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Dinámica de algunos virus respiratorios en dos granjas porcinas tecnificadas en el trópico alto colombiano

dc.rights.licenseAtribución-NoComercial-SinDerivadas 4.0 Internacional
dc.contributor.advisorJaime Correa, Jairo Aureliano
dc.contributor.authorTarazona Manrique, Luis Edgar
dc.date.accessioned2024-04-08T19:03:14Z
dc.date.available2024-04-08T19:03:14Z
dc.date.issued2024-03-13
dc.identifier.urihttps://repositorio.unal.edu.co/handle/unal/85879
dc.descriptionilustraciones, diagramas
dc.description.abstractEl complejo respiratorio porcino (PCR) es una entidad multifactorial que afecta negativamente a los cerdos en todas las etapas productivas, su variado comportamiento requiere un estudio particular en cada país y región geográfica. Este trabajo tuvo como objetivo determinar la dinámica de algunos virus respiratorios relacionados con la PCR en dos fincas del trópico alto colombiano, una positiva para PRRSV y la otra negativa. Se realizó un estudio longitudinal entre marzo y septiembre de 2022. Se hicieron tres muestreos bimensuales por granja donde se colectaron muestras de fluidos orales, piso de maternidad, glándulas mamarias, comederos, objetos de enriquecimiento ambiental y suero de cerdos ubicados en las etapas de predestete, preceba, finalización. Se hizo detección de virus asociados con el CRP (PPRSV, SIV-A, PCV2, PCV3, PPV2 y PPIV-1) por técnicas de PCR y RT-PCT junto con qPCR. Se encontró que los virus circulan en las dos granjas en todas las etapas productivas de forma de mono y copresencias, siendo las de tipo doble y triple las más frecuentes y se asociaron con la presencia de PRRSV. Se evaluaron y asociaron variables climáticas con la dinámica viral encontrándose que estas se correlacionaron con la presencia de virus como PCV3 y PPV2, este último incrementó la mortalidad y retrasó el crecimiento de los cerdos. Los análisis de secuenciación de PPV2 muestran que pertenece al clado 1. Este es un estudio pionero para el país que contribuye a entender cómo se comportan los virus asociados al CRP en condiciones propias de Colombia como lo es en el trópico alto (entre 1500 y 2000 msnm), ese comportamiento se complementa estableciendo las copresencias y coinfecciones virales más importantes abriendo toda una línea de investigación sobre la temática del efecto de simultaneidad infecciosa en las granjas de producción de cerdo en Colombia. (Texto tomado de la fuente).
dc.description.abstractThe porcine respiratory complex (PRC) is a multifactorial entity that negatively affects pigs in all productive stages; its varied behavior requires a particular study in each country and geographical region. This work aimed to determine the dynamics of some respiratory viruses related to PRC in two farms in the Colombian high tropics, one positive for PRRSV and the other negative. A longitudinal study was carried out between March and September 2022. Three bimonthly samplings were carried out per farm, collecting samples of oral fluids, farrowing floor, mammary glands, feeders, environmental enrichment objects, and serum from pigs in the pre-weaning, pre-fattening, and fattening stages. PCR and RT-PCT techniques and qPCR detected viruses associated with PRC (PPRSV, SIV-A, PCV2, PCV3, PPV2, and PPIV-1). It was found that the viruses circulating in the two farms in all the productive stages in a mono and co-presence form, being the double and triple types the most frequent and were associated with the presence of PRRSV. Climatic variables were evaluated and associated with viral dynamics, finding that these correlated with viruses such as PCV3 and PPV2. The latter increased mortality and retarded the growth of pigs. Sequencing analyses of PPV2 show that it belongs to clade 1. This pioneering study for the country contributes to understanding how CRP-associated viruses behave in Colombian conditions, such as in the high tropics (between 1,500 and 2,000 masl). This behavior is complemented by establishing viral co-presences and coinfections. Most important, opening a whole line of research on the effect of infectious simultaneity in pig production farms in Colombia.
dc.description.sponsorshipLa universidad financió el proyecto a través de convocatorias internas de apoyo a tesis de maestría
dc.format.extentxiv, 160 páginas
dc.format.mimetypeapplication/pdf
dc.language.isospa
dc.publisherUniversidad Nacional de Colombia
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc590 - Animales::599 - Mamíferos
dc.subject.ddc630 - Agricultura y tecnologías relacionadas::636 - Producción animal
dc.titleDinámica de algunos virus respiratorios en dos granjas porcinas tecnificadas en el trópico alto colombiano
dc.typeTrabajo de grado - Maestría
dc.type.driverinfo:eu-repo/semantics/masterThesis
dc.type.versioninfo:eu-repo/semantics/acceptedVersion
dc.publisher.programBogotá - Medicina Veterinaria y de Zootecnia - Maestría en Salud y Producción Animal
dc.contributor.researchgroupCentro de investigación en infectología e inmunología veterinaria - CI3V
dc.coverage.countryColombia
dc.coverage.tgnhttp://vocab.getty.edu/page/tgn/1000050
dc.description.degreelevelMaestría
dc.description.degreenameMagíster en Salud Animal o Magíster en Producción Animal
dc.description.methodsSe realizó un estudio de tipo longitudinal, estratificado y comparativo en dos granjas tecnificadas ubicadas en el trópico alto colombiano durante el período comprendido entre marzo y septiembre del 2022. El estudio se realizó en el departamento de Cundinamarca, las dos granjas están ubicadas en municipios distintos, pero a altitudes similares, así: i) granja 1 (G1): ubicada a una altitud de 1350 msnm; y, ii) granja 2 (G2): ubicada a 1600 msnm, es decir, las dos están en la altura conocida como trópico alto colombiano (entre 1000 y 2000 msnm). La distancia entre las granjas y el laboratorio de virología y biología molecular de la Facultad de Medicina Veterinaria y de Zootecnia de la Universidad Nacional de Colombia no superaba los 60 kilómetros; y entre ellas, la distancia era de 91.5 kilómetros. Las granjas fueron seleccionadas por conveniencia (teniendo en cuenta el deseo manifiesto por los propietarios y veterinarios de participar en el estudio). Las dos granjas manejaban un sistema productivo de ciclo completo con sistema de flujo continuo (G1) y en bandas (G2) y contaban con un inventario total de 150 madres cada una; el fin productivo de las dos granjas es la cría, levante y finalización de cerdos destinados al faenado para consumo humano. El factor diferencial entre granjas fue que G1 tiene un registro histórico de la ausencia de PRRSV por diagnostico serológico (ELISA) mientras que la G2 tiene un registro histórico de presencia de PRRSV también por pruebas serológicas. Es importante señalar que en Colombia no se vacuna contra PRRSV Tomando en cuenta el sistema productivo de ciclo completo la población a evaluar fue dividida en cuatro diferentes etapas, así: (i) cerdos en pre-destete (PD: menos de 21 días de vida), (ii) pre-ceba (PC: 7 - 10 semanas de edad), (iii) ceba (CC: 12 - 15 semanas de edad) y (iv) finalización (CF: 18 - 22 semanas de edad); lo anterior basados en estudios previos del grupo de investigación (Mendoza, 2015) y lo recomendado por (Vilalta et al., 2018; 2019). Se realizó un muestreo longitudinal estratificado con una periodicidad bimensual por seis meses, es decir tres muestreos por granja. En G1, el primer muestreo fue el 14/03/2022, el segundo muestreo ocurrió el 08/06/2022 y se finalizó el 23/08/2022. Por su parte, el muestreo en G2 fue: inició el 22/04/2022, segundo muestreo el 29/06/2022 y el tercer muestreo en 07/09/2022. Dentro de los periodos climáticos de Colombia, los dos primeros muestreos para cada granja ocurrieron durante el primer período de lluvias que tiene el país (marzo-junio); mientras que el tercer muestreo se hizo durante el denominado período seco (ausencia de lluvias, agosto - septiembre). Se determinó un mínimo de tiempo entre cada muestreo entre granjas de 15 días con el fin de prevenir la contaminación de patógenos de una a la otra, así mismo, los materiales fueron preparados para cada muestreo y ningún sobrante de material utilizado en una granja fue llevado a la otra. Es necesario aclarar que no se evaluaron los mismos cerdos en cada muestreo en cada granja, sino que se tomaron muestras de animales que se encontraban en la etapa de desarrollo planteada para el muestro Los virus que se evaluaron en cada granja fueron: virus del síndrome respiratorio y reproductivo porcino (PRRSV), circovirus porcino tipo 2 (PCV2), circovirus porcino tipo 3 (PCV3), el virus de la influenza porcina (SIV) tipo A, Parvovirus porcino 2 (PPV2) y Parainfluenza porcina tipo 1 (PPIV-1). PCR A PUNTO FINAL: esta técnica se empleó para la detección molecular de SIV-A y PPV2 utilizando la enzima Taq Polimerasa Recombinante de Invitrogen® utilizando el siguiente protocolo: denaturación inicial a 94 °C durante 1 min, seguido por 38 ciclos cada uno con una denaturación a 94 °C durante 5 min, alineación a 57 °C durante 30 segundos (seg) y, extensión a 72 °C durante 1 min. La obtención de los fragmentos genómicos para secuenciación de los virus de las muestras positivas se realizó utilizando el siguiente protocolo: denaturación inicial a 98°C durante 1 min, seguido por 35 ciclos, cada uno con una denaturación a 98°C durante 15 seg, alineación a 57°C durante 30 seg y, extensión a 72°C durante 40 seg, utilizando la enzima Q5 High-Fidelity DNA polymerase. Esto se realizó en el equipo Bio-Rad C-1000 Touch. Luego de la PCR, los amplicones fueron visualizados en un gel de agarosa al 1.5%. qPCR: esta técnica se empleó para la detección molecular y la cuantificación de cargas de PRRSV, PCV2, PCV3, SIV-A y PPIV-1. Cada uno de estos agentes fue detectado utilizando primers y sondas específicas utilizadas y validadas en el laboratorio. El protocolo utilizado fue el siguiente: denaturación inicial a 95 °C durante 10 min, seguida de 42 ciclos con una denaturación a 95 °C por 15 seg, alineación a 60°C por 45 seg, extensión (detección de fluorescencia) a 72°C por 2 segundos a 530- 560 nm, y un ciclo final de 40°C por 30 seg. Empleando primers para secuenciación, se intentó la secuenciación parcial de genomas virales: PRRSV (ORF-5); PCV2 (Cap), PCV3 (Cap), SIV-A (H1-H3-H5-N1-N2), PPV2 (VP2). Con esto se realizó genotipificación de los virus encontrados. Esta secuenciación se realizó empleando el método de Sanger en el Servicio de Secuenciación y Análisis Molecular – SsiGMoL de la Universidad Nacional de Colombia. Las secuencias obtenidas fueron curadas empleando el programa Bioedit® y se realizó filogenia empleando MEGA7® comparando las secuencias obtenidas con las reportadas en las bases de datos (GenBank) utilizando el método estadístico “Maximun Likelihood”, con el método de filogenia de “Bootstrap” con 1000 réplicas, el método de inferencia Tamura-3 y el análisis “Gamma Distributed With Invariant Sites (G+I)”. La modificación final del árbol se realizó utilizando el programa FigTree versión 1.4.4. Con los resultados obtenidos se determinó el porcentaje de positividad de cada virus en cada granja, como también el porcentaje de positividad de cada una de las diferentes copresencias virales posibles. Aquí es importante aclarar que el termino rutinario empleado para la presencia simultánea de patógenos en un individuo es “coinfección”. En el presente estudio, la mayoría de las muestras fueron de tipo ambiental (fluidos orales, pisos de parideras, glándula mamaria, enriquecimiento ambiental y comederos), excepto los sueros. Consideramos que el termino coinfección no es el apropiado ya que al encontrar los virus en el ambiente no es indicativo de que los cerdos estén infectados. Así, decidimos emplear el término “copresencia” a lo largo del texto excepto cuando se hable particularmente de sueros donde se empleará el termino coinfección. Se establecieron diferencias entre los grupos evaluados tanto dentro de granja como entre granjas mediante estadística descriptiva. Igualmente, se analizó con estadística descriptiva el comportamiento de cada virus y copresencias a través del tiempo (evolución en el tiempo). Los análisis de asociación entre las variables climáticas y presencia de virus como también con las variables productivas se analizaron mediante correlación múltiple utilizando el método de Pearson con un nivel de significancia del 95%. Los análisis de resultados fueron asistidos por el analista estadígrafo del grupo de investigación y las gráficas fueron realizadas utilizando el programa Excel® para Windows 2010 y RStudio. El proyecto contó con el aval del Comité de Bioética de la Facultad de Medicina Veterinaria y de Zootecnia de la Universidad Nacional de Colombia, Sede Bogotá (Acta No. CB-FMVZ-UN-009-2022). La ejecución de este proyecto está sustentada según lo expuesto en la resolución 8430 de 1993 “Por la cual se establecen las normas científicas, técnicas y administrativas para la investigación en salud”. La ley 84 de 1989 “Por la cual se adopta el estatuto nacional de protección de los animales y se crean unas contravenciones y se regula lo referente a su procedimiento y competencia”, ley 576 del 2000 “Por la cual se expide el Código de Ética para el ejercicio profesional de la medicina veterinaria, la medicina veterinaria y zootecnia y la zootecnia”, y la ley 1774 de 2016 “por medio de la cual se modifican el código civil, la ley 84 de 1989”. Por último, el Código Penal, artículo 1 “Los animales como seres sintientes no son cosas, recibirán especial protección contra el sufrimiento y el dolor, en especial, el causado directa o indirectamente por los humanos, por lo cual en la presente ley se tipifican como punibles algunas conductas relacionadas con el maltrato a los animales, y se establece un procedimiento sancionatorio de carácter policivo y judicial”.
dc.description.researchareaVirología veterinaria
dc.identifier.instnameUniversidad Nacional de Colombia
dc.identifier.reponameRepositorio Institucional Universidad Nacional de Colombia
dc.identifier.repourlhttps://repositorio.unal.edu.co/
dc.publisher.facultyFacultad de Medicina Veterinaria y de Zootecnia
dc.publisher.placeBogotá, Colombia
dc.publisher.branchUniversidad Nacional de Colombia - Sede Bogotá
dc.relation.indexedAgrosavia
dc.relation.indexedAgrovoc
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dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.subject.agrovocVirus de los animales
dc.subject.agrovocAnimal viruses
dc.subject.agrovocEnfermedades respiratorias
dc.subject.agrovocrespiratory diseases
dc.subject.agrovocProducción animal
dc.subject.agrovocanimal production
dc.subject.proposalAnálisis molecular
dc.subject.proposalComplejo respiratorio porcino
dc.subject.proposalDiagnóstico
dc.subject.proposalPatógenos respiratorios
dc.subject.proposalVirología
dc.subject.proposalMolecular analysis
dc.subject.proposalPorcine respiratory complex
dc.subject.proposalDiagnosis
dc.subject.proposalRespiratory Pathogens
dc.subject.proposalVirology
dc.title.translatedDynamics of some respiratory viruses in two techinified pig farms in the Colombian high tropics
dc.type.coarhttp://purl.org/coar/resource_type/c_bdcc
dc.type.coarversionhttp://purl.org/coar/version/c_ab4af688f83e57aa
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oaire.accessrightshttp://purl.org/coar/access_right/c_abf2
oaire.fundernameUniversidad Nacional de Colombia
dcterms.audience.professionaldevelopmentInvestigadores
dc.contributor.orcidhttps://orcid.org/0000-0003-2819-0582
dc.contributor.cvlachttps://scienti.minciencias.gov.co/cvlac/visualizador/generarCurriculoCv.do?cod_rh=0000169270
dc.contributor.scopushttp://www.scopus.com/inward/authorDetails.url?authorID=57219535130&partnerID=MN8TOARS
dc.contributor.researchgatehttps://www.researchgate.net/profile/Luis-Tarazona-Manrique
dc.contributor.googlescholarhttps://scholar.google.es/citations?user=ZLYqyhoAAAAJ&hl=es


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