Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation

Autores
Decker Franco, Cecilia; Romero, Sandra Raquel; Ferrari, Alejandro; Schnittger, Leonhard; Florin-Christensen, Mónica
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The protozoon Sarcocystis aucheniae is the causative agent of South American camelid (SAC) sarcocystosis. Infections are characterized by the presence of cysts in muscles which are in size and appearance similar to rice grains. As consumption of insufficiently cooked infected meat produces gastroenteritis, cyst-containing SAC meat is confiscated by sanitary authorities or depreciated with serious economic consequences for SAC breeders. In this work, a duplex semi-nested PCR was designed to simultaneously detect parasite and llama DNA in host blood samples. Species-specific regions of S. aucheniae 18S rRNA gene and Lama glama 16S mitochondrial gene were amplified, yielding bands of 583 and 257 bp, respectively, and separated by gel electrophoresis. The method proved to be highly sensitive, with a detection limit lower than one parasite per milliliter blood, and the inclusion of primers to detect llama-specific DNA resulted useful as a methodological control. Blood samples collected from llamas of Argentina and Bolivia (n = 225) were analyzed using this method, and 18.7 % resulted positive for S. aucheniae. No correlation was found between PCR results and llama age, sex or the finding of macroscopic cysts in meat after slaughter. Lack of molecular detection in the blood of some llamas harboring macrocysts suggests that parasite circulation in the bloodstream after encystment is under the detection threshold of the test or even absent, while PCR positive results in cyst-infested animals suggests that prior exposure to the parasite does not impede subsequent infections. The described method can be useful to detect active foci of infection, to assess the effectiveness of parasiticide treatments, and for the surveillance and tracing of definitive hosts.
Instituto de Patobiología
Fil: Decker Franco, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Romero, Sandra Raquel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación y Desarrollo Tecnológico para la Agricultura Familiar Región NOA; Argentina
Fil: Ferrari, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; Argentina
Fil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
Heliyon 4 : e00928. (2018)
Materia
Llama
Sangre
PCR
Infestación
Infecciones por Protozoos
Blood
Infestation
Protozoal Infections
Sarcocystis Aucheniae
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/4436
Acceder
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oai_identifier_str oai:localhost:20.500.12123/4436
network_acronym_str INTADig
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network_name_str INTA Digital (INTA)
spelling Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestationDecker Franco, CeciliaRomero, Sandra RaquelFerrari, AlejandroSchnittger, LeonhardFlorin-Christensen, MónicaLlamaSangrePCRInfestaciónInfecciones por ProtozoosBloodInfestationProtozoal InfectionsSarcocystis AucheniaeThe protozoon Sarcocystis aucheniae is the causative agent of South American camelid (SAC) sarcocystosis. Infections are characterized by the presence of cysts in muscles which are in size and appearance similar to rice grains. As consumption of insufficiently cooked infected meat produces gastroenteritis, cyst-containing SAC meat is confiscated by sanitary authorities or depreciated with serious economic consequences for SAC breeders. In this work, a duplex semi-nested PCR was designed to simultaneously detect parasite and llama DNA in host blood samples. Species-specific regions of S. aucheniae 18S rRNA gene and Lama glama 16S mitochondrial gene were amplified, yielding bands of 583 and 257 bp, respectively, and separated by gel electrophoresis. The method proved to be highly sensitive, with a detection limit lower than one parasite per milliliter blood, and the inclusion of primers to detect llama-specific DNA resulted useful as a methodological control. Blood samples collected from llamas of Argentina and Bolivia (n = 225) were analyzed using this method, and 18.7 % resulted positive for S. aucheniae. No correlation was found between PCR results and llama age, sex or the finding of macroscopic cysts in meat after slaughter. Lack of molecular detection in the blood of some llamas harboring macrocysts suggests that parasite circulation in the bloodstream after encystment is under the detection threshold of the test or even absent, while PCR positive results in cyst-infested animals suggests that prior exposure to the parasite does not impede subsequent infections. The described method can be useful to detect active foci of infection, to assess the effectiveness of parasiticide treatments, and for the surveillance and tracing of definitive hosts.Instituto de PatobiologíaFil: Decker Franco, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romero, Sandra Raquel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación y Desarrollo Tecnológico para la Agricultura Familiar Región NOA; ArgentinaFil: Ferrari, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaElsevier2019-02-13T14:46:53Z2019-02-13T14:46:53Z2018info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/4436https://www.heliyon.com/article/e009282405-8440https://doi.org/10.1016/j.heliyon.2018.e00928Heliyon 4 : e00928. (2018)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-04T09:47:48Zoai:localhost:20.500.12123/4436instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:47:48.977INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
title Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
spellingShingle Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
Decker Franco, Cecilia
Llama
Sangre
PCR
Infestación
Infecciones por Protozoos
Blood
Infestation
Protozoal Infections
Sarcocystis Aucheniae
title_short Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
title_full Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
title_fullStr Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
title_full_unstemmed Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
title_sort Detection of Sarcocystis aucheniae in blood of llama using a duplex semi-nested PCR assay and its association with cyst infestation
dc.creator.none.fl_str_mv Decker Franco, Cecilia
Romero, Sandra Raquel
Ferrari, Alejandro
Schnittger, Leonhard
Florin-Christensen, Mónica
author Decker Franco, Cecilia
author_facet Decker Franco, Cecilia
Romero, Sandra Raquel
Ferrari, Alejandro
Schnittger, Leonhard
Florin-Christensen, Mónica
author_role author
author2 Romero, Sandra Raquel
Ferrari, Alejandro
Schnittger, Leonhard
Florin-Christensen, Mónica
author2_role author
author
author
author
dc.subject.none.fl_str_mv Llama
Sangre
PCR
Infestación
Infecciones por Protozoos
Blood
Infestation
Protozoal Infections
Sarcocystis Aucheniae
topic Llama
Sangre
PCR
Infestación
Infecciones por Protozoos
Blood
Infestation
Protozoal Infections
Sarcocystis Aucheniae
dc.description.none.fl_txt_mv The protozoon Sarcocystis aucheniae is the causative agent of South American camelid (SAC) sarcocystosis. Infections are characterized by the presence of cysts in muscles which are in size and appearance similar to rice grains. As consumption of insufficiently cooked infected meat produces gastroenteritis, cyst-containing SAC meat is confiscated by sanitary authorities or depreciated with serious economic consequences for SAC breeders. In this work, a duplex semi-nested PCR was designed to simultaneously detect parasite and llama DNA in host blood samples. Species-specific regions of S. aucheniae 18S rRNA gene and Lama glama 16S mitochondrial gene were amplified, yielding bands of 583 and 257 bp, respectively, and separated by gel electrophoresis. The method proved to be highly sensitive, with a detection limit lower than one parasite per milliliter blood, and the inclusion of primers to detect llama-specific DNA resulted useful as a methodological control. Blood samples collected from llamas of Argentina and Bolivia (n = 225) were analyzed using this method, and 18.7 % resulted positive for S. aucheniae. No correlation was found between PCR results and llama age, sex or the finding of macroscopic cysts in meat after slaughter. Lack of molecular detection in the blood of some llamas harboring macrocysts suggests that parasite circulation in the bloodstream after encystment is under the detection threshold of the test or even absent, while PCR positive results in cyst-infested animals suggests that prior exposure to the parasite does not impede subsequent infections. The described method can be useful to detect active foci of infection, to assess the effectiveness of parasiticide treatments, and for the surveillance and tracing of definitive hosts.
Instituto de Patobiología
Fil: Decker Franco, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Romero, Sandra Raquel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación y Desarrollo Tecnológico para la Agricultura Familiar Región NOA; Argentina
Fil: Ferrari, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; Argentina
Fil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description The protozoon Sarcocystis aucheniae is the causative agent of South American camelid (SAC) sarcocystosis. Infections are characterized by the presence of cysts in muscles which are in size and appearance similar to rice grains. As consumption of insufficiently cooked infected meat produces gastroenteritis, cyst-containing SAC meat is confiscated by sanitary authorities or depreciated with serious economic consequences for SAC breeders. In this work, a duplex semi-nested PCR was designed to simultaneously detect parasite and llama DNA in host blood samples. Species-specific regions of S. aucheniae 18S rRNA gene and Lama glama 16S mitochondrial gene were amplified, yielding bands of 583 and 257 bp, respectively, and separated by gel electrophoresis. The method proved to be highly sensitive, with a detection limit lower than one parasite per milliliter blood, and the inclusion of primers to detect llama-specific DNA resulted useful as a methodological control. Blood samples collected from llamas of Argentina and Bolivia (n = 225) were analyzed using this method, and 18.7 % resulted positive for S. aucheniae. No correlation was found between PCR results and llama age, sex or the finding of macroscopic cysts in meat after slaughter. Lack of molecular detection in the blood of some llamas harboring macrocysts suggests that parasite circulation in the bloodstream after encystment is under the detection threshold of the test or even absent, while PCR positive results in cyst-infested animals suggests that prior exposure to the parasite does not impede subsequent infections. The described method can be useful to detect active foci of infection, to assess the effectiveness of parasiticide treatments, and for the surveillance and tracing of definitive hosts.
publishDate 2018
dc.date.none.fl_str_mv 2018
2019-02-13T14:46:53Z
2019-02-13T14:46:53Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/4436
https://www.heliyon.com/article/e00928
2405-8440
https://doi.org/10.1016/j.heliyon.2018.e00928
url http://hdl.handle.net/20.500.12123/4436
https://www.heliyon.com/article/e00928
https://doi.org/10.1016/j.heliyon.2018.e00928
identifier_str_mv 2405-8440
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Heliyon 4 : e00928. (2018)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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