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Caracterización de Clostridium perfringens circulante en muestras fecales de humanos y animales en el altiplano Cundiboyacense Colombiano

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dc.contributor.advisorMuñoz Díaz, Claudia Marina
dc.contributor.advisorRamírez González, Juan David
dc.contributor.gruplacGrupo de Investigaciones Microbiológicas UR (GIMUR)
dc.creatorCamargo Mancipe, Anny Jineth
dc.creator.degreeDoctor en Ciencias Biomédicas y Biológicas
dc.creator.degreeLevelDoctorado
dc.creator.degreetypeFull time
dc.date.accessioned2024-07-10T13:16:51Z
dc.date.available2024-07-10T13:16:51Z
dc.date.created2024-06-18
dc.descriptionClostridium perfringens es una bacteria anaerobia Gram positiva, formadora de esporas y productora de toxinas, que puede encontrarse en el tracto intestinal humano y animal, así como en alimentos, suelo y agua en forma esporulada. Esta bacteria es reconocida como causante de brotes de intoxicación alimentaria, necrosis intestinal y mionecrosis grave en humanos y animales, ocupando el tercer lugar en incidencia de enfermedades transmitidas por alimentos (ETA) en los Estados Unidos. Estudios recientes han revelado la presencia de aislamientos toxigénicos de C. perfringens, portadores de la toxina perfingolisina O (PFOA) en individuos sanos. Esta toxina se relacionó con toxicidad celular, hemólisis completa, respuestas proinflamatorias y una mayor capacidad de esporulación. La combinación de es estas características biológicas en aislamientos de C. perfringens se correlacionan con un avance desalentador de la patología intestinal, estando asociado con infección/enfermedad y una mayor propagación entre hospederos. Asimismo, se ha observado un aumento en los factores de virulencia y marcadores de resistencia a los antibióticos (MRA) que transporta esta especie bacteriana, lo que representa un desafío considerable tanto a nivel clínico como de salud pública a escala global. A pesar de la relevancia para la salud pública, es evidente que en Colombia y en Suramérica existe un conocimiento limitado sobre esta bacteria. Por lo tanto, es crucial avanzar en la descripción de su arquitectura genómica, así como de sus factores de virulencia y resistencia antibiótica. Estos conocimientos son fundamentales para mejorar las estrategias de prevención y manejo de las enfermedades causadas por C. perfringens. Por lo tanto, esta tesis doctoral tuvo como objetivo describir la frecuencia de detección, diversidad genética y principales factores de virulencia de C. perfringens en muestras fecales de humanos y animales en el altiplano Cundiboyacense Colombiano. La tesis se dividió en tres capítulos: El Capítulo I se enfocó en determinar la frecuencia de detección de C. perfringens en humanos y animales en el altiplano Cundiboyacense colombiano mediante pruebas moleculares. Las muestras fecales de humanos con y sin síntomas gastrointestinales, así como muestras fecales de diversas especies animales, fueron colectadas en los Departamentos de Cundinamarca y Boyacá. Se realizó la extracción de ADN y PCR dirigida al gen 16S-rRNA y al gen de la toxina alfa (cpa) para la detección de C. perfringens. Los resultados revelaron una alta frecuencia de detección de C. perfringens a nivel general en animales domésticos y cerdos. En humanos la toxina beta 2 de C. perfringens (CPB2) se asoció con la presencia de diabetes, lo que sugiere interacciones con el sistema inmunológico del huésped. El Capítulo II tuvo como objetivo describir la estructura genética poblacional y los factores de virulencia de C. perfringens utilizando genomas públicos para evaluar la diversidad genética, linajes y toxinotipos circulantes en diferentes hospederos, así como los principales factores de virulencia a nivel global. Los análisis bioinformáticos revelaron que la mayoría de los genomas provienen de países desarrollados como EE. UU., Francia y China, cuyos aislamientos fueron establecidos principalmente de alimentos, aves y humanos. La clasificación filogenética mostró rutas de dispersión entre diferentes hospederos, además se encontró una alta frecuencia de toxinas como toxina alfa (CPA) y enterotoxina (CPE), así como el incremento en la detección de MRA asociados a tetraciclinas y macrólidos. Por otra parte, se detectó una escasa representación de aislamientos en países en desarrollo, destacando la necesidad de investigaciones locales para comprender mejor la diversidad genética y los mecanismos de transmisión y virulencia de este patógeno. El propósito del Capítulo III fue caracterizar la arquitectura genómica, factores de virulencia y MRA de aislamientos colombianos de C. perfringens, obtenidos de muestras positivas de humanos y animales recolectadas en el Capítulo I. Además, se llevaron a cabo pruebas fenotípicas de hemólisis, inhibición del crecimiento celular, esporulación y susceptibilidad a antibióticos en un grupo representativo de aislamientos. El criterio de selección de los aislamientos se basó en su capacidad para producir la toxina PFOA, una toxina formadora de poros que juega un papel crucial en la patogenia intestinal, con el objetivo de evaluar su impacto biológico. El análisis microgeográfico de aislamientos obtenidos de diversas fuentes en una región central de Colombia, reveló una amplia diversidad genética y posibles eventos de dispersión entre humanos, y animales domésticos como perros y gatos. La presencia de toxinas como PFOA en aislamientos de individuos asintomáticos plantea un potencial riesgo de infección en dicha población, debido a su asociación con hemolisis, inhibición del crecimiento celular y mayor capacidad de esporulación. Además, se observó una reducción de la susceptibilidad a varios antibióticos, incluyendo gentamicina, eritromicina, metronidazol y tetraciclina. En resumen, este estudio ha enriquecido el entendimiento de C. perfringens en Colombia al proporcionar datos de epidemiología molecular y genómica de C. perfringens circulante en la región de análisis. Además, ofreció el primer reporte de genomas obtenidos de gatos a nivel mundial e incluyó el mayor número de genomas registrados en América Latina hasta la fecha. Nuestros hallazgos permitieron fortalecer las acciones de promoción de la salud y prevención de la transmisión de enfermedades infecciosas a través de campañas educativas con la comunidad y las entidades de salud, con implicaciones significativas para mejorar la calidad de vida de poblaciones vulnerables.
dc.description.abstractClostridium perfringens is a Gram-positive, spore-forming, toxin-producing, anaerobic, Gram-positive bacterium that can be found in the human and animal intestinal tract, as well as in food, soil and water in sporulated form. This bacterium is known to cause outbreaks of food poisoning, intestinal necrosis and severe myonecrosis in humans and animals, ranking third in incidence of foodborne illness (FBD) in the United States. Recent studies have revealed the presence of toxigenic isolates of C. perfringens, carriers of the toxin perfingolysin O (PFOA) in healthy individuals. This toxin was associated with cell toxicity, complete hemolysis, proinflammatory responses and increased sporulation capacity. The combination of these biological characteristics in C. perfringens isolates correlates with a discouraging progression of intestinal pathology, being associated with infection/disease and increased host-to-host spread. Furthermore, an increase in virulence factors and antibiotic resistance markers (ARMs) carried by this bacterial species has been observed, representing a considerable challenge both clinically and in public health on a global scale. Despite the relevance for public health, it is evident that in Colombia and South America there is limited knowledge about this bacterium. Therefore, it is crucial to advance in the description of its genomic architecture, as well as its virulence and antibiotic resistance factors. This knowledge is fundamental to improve prevention and management strategies for diseases caused by C. perfringens. Therefore, this doctoral thesis aimed to describe the frequency of detection, genetic diversity and main virulence factors of C. perfringens in fecal samples from humans and animals in the Cundiboyacense highlands of Colombia. The thesis was divided into three chapters: Chapter I focused on determining the frequency of detection of C. perfringens in humans and animals in the Colombian Cundiboyacense highlands using molecular tests. Fecal samples from humans with and without gastrointestinal symptoms, as well as fecal samples from various animal species, were collected in the Departments of Cundinamarca and Boyacá. DNA extraction and PCR targeting the 16S-rRNA gene and the alpha toxin gene (cpa) were performed for the detection of C. perfringens. The results revealed a high frequency of detection of C. perfringens at the general level in domestic animals and pigs. In humans, C. perfringens beta 2 toxin (CPB2) was associated with the presence of diabetes, suggesting interactions with the host immune system. Chapter II aimed to describe the population genetic structure and virulence factors of C. perfringens using public genomes to assess genetic diversity, lineages and circulating toxinotypes in different hosts, as well as the main virulence factors at the global level. Bioinformatics analysis revealed that most of the genomes came from developed countries such as USA, France and China, whose isolates were mainly established from food, poultry and humans. Phylogenetic classification showed dispersal routes between different hosts, and a high frequency of toxins such as alpha toxin (CPA) and enterotoxin (CPE) was found, as well as an increase in the detection of MRA associated with tetracyclines and macrolides. On the other hand, an underrepresentation of isolates from developing countries was detected, highlighting the need for local research to better understand the genetic diversity and the mechanisms of transmission and virulence of this pathogen. The purpose of Chapter III was to characterize the genomic architecture, virulence factors and MRA of Colombian isolates of C. perfringens, obtained from positive samples from humans and animals collected in Chapter I. In addition, phenotypic tests for hemolysis, cell growth inhibition, sporulation and antibiotic susceptibility were performed on a representative group of isolates. The selection criteria for isolates were based on their ability to produce PFOA toxin, a pore-forming toxin that plays a crucial role in intestinal pathogenesis, in order to assess their biological impact. Microgeographic analysis of isolates obtained from different sources in a central region of Colombia revealed a wide genetic diversity and possible dispersal events among humans and domestic animals such as dogs and cats. The presence of toxins such as PFOA in isolates from asymptomatic individuals poses a potential risk of infection in this population, due to their association with hemolysis, cell growth inhibition and increased sporulation capacity. In addition, reduced susceptibility to several antibiotics, including gentamicin, erythromycin, metronidazole and tetracycline, was observed. In summary, this study has enriched the understanding of C. perfringens in Colombia by providing molecular epidemiology and genomic data of circulating C. perfringens in the region of analysis. In addition, it provided the first report of genomes obtained from cats worldwide and included the largest number of genomes recorded in Latin America to date. Our findings allowed us to strengthen health promotion actions and prevention of infectious disease transmission through educational campaigns with the community and health entities, with significant implications for improving the quality of life of vulnerable populations.
dc.format.extent170 pp
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.48713/10336_42986
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/42986
dc.language.isospa
dc.publisherUniversidad del Rosario
dc.publisher.departmentEscuela de Medicina y Ciencias de la Salud
dc.publisher.programDoctorado en Ciencias Biomédicas y Biológicas
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dc.source.instnameinstname:Universidad del Rosario
dc.source.reponamereponame:Repositorio Institucional EdocUR
dc.subjectClostridium perfringens
dc.subjectDetección
dc.subjectColonización
dc.subjectColombia
dc.subjectAltiplano Cundiboyacense Colombiano
dc.subject.keywordClostridium perfringens
dc.subject.keywordDetection
dc.subject.keywordColonization
dc.subject.keywordCundiboyacense Colombian Highlands
dc.titleCaracterización de Clostridium perfringens circulante en muestras fecales de humanos y animales en el altiplano Cundiboyacense Colombiano
dc.title.TranslatedTitleCharacterization of circulating Clostridium perfringens in fecal samples from humans and animals in the Cundiboyacense highlands of Colombia.
dc.typedoctoralThesis
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dc.type.hasVersioninfo:eu-repo/semantics/acceptedVersion
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local.department.reportEscuela de Medicina y Ciencias de la Salud
local.regionesBogotá
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Doctorado en Ciencias Biomédicas y Biológicas