Escherichia coli ST471 Producing VIM-4 Metallo-ß-Lactamase in Colombia.

An Escherichia coli isolate sequence-type 471 (ST471) producing Verona integron-encoded metallo-ß-lactamases (VIM)-4 was recovered from a rectal swab in a patient without travel records with osteomyelitis in Colombia. The isolate carried a class 1 integron-borne blaVIM-4 gene with a 170-bp duplication in the 3' end of the gene, preceded by an aac(6')-Ib gene. The genetic environment of blaVIM-4, blaCMY-2, and sul2 genes showed similarities to the backbone of pKKp4, an IncA/C-type plasmid from a Klebsiella pneumoniae strain carrying blaVIM-4 recovered in Kuwait. This is the first report of blaVIM-4 in Enterobacterales in South America. Our results suggest that blaVIM-4 gene was found on an IncA/C-type plasmid that could play a role in the spread of VIM-4 carbapenemase in Colombia.

Genetic Diversity of Multidrug-Resistant Pseudomonas aeruginosa Isolates Carrying bla VIM-2 and bla KPC-2 Genes That Spread on Different Genetic Environment in Colombia.

Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen with an increase in the frequency of infections caused by multidrug resistant (MDR) and extensively drug resistant (XDR) strains, limiting the available therapeutic options. The most troublesome resistance is the acquisition and production of carbapenemases such as Verona integron-encoded metallo-ß-lactamases (VIM), the most frequent and widespread, and the Klebsiella pneumoniae carbapenemases (KPC), which has continuously spread in the last decade. Its dissemination is linked to their location on mobile genetic elements (MGEs). In Colombia, VIM and KPC have been increasing in its frequency showing major successful dissemination. In this article, we molecularly characterized and analyzed the genetic context of bla VIM and bla KPC in carbapenem-resistant P. aeruginosa (CRPA) isolates from infected and colonized patients in two tertiary-care hospitals, one in Medellín and the other in a municipality close to Medellín, both areas with high carbapenemase endemicity in Colombia (2013-2015). Using whole-genome sequencing (WGS), we identified a remarkable variety of genetic backgrounds in these MDR P. aeruginosa isolates carrying bla KPC- 2 and bla VIM- 2. There were a diversity of class 1 integron and variations in the gene cassettes associated to bla VIM- 2, as well as a possible event of spread of bla KPC- 2 mediated by a plasmid that contained part of Tn4401b in one infection case. The dissemination of bla VIM- 2 and bla KPC- 2 in P. aeruginosa in this area in Colombia has been strongly influenced by successful international clones, carrying these genes and additional determinants of resistance on MGEs, accompanied by gene rearrangement under an antimicrobial selection pressure. These findings emphasize the need to implement control strategies based on rational antibiotic use.

Prevalence of Chlamydia trachomatis in an asymptomatic female population attending cervical cytology services of three healthcare centers in Medellín, Colombia / Prevalencia de Chlamydia trachomatis en la población femenina asintomática atendida en los servicios de citología cervical de tres instituciones prestadoras de servicios de salud en Medellín, Colombia.

Introduction: The sexually transmitted infection caused by Chlamydia trachomatis is the most common of bacterial etiology in the world. Although it can be asymptomatic in most cases, it can produce several reproductive health problems in women such as cervicitis, endometritis, and salpingitis. Despite its importance, the epidemiological data on this infection is insufficient in Medellín. Objective: To determine the prevalence of C. trachomatis in women from Medellín, identify possible risk factors, and evaluate its relationship with the human papillomavirus infection (HPV). Materials and methods: We conducted a multicenter cross-sectional study to detect C. trachomatis infection in 1,282 women using PCR and the LightMix 480 HT CT/NG™ commercial kit (Roche, Basel, Switzerland). Results: The total prevalence of C. trachomatis infection was 4.1% (95% CI: 2.9-5.3). We found an association between the presence of the infection and age, cigarette consumption, and the use of hormonalcontraceptives. Conclusion: The prevalence of the infection is similar to that reported in other parts of the world with young women being the most affected population. No relation was found with the presence of HPV.

Introducción. La infección de transmisión sexual causada por Chlamydia trachomatis es la más frecuente de etiología bacteriana en el mundo. Aunque puede ser asintomática en la mayoría de los casos, C. trachomatis puede generar diversos problemas de salud reproductiva en mujeres, como cervicitis, endometritis y salpingitis. A pesar de su importancia, en Medellín no se cuenta con suficientes datos epidemiológicos sobre esta infección. Objetivo. Determinar la prevalencia de C. trachomatis en mujeres de Medellín, determinar los posibles factores de riesgo y evaluar la relación con la infección por el virus de papiloma humano (HPV). Materiales y métodos. Se hizo un estudio transversal multicéntrico para detectar la infección por C. trachomatis en 1.282 mujeres mediante reacción en cadena de la polimerasa (PCR) convencional y el estuche comercial LightMix 480 HT CT/NG™ (Roche, Basilea, Suiza). Resultados. La prevalencia total de la infección por C. trachomatis fue de 4,1 % (IC95% 2,9-5,3). Se encontró una relación significativa de la infección con la edad, el consumo de cigarrillo y el uso de anticonceptivos hormonales. Conclusión. La prevalencia de la infección es similar a la reportada en otros lugares del país y del mundo, siendo las mujeres más jóvenes las más afectadas. En cuanto a la presentación del HPV, no se encontró ningún tipo de relación con C. trachomatis.

Dynamics of blaKPC-2 Dissemination from Non-CG258 Klebsiella pneumoniae to Other Enterobacterales via IncN Plasmids in an Area of High Endemicity.

Carbapenem-resistant Enterobacterales (CRE) pose a significant threat to global public health. The most important mechanism for carbapenem resistance is the production of carbapenemases. Klebsiella pneumoniae carbapenemase (KPC) represents one of the main carbapenemases worldwide. Complex mechanisms of blaKPC dissemination have been reported in Colombia, a country with a high endemicity of carbapenem resistance. Here, we characterized the dynamics of dissemination of blaKPC gene among CRE infecting and colonizing patients in three hospitals localized in a highly endemic area of Colombia (2013 and 2015). We identified the genomic characteristics of KPC-producing Enterobacterales recovered from patients infected/colonized and reconstructed the dynamics of dissemination of blaKPC-2 using both short and long read sequencing. We found that spread of blaKPC-2 among Enterobacterales in the participating hospitals was due to intra- and interspecies horizontal gene transfer (HGT) mediated by promiscuous plasmids associated with transposable elements that was originated from a multispecies outbreak of KPC-producing Enterobacterales in a neonatal intensive care unit. The plasmids were detected in isolates recovered in other units within the same hospital and nearby hospitals. The gene "epidemic" was driven by IncN-pST15-type plasmids carrying a novel Tn4401b structure and non-Tn4401 elements (NTEKPC) in Klebsiella spp., Escherichia coli, Enterobacter spp., and Citrobacter spp. Of note, mcr-9 was found to coexist with blaKPC-2 in species of the Enterobacter cloacae complex. Our findings suggest that the main mechanism for dissemination of blaKPC-2 is HGT mediated by highly transferable plasmids among species of Enterobacterales in infected/colonized patients, presenting a major challenge for public health interventions in developing countries such as Colombia.

Prevalencia de Chlamydia trachomatis en la población femenina asintomática atendida en los servicios de citología cervical de tres instituciones prestadoras de servicios de salud en Medellín, Colombia / Prevalence of Chlamydia trachomatis in an asymptomatic female population attending cervical cytology services of three healthcare centers in Medellín, Colombia

Introduction:The sexually transmitted infection caused by Chlamydia trachomatis is the most common of bacterial etiology in the world. Although it can be asymptomatic in most cases, it can produce several reproductive health problems in women such as cervicitis, endometritis, and salpingitis. Despite its importance, the epidemiological data on this infection is insufficient in Medellín. Objective: To determine the prevalence of C. trachomatis in women from Medellín, identify possible risk factors, and evaluate its relationship with the human papillomavirus infection (HPV). Materials and methods: We conducted a multicenter cross-sectional study to detect C. trachomatis infection in 1,282 women using PCR and the LightMix 480 HT CT/NG™ commercial kit (Roche, Basel, Switzerland). Results: The total prevalence of C. trachomatis infection was 4.1% (95% CI: 2.9-5.3). We found an association between the presence of the infection and age, cigarette consumption, and the use of hormonal contraceptives. Conclusion: The prevalence of the infection is similar to that reported in other parts of the world with young women being the most affected population. No relation was found with the presence of HPV.

Introducción. La infección de transmisión sexual causada por Chlamydia trachomatises la más frecuente de etiología bacteriana en el mundo. Aunque puede ser asintomática en la mayoría de los casos, C. trachomatis puede generar diversos problemas de salud reproductiva en mujeres, como cervicitis, endometritis y salpingitis. A pesar de su importancia, en Medellín no se cuenta con suficientes datos epidemiológicos sobre esta infección. Objetivo. Determinar la prevalencia de C. trachomatis en mujeres de Medellín, determinar los posibles factores de riesgo y evaluar la relación con la infección por el virus de papiloma humano (HPV). Materiales y métodos. Se hizo un estudio transversal multicéntrico para detectar la infección por C. trachomatis en 1.282 mujeres mediante reacción en cadena de la polimerasa (PCR) convencional y el estuche comercial LightMix 480 HT CT/NG™ (Roche, Basilea, Suiza). Resultados. La prevalencia total de la infección por C. trachomatis fue de 4,1 % (IC95%2,9-5,3). Se encontró una relación significativa de la infección con la edad, el consumo de cigarrillo y el uso de anticonceptivos hormonales. Conclusión. La prevalencia de la infección es similar a la reportada en otros lugares del país y del mundo, siendo las mujeres más jóvenes lasmás afectadas. En cuanto a la presentación del HPV, no se encontró ningún tipo de relación con C. trachomatis.

Distribution and molecular characterization of beta-lactamases in Gram-negative bacteria in Colombia, 2001-2016 / Distribución y caracterización molecular de betalactamasas en bacterias Gram negativas en Colombia, 2001-2016

Beta-lactamases are enzymes with hydrolytic activity over beta-lactam antibiotics and they are the main resistance mechanism in Gram-negative bacteria. Extended-spectrum beta-lactamases (ESBL), AmpC, and carbapenemases have the greatest clinical and epidemiological impact in hospital settings. The increasing frequency and worldwide spread of these enzymes have limited the therapeutic options in hospital-acquired infections and those originating in the community. In Colombia, surveillance networks and research groups began studying them in the late 90s. Different variants of these enzymes have been molecularly characterized and their high prevalence and dissemination in medium and high complexity hospitals, along with a high clinical impact, have been reported. Furthermore, many studies in Colombia have evidenced high endemicity for some of these beta-lactamases, which requires an urgent implementation of antimicrobial stewardship programs in order to preserve the few therapeutic options and infection control strategies to prevent and limit their dissemination. In this publication, we carried out a review of the different enzyme variants, geographic distribution, and molecular characterization of these beta-lactamases in Colombia. Additionally, we describe the available information in the literature regarding studies conducted between the late 1990s and 2016, which provide an overview of the beta-lactamases circulating in different regions of Colombia, their increase over time, and their clinical implications.

Las betalactamasas, enzimas con capacidad hidrolítica frente a los antibióticos betalactámicos, son responsables del principal mecanismo de resistencia en bacterias Gram negativas; las de mayor impacto clínico y epidemiológico en los hospitales, son las betalactamasas de espectro extendido (BLEE), las de tipo AmpC y las carbapenemasas. El incremento en su frecuencia y su diseminación a nivel mundial ha limitado cada vez más las opciones terapéuticas tanto en infecciones adquiridas en los hospitales como las que se generan en la comunidad. En Colombia, las redes de vigilancia y los grupos de investigación iniciaron su estudio desde finales de los años 90 y, así, se logró la caracterización molecular de las diferentes variantes; además, se reportó una gran prevalencia y diseminación en los hospitales de mediana y alta complejidad, y se describió el impacto clínico de las infecciones que causan. Dichos estudios han evidenciado el alto grado de endemia de algunas de estas betalactamasas y, en consecuencia, la necesidad de una inmediata implementación de programas para inducir el uso prudente de los antibióticos y de medidas de vigilancia, que permitan controlar y prevenir su diseminación, con el fin de disminuir la morbimortalidad en los pacientes y preservar las opciones terapéuticas disponibles en la actualidad. En esta revisión, se recopiló la información sobre las variantes, la distribución geográfica y la caracterización molecular de las betalactamasas en Colombia, así como los estudios llevados a cabo desde finales de la década de 90 hasta el 2016.

Distribución y caracterización molecular de betalactamasas en bacterias Gram negativas en Colombia, 2001-2016 / Distribution and molecular characterization of beta-lactamases in Gram-negative bacteria in Colombia, 2001-2016

Resumen Las betalactamasas, enzimas con capacidad hidrolítica frente a los antibióticos betalactámicos, son responsables del principal mecanismo de resistencia en bacterias Gram negativas; las de mayor impacto clínico y epidemiológico en los hospitales, son las betalactamasas de espectro extendido (BLEE), las de tipo AmpC y las carbapenemasas. El incremento en su frecuencia y su diseminación a nivel mundial ha limitado cada vez más las opciones terapéuticas tanto en infecciones adquiridas en los hospitales como las que se generan en la comunidad. En Colombia, las redes de vigilancia y los grupos de investigación iniciaron su estudio desde finales de los años 90 y, así, se logró la caracterización molecular de las diferentes variantes; además, se reportó una gran prevalencia y diseminación en los hospitales de mediana y alta complejidad, y se describió el impacto clínico de las infecciones que causan. Dichos estudios han evidenciado el alto grado de endemia de algunas de estas betalactamasas y, en consecuencia, la necesidad de una inmediata implementación de programas para inducir el uso prudente de los antibióticos y de medidas de vigilancia, que permitan controlar y prevenir su diseminación, con el fin de disminuir la morbimortalidad en los pacientes y preservar las opciones terapéuticas disponibles en la actualidad. En esta revisión, se recopiló la información sobre las variantes, la distribución geográfica y la caracterización molecular de las betalactamasas en Colombia, así como los estudios llevados a cabo desde finales de la década de 90 hasta el 2016, lo cual permitió tener un panorama de las betalactamasas que circulan en diferentes regiones, su incremento en el tiempo y sus implicaciones clínicas.

Abstract Beta-lactamases are enzymes with hydrolytic activity over beta-lactam antibiotics and they are the main resistance mechanism in Gram-negative bacteria. Extended-spectrum beta-lactamases (ESBL), AmpC, and carbapenemases have the greatest clinical and epidemiological impact in hospital settings. The increasing frequency and worldwide spread of these enzymes have limited the therapeutic options in hospital-acquired infections and those originating in the community. In Colombia, surveillance networks and research groups began studying them in the late 90s. Different variants of these enzymes have been molecularly characterized and their high prevalence and dissemination in medium and high complexity hospitals, along with a high clinical impact, have been reported. Furthermore, many studies in Colombia have evidenced high endemicity for some of these beta-lactamases, which requires an urgent implementation of antimicrobial stewardship programs in order to preserve the few therapeutic options and infection control strategies to prevent and limit their dissemination. In this publication, we carried out a review of the different enzyme variants, geographic distribution, and molecular characterization of these beta-lactamases in Colombia. Additionally, we describe the available information in the literature regarding studies conducted between the late 1990s and 2016, which provide an overview of the beta-lactamases circulating in different regions of Colombia, their increase over time, and their clinical implications.

High genetic polymorphism of relapsing P. vivax isolates in northwest Colombia.

Genetic diversity of Plasmodium populations has been more extensively documented in Colombia for Plasmodium falciparum than for Plasmodium vivax. Recently, highly variable microsatellite markers have been described and used in population-level studies of genetic variation of P. vivax throughout the world. We applied this approach to understand the genetic structure of P. vivax populations and to identify recurrence-associated haplotypes. In this, three microsatellite markers of P. vivax were amplified and the combined size of the fragments was used to establish genotypes. Patients from an ongoing treatment efficacy trial who were kept either in endemic or non-endemic regions in the northwest of Colombia were included in the study. In total 58 paired clinical isolates, were amplified. A total of 54 haplotypes were observed among the two regions. Some haplotypes were exclusive to the endemic region where the highest degree of polymorphism was detected. In addition, we confirmed the different genotypes of recurrent-relapsing and primary infection isolates suggesting the activation of heterologous hypnozoite populations. We conclude that analysis of the three microsatellites is a valuable tool to establish the genetic characteristics of P. vivax populations in Colombia.

Alta frecuencia de mutaciones puntuales en pfcrt de Plasmodium falciparum y emergencia de nuevos haplotipos mutantes en Colombia / Plasmodium falciparum: high frequency of pfcrt point mutations and emergence of new mutant haplotypes in Colombia

Introducción. Los estudios en epidemiología molecular de resistencia a antipalúdicos constituyen una herramienta útil para comprender eventos involucrados en la falla al tratamiento y la resistencia en paludismo por Plasmodium falciparum en Colombia. Diversos autores han informado sobre la eficacia de algunos marcadores moleculares para predecir resistencia a fármacos en P. falciparum y el gen pfcrt ha sido ampliamente caracterizado en este contexto. Objetivo. Estudiar la frecuencia de mutaciones en el gen pfcrt de P. falciparum y su asociacióncon falla al tratamiento con cloroquina, mefloquina, amodiaquina y sulfadoxina/pirimetamina, en dos regiones muy endémicas para paludismo del noroeste de Colombia: Turbo y Bajo Cauca. Materiales y métodos. Una muestra representativa de pacientes con paludismo por P. falciparum no complicado fue seleccionada de cada localidad para la evaluación de la respuesta al tratamiento y la determinación del estado de los codones 72, 74, 75 y 76 de pfcrt, usando una aproximación basada en PCR-RFLP. Resultados. Se confirmó una alta frecuencia de falla al tratamiento con cloroquina (82 por ciento) y amodiaquina (29 por ciento), mientras que la mefloquina y la rapia combinada fueron eficaces para eliminar la infección. La presencia de la mutación T76 en pfcrt fue confirmada en todas las 172 muestras; el haplotipo más común fue CMNT (67 por ciento). Conclusiones. No se observó asociación significativa entre un haplotipo particular y la respuesta al tratamiento en cualquiera de los grupos. Se reporta por primera vez en Colombia la presencia de dos haplotipos, SMET y SMNT; se encontraron alelos mutantes y silvestres simultáneamente en 12 por ciento de las muestras.

Plasmodium falciparum: high frequency of pfcrt point mutations and emergence of new mutant haplotypes in Colombia.

INTRODUCTION: Studies on the molecular epidemiology of antimalarial resistance constitute a useful tool to understand the events underlying treatment failure and resistance in falciparum malaria in Colombia. Several authors have reported on the efficacy of some molecular markers to predict drug resistance in Plasmodium falciparum. The P. falciparum pfcrt gene has been widely characterized in this context. OBJECTIVE: The frequency of pfcrt gene mutations in P. falciparum were associated with treatment failure to the antimalarials chloroquine, mefloquine, amodiaquine and sulfadoxine/pyrimethamine. MATERIALS AND METHODS: A representative sample of 172 patients with non-complicated falciparum malaria was selected from two highly malaria-endemic areas of northeastern Colombia, the Turbo and Bajo Cauca regions. These patients were assessed for treatment response together with the status of codons 72, 74, 75 and 76 in the pfcrt gene using a PCR-RFLP approach. RESULTS: A high frequency of treatment failure to chloroquine (82%) and to amodiaquine (29%) was confirmed, whereas mefloquine and combined therapy remained effective. The presence of the T76 mutation in pfcrt was confirmed in all samples. The most common haplotype was CMNT (67%). CONCLUSIONS: No significant association was confirmed between specific haplotypes and the treatment response in any of the treatment groups. Two haplotypes, SMET and SMNT, were reported for the first time in Colombia. Twelve percent of the samples carried both mixed mutant and wild-type alleles.

Nosocomial Plasmodium falciparum infections confirmed by molecular typing in Medellín, Colombia.

Three cases of nosocomial malaria are reported from patients of the Internal Medicine Ward of a tertiary University teaching hospital in Medellin, Colombia. Epidemiological research, based on entomological captures, medical records review and interviews of nursery staff about patient care practices potentially involving contact with blood, were carried out. Molecular characterization of Plasmodium falciparum was based on the amplification of MSP1, MSP2 and GLURP genes. This method enabled confirmation of the same P. falciparum genotype in all three patients as well as in a fourth one (index case). The presence of nosocomial malaria was confirmed and it was concluded that the most likely source of transmission was through multi-dose preparations of heparin applied to heparin locks.