Piedad Agudelo-Flórez

Prevalence of Leptospira spp. in Urban Rodents from a Groceries Trade Center of Medellín, Colombia

Leptospirosis is a widely distributed zoonosis, and rats are its most common source of infection. Our goal was to determine the frequency for Leptospira infection in rodents in a farmers market in the city of Medellin. We performed a descriptive transversal study sampling 254 rodents. Rodents were bled and killed, and kidneys samples were taken. Supernatants of macerated kidneys were cultured on Fletcher medium. Microagglutination tests (MATs) with 11 serovars were also carried out in rat serum, and a polymerase chain reaction (PCR) specific for pathogenic species was used to test each bacterial culture. All animals were identified as Rattus norvegicus; 25% and 20% were positive by MAT and culture, respectively. PCR tests of 12 isolates were positive for pathogenic serovars, and 4 of them were confirmed as L. interrogans by sequencing. These data show the role of this natural carrier and shedder of pathogenic leptospires in the epidemiology of urban leptospirosis in Colombia.

An outbreak of severe leptospirosis in capuchin (Cebus) monkeys

Naturally acquired acute leptospirosis in monkeys is uncommon. This study reports an outbreak of severe leptospirosis among 52 capuchin (Cebus) monkeys that had been rescued from homes and housed in a wildlife rehabilitation center in Colombia in 2007. Case confirmation consisted of Leptospira isolation followed by a polymerase chain reaction targeting the LipL32 gene. The attack and mortality rates were 71% and 27%, respectively. Sixteen cases were confirmed. Necropsy revealed diffuse jaundice and pulmonary hemorrhage. Multi-locus sequence typing identified the agent to be Leptospira interrogans sequence type 17, indicating rats as the source of infection. An environmental survey confirmed rodent infestation as the cause of the outbreak. The extent of Leptospira transmission between humans and monkeys is unknown. Improper husbandry of non-human primates could create new reservoirs and transmission routes for Leptospira threatening conservation efforts and public health.

Situación de la leptospirosis en el Urabá antioqueño colombiano: estudio seroepidemiológico y factores de riesgo en población general urbana

Leptospirosis is a widespread zoonosis in tropical regions. The prevalence is unknown in the Colombian region of Uraba. A cross sectional study was conducted from March to October 2000 in order to determine the prevalence of Leptospira spp. antibodies and describe risk factors in nine counties in the region. The sample consisted of 582 individuals, who answered a questionnaire and had blood samples drawn to determine risk factors. Detection of Leptospira spp. antibodies was based on indirect inmunofluorescence and microagglutination. Seroprevalence was 12.5% (95%CI: 10.01-15.5). No differences were observed according to race, gender, occupation, age, living conditions, or time of residence in the area. L .interrogans serovar Grippotyphosa was the most prevalent species, identified in 53 individuals. Titers were > 1:400 in 38 seropositive individuals. In conclusion, there is a high prevalence of Leptospira spp. antibodies in the area, where it is thus necessary to establish control measures to decrease the risk of environmental exposure to leptospirosis.

Genetic Evidence of Hantavirus Infections in Wild Rodents from Northwestern Colombia

This report builds on recent serological evidence for the presence of hantavirus in northern Colombia by providing sequence-specific and phylogenetic data of hantavirus infections in wild rodents. From August 2007 to August 2008, 354 rodent specimens representing four families were collected in the northwestern Antioquia region of Colombia. Antibodies reactive to Sin Nombre virus and Maciel virus antigens by IgG enzyme-linked immunosorbent assay were found in 15 of 109 (14%) Cherries cane rats (Zygodontomys cherriei), the only sigmodontinae rodents captured. Lung tissue samples from 11 of the 15 seropositive rodents were RT-polymerase chain reaction positive for hantavirus RNA, using primers for the S and M genome segments. Eight of these amplicons were sequenced and phylogenetic analyses indicated RNA of a hantavirus closely related to Calabazo virus, previously found in Panama. This is the first report of the genetic characterization of a hantavirus in rodents in Colombia.

Cross-Sectional Study of Leptospira Seroprevalence in Humans, Rats, Mice, and Dogs in a Main Tropical Sea-Port City

Samples were collected from 128 symptomatic humans, 83 dogs, 49 mice, and 20 rats (Rattus rattus: 16; Rattus norvegicus: 4) in neighborhoods where human leptospirosis have been reported within the principal sea-port city of Colombia. Seroprevalences were assessed against 19 pathogenic, 1 intermediate pathogenic, and 1 saprophytic Leptospira serogroups. Pathogenic Leptospira were confirmed using conventional Leptospira-specific polymerase chain-reaction and pulsed-field gel electrophoresis analysis was used for serovar identification. Seroprevalences of 20.4%, 12.5%, 25.0%, 22.9%, and 12.4% were obtained against one to seven different serogroups in mice, R. rattus, R. norvegicus, dogs, and humans, respectively. The DNA was confirmed to be from pathogenic Leptospira by detecting the lipL32 gene in 12.5%, 3.7%, and 0.03% of the R. rattus, dog, and human samples, respectively. The first genetically typed Colombian isolate was obtained from a rat and identified as Leptospira interrogans serovar Icterohaemorrhagiae/Copenhageni.

APLICACIÓN DE LAS PRUEBAS DE PCR CONVENCIONAL SIMPLE Y MÚLTIPLE PARA LA IDENTIFICACIÓN DE AISLAMIENTOS DE Leptospira spp. EN COLOMBIA

Serological identification of Leptospira ssp isolates is difficult to achieve. Thus, molecular testing may be of great interest thanks to its high discrimination power, reproducibility and easy interpretation. Objective. To implement and validate conventional and multiplex PCR methods (using primers directed against lipl32 and secY/flaB genes, respectively). To assess the capacity of PCR methods to identify pathogenic and saprophytic species of Leptospira ssp. Material and methods. 22 international reference strains and 12 colombian isolates were used. DNA was extracted with a commercial kit (Wizard). Specificity and sensitivity of both PCR methods were evaluated. Results. The maximum dilution of DNA samples allowing the detection of Leptospira ssp was determined to be 1:10000 for the PCR lipL32 and 1:100/1:1000 for the multiplex PCR secY/flaB. Both PCR didn’t detect DNA from microorganisms unrelated to Leptospira ssp. The lipL32 PCR specifically amplified a 423bp fragment from all pathogenic Leptospira reference strains, while the secY/flaB PCR amplified both 285bp (secY) and 793bp (flaB) fragments from 18 reference strains. The lipL32 PCR detected 7/12 colombian isolates, while secY/flaB PCR detected both secY and flaB genes from 6/12 isolates. Conclusions. Best results were obtained with the lipL32 PCR, which displayed a better sensitivity and a better capacity to detect different strains than the multiplex PCR. The secY primers showed a poor specificity to pathogenic species and a poor sensitivity. Thus, lipL32 primers show high potential for molecular diagnosis of Leptospira spp in clinical and environmental samples.

Conocimiento y prácticas sobre teniasis-cisticercosis en una comunidad colombiana

Objetivo Determinar los conocimientos y practicas sobre teniasis-cisticercosis y la frecuencia de anticuerpos contra Taenia solium en habitantes de la localidad de Andagoya, Colombia. Metodos Se realizo un estudio cualitativo-cuantitativo. Las poblaciones de estudio fueron los criadores de cerdos y sus familias, poblacion local y cerdos. Resultados La poblacion tiene un conocimiento parcial del complejo teniasis-cisticercosis. Identifica la cisticercosis como una enfermedad solo del cerdo y no del humano, considera la teniasis como una enfermedad de transmision fecal y no ocasionada por el consumo de carne de cerdo con cisticercosis. La crianza de cerdos no se hace en confinamiento y aunque se conocen los habitos higienicos para el control de enfermedades parasitarias su cumplimiento no es adecuado. La presencia de anticuerpos contra T. solium en los criadores de cerdos y sus familiares fue del 8,7 % y en los cerdos del 20,9 %. Conclusion Se debe desarrollar un programa educativo sobre teniasis-cisticercosis que permita sensibilizar a la poblacion para el conocimiento y la aplicacion de medidas de control.

High-Resolution Melting Curve Analysis of the 16S Ribosomal Gene to Detect and Identify Pathogenic and Saprophytic Leptospira species in Colombian Isolates

AbstractIt is important to identify the circulating Leptospira agent to enhance the performance of serodiagnostic tests by incorporating specific antigens of native species, develop vaccines that take into account the species/serovars circulating in different regions, and optimize prevention and control strategies. The objectives of this study were to develop a polymerase chain reaction (PCR)-high-resolution melting (HRM) assay for differentiating between species of the genus Leptospira and to verify its usefulness in identifying unknown samples to species level. A set of primers from the initial region of the 16S ribosomal gene was designed to detect and differentiate the 22 species of Leptospira. Eleven reference strains were used as controls to establish the reference species and differential melting curves. Twenty-five Colombian Leptospira isolates were studied to evaluate the usefulness of the PCR-HRM assay in identifying unknown samples to species level. This identification was confirmed by sequencing and phylogenetic analysis of the 16S ribosomal gene. Eleven Leptospira species were successfully identified, except for Leptospira meyeri/Leptospira yanagawae because the sequences were 100% identical. The 25 isolates from humans, animals, and environmental water sources were identified as Leptospira santarosai (twelve), Leptospira interrogans (nine), and L. meyeri/L. yanagawae (four). The species verification was 100% concordant between PCR-HRM and phylogenetic analysis of the 16S ribosomal gene. The PCR-HRM assay designed in this study is a useful tool for identifying Leptospira species from isolates.

Evidencia serológica de circulación de Leptospira spp en Rattus norvegicus naturalmente expuestos en una zona urbana colombiana

Objetivo Determinar la frecuencia de anticuerpos dirigidos contra especies de Leptospira patogenas en reservorios naturales, con el proposito de establecer evidencia de su papel potencial en la dispersion ambiental de Leptospira y por consiguiente su actuacion como agente diseminador de la infeccion tanto al humano como a otras especies susceptibles. Materiales y metodos El muestreo se llevo a cabo en la Plaza Minorista durante el periodo comprendido entre agosto de 2006 y abril de 2007, en el cual se capturaron 254 roedores Rattus norvegicus. Se obtuvo sangre de estos por puncion cardiaca y el suero resultante se proceso con la prueba de Microaglutinacion. Resultados El analisis serologico para verificar las serovariedades circulantes de Leptospira spp, dio como resultado que 64 roedores (25,2 %, 95 % CI=19,5-30,1) tuvieron titulos positivos para al menos una de las 11 serovariedades probadas. Conclusiones Los datos muestran que no solo la serovariedad Icterohaemorrhagiae esta asociada con esta especie reservoria; tambien lo estan otras serovariedades como Grippothyphosa y Canicola. El estudio permitio determinar la frecuencia de anticuerpos contra especies patogenas de Leptospira para reservorios procedentes de una zona urbana colombiana. Estos datos son relevantes para las entidades de salud publica por constituir la base para la implementacion de campanas de control adecuadas para esta zona del pais y como modelo de otros estudios similares en otras ciudades colombianas.OBJECTIVE Assessing the frequency of antibodies directed against pathogenic Leptospira species found in natural reservoirs for establishing evidence of their potential role in the environmental dispersion of Leptospira and consequent dissemination of the infection to humans as well as to other susceptible species. MATERIAL AND METHODS A survey was carried out in the Plaza Minorista from August 2006 to April 2007 in which 254 rats (Rattusnorvegicus) were captured. Blood was obtained from these rodents by cardiac puncture and the resulting serum was used for microagglutination tests. RESULTS Serological analysis for verifying Leptospira spp circulating serovars resulted in 64 rodents (25.2 %;19.5-30.1 95 %CI) having positive antibody titres for at least 11 of the serovars tested. CONCLUSIONS Frequency data regarding the antibodies so detected showed that the Icterohaemorrhagiae serovar was not the only one possibly associated with this reservoir species, but also with others such as the Grippothyphosa and Canícolaserovars. The study determined the frequency of antibodies against pathogenic Leptospira species for reservoirs from an urban area in Colombia. This data is relevant for public health authorities and might constitute the basis for implementing appropriate control campaigns for this area of the country and, likewise, this work could serve as a model for similar studies in other Colombian cities.

Evidencia serológica retrospectiva de infecciones por Leptospira spp., dengue, hantavirus y arenavirus en indígenas Emberá-Katío, Colombia

We have developed a study in the Embera-Katio indigenous community and we found that the seropositivity for IgG antibodies to Leptospira spp. was 18.1%; to dengue was 61.1%; to arenavirus was 3.1% and to hantavirus was 1.5%. We conclude that this indigenous community live in conditions that promote the reemerging and emerging, infections.