Publicación:
Estudio sobre la patogenicidad de Candidatus Rickettsia colombiensis

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dc.contributor.advisorMáttar Velilla, Salim
dc.contributor.authorMiranda Regino, Jorge
dc.contributor.juryCastro, Lyda
dc.contributor.juryMonsalve Buriticá, Santiago
dc.contributor.juryBermúdez Sergio
dc.date.accessioned2025-10-07T13:32:32Z
dc.date.available2025-10-07T13:32:32Z
dc.date.issued2025-10-06
dc.description.abstractLas Rickettsiosis son zoonosis transmitidas por garrapatas, y existe preocupación por el aumento de casos y en particular por la aparición de nuevas especies del grupo de las fiebres manchadas (GFM). Candidatus Rickettsia colombiensis es una nueva especie del GFM, de patogenicidad desconocida, pero relacionada filogenéticamente con especies patógenas. Por otro lado, la reactividad serológica cruzada entre especies de rickettsia es bien conocida, en ese sentido, Ca. R. colombiensis podría ser útil para el diagnóstico de rickettsiosis causadas por rickettsias del GFM. Objetivo. Determinar el potencial patógeno de Candidatus Rickettsia colombiensis y analizar la reactividad cruzada frente a otras especies rickettsia del GFM. Metodos. Ca. R. colombiensis fue aislado de Amblyomma dissimile mediante Shell vial. Posteriormente, cinco Syriam hámsters (M. auratus) machos fueron inoculados vía intraperitoneal (IP) y 5 por vía intradérmica (ID) con 1×106 células Vero infectadas con Ca. R. colombiensis. Un animal control fue usado para cada grupo. El estado de salud de los animales fue evaluado diariamente, las necropsias se realizaron los días 5, 10, 15 y 16 DPI, muestras de suero para inmunofluorescencia indirecta y tejidos para qPCR e histopatológia, fueron procesadas. La reactividad cruzada de los anticuerpos en los hamsteres y sueros humanos infectados por Rickettsia sp. del GFM fue establecido utilizando las láminas antigenadas con R. rickettsii, R. parkeri y Ca. R. colombiensis. Para caracterizar el genoma se tomaron los datos crudos de la secuenciación y se realizó un reensamblaje y una nueva anotación del genoma. Posteriormente, se realizó un analisis filogenetico y se identificaron los genes de virulencia mediante VFanalyzer y por comparativa genómica. A cada gen se le realizó un análisis de la secuencia nucleotídica y una modelación de la estructura 3D para las proteínas OmpA, OmpB, Sca1 y Sca2 en SWISS-MODEL. Resultados. Todos los animales se mantuvieron sanos y no mostraron cambios en los parámetros fisiológicos ni histopatologicos. DNA de Rickettsia fue negativo en los tejidos. Todos los animales inoculados tuvieron anticuerpos IgG anti- Ca. R. colombiensis con títulos de 1:64 hasta 1:1024. Los controles fueron negativos. Ninguno de los sueros presentó anticuerpos contra R. rickettsii y R. parkeri. El 56% (84/150) de los sueros humanos, tuvieron anticuerpos IgG contra Ca. R colombiensis. Pero cuando se evaluaron sueros con títulos iguales o superiores a 1:128, el 100% fueron seroreactivas. El reensamblaje produjo un genoma de (~1,4 Mbp) y cuatro contigs plasmídicos. En total 1852 genes, de los cuales 1200 son codificantes y 652 (35%) pseudogenes, el contenido de GC 32,4 % y la presencia de cuatro plásmidos. El análisis filogenético demostró que Ca. R. colombiensis se encuentra en el subclado GFMIb que contienen R. tamurae, R. monacensis y Rickettsia endosimbionte de Ixodes pacificus con soportes de rama de 100%. El VFanalyzer y la genómica comparativa muestran la ausencia de los genes rickA, RARP-2, VapC y la pseudogenizacion del gen ralf y RisK1. La estructura 3D de las proteínas OmpA y OmpB estuvieron acorde a los modelos del PDB, mientras que los modelos de Sca1 y Sca2 presentaron una baja calidad. Conclusión. Ca. R. colombiensis causo una infección subclínica en los hámsters y sugiere la posibilidad de infectar otros mamíferos. El análisis filogenético demostró que Ca. R. colombiensis se encuentra en el subclado GFMIb que contienen R. tamurae, R. monacensis y Rickettsia endosimbionte. Los hallazgos clínicos, patológicos y moleculares concluyen que no es una especie patógena. La reactividad cruzada podría ser una ventaja para realizar el diagnostico de rickettsiosis con antígenos de Ca. R. colombiensisspa
dc.description.abstractRickettsioses are zoonoses transmitted primarily by ticks. There is concern about the increase in cases and, in particular, the emergence of new species of the spotted fever group (SFG). Candidatus Rickettsia colombiensis is a new species of the SFG, of unknown pathogenicity, but phylogenetically related to pathogenic species. On the other hand, serological cross-reactivity between rickettsia species is well known; therefore, Ca. R. colombiensis could be useful for the diagnosis of rickettsioses caused by SFG rickettsias. Objective. To determine the pathogenic potential of Ca. R. colombiensis, a new species of the SFG in Syrian hamsters (M. auratus), and to analyze cross-reactivity against other SFG rickettsia species in human and Syrian hamster sera. Methods. Ca. R. colombiensis was isolated from Amblyomma dissimile by Shell vial. Subsequently, five male hamsters were inoculated intraperitoneally (IP) and 5 intradermally (ID) with 1 × 106 Vero cells infected with Ca. R. colombiensis. One control animal was used for each group. The health status of the animals was assessed daily, necropsies were performed on days 5, 10, 15 and 16 DPI, serum samples for indirect immunofluorescence and tissues for qPCR and histopathology were processed. Cross-reactivity of antibodies in hamsters and human sera infected with Rickettsia sp. of SFG was established using slides antigenized with R. rickettsii, R. parkeri and Ca. R. colombiensis. To characterize the genome, raw sequencing data were taken and genome reassembly and re-annotation were performed. Subsequently, a phylogenetic analysis was performed and virulence genes were identified using VFanalyzer and by genomic comparison. Nucleotide sequence analysis for each gene and 3D structure modeling the OmpA, OmpB, Sca1, and Sca2 proteins were performed in SWISS-MODEL. Results. All animals remained healthy and showed no changes in physiological or histopathological parameters. Rickettsia DNA was negative in tissues. All inoculated animals had anti-Ca. R. colombiensis IgG antibodies with titers ranging from 1:64 to 1:1024. Controls were negative. None of the sera presented antibodies against R. rickettsii and R. parkeri. 56% (84/150) of the human sera had IgG antibodies against Ca. R. colombiensis. However, when sera with titers equal to or greater than 1:128 were evaluated, 100% were seroreactive. The reassembly produced a genome of (~1.4 Mbp) and four plasmid contigs. A total of 1852 genes were identified, of which 1200 were coding and 652 (35%) were pseudogenes, with a GC content of 32.4% and the presence of four plasmids. Phylogenetic analysis showed that Ca. R. colombiensis is located in the GFMIb suclade containing R. tamurae, R. monacensis and Rickettsia endosymbiont of Ixodes pacificus with 100% branch support. VFanalyzer and comparative genomics showed the absence of the genes rickA, RARP-2, VapC and the pseudogenization of ralf and RisK1. The 3D structures of the OmpA and OmpB proteins were in agreement with the PDB models, whereas the Sca1 and Sca2 models presented a low quality. Conclusion. Ca. R. colombiensis causes a subclinical infection in hamsters and suggests the possibility of infecting other mammals. However, clinical, pathological, and molecular findings conclude that it is not a pathogenic species. Cross-reactivity could be an advantage for diagnosing rickettsiosis with Ca. R. colombiensis antigenseng
dc.description.degreelevelDoctorado
dc.description.degreenameDoctor(a) en Microbiología y Salud Tropical
dc.description.modalityTrabajos de Investigación y/o Extensión
dc.description.tableofcontentsINDICE DE FIGURASspa
dc.description.tableofcontentsINDICE DE TABLASspa
dc.description.tableofcontentsINDICE DE ANEXOSspa
dc.description.tableofcontentsRESUMENspa
dc.description.tableofcontentsABSTRACTspa
dc.description.tableofcontentsINTRODUCCIÓNspa
dc.description.tableofcontentsPLANTEAMIENTO DEL PROBLEMAspa
dc.description.tableofcontentsMARCO TEORICOspa
dc.description.tableofcontentsClasificación de Rickettsiaespa
dc.description.tableofcontentsCandidatus a Rickettsia patógenos de humanos.spa
dc.description.tableofcontentsCandidatus a Rickettsia como posible patógenos humanosspa
dc.description.tableofcontentsCandidatus a Rickettsia reportados en animalesspa
dc.description.tableofcontentsPatogenesis de Rickettsiaespa
dc.description.tableofcontentsInmunidad del hospedero a la infecciónspa
dc.description.tableofcontentsOBJETIVOSspa
dc.description.tableofcontentsHIPÓTESISspa
dc.description.tableofcontentsMETODOLOGIAspa
dc.description.tableofcontentsTipo de estudio y aspectos eticosspa
dc.description.tableofcontentsMetodología Objetivo específico 1. Establecer la infección y los cambios patológicos en Syriam hamster (Mesocricetus auratus) inoculados con Candidatus Rickettsia colombiensisspa
dc.description.tableofcontentsMetodología Objetivo específico 2. Establecer la reactividad cruzada de los anticuerpos producidos en los Syriam hamster (M. auratus) infectados con Ca. R. colombiensis contra otras especies de rickettsia del GFM circulantes en la regiónspa
dc.description.tableofcontentsMetodología Objetivo específico 3. Analizar la reactividad cruzada de los anticuerpos producidos en los Syriam hamster (M. auratus) infectados con Ca. R. colombiensis contra otras especies de rickettsia circulantes en la regiónspa
dc.description.tableofcontentsMetodología Objetivo específico 4. Caracterización del genoma Candidatus Rickettsia colombiensis (anteriormente Candidatus Rickettsia colombianensi)spa
dc.description.tableofcontentsRESULTADOSspa
dc.description.tableofcontentsResultados del Objetivo específico 1spa
dc.description.tableofcontentsResultados del Objetivo 2spa
dc.description.tableofcontentsResultados del Objetivo 3spa
dc.description.tableofcontentsResultados objetivo 4spa
dc.description.tableofcontentsDISCUSIÓNspa
dc.description.tableofcontentsCONCLUSIONESspa
dc.description.tableofcontentsRECOMENDACIONES Y PERSPECTIVASspa
dc.description.tableofcontentsREFERENCIASspa
dc.description.tableofcontentsANEXOSspa
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/9560
dc.language.isospa
dc.publisherUniversidad de Córdoba
dc.publisher.facultyFacultad de Medicina Veterinaria y Zootecnia
dc.publisher.placeMontería, Córdoba, Colombia
dc.publisher.programDoctorado en Microbiología y Salud Tropical
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dc.rightsCopyright Universidad de Córdoba, 2025
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.rights.coarhttp://purl.org/coar/access_right/c_abf2
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.subject.keywordsPathogeniceng
dc.subject.keywordsAnimal modeleng
dc.subject.keywordsCross-reactivityeng
dc.subject.keywordsRickettsiaeng
dc.subject.keywordsIndirect fluorescent antibody techniqueeng
dc.subject.keywordsGenomeeng
dc.subject.proposalPatogenicidadspa
dc.subject.proposalModel animalspa
dc.subject.proposalReactividad cruzadaspa
dc.subject.proposalRickettsiaspa
dc.subject.proposalInmunofluorescencia indirectaspa
dc.subject.proposalGenomaspa
dc.titleEstudio sobre la patogenicidad de Candidatus Rickettsia colombiensisspa
dc.typeTrabajo de grado - Doctorado
dc.type.coarhttp://purl.org/coar/resource_type/c_db06
dc.type.coarversionhttp://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.contentText
dc.type.driverinfo:eu-repo/semantics/doctoralThesis
dc.type.redcolhttp://purl.org/redcol/resource_type/TD
dc.type.versioninfo:eu-repo/semantics/acceptedVersion
dspace.entity.typePublication
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