Publicación:
Resistencia a insecticidas en Aedes aegypti del departamento de Córdoba: estatus, mecanismos moleculares y bioquímicos e implicaciones en salud pública

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dc.contributor.advisorHoyos López, Richard Onalbis
dc.contributor.advisorMaestre Serrano, Ronald Yesid
dc.contributor.advisorPareja Loaiza, Paula Ximena
dc.contributor.authorAtencia Pineda, María Claudia
dc.contributor.juryContreras Gutiérrez, María Angélica
dc.contributor.juryLaurito, Magdalena
dc.contributor.juryDávila Barboza, Jesús Antonio
dc.date.accessioned2025-08-25T15:40:42Z
dc.date.available2025-08-25T15:40:42Z
dc.date.issued2025-08-25
dc.description.abstractIntroducción: El control vectorial es hasta el momento la estrategia más eficaz para disminuir las densidades de insectos y reducir los casos de las enfermedades trasmitidas por vector (ETV). Los insecticidas son la piedra angular de los programas de control vectorial y su uso esta especialmente indicado durante brotes y epidemias. Sin embargo, el uso incorrecto de los insecticidas ha ocasionado la selección de poblaciones resistentes a los insecticidas de uso en salud pública. Por tal motivo antes de iniciar una intervención con insecticidas se debe conocer el estado de susceptibilidad de las poblaciones del vector a intervenir para que el control sea eficaz. El departamento de Córdoba es una de las entidades territoriales en Colombia donde más se presentan casos de dengue. Por esta razón, se utiliza durante las epidemias los insecticidas para interrumpir la transmisión y disminuir las densidades del vector Aedes aegypti. Desafortunadamente en el departamento de Córdoba no se tiene línea base de susceptibilidad a los insecticidas para las poblaciones del vector en aproximadamente el 90% de sus municipios. Esta poca información relacionada con el estado de susceptibilidad a insecticidas piretroides fue la iniciativa para el desarrollo de este trabajo. Objetivo: Evaluar el estado de la susceptibilidad a insecticidas organofosforados y piretroides y los mecanismos enzimáticos y moleculares asociados en poblaciones de Ae. aegypti del departamento de Córdoba. Métodos: Se evalúo la susceptibilidad al larvicida temefos (DD: 0,0012 µg/µl) con la metodología de la OMS y a los organofosforados malatión (50 μg/botella), fenitrotión (50 μg/botella), pirimifos-metil (75 μg/botella) con la metodología de las botellas impregnadas de los CDC y a los insecticidas piretroides permetrina (DD: 0,4% y DD: 15 µg/botella), lambdacialotrina (DD: 0,05% y DD: 10 µg/botella) y deltametrina (DD: 0,03% y DD: 10 µg/botella) con la metodología de los tubos con papeles impregnados de la OMS y la técnica del CDC. Las poblaciones que mostraron resistencia a las dosis diagnosticas establecidas para los organofosforados y piretroides con la metodología CDC se evalúo la intensidad de la resistencia 2 veces la dosis diagnostica, 5 veces y 10 veces. Se realizaron ensayos bioquímicos para detectar alteraciones en la actividad de las enzimas α, β-esterasas, GST, OFM y iAChE como posibles mecanismos de resistencia metabólica y para la identificación de las mutaciones Kdr F1534C, V101I y V410L se realizó una PCR alelo específica para la genotipificación de los mosquitos identificados como fenotípicamente resistentes y susceptibles a los piretroides evaluados por las técnicas OMS y CDC. Se determino la frecuencia alélica y genotípica para cada mutación y la asociación de los haplotipos trilocus con la resistencia a los insecticidas piretroides mediante odds ratio. Para la detección de las mutaciones en el gen Ace-1 que pudieran conferir resistencia a los organofosforados se realizó una PCR convencional con la cual se amplifico un fragmento de aproximadamente 500 pb del Exón 5 del gen, en 40 individuos de cada población y se obtuvieron las secuencias de ADN mediante secuenciación por el método de Sanger. Resultados: Se observó susceptibilidad a los insecticidas temefos, malatión en todas las poblaciones, susceptibilidad a pirimifos-metil en 7 poblaciones y resistencia generalizada al organofosforado fenitrotión. Se encontró resistencia a los insecticidas piretroides permetrina, lambdacialotrina y deltametrina por la metodología OMS y resistencia a permetrina en nueve poblaciones y a lambdacialotrina en Montería con la metodología del CDC. Se encontraron alterados los niveles de actividad de las enzimas α, β-esterasas, GST y iAChE en algunas poblaciones, sin embargo, no se encontró asociación entre la actividad enzimática y el fenotipo de resistencia. Se identificaron las mutaciones Kdr F1534C, V1016I y V410L con frecuencias alélicas que oscilaron entre 0.94 – 1.0, 0.01 – 0.6, 0.01 – 0.59 respectivamente. El haplotipo kdr triple mutante CIL se asoció con la resistencia a permetrina, lambdacialotrina y deltametrina, mientras que el haplotipo CVL se asoció con la resistencia a deltametrina. No se encontraron mutaciones en el gen Ace-1 relacionadas con la resistencia a organofosforados. Conclusiones: Los resultados obtenidos contribuyen conocer el estado actual de la susceptibilidad a insecticidas en las zonas evaluadas y proveen información para la vigilancia de la resistencia en estos municipios, lo que resulta útil para el fortalecimiento de las políticas públicas en el control vectorial.spa
dc.description.abstractBackground: Vector control is currently the most effective strategy for reducing insect densities and reducing the incidence of vector-borne diseases (VBDs). Insecticides are the cornerstone of vector control programs and are especially recommended during outbreaks and epidemics. However, the incorrect use of insecticides has led to the selection of populations resistant to public health insecticides. Therefore, before initiating an intervention with insecticides, the susceptibility status of the target vector populations must be known to ensure effective control. The department of Córdoba is one of the Colombian territories with the highest incidence of dengue fever. For this reason, insecticides are used during epidemics to interrupt transmission and reduce the density of the Aedes aegypti vector. Unfortunately, in the department of Córdoba, there is no baseline insecticide susceptibility for vector populations in approximately 90% of its municipalities. This limited information regarding susceptibility to pyrethroid insecticides was the impetus for this study. Objective: To evaluate the susceptibility status to organophosphate and pyrethroid insecticides and the associated enzymatic and molecular mechanisms in Ae. aegypti populations in the department of Córdoba. Methods: Susceptibility to the larvicide temephos (DD: 0.0012 µg/µl) was assessed using the WHO methodology and to the organophosphates malathion (50 µg/bottle), fenitrothion (50 µg/bottle), pirimiphos-methyl (75 µg/bottle) using the CDC soaked bottle methodology and to the pyrethroid insecticides permethrin (DD: 0.4% and DD: 15 µg/bottle), lambda-cyhalothrin (DD: 0.05% and DD: 10 µg/bottle) and deltamethrin (DD: 0.03% and DD: 10 µg/bottle) using the WHO soaked paper tube methodology and the CDC technique. The populations that showed resistance to the diagnostic doses established for organophosphates and pyrethroids using the CDC methodology were evaluated for the intensity of resistance at 2 times, 5 times, and 10 times the diagnostic dose. Biochemical assays were performed to detect alterations in the activity of the enzymes α, β-esterases, GST, OFM and iAChE as possible mechanisms of metabolic resistance and to identify the Kdr F1534C, V101I and V410L mutations, an allele-specific PCR was performed for genotyping the mosquitoes identified as phenotypically resistant and susceptible to pyrethroids evaluated by the WHO and CDC techniques. The allele and genotype frequencies for each mutation were determined, as well as the association of trilocus haplotypes with resistance to pyrethroid insecticides using odds ratios. To detect mutations in the Ace-1 gene that could confer resistance to organophosphates, a conventional PCR was performed, amplifying a fragment of approximately 500 bp from exon 5 of the gene in 40 individuals from each population, and DNA sequences were obtained by Sanger sequencing. Results: Susceptibility to the insecticides temephos and malathion was observed in all populations, susceptibility to pirimiphos-methyl in seven populations, and widespread resistance to the organophosphate fenitrothion. Resistance to the pyrethroid insecticides permethrin, lambda-cyhalothrin, and deltamethrin was found using the WHO methodology, while resistance to permethrin was found in nine populations and to lambda-cyhalothrin in Montería using the CDC methodology. Altered activity levels of α-, β-esterase, GST, and iAChE enzymes were found in some populations; however, no association was found between enzyme activity and the resistance phenotype. Kdr mutations F1534C, V1016I, and V410L were identified with allele frequencies ranging from 0.94–1.0, 0.01–0.6, and 0.01–0.59, respectively. The triple mutant kdr haplotype CIL was associated with resistance to permethrin, lambda-cyhalothrin, and deltamethrin, while the CVL haplotype was associated with resistance to deltamethrin. No mutations in the Ace-1 gene were found to be associated with organophosphate resistance. Conclusions: The results obtained contribute to understanding the current status of insecticide susceptibility in the evaluated areas and provide information for resistance surveillance in these municipalities, which is useful for strengthening public policies for vector control.eng
dc.description.degreelevelDoctorado
dc.description.degreenameDoctor(a) en Microbiología y Salud Tropical
dc.description.modalityTrabajos de Investigación y/o Extensión
dc.description.tableofcontentsRESUMENspa
dc.description.tableofcontentsABSTRACTspa
dc.description.tableofcontentsINTRODUCCIÓNspa
dc.description.tableofcontentsPLANTEAMIENTO DEL PROBLEMAspa
dc.description.tableofcontentsJUSTIFICACIÓNspa
dc.description.tableofcontentsMARCO CONCEPTUALspa
dc.description.tableofcontentsANTECEDENTESspa
dc.description.tableofcontentsHIPÓTESISspa
dc.description.tableofcontentsOBJETIVOSspa
dc.description.tableofcontentsCAPITULO I: DETERMINAR EL ESTADO DE LA SUSCEPTIBILIDAD Y LA INTENSIDAD DE LA RESISTENCIA A INSECTICIDAS ORGANOFOSFORADOS y PIRETROIDES EN DIEZ POBLACIONES DE Aedes aegypti DEL DEPARTAMENTO DE CÓRDOBA.spa
dc.description.tableofcontentsMETODOLOGÍAspa
dc.description.tableofcontentsRESULTADOSspa
dc.description.tableofcontentsDISCUSIÓNspa
dc.description.tableofcontentsCAPITULO II: IDENTIFICAR MECANISMOS ENZIMÁTICOS DE RESISTENCIA ASOCIADOS A INSECTICIDAS DE TIPO ORGANOFOSFORADO Y PIRETROIDE EN LAS POBLACIONES DE Aedes aegypti OBJETO DE ESTUDIO.spa
dc.description.tableofcontentsMETODOLOGIAspa
dc.description.tableofcontentsRESULTADOSspa
dc.description.tableofcontentsDISCUSIÓNspa
dc.description.tableofcontentsCAPITULO III: DETECTAR LAS MUTACIONES kdr F1534C, V1016I, V410L Y MUTACIONES EN EL GEN Ace-1, SUS FRECUENCIAS ALÉLICAS Y GENOTÍPICAS Y EVALUAR SU ASOCIACIÓN CON LA RESISTENCIA OBSERVADA.spa
dc.description.tableofcontentsMETODOLOGIAspa
dc.description.tableofcontentsRESULTADOSspa
dc.description.tableofcontentsDISCUSIÓNspa
dc.description.tableofcontentsDISCUSIÓN GENERALspa
dc.description.tableofcontentsCONCLUSIONESspa
dc.description.tableofcontentsRECOMENDACIONESspa
dc.description.tableofcontentsREFERENCIASspa
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/9536
dc.language.isospa
dc.publisherUniversidad de Córdoba
dc.publisher.facultyFacultad de Medicina Veterinaria y Zootecnia
dc.publisher.placeBerástegui, 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.keywordsAedes aegyptieng
dc.subject.keywordsInsecticide resistanceeng
dc.subject.keywordsKdr mutationseng
dc.subject.keywordsDetoxification enzymeseng
dc.subject.keywordsCórdobaeng
dc.subject.proposalAedes aegyptispa
dc.subject.proposalResistencia a insecticidasspa
dc.subject.proposalMutaciones kdrspa
dc.subject.proposalEnzimas de detoxificaciónspa
dc.subject.proposalCórdobaspa
dc.titleResistencia a insecticidas en Aedes aegypti del departamento de Córdoba: estatus, mecanismos moleculares y bioquímicos e implicaciones en salud públicaspa
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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