Presence of Pathogenic Leptospira spp. in an Urban Slum of the Colombian Caribbean: A One Health Approach

Introducción: La leptospirosis es una zoonosis que tiene alto impacto en la salud de las personas y los animales, especialmente en áreas tropicales y subtropicales. Esta enfermedad es causada por el patógeno Leptospira spp. y es transmitida principalmente por los roedores. Objetivo: Describir la presencia de Leptospira patógena y los posibles factores de riesgo de leptospirosis en un sector marginal de Colombia, con un enfoque One-Health. Métodos: Se llevó acabo un estudio exploratorio en un sector marginal de Soledad, municipio situado en la costa norte de Colombia. Se tomaron muestras de sangre de 83 sujetos. Se analizaron variables sociodemográficas, clínicas y ecológicas relacionadas con la leptospirosis. La presencia de anticuerpos IgM anti-leptospira en el suero humano fue detectado mediante la prueba ELISA. También, se tomaron muestras de tejido renal de 53 roedores sinantrópicos para identificar Leptospira spp. patógenas mediante PCR convencional a través del uso de cebadores específicos. Resultados: Se detectaron anticuerpos IgM-anti-leptospira en el 30,12 por ciento de los sujetos de estudio. La Leptospira spp. patógena fue identificada en el 7,55 por ciento de los roedores analizados. En la muestra seleccionada se encontró asociación de casos seropositivos con ser comerciantes, ama de casa y estar en contacto con cerdos. Las condiciones higiénico-sanitarias subóptimas también fueron evidentes en el área de estudio. Conclusiones: La circulación de Leptospira spp. patógena y la exposición a factores de riesgo humanos y ecológicos es elevada en el sector marginal (área de pobreza) del Caribe colombiano. Se recomienda dirigir las intervenciones en la interfaz hombre-animal-ambiente de acuerdo con el paradigma One-Health; se debe considerar la extrema pobreza como un factor determinante para la ocurrencia de la leptospirosis(AU)

Objective: This study aims to describe the presence of pathogenic Leptospira and potential risk factors for leptospirosis in a marginal sector of the Colombian Caribbean, with a One Health approach. Methods: an exploratory study was carried out in a marginal sector of Soledad, a municipality located in the north coast of Colombia. Blood samples were taken from 83 subjects, who were also questioned about sociodemographic, clinical and ecological variables related to leptospirosis. The presence of IgM Anti-Leptospira antibodies in human serum was performed by ELISA. A total 53 synanthropic rodents were also captured using Sherman traps. Renal tissue samples were taken from rodents to identify pathogen Leptospira spp. by conventional PCR using specific primers. Results: IgM-anti-Leptospira was present in 30.12 percent of study subjects and pathogenic Leptospira spp. was identified in 7.55 percent of captured rodents. In the selected sample we found an association of positive cases with being a merchant, housewife and being in contact with pigs. Suboptimal hygienic-sanitary conditions were also evident in the study area. Conclusions: Our results show the circulation of pathogenic Leptospira spp. and exposure to human and ecological risk factors in a marginal sector (slum) of the Colombian Caribbean. We suggest to direct interventions in the human-animal-environment interface according to the One Health paradigm, considering extreme poverty as a determining factor for Leptospirosis occurrence(AU)

Impact of the International Nosocomial Infection Control Consortium (INICC) multidimensional hand hygiene approach in three cities of Colombia.

OBJECTIVE: To assess the feasibility and effectiveness of the International Nosocomial Infection Control Consortium (INICC) multidimensional hand hygiene (HH) approach in Colombia, and analyze predictors of poor HH compliance. METHODS: An observational, prospective, interventional, before-and-after study was conducted from May 2003 through September 2010 in 10 intensive care units (ICUs) of six hospitals in three cities. The study was divided into two periods: a baseline and a follow-up period. Observations for HH compliance were done in each ICU during randomly selected 30-min periods. The multidimensional HH approach included: (1) administrative support, (2) supplies availability, (3) education and training, (4) reminders in the workplace, (5) process surveillance, and (6) performance feedback. RESULTS: A total of 13 187 opportunities for HH were observed. Overall HH compliance increased from 50% to 77% (relative risk 1.55, 95% confidence interval 1.43-1.68; p=0.0001). Multivariate and univariate analyses showed that several variables were significantly associated with poor HH compliance: males vs. females (67% vs. 77%; p=0.0001), physicians vs. nurses (59% vs. 78%; p<0.0001), and adult vs. pediatric ICUs (76% vs. 42%; p<0.001), among others. CONCLUSIONS: Adherence to HH was increased by 55% with the INICC approach. Programs targeted at improving HH in variables found to be predictors of poor compliance should be implemented.

Mutaciones puntuales en el gen de la dihidropteroato sintetasa de Plasmodium falciparum en Bolívar, Colombia / Point mutations in the dihydropteroate synthetase gene of Plasmodium falciparum in Bolivar, Colombia

Introducción: la resistencia antimalárica dificulta el control del paludismo en Colombia. La vigilancia molecular de mutaciones puntuales en blancos terapéuticos es fundamental en el estudio de la resistencia a los antimaláricos. Objetivo: identificar mutaciones puntuales en el gen de la dihidropteroato sintetasa de Plasmodium falciparum (pfdhps), asociadas con resistencia in vitro a sulfadoxina. Métodos: la fuente de ADN de Plasmodium falciparum consistió en láminas de gota gruesa de 55 individuos con infección malárica, reportados en el departamento de Bolívar, Colombia. El ADN se extrajo con solución de Chelex-100 al 5 %. Las mutaciones se identificaron mediante PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) y secuenciación. Resultados: 17 muestras (31 %) amplificaron un fragmento de 438 pb de pfdhps. La PCR-RFLP mostró frecuencia del genotipo mutante G-437 en 65 % de los amplificados, el mixto A/G-437 en 29 % y el silvestre A-437 en 6 %. Los alelos mutantes G-437, F-436 y el alelo silvestre K-540 se identificaron en todas las muestras secuenciadas. Conclusiones: este es el primer reporte de mutaciones puntuales en el gen dhps de Plasmodium falciparum en el departamento de Bolívar, Colombia, lo cual contribuye al conocimiento de la resistencia a los antimaláricos en el Caribe colombiano.

Introduction: antimalarial drug resistance hinders the control of malaria in Colombia. The molecular surveillance of point mutations in therapeutic targets is essential in the study of antimalarial drugs. Objective: to identify point mutations at dihydropteroate synthetase gene of Plasmodium falciparum (pfdhps) associated with sulfadoxine resistance. Methods: source of P. falciparum DNA was blood thick smears of 55 individuals with malaria infection, reported in Bolivar, Colombia. The DNA was extracted with 5 %. Chelex-100 solution. The mutations were identified by PCR-RFLP and sequencing. Results: seventeen samples (31 %) amplified a 438 bp fragment of pfdhps. The PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) showed a frequency of mutant genotype (G-437) in 65 % of amplicons, mixed genotype (A/G-437) in 29 % and wild genotype (A/G-437) in 6 %. The mutant alleles G-437, F-436 and the wild allele K-540 were identified in all sequenced samples. Conclusions: this is the first report of point mutations in the P. falciparum dhps gene in Bolivar, Colombia. This result contributes to the knowledge of antimalarial drug resistance in the Colombian Caribbean region.

Findings of the International Nosocomial Infection Control Consortium (INICC), Part I: Effectiveness of a multidimensional infection control approach on catheter-associated urinary tract infection rates in pediatric intensive care units of 6 developing countries.

DESIGN: A before-after prospective surveillance study to assess the impact of a multidimensional infection control approach for the reduction of catheter-associated urinary tract infection (CAUTI) rates. SETTING: Pediatric intensive care units (PICUs) of hospital members of the International Nosocomial Infection Control Consortium (INICC) from 10 cities of the following 6 developing countries: Colombia, El Salvador, India, Mexico, Philippines, and Turkey. PATIENTS: PICU inpatients. METHODS: We performed a prospective active surveillance to determine rates of CAUTI among 3,877 patients hospitalized in 10 PICUs for a total of 27,345 bed-days. The study was divided into a baseline period (phase 1) and an intervention period (phase 2). In phase 1, surveillance was performed without the implementation of the multidimensional approach. In phase 2, we implemented a multidimensional infection control approach that included outcome surveillance, process surveillance, feedback on CAUTI rates, feedback on performance, education, and a bundle of preventive measures. The rates of CAUTI obtained in phase 1 were compared with the rates obtained in phase 2, after interventions were implemented. RESULTS: During the study period, we recorded 8,513 urinary catheter (UC) days, including 1,513 UC-days in phase 1 and 7,000 UC-days in phase 2. In phase 1, the CAUTI rate was 5.9 cases per 1,000 UC-days, and in phase 2, after implementing the multidimensional infection control approach for CAUTI prevention, the rate of CAUTI decreased to 2.6 cases per 1,000 UC-days (relative risk, 0.43 [95% confidence interval, 0.21-1.0]), indicating a rate reduction of 57%. CONCLUSIONS: Our findings demonstrated that implementing a multidimensional infection control approach is associated with a significant reduction in the CAUTI rate of PICUs in developing countries.

Determinación de la mutación K76T de la proteína transportadora resistente a Cloroquina de Plasmodium falciparum (PFCRT) en individuos con malaria en una zona de transmisión moderada del Caribe colombiano / Determination of K76T mutation in the Plasmodium falciparum Chloroquine resistance transporter (PFCRT) in people with malaria in a zone of moderate transmission of the Colombian Caribbean

Introducción: La resistencia de Plasmodium falciparum a los antimalaricos, está relacionada con mutaciones puntuales en proteínas esenciales en la biología del parásito. La mutación K76T en la proteína transportadora resistente a Cloroquina (CRT) es un marcador molecular de resistencia a este fármaco. En Colombia se ha reportado una frecuencia del 100% a esta mutación. Objetivo: Determinar la frecuencia de la mutación K76T de CRT en Plasmodium falciparum, infectando sangre de individuos con malaria adquirida en una zona del Caribe Colombiano con transmisión moderada. Materiales y métodos: El ADN del parásito se extrajo de láminas diagnósticas usando Chelex-100. Un fragmento de 148 pb de Pfcrt se amplificó por PCR, la mutación K76T fue determinada por análisis de Polimorfismos en Longitud de Fragmentos de Restricción (RFLP) usando la endonucleasa Apo I, los productos fueron separados por electroforesis en geles de poliacrilamida (PAGE). Resultados: Se estudiaron 66 muestras de individuos con malaria adquirida en seis municipios del Magdalena. Se logró amplificación de Pfcrt en 56 y la mutación T76 fue detectada en 55 (98,2%). La presencia de una muestra con el alelo K76 silvestre indicaría la existencia de reversión de la mutación K76T en Colombia, este fenómeno se ha observado en algunos países del continente africano, donde se reportó recuperación de la sensibilidad a Cloroquina. Conclusión: Este estudio es un primer acercamiento hacia el conocimiento de la resistencia a los antimaláricos en zonas de transmisión moderada en Colombia.

Introduction: The antimalarial drugs resistance from Plasmodium falciparum is related to point mutations in the essential proteins to the parasite biology. The K76T mutation in the Plasmodium falciparum Chloroquine resistance transporter (PFCRT) is a molecular marker resistance to Chloroquine. In Colombia the frequency of this mutation is 100%. Objective: To determine the frequency of K76T mutation in the Plasmodium falciparum CRT, infecting blood from people with malaria in the Colombia Caribbean zone with moderate transmission. Materials and methods: The DNA of the parasite was extracted from diagnostic slide using Chelex-100. A fragment of 148 pb of Pfcrt was amplified by PCR, the K76T mutation was evaluated by Restriction Fragment Length Polymorphism (RFLP) using endonuclease Apo I, the products were separated by polyacrylamide gel electrophoresis (PAGE). Results: Sixty six samples from people with malaria of six municipalities of Magdalena were studied. Fifty six DNA samples amplified of Pfcrt gene, T76 mutation was detected in 55 (98.2%). Occurrence of one sample carried the wild allele K76 would indicated the reversion of K76T mutation in Colombia, this phenomenon has been observed in some countries of the African continent, where recovery of sensitivity to Chloroquine was reported. Conclusions: This study is the first approach towards the knowledge of the antimalarials resistance in zones with moderate transmission in Colombia.