercero fueron centrifugados con una gradiente de densidad discontinua de bromuro de potasio obteniéndose una separación de 99%. Este último procedimiento permitió la obtención de ooquistes de Sa en cantidades relevantes, purificados y sin pérdida de la viabilidad. Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria 2013-09-16 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article application/pdf https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/2590 10.15381/rivep.v24i3.2590 Revista de Investigaciones Veterinarias del Perú; Vol. 24 No. 3 (2013); 396-403 Revista de Investigaciones Veterinarias del Perú; Vol. 24 Núm. 3 (2013); 396-403 1682-3419 1609-9117 spa https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/2590/2267 Derechos de autor 2013 Fiorella Zacarías S., Rosa Sam T., Daphne Ramos D., Orlando Lucas A., Juan Lucas L. https://creativecommons.org/licenses/by-nc-sa/4.0 |
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Zacarías S., Fiorella Sam T., Rosa Ramos D., Daphne Lucas A., Orlando Lucas L., Juan |
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Zacarías S., Fiorella Sam T., Rosa Ramos D., Daphne Lucas A., Orlando Lucas L., Juan TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS |
| author_facet |
Zacarías S., Fiorella Sam T., Rosa Ramos D., Daphne Lucas A., Orlando Lucas L., Juan |
| author_sort |
Zacarías S., Fiorella |
| title |
TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS |
| title_short |
TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS |
| title_full |
TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS |
| title_fullStr |
TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS |
| title_full_unstemmed |
TECHNIQUES FOR THE ISOLATION AND PURIFICATION OF SARCOCYSTIS AUCHENIAE OOCYSTS FROM SMALL INTESTINE OF EXPERIMENTALLY INFECTED DOGS |
| title_sort |
techniques for the isolation and purification of sarcocystis aucheniae oocysts from small intestine of experimentally infected dogs |
| description |
Procedures were evaluated for the rapid and efficient purification of oocysts and esporocysts of Sarcocystis aucheniae (Sa) from the small intestine of experimentally infected dogs with 400 Sa macrocysts obtained from meat of naturally infected alpacas. The mucosa of the small intestine was removed. Samples were washed by centrifugation with PBS, homogenized and centrifuged. Large quantities of oocysts attached to the intestinal microvilli were obtained. In the first procedure, an ezimatic treatment with 5% tripsin was used resulting in 50% oocyst separation. In the second procedure samples were treated with 2.6% sodium hypoclorite solution achieving 80% separation. In the third procedure, samples were centrifuged with a discontinuous density gradient of potassium bromide rendering 99% oocyst separation. The later procedure allowed obtaining relevant quantities of purified Sa oocysts and without loss of viability. |
| publisher |
Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria |
| publishDate |
2013 |
| url |
https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/2590 |
| work_keys_str_mv |
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