Publicación:
Estudio molecular de especies de Borrelia y Coxiella en garrapatas Ixodidae del departamento del Atlántico

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dc.audience
dc.contributor.advisorLópez Mejia, Yesica
dc.contributor.advisorBadillo Viloria, María Auxiliadora
dc.contributor.authorRosa Jaramillo, Steffania De la
dc.contributor.juryContreras Cogollo, Verónica
dc.contributor.juryMiranda, Jorge
dc.date.accessioned2024-01-31T14:54:12Z
dc.date.available2026-12-26
dc.date.available2024-01-31T14:54:12Z
dc.date.issued2024-01-30
dc.description.abstractLas garrapatas son ectoparásitos hematófagos que infestan a la mayoría de los vertebrados. Clasificadas en tres familias: Argasidae (garrapatas blandas), Ixodidae (garrapatas duras). Algunas especies de la familia Ixodidae son vectores de diversos patógenos, siendo una preocupación para la salud humana y animal. Dentro de las principales bacterias zoonóticas transmitidas por garrapatas se encuentras especies de Borrelia sp, Coxiella sp., Rickettsia rickettsii, Anaplasma phagocitophylum, Ehrlichia chaffeensis, entre otras. En Colombia se han identificado 58 especies de garrapatas; 43 de la familia Ixodidae) entre las que se resaltan Rhipicephalus microplus, Rhipicephalus sanguineus, Amblyomma complejo cajennense, Amblyomma dissimile, Dermacentor nitens, entre otros. El conocimiento sobre las especies de Borrelia y Coxiella que circulan en garrapatas duras en el departamento del Atlántico, es crucial, para evaluar el riesgo de infecciones e identificar nuevas especies. Objetivo. Estudiar la distribución molecular de especies de Borrelia y Coxiella en garrapatas Ixodidae recolectadas de animales domésticos, silvestres y de vida libre en el departamento del Atlántico, Colombia. Materiales y métodos. Se realizó un estudio de corte transversal descriptivo a partir de 3045 garrapatas. Las muestras fueron recolectadas entre enero de 2021 a noviembre de 2022 en 18 municipios de las cinco subregiones del departamento del Atlántico, Colombia. Las garrapatas se identificaron a nivel de especie mediante claves morfológicas y se agruparon en 509 grupos. Los grupos de garrapatas se maceraron y se empleó 200μl del macerado para la extracción. La extracción de ADN se llevó a cabo utilizando el kit comercial GeneJET Genomic DNA Purification Kit (Thermo Scientific, Waltham, Massachusetts, EE. UU). Todos los grupos se analizaron para detectar la presencia de Borrelia spp. y Coxiella sp. mediante una Reacción en Cadena de la Polimerasa en tiempo real (qPCR) utilizando cebadores forward Bor16S3F y reverse Bor16S3R y una sonda Probe Bor16S3P dirigidos al gen 16S rRNA para especies de Borrelia y cebadores forward Trans-1F e reverse Trans-2R y una sonda Probe Coxiella sp. TM ([6FAM]) dirigidos a la secuencia de inserción IS1111 para Coxiella. Las muestras con punto de corte Cq <33 se sometieron a PCR anidadas y convencionales semianidadas para amplificar fragmentos de los genes 16S rRNA y flaB para Borrelia sp. Los fragmentos de secuencias se ensamblaron empleando el software MEGAX y los análisis filogenéticos se realizaron con IQ-TREE2 version 2.2.2.6. Resultados. Las garrapatas fueron identificadas morfológicamente como Amblyomma dissimile (5,02%), Amblyomma patinoi (0,39%), Amblyomma sp. (0,09%), Dermacentor nitens (35,9 %), Rhipicephalus microplus (51,2%), Rhipicephalus sanguineus sensu lato (7,4%). Se confirmó la identidad de Amblyomma patinoi en el departamento del Atlántico. Un total de 38 grupos (7,5%; IC95% 5.3-10.1; p= <0,001) analizados resultaron positivos para Borrelia sp. Los grupos analizados para la secuencia de inserción IS1111 de Coxiella sp. ninguno resultó positivo. El análisis BlastN de los fragmentos de los genes 16S rRNA y flaB mostro que las secuencias compartían similitud entre un 98,87 y 100% con Borrelia theileri y del 99.52% con Borrelia sp. depositada en GenBank. Los análisis filogenéticos por el método de máxima verosimilitud del gen 16S rRNA mostraron que la mayoría de las secuencias de Borrelia spp. se agrupaban a con secuencia de Borrelia theileri, y una secuencia de Borrelia sp. detectada en Amblyomma dissimile se agrupaba al clado de Borrelia asociado a reptiles (REP). Conclusiones: Los resultados filogenéticos muestran la distribución geográfica de B. theileri y Borrelia sp. asociada a reptiles recolectadas de animales domésticos, vida silvestre y garrapatas de vida libre en las cinco subregiones del departamento. La frecuencia de Borrelia theileri y su distribución en las cinco subregiones del departamento del Atlántico, sugiriendo que existe un riesgo potencial de borreliosis animal en estas áreas. Estos hallazgos son el primer informe sobre la presencia de Borrelia sp. en Amblyomma dissimile de garrapatas de reptiles en Colombia que se suman a la distribución geográfica de Borrelia spp. a América.spa
dc.description.abstractTicks are hematophagous ectoparasites that infest most vertebrates. They are classified into three families: Argasidae (soft ticks), and Ixodidae (hard ticks). Some species of the Ixodidae family are vectors of various pathogens, being a concern for human and animal health. Among the main zoonotic bacteria transmitted by ticks are species of Borrelia sp., Coxiella sp., Rickettsia rickettsii, Anaplasma phagocitophylum, and Ehrlichia chaffeensis, among others. In Colombia, 58 species of ticks have been identified; 43 of the family Ixodidae) among which Rhipicephalus microplus, Rhipicephalus sanguineus, Amblyomma complejo cajennense, Amblyomma dissimile, Dermacentor nitens, among others, stand out. Knowledge about the species of Borrelia and Coxiella that circulate in hard ticks in the Department of Atlántico is crucial to assessing the risk of infections and identifying new species. Objective. To study the molecular distribution of Borrelia and Coxiella species in Ixodidae ticks collected from domestic, wild, and free-living animals in the department of Atlántico, Colombia. Materials and methods. A descriptive cross-sectional study was conducted using 3045 ticks. The samples were collected between January 2021 and November 2022 in 18 municipalities in the five subregions of the department of Atlántico, Colombia. Ticks were identified at the species level by morphological cues and grouped into 509 pooles. The pooles of ticks were macerated and 200μl of the macerate was used for extraction. DNA extraction was performed using the commercial GeneJET Genomic DNA Purification Kit (Thermo Scientific, Waltham, Massachusetts, USA). All pooles were tested for the presence of Borrelia sp. and Coxiella sp. by a real-time Polymerase Chain Reaction (qPCR) using Bor16S3F forward and Bor16S3R forward primers and a Bor16S3P Probe targeting the 16S rRNA gene for Borrelia species and Trans-1F and reverse Trans-2R forward primers and a Coxiella Probesp. TM ([6FAM]) targeting the IS1111 insertion sequence for Coxiella. Samples with Cq <33 cut-off point were subjected to nested and conventional semi-nested PCR to amplify fragments of the 16S rRNA and flaB genes for Borrelia sp. Sequence fragments were assembled using MEGAX software and phylogenetic analyses were performed using IQ-TREE2 version 2.2.2.6. Results. Ticks were morphologically identified as Amblyomma dissimile (5.02%), Amblyomma patinoi (0.39%), Amblyomma sp. (0.09%), Dermacentor nitens (35.9%), Rhipicephalus microplus (51.2%), Rhipicephalus sanguineus (7.4%). The identity of Amblyomma patinoi was confirmed in the Department of Atlantic. A total of 38 pooles (7.5%; 95% CI 5.3-10.1; p= <0.001) tested positive for Borrelia sp. None of the pooles tested for the IS1111 insertion sequence of Coxiella sp. were positive. BlastN analysis of the fragments of the 16S rRNA and flaB genes showed that the sequences shared 98.87 to 100% similarity with Borrelia theileri and 99.52% with Borrelia sp. deposited in GenBank. Phylogenetic analyses by the maximum likelihood method of the 16S rRNA gene showed that most of the sequences of Borrelia spp. were grouped with a sequence of Borrelia theileri, and a sequence of Borrelia sp. detected in Amblyomma dissimile was grouped with the reptile-associated clade of Borrelia (REP). Conclusions: The phylogenetic results show the geographic distribution of B. theileri and Borrelia sp. associated with reptiles collected from domestic animals, wildlife, and free-living ticks in the five subregions of the department. The frequency of Borrelia theileri and its distribution in the five subregions of the Department of Atlantic, suggests that there is a potential risk of animal borreliosis in these areas. These findings are the first report on the presence of Borrelia sp. in Amblyomma dissimile from reptile ticks in Colombia that add to the geographic distribution of Borrelia spp. to the Americas.eng
dc.description.degreelevelMaestría
dc.description.degreenameMagíster en Microbiología Tropical
dc.description.modalityTrabajos de Investigación y/o Extensión
dc.description.tableofcontents2 INTRODUCCIÓN 14spa
dc.description.tableofcontents3. OBJETIVOS 16spa
dc.description.tableofcontents3.1. OBJETIVO GENERAL 16spa
dc.description.tableofcontents3.2. OBJETIVOS ESPECIFICOS 16spa
dc.description.tableofcontents4. ESTADO DEL ARTE 17spa
dc.description.tableofcontents4.1. Garrapatas: aspectos generales 17spa
dc.description.tableofcontents4.1.1. Clasificación Taxonómica 17spa
dc.description.tableofcontents4.1.2. Morfología 18spa
dc.description.tableofcontents4.1.3. Ciclo de vida 18spa
dc.description.tableofcontents4.1.4. Ecología 20spa
dc.description.tableofcontents4.1.5. Implicaciones en la salud humana y animal 22spa
dc.description.tableofcontents4.2. Género Borrelia 23spa
dc.description.tableofcontents4.2.1. Generalidades 24spa
dc.description.tableofcontents4.2.2. Estructura y organización del genoma de Borrelia. 24spa
dc.description.tableofcontents4.2.3. Patogénesis y Ciclo de Vida 27spa
dc.description.tableofcontents4.2.4. Epidemiología y distribución geográfica 28spa
dc.description.tableofcontents4.3. Coxiella 31spa
dc.description.tableofcontents4.3.1. Generalidades 31spa
dc.description.tableofcontents4.3.2. Estructura y organización del genoma de Coxiella. 32spa
dc.description.tableofcontents4.3.3. Patogénesis y ciclos de vida 34spa
dc.description.tableofcontents4.3.4. Epidemiología y distribución geográfica. 34spa
dc.description.tableofcontents5. MÉTODOS 37spa
dc.description.tableofcontents5.2. Área de estudio 37spa
dc.description.tableofcontents5.3. Población de estudio y tamaño de muestra. 37spa
dc.description.tableofcontents5.4. Identificación morfológica de garrapatas 39spa
dc.description.tableofcontents5.5. Detección molecular de Borrelia sp. Por qPCR. 39spa
dc.description.tableofcontents5.6. Análisis filogenético de secuencias 41spa
dc.description.tableofcontents5.7. Implementación de Nextstrain para Borrelia. 41spa
dc.description.tableofcontents5.8. Tratamiento estadístico de datos 41spa
dc.description.tableofcontents5.9. Consideraciones éticas. 42spa
dc.description.tableofcontents6. RESULTADOS 43spa
dc.description.tableofcontents6.1. Garrapatas identificadas en el área de estudio. 43spa
dc.description.tableofcontents6.2. Detección molecular de Borrelia sp. y Coxiella sp. en las garrapatas colectadas. 46spa
dc.description.tableofcontents6.3. Análisis filogenético 47spa
dc.description.tableofcontents6.4. Distribución de especies de Borrelia en el departamento del Atlántico. 50spa
dc.description.tableofcontents7. DISCUSIÓN 54spa
dc.description.tableofcontents8. CONCLUSIONES 59spa
dc.description.tableofcontents9. RECOMENDACIONES 60spa
dc.description.tableofcontents10. REFERENCIAS 61spa
dc.description.tableofcontents11. ANEXOS 72spa
dc.format.mimetypeapplication/pdf
dc.identifier.instnameUniversidad de Córdoba
dc.identifier.reponameRepositorio universidad de Córdoba
dc.identifier.repourlhttps://repositorio.unicordoba.edu.co
dc.identifier.urihttps://repositorio.unicordoba.edu.co/handle/ucordoba/8157
dc.language.isospa
dc.publisherUniversidad de Córdoba
dc.publisher.facultyFacultad de Medicina Veterinaria y Zootecnia
dc.publisher.placeMontería, Córdoba, Colombia
dc.publisher.programMaestría en Microbiología Tropical
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dc.rightsCopyright Universidad de Córdoba, 2024
dc.rights.accessrightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.coarhttp://purl.org/coar/access_right/c_f1cf
dc.rights.licenseAtribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.sourceUniversidad de Córdoba
dc.subject.keywordsBorrelia sp
dc.subject.keywordsCoxiella sp
dc.subject.keywordsTicks
dc.subject.keywordsColombia
dc.subject.proposalBorrelia sp
dc.subject.proposalCoxiella sp
dc.subject.proposalGarrapatas
dc.subject.proposalColombia
dc.titleEstudio molecular de especies de Borrelia y Coxiella en garrapatas Ixodidae del departamento del Atlánticospa
dc.typeTrabajo de grado - Maestría
dc.type.coarhttp://purl.org/coar/resource_type/c_bdcc
dc.type.coarversionhttp://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.contentText
dc.type.driverinfo:eu-repo/semantics/masterThesis
dc.type.redcolhttp://purl.org/redcol/resource_type/TM
dc.type.versioninfo:eu-repo/semantics/acceptedVersion
dspace.entity.typePublication
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