Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia

ABSTRACT: Introduction: Paratuberculosis is an economically important, chronic, and incurable disease in ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). Since the effects of the disease represent a major source of losses for the milk-producing sector worldwide, it is importan...

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Autores:
Correa Valencia, Nathalia María del Pilar
Tipo de recurso:
Doctoral thesis
Fecha de publicación:
2019
Institución:
Universidad de Antioquia
Repositorio:
Repositorio UdeA
Idioma:
eng
OAI Identifier:
oai:bibliotecadigital.udea.edu.co:10495/27581
Acceso en línea:
http://hdl.handle.net/10495/27581
Palabra clave:
Paratuberculosis
Cattle diseases
Mycobacterium avium paratuberculosis
Veterinary epidemiology
Veterinary serology
Enfermedades del ganado
Epidemiología veterinaria
Serología veterinaria
Enfermedad de Johne
Johne´s disease
https://lccn.loc.gov/sh88023291
https://lccn.loc.gov/sh85070623
https://lccn.loc.gov/sh85021307
https://lccn.loc.gov/sh85143077
https://lccn.loc.gov/sh85143029
Rights
openAccess
License
https://creativecommons.org/licenses/by-nc-sa/2.5/co/
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oai_identifier_str oai:bibliotecadigital.udea.edu.co:10495/27581
network_acronym_str UDEA2
network_name_str Repositorio UdeA
repository_id_str
dc.title.spa.fl_str_mv Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
dc.title.alternative.spa.fl_str_mv Epidemiología de Mycobacterium avium subsp. paratuberculosis (MAP) en la región lechera del Norte de Antioquia, Colombia.
title Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
spellingShingle Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
Paratuberculosis
Cattle diseases
Mycobacterium avium paratuberculosis
Veterinary epidemiology
Veterinary serology
Enfermedades del ganado
Epidemiología veterinaria
Serología veterinaria
Enfermedad de Johne
Johne´s disease
https://lccn.loc.gov/sh88023291
https://lccn.loc.gov/sh85070623
https://lccn.loc.gov/sh85021307
https://lccn.loc.gov/sh85143077
https://lccn.loc.gov/sh85143029
title_short Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
title_full Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
title_fullStr Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
title_full_unstemmed Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
title_sort Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, Colombia
dc.creator.fl_str_mv Correa Valencia, Nathalia María del Pilar
dc.contributor.advisor.none.fl_str_mv Fernández Silva, Jorge Arturo
dc.contributor.author.none.fl_str_mv Correa Valencia, Nathalia María del Pilar
dc.subject.lcsh.none.fl_str_mv Paratuberculosis
Cattle diseases
Mycobacterium avium paratuberculosis
Veterinary epidemiology
Veterinary serology
Enfermedades del ganado
Epidemiología veterinaria
Serología veterinaria
topic Paratuberculosis
Cattle diseases
Mycobacterium avium paratuberculosis
Veterinary epidemiology
Veterinary serology
Enfermedades del ganado
Epidemiología veterinaria
Serología veterinaria
Enfermedad de Johne
Johne´s disease
https://lccn.loc.gov/sh88023291
https://lccn.loc.gov/sh85070623
https://lccn.loc.gov/sh85021307
https://lccn.loc.gov/sh85143077
https://lccn.loc.gov/sh85143029
dc.subject.proposal.spa.fl_str_mv Enfermedad de Johne
Johne´s disease
dc.subject.lcshuri.none.fl_str_mv https://lccn.loc.gov/sh88023291
https://lccn.loc.gov/sh85070623
https://lccn.loc.gov/sh85021307
https://lccn.loc.gov/sh85143077
https://lccn.loc.gov/sh85143029
description ABSTRACT: Introduction: Paratuberculosis is an economically important, chronic, and incurable disease in ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). Since the effects of the disease represent a major source of losses for the milk-producing sector worldwide, it is important to define its impact and epidemiological dynamics both at regional and country-level. Objective: This cross-sectional study aimed to determine MAP herd-level prevalence, the herd level risk factors associated, the circulating genotypes, and to describe the spatial distribution and the environmental variables related to MAP in dairy herds of the Northern region of Antioquia, Colombia. Methods: Study herds (n = 386) located in 62 different districts from six municipalities were randomly selected amongst 7,794 dairies registered in the foot-and-mouth disease vaccination records from 2015. The sampling strategy considered proportional allocation, both at municipality and district-level. Participant herds were visited once between June and October 2016 to collect composite environmental samples and to complete a risk assessment questionnaire. The study herds were clasified in two groups, according to their productive system, those with under mechanical milking parlor and pasture grazing-based systems, and those under with in-paddock milking facilities. Each composite environmental sample contained material from six different sites of concentration of adult cattle and/or high traffic areas (e.g. areas surrounding waterers and feeders, areas surrounding the current mobile milking-unit places) and, depending on the production system, a second composite was collected from the perimeter of the manure storage lagoon (containing manure from the milking parlor). Identification of MAP was achieved using a duplex IS900-qPCR (Bactotype MAP PCR Kit®, Qiagen). A herd was considered as MAP infected if the environmental sample was positive in the qPCR. Information about the general characteristics of the herd, management practices, and knowledge about the disease was collected using the risk-assessment questionnaire. The information on risk factors was analyzed using a multivariable logistic regression model. Environmental samples from the 25 MAP-qPCR positive dairy herds were cultured by duplicate in Herrold´s egg yolk medium with mycobactin J to obtain isolates. Suspicious colonies were confirmed by MAP-qPCR. Positive DNA was sub-typed using mycobacterial interspersed repetitive units-variable number of tandem repeat (MIRU-VNTR) and multilocus short sequence repeats (MLSSR) techniques to analyze the genetic difference(s) between the isolates. To describe the spatial distribution and the environmental variables related to MAP, rainfall trends, day and nighttime surface temperature, and vegetation cover index were taken as environmental references of the physical background of the study area. Results: We found a herd-level prevalence of 4.1% (12/292; 95% CI: 1.8-6.4) and “having a history of mixed farming of cattle with other ruminants (i.e. sheep, goats) in the last 2 years” as a risk factor for MAP infection (OR = 3.9; 95% CI: 1.2-13.2) in 292 dairies under mechanical milking parlor and pasture grazing-based systems. On the other hand, we found a prevalence of 14.9% (14/94; 95% CI: 7.7-22.1) and “having other than Holstein breeds were predominant” (namely, Jersey, Jersey×Holstein crossbreeds, and Jersey×Swedish red crossbreeds) as a risk factor in 94 dairies with in-paddock milking facilities (OR = 3.7; 95% CI: 1.1-15.2). Sub-typing revealed two different genotypes by MIRU-VNTR (INMV 2 and INMV 36). MLSSR was carried out to increase the discriminatory power from what was obtained by MIRU-VNTR, but no differences were observed among the isolates recovered. According to the spatial distribution and the environmental analysis, an overall high rainfall regime was found in the study area. The daytime and nighttime surface temperature showed important variations during sampling months. No evidence of management of the vegetation cover was found. Conclusions: Our study reported the MAP prevalence in dairy herds from a representative dairy region in the Province of Antioquia (Colombia) and the possible relationship between MAP environmental positivity with the history of mixed farming of cattle with other susceptible ruminants and with the predominant breed of cattle in the herd. MAP genotypes INMV 2 and INMV 36 circulate in the study region. According to the spatial distribution and the environmental analysis, our study referred to an exploratory, non-experimental observational study carried out on an uncontrolled tropical and a real dynamic environment. Our purpose was to describe the general conditions of the environmental context where the detection of positive herds is most likely to happen, considering the same (or a very approximate) sample collection and handling, and molecular detection method. In general, our study represents an important approach to the knowledge on MAP epidemiological status in the Colombian dairy population of study.
publishDate 2019
dc.date.issued.none.fl_str_mv 2019
dc.date.accessioned.none.fl_str_mv 2022-04-18T20:51:32Z
dc.date.available.none.fl_str_mv 2022-04-18T20:51:32Z
dc.type.spa.fl_str_mv info:eu-repo/semantics/doctoralThesis
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dc.type.redcol.spa.fl_str_mv https://purl.org/redcol/resource_type/TD
dc.type.local.spa.fl_str_mv Tesis/Trabajo de grado - Monografía - Doctorado
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dc.identifier.uri.none.fl_str_mv http://hdl.handle.net/10495/27581
url http://hdl.handle.net/10495/27581
dc.language.iso.spa.fl_str_mv eng
language eng
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dc.format.mimetype.spa.fl_str_mv application/pdf
dc.publisher.group.spa.fl_str_mv CENTAURO
dc.publisher.place.spa.fl_str_mv Medellín, Colombia
institution Universidad de Antioquia
bitstream.url.fl_str_mv http://bibliotecadigital.udea.edu.co/bitstream/10495/27581/2/CorreaNathalia_2019_Epidemiology_MAP_Antioquia.pdf
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spelling Fernández Silva, Jorge ArturoCorrea Valencia, Nathalia María del Pilar2022-04-18T20:51:32Z2022-04-18T20:51:32Z2019http://hdl.handle.net/10495/27581ABSTRACT: Introduction: Paratuberculosis is an economically important, chronic, and incurable disease in ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). Since the effects of the disease represent a major source of losses for the milk-producing sector worldwide, it is important to define its impact and epidemiological dynamics both at regional and country-level. Objective: This cross-sectional study aimed to determine MAP herd-level prevalence, the herd level risk factors associated, the circulating genotypes, and to describe the spatial distribution and the environmental variables related to MAP in dairy herds of the Northern region of Antioquia, Colombia. Methods: Study herds (n = 386) located in 62 different districts from six municipalities were randomly selected amongst 7,794 dairies registered in the foot-and-mouth disease vaccination records from 2015. The sampling strategy considered proportional allocation, both at municipality and district-level. Participant herds were visited once between June and October 2016 to collect composite environmental samples and to complete a risk assessment questionnaire. The study herds were clasified in two groups, according to their productive system, those with under mechanical milking parlor and pasture grazing-based systems, and those under with in-paddock milking facilities. Each composite environmental sample contained material from six different sites of concentration of adult cattle and/or high traffic areas (e.g. areas surrounding waterers and feeders, areas surrounding the current mobile milking-unit places) and, depending on the production system, a second composite was collected from the perimeter of the manure storage lagoon (containing manure from the milking parlor). Identification of MAP was achieved using a duplex IS900-qPCR (Bactotype MAP PCR Kit®, Qiagen). A herd was considered as MAP infected if the environmental sample was positive in the qPCR. Information about the general characteristics of the herd, management practices, and knowledge about the disease was collected using the risk-assessment questionnaire. The information on risk factors was analyzed using a multivariable logistic regression model. Environmental samples from the 25 MAP-qPCR positive dairy herds were cultured by duplicate in Herrold´s egg yolk medium with mycobactin J to obtain isolates. Suspicious colonies were confirmed by MAP-qPCR. Positive DNA was sub-typed using mycobacterial interspersed repetitive units-variable number of tandem repeat (MIRU-VNTR) and multilocus short sequence repeats (MLSSR) techniques to analyze the genetic difference(s) between the isolates. To describe the spatial distribution and the environmental variables related to MAP, rainfall trends, day and nighttime surface temperature, and vegetation cover index were taken as environmental references of the physical background of the study area. Results: We found a herd-level prevalence of 4.1% (12/292; 95% CI: 1.8-6.4) and “having a history of mixed farming of cattle with other ruminants (i.e. sheep, goats) in the last 2 years” as a risk factor for MAP infection (OR = 3.9; 95% CI: 1.2-13.2) in 292 dairies under mechanical milking parlor and pasture grazing-based systems. On the other hand, we found a prevalence of 14.9% (14/94; 95% CI: 7.7-22.1) and “having other than Holstein breeds were predominant” (namely, Jersey, Jersey×Holstein crossbreeds, and Jersey×Swedish red crossbreeds) as a risk factor in 94 dairies with in-paddock milking facilities (OR = 3.7; 95% CI: 1.1-15.2). Sub-typing revealed two different genotypes by MIRU-VNTR (INMV 2 and INMV 36). MLSSR was carried out to increase the discriminatory power from what was obtained by MIRU-VNTR, but no differences were observed among the isolates recovered. According to the spatial distribution and the environmental analysis, an overall high rainfall regime was found in the study area. The daytime and nighttime surface temperature showed important variations during sampling months. No evidence of management of the vegetation cover was found. Conclusions: Our study reported the MAP prevalence in dairy herds from a representative dairy region in the Province of Antioquia (Colombia) and the possible relationship between MAP environmental positivity with the history of mixed farming of cattle with other susceptible ruminants and with the predominant breed of cattle in the herd. MAP genotypes INMV 2 and INMV 36 circulate in the study region. According to the spatial distribution and the environmental analysis, our study referred to an exploratory, non-experimental observational study carried out on an uncontrolled tropical and a real dynamic environment. Our purpose was to describe the general conditions of the environmental context where the detection of positive herds is most likely to happen, considering the same (or a very approximate) sample collection and handling, and molecular detection method. In general, our study represents an important approach to the knowledge on MAP epidemiological status in the Colombian dairy population of study.RESUMEN: Introducción: La paratuberculosis es una enfermedad económicamente importante, crónica e incurable en rumiantes, causada por Mycobacterium avium subsp. paratuberculosis (MAP). Dado que los efectos de la enfermedad representan una fuente importante de pérdidas para el sector productor de leche en todo el mundo, es importante definir su impacto y la dinámica epidemiológica tanto a nivel regional como nacional. Objetivo: El presente estudio transversal tuvo como objetivo determinar la prevalencia de MAP a nivel de hato, los factores de riesgo asociados, los genotipos circulantes y describir la distribución espacial y las variables ambientales relacionadas con MAP en hatos lecheros de la región norte de Antioquia. Colombia. Métodos: Los hatos de estudio (n = 386), ubicados en 62 veredas diferentes de seis municipios fueron seleccionados al azar entre 7,794 lecherías registradas en los registros de vacunación contra la fiebre aftosa en 2015. La estrategia de muestreo consideró la asignación proporcional, tanto a nivel de municipio como de vereda. Los hatos participantes fueron visitados una vez entre junio y octubre de 2016 para recolectar muestras ambientales en pool y completar un cuestionario de evaluación de factores de riesgo. Los hatos de estudio se clasificaron en dos grupos, de acuerdo con su sistema productivo, aquellos con sistema de ordeño mecánico y sistema de pastoreo en potrero, y aquellos bajo sistema de ordeño en sala Cada muestra ambiental en pool contenía material de seis sitios diferentes de concentración de ganado adulto y/o áreas de alto tráfico (e.g. áreas que rodean los bebederos y comederos, áreas que rodean los lugares de las unidades de ordeño móviles) y, dependiendo del sistema de producción, un segundo pool se recolectó en el perímetro del tanque estercolero (que contenía estiércol de la sala de ordeño). La identificación de MAP se logró utilizando un PCR en tiempo real dúplex (IS900-qPCR; Bactotype MAP PCR Kit®, Qiagen). Un hato se consideró como infectado por MAP si la muestra ambiental era positiva a qPCR. La información sobre las características generales del hato, las prácticas de manejo y el conocimiento sobre la enfermedad se recopilaron mediante el cuestionario de evaluación de factores de riesgo. La información sobre los factores de riesgo se analizó mediante un modelo de regresión logística multivariable. Las muestras ambientales de los 25 hatos positivos por MAP-qPCR se cultivaron por duplicado en medio de Herrold con yema de huevo con micobactina J para obtener aislamientos. Las colonias sospechosas fueron confirmadas por MAP-qPCR. El ADN positivo se subtipó utilizando un número variable de unidades repetitivas intercaladas de micobacterias de repetición en tándem (MIRU-VNTR) y repeticiones de secuencia corta multilocus (MLSSR) para analizar las diferencias genéticas entre los aislamientos. Para describir la distribución espacial y las variables ambientales relacionadas con MAP, las tendencias de precipitación, la temperatura de la superficie diurna y nocturna y el índice de cobertura de vegetación se tomaron como referencias ambientales del fondo físico del área de estudio. Resultados: Encontramos una prevalencia a nivel de hato de 4,1% (12/292; IC 95%: 1,8-6,4) y "tener un historial de cría mixta de ganado con otros rumiantes (i.e. ovejas, cabras) en los últimos 2 años" como factor de riesgo para la infección por MAP (OR = 3,9; IC 95%: 1,2-13,2) en 292 hatos bajo sistemas de ordeño mecánico y sistemas basados en pastoreo. Por otro lado, encontramos una prevalencia de 14,9% (14/94; IC 95%: 7,7-22,1) y "raza predominante diferente a Holstein en el hato" (i.e. Jersey, cruces de Jersey×Holstein y cruces de Jersey×Rojo sueco) como factor de riesgo en 94 hatos con instalaciones de ordeño en sala (OR = 3,7; IC del 95%: 1,1-15,2). El subtipado reveló dos genotipos diferentes por MIRU-VNTR (INMV 2 e INMV 36). MLSSR se llevó a cabo para aumentar el poder discriminatorio de lo que se obtuvo por MIRU-VNTR, pero no se observaron diferencias entre los aislamientos recuperados. De acuerdo con la distribución espacial y el análisis ambiental, se encontró un régimen general de alta precipitación en el área de estudio. La temperatura de la superficie diurna y nocturna mostró importantes variaciones durante los meses de muestreo. No se encontró evidencia de manejo de la cubierta vegetal. Conclusiones: Nuestro estudio reportó la prevalencia de MAP en hatos de una región lechera representativa en la Provincia de Antioquia (Colombia) y la posible relación entre la positividad ambiental a MAP con la historia de la cría de ganado bovino con otros rumiantes susceptibles y con la raza predominante del ganado en el hato. Los genotipos de MAP INMV 2 e INMV 36 circulan en la región de estudio. De acuerdo con la distribución espacial y el análisis ambiental, nuestro estudio se refirió a un estudio observacional no experimental, exploratorio, realizado en un entorno tropical no controlado y en un entorno real dinámico. Nuestro propósito fue describir las condiciones generales del contexto ambiental donde es más probable que ocurra la detección de hatos positivos, considerando la misma recolección y manejo de la muestra (o una muy aproximada) y método de detección molecular. En general, nuestro estudio representa un enfoque importante del conocimiento sobre el estado epidemiológico del MAP en la población colombiana lechera de estudio.145application/pdfenginfo:eu-repo/semantics/draftinfo:eu-repo/semantics/doctoralThesishttp://purl.org/coar/resource_type/c_db06https://purl.org/redcol/resource_type/TDTesis/Trabajo de grado - Monografía - Doctoradohttp://purl.org/coar/version/c_b1a7d7d4d402bcceinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by-nc-sa/4.0/ParatuberculosisCattle diseasesMycobacterium avium paratuberculosisVeterinary epidemiologyVeterinary serologyEnfermedades del ganadoEpidemiología veterinariaSerología veterinariaEnfermedad de JohneJohne´s diseasehttps://lccn.loc.gov/sh88023291https://lccn.loc.gov/sh85070623https://lccn.loc.gov/sh85021307https://lccn.loc.gov/sh85143077https://lccn.loc.gov/sh85143029Epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP) in the Northern dairy region of Antioquia, ColombiaEpidemiología de Mycobacterium avium subsp. paratuberculosis (MAP) en la región lechera del Norte de Antioquia, Colombia.CENTAUROMedellín, ColombiaDoctor en Ciencias VeterinariasDoctoradoFacultad de Ciencias Agrarias. Doctorado en Ciencias VeterinariasUniversidad de AntioquiaORIGINALCorreaNathalia_2019_Epidemiology_MAP_Antioquia.pdfCorreaNathalia_2019_Epidemiology_MAP_Antioquia.pdfTesis doctoralapplication/pdf3553718http://bibliotecadigital.udea.edu.co/bitstream/10495/27581/2/CorreaNathalia_2019_Epidemiology_MAP_Antioquia.pdfea280a6ac256b9e13f4c4d76a0a9825dMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://bibliotecadigital.udea.edu.co/bitstream/10495/27581/4/license.txt8a4605be74aa9ea9d79846c1fba20a33MD54CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8712http://bibliotecadigital.udea.edu.co/bitstream/10495/27581/3/license_rdffd0548b8694973befb689f3e7a707f1dMD5310495/27581oai:bibliotecadigital.udea.edu.co:10495/275812022-04-21 09:52:15.501Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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