Genetic Relatedness Among Escherichia coli Pathotypes Isolated from Food Products for Human Consumption in Cartagena, Colombia.

Foodborne pathogens are a leading cause of mild-to-severe gastrointestinal illnesses worldwide. Escherichia coli pathotypes have been known to cause gastrointestinal illnesses in children less than 5 years old in Colombia. However, insufficient information is available on the prevalence of E. coli contamination of food products and the kind of E. coli food product reservoirs. The two objectives of this study were designed to address this issue. The first objective was to ascertain coliform, E. coli, and pathogenic E. coli contamination of food products readily available for human consumption in Cartagena, Colombia. The second objective was to evaluate the relationship between pathogenic E. coli isolated from food products and those isolated from cases of diarrhea in children. Food product samples consisting of pasteurized milk, unpasteurized fruit juice, ground beef, cheese, and vegetables were obtained at four retail stores. The food samples were cultured in liquid media and tested for the presence of coliforms and E. coli. E. coli isolates were tested by polymerase chain reaction for the presence of pathogenic E. coli. Coliforms, E. coli, and E. coli intestinal pathotypes contamination were detected in 88.4%, 53%, and 2.1% of food product samples, respectively. Ground beef and cheese were the only food samples contaminated with E. coli intestinal pathotypes including enteropathogenic (EPEC), Shiga toxin-producing (STEC), and enterotoxigenic E. coli (ETEC). Closed multilocus sequencing typing relationships between diarrheagenic E. coli isolates from food products and from individuals with diarrhea suggest that food products readily available at public markets in Cartagena can transmit ETEC and possibly EPEC and STEC. We demonstrated that a high proportion of food products for human consumption available at public markets in Cartagena are contaminated with coliforms, E. coli, and E. coli intestinal pathogens. Furthermore, food products containing E. coli intestinal pathogens may be involved in the transmission of foodborne illnesses among children in Cartagena, Colombia.

[Genetic profiling of Giardia intestinalis by polimerase chain in human and dogs samples of Colombian Caribean Coast]. / Caracterización genética por reacción en cadena de la polimerasa de Giardia intestinalis en muestras de humanos y perros del Caribe colombiano.

INTRODUCTION: Giardia intestinalis (G. Intestinalis) is a protozoan that causes diarrheal disease and malabsorption syndrome in humans and other mammals. It presents a high genetic diversity evidenced in the recognition of 7 genotypes (A-G). Genotypes A and B are commonly associated to humans and domestic animals such as dogs. The aim of this study was to conduct a preliminary genetic characterization of G. intestinalis in humans and dogs from two cities on the Caribbean coast of Colombia. METHODS: Sampling areas were selected according to the highest numbers of acute diarrheal disease. Stool samples were collected from children under 7 years old, with positive medical tests for G. intestinalis. Cysts were purified by sucrose gradient and DNA samples were isolated by extraction with organic solvents. Molecular characterization was performed by amplifying the gene triose phosphate isomerase (tpi) by using a semi-nested PCR. RESULTS: A total of 202 samples of DNA were obtained; of these, 111 were positive in coproparasitological analysis (13 dogs and 98 children). Genotype distribution in positive samples was: 5.1% belonged to genotype A and 92.3% to genotype B. Genotype B was present in humans and animals. CONCLUSIONS: The most common genotype in both human and animal samples was genotype B, suggesting a zoonotic transmission cycle.

Detection of Escherichia coli enteropathogens by multiplex polymerase chain reaction from children's diarrheal stools in two Caribbean-Colombian cities.

Acute diarrheal disease is a leading cause of childhood morbidity and mortality in the developing world and Escherichia coli intestinal pathogens are important causative agents. Information on the epidemiology of E. coli intestinal pathogens and their association with diarrheal disease is limited because no diagnostic testing is available in countries with limited resources. To evaluate the prevalence of E. coli intestinal pathogens in a Caribbean-Colombian region, E. coli clinical isolates from children with diarrhea were analyzed by a recently reported two-reaction multiplex polymerase chain reaction (Gomez-Duarte et al., Diagn Microbiol Infect Dis 2009;63:1-9). The phylogenetic group from all E. coli isolates was also typed by a single-reaction multiplex polymerase chain reaction. We found that among 139 E. coli strains analyzed, 20 (14.4%) corresponded to E. coli diarrheagenic pathotypes. Enterotoxigenic, shiga-toxin-producing, enteroaggregative, diffuse adherent, and enteropathogenic E. coli pathotypes were detected, and most of them belonged to the phylogenetic groups A and B1, known to be associated with intestinal pathogens. This is the first report on the molecular characterization of E. coli diarrheogenic isolates in Colombia and the first report on the potential role of E. coli in childhood diarrhea in this geographic area.

Phenotypic and genotypic characterization of enterotoxigenic Escherichia coli clinical isolates from northern Colombia, South America.

Enterotoxigenic Escherichia coli (ETEC) are major causes of childhood diarrhea in low and middle income countries including Colombia, South America. To understand the diversity of ETEC strains in the region, clinical isolates obtained from northern Colombia children were evaluated for multiple locus sequencing typing, serotyping, classical and nonclassical virulence genes, and antibiotic susceptibility. Among 40 ETEC clinical isolates evaluated, 21 (52.5%) were positive for LT gene, 13 (32.5%) for ST gene, and 6 (15%) for both ST and LT. The most prevalent colonization surface antigens (CS) were CS21 and CFA/I identified in 21 (50%) and 13 (32.5%) isolates, respectively. The eatA, irp2, and fyuA were the most common nonclassical virulence genes present in more than 60% of the isolates. Ampicillin resistance (80% of the strains) was the most frequent phenotype among ETEC strains followed by trimethoprim-sulfamethoxazole resistance (52.5%). Based on multiple locus sequencing typing (MLST), we recognize that 6 clonal groups of ETEC clinical isolates circulate in Colombia. ETEC clinical isolates from children in northern Colombia are highly diverse, yet some isolates circulating in the community belong to well-defined clonal groups that share a unique set of virulence factors, serotypes, and MLST sequence types.

Enterotoxigenic Escherichia coli associated with childhood diarrhoea in Colombia, South America.

INTRODUCTION: Intestinal Escherichia coli pathogens are major causes of diarrhoeal disease in children under five years of age worldwide. The aim of this study is to evaluate the association of E. coli pathotypes with childhood diarrhoea in Colombia. METHODOLOGY: A case-control study was conducted in 815 samples from children younger than five years of age in Cartagena, Colombia (466 cases and 349 controls). Controls were randomly selected 1:1 to cases, to obtain 349 cases and 349 controls. RESULTS: This study revealed that 27 (7.44%) cases and 12 (3.43%) controls were positives for any of the E. coli pathotypes. The difference observed was statistically significant indicating that E. coli pathotypes were associated with cases of childhood diarrhoea. Enterotoxigenic E. coli (ETEC) was the most common pathotype associated with childhood diarrhoea. Additional E. coli pathotypes were also identified. CONCLUSIONS: We conclude that after the adjustment by age, sex and socioeconomic stratum, the odds ratio obtained by logistic regression shows an association between infection with ETEC and childhood diarrhoea.

Rotavirus type A and other enteric pathogens in stool samples from children with acute diarrhea on the Colombian northern coast.

The present study, conducted from March 1998 to July 2000, determined the etiology of acute diarrhea in 253 young children and infants from Cartagena and Sincelejo, Colombia. In 253 stool samples, the following enteric pathogens were recovered: rotavirus type A (36.6%) as the major agent, Salmonella spp (9.0%), Shigella spp (8.0%), enteric pathogenic Escherichia coli (6.0%), enteric hemorragic Esc. coli (2.8%), Providencia alcalifaciens (2.8%), Aeromonas hydrophila (2.0%), Yersinia enterocolitica (0.8%), Entamoeba hystolitica (10%), Giardia lamblia (4%), Endolimax nana (3.2%), Ascaris lumbricoides (2.8%), Ent. coli (1.2%), Balantidium coli (0.8%), Blastocystis hominis (0.8%), Dypilidium caninum (0.4%) and hook worm sp. (0.4%). Infection with more than one pathogen occurred in 96 (37.9%) patients. Rotavirus and enteric pathogenic Esc. coli were frequent. Concurrent infection by more than one parasite occurred in 18.6% of the infants. Most rotavirus infections (76.7%) occurred in infants under 12 months. Vomiting, severe dehydration and fever were frequent in children with rotavirus infection. At least one fecal marker of inflammatory diarrhea was registered in patients with bacterial infection. To our knowledge, this is first report of P. alcalifaciens associated with infantile diarrhea in Colombia and the first description of Esc. coli O157:H7 and Y. enterocolitica in our region.

G and P genotypes of rotavirus circulating among children with diarrhea in the Colombian northern coast.

A study on the prevalence of rotavirus G and P genotypes was carried out based on 253 stool specimens obtained from children living in the Colombia northern coast region who were less than 3-years-old and who suffered from acute diarrhea. A previous study had detected the presence of rotavirus A in 90 (36.5%) of the 246 samples tested by enzyme immunoassay (EIA), and these strains were investigated in the present study. Of these, 50 strains yielded an RNA electropherotype, most of which (80.0%) had long profiles and 20.0% of which had short profiles. Genotyping of 84 positive samples indicated that 67.9% of the strains could be typed. G1 (57.9%), was the most predominant VP7 genotype, followed by G3 (21.1%), G9 (15.8%) and G2 (5.3%). Among the VP4 genotypes, P[4] (49.1%) was the most prevalent, followed by P[6] 36.4% and P[8] (14.5%). Neither G4 nor G8 nor P[9] types were detected. The most common G-P combinations were G3 P[4] (8.8%) and G9 P[6] (7.0%), followed by G1 P[4] and G1 P[8] (5.3% each). All G1 P[8] strains showed long RNA profiles, whereas G3 P[4] and G9 P[6] displayed both long and short patterns. Mixed infections involved 21.0% of strains. There was a marked diversity among strains collected, and novel strains, including G9, as well as other atypical combinations of G and P genotypes, such as G9 P[6] and G3 P[4], were found.

Infecciones parasitarias intestinales en pacientes positivos para el virus de la inmunodeficiencia humana (VIH) en la ciudad de Cartagena de Indias, Colombia / Intestinal parasitic infections in HIV positive patients in Cartagena de Indias, Colombia

Objetivo: evaluar la prevalencia de parásitos intestinales en heces de pacientes de Programa de Prevención de Enfermedades Sexualmente Transmitidas VIH - SIDA, del Distrito Integrado de Salud (DADIS) en Cartagena de Indias. Materiales y métodos: se realizó un estudio descriptivo transversal en 114 muestras fecales de 38 pacientes diagnosticados con VIH tomando heces seriadas para coprológico dirigido, concentración, coloración Ziehl Neelsen modificado y cromotropo modificado (Weber). Para el análisis estadístico se aplicó PHARMA con pruebas de X² de independencia. Resultados: de 38 pacientes del estudio 29(76.3 por ciento) fueron masculinos, 9(23.7 por ciento) femeninos y 3(7.9 por ciento) homosexuales. Rango etáreo 10 - 63 años (x: 32 años). Un paciente (2.6 por ciento) adquirió el VIH vía congénita y 4 (10.5 por ciento) eran extranjeros. El 55.3 por ciento se describió así: Cryptosporidium parvum 23.7 por ciento Blastocystis hominis 18.4 por ciento, Giardia lamblia 7.9 por ciento, Isospora belli 7.9 por ciento, Myxospora 5.3 por ciento, Microsporidios 5.3 por ciento, Entamoeba coli 5.3 por ciento, Strongyloides stercolaris 5.3 por ciento, Chilomastix mesnili 5.3 por ciento, Cyclospora cayetanensis 2.6 por ciento 2.6 por ciento, Trichuris trichiura 2.6 por iento y Ascarís lumbricoides 2.6 por ciento. 44.7 por ciento fue negativo. El OR fue de 14(p<0.05) para la asociación estadíos de la infección - parásitos. Conclusiones: se detectaron altas frecuencias de protozoarios esporulados y Blastocystis hominis. Los primeros pueden explicarse por factores climáticos y ambientales que favorecen la maduración de ooquistes infectantes. No hubo diferencias significativas de parásitos en muestras seriadas. Estos resultados confirman la importancia de parásitos esporulados haciéndose necesaria su detección en diarrea crónica

Rotavirus type A and other enteric pathogens in stool samples from children with acute diarrhea in the Colombian northern coast / Rotavirus de tipo A y otros patógenos entéricos en muestras de heces de niños con diarrea aguda en la costa norte colombiana

The present study, conducted from March 1998 to July 2000, determined the etiology of acute diarrhea in 253 young children and infants from Cartagena and Sincelejo, Colombia. In 253 stool samples, the following enteric pathogens were recovered: rotavirus type A (36.6%) as the major agent, Salmonella spp (9.0%), Shigella spp (8.0%), enteric pathogenic Escherichia coli (6.0%), enteric hemorragic Esc. coli (2.8%), Providencia alcalifaciens (2.8%), Aeromonas hydrophila (2.0%), Yersinia enterocolitica (0.8%), Entamoeba hystolitica (10%), Giardia lamblia (4%), Endolimax nana (3.2%), Ascaris lumbricoides (2.8%), Ent. coli (1.2%), Balantidium coli (0.8%), Blastocystis hominis (0.8%), Dypilidium caninum (0.4%) and hook worm sp. (0.4%). Infection with more than one pathogen occurred in 96 (37.9%) patients. Rotavirus and enteric pathogenic Esc. coli were frequent. Concurrent infection by more than one parasite occurred in 18.6% of the infants. Most rotavirus infections (76.7%) occurred in infants under 12 months. Vomiting, severe dehydration and fever were frequent in children with rotavirus infection. At least one fecal marker of inflammatory diarrhea was registered in patients with bacterial infection. To our knowledge, this is first report of P. alcalifaciens associated with infantile diarrhea in Colombia and the first description of Esc. coli O157:H7 and Y. enterocolitica in our region (AU)

El presente estudio, llevado a cabo entre marzo de 1998 y julio de 2000, determinó la etiología de diarrea aguda en 253 niños y bebés de Cartagena y Sincelejo (Colombia). En 253 muestras de heces se encontraron los siguientes patógenos entéricos: rotavirus de tipo A (36,6 por ciento), que era el patógeno más frecuente, Salmonella spp. (9,0 por ciento), Shigella spp. (8,0 por ciento), Escherichia coli patógena entérica (6,0 por ciento), E. coli hemorrágica entérica (2,8 por ciento), Providencia alcalifaciens (2,8 por ciento), Aeromonas hydrophila (2,0 por ciento), Yersinia enterocolitica (0,8 por ciento), Entamoeba hystolitica (10 por ciento), Giardia lamblia (4 por ciento), Endolimax nana (3,2 por ciento), Ascaris lumbricoides (2,8 por ciento), Entamoeba coli (1,2 por ciento), Balantidium coli (0,8 por ciento), Blastocystis hominis (0,8 por ciento), Dypilidium caninum (0,4 por ciento) y y alguna especie del gusano uncinaria (0,4 por ciento). En 96 pacientes (37,9 por ciento) había infección por más de un patógeno. Los más frecuentes eran rotavirus y E. coli patógena entérica. En un 18,6 por ciento de los niños se daba infección concurrente con más de un parásito. La mayoría de las infecciones por rotavirus (76,7 por ciento) se daban en niños menores de 12 meses. Los síntomas más frecuentes en niños infectados por rotavirus eran vómitos, deshidratación grave y fiebre. Se registró un marcador fecal de diarrea inflamatoria en pacientes con infecciones bacterianas. Según nuestros datos, esta es la primera vez que se encuentra Providencia alcalifaciens asociada a diarrea infantil en Colombia, y la primera descripción de E. coli O157:H7 y Yersinia enterocolitica en nuestra región. (AU)

G and P genotypes of rotavirus circulating among children with diarrhea in the Colombian northern coast / Genotipos G y P de rotavirus encontrados en niños aquejados de diarrea en la costa norte de Colombia

A study on the prevalence of rotavirus G and P genotypes was carried out based on 253 stool specimens obtained from children living in the Colombia northern coast region who were less than 3-years-old and who suffered from acute diarrhea. A previous study had detected the presence of rotavirus A in 90 (36.5%) of the 246 samples tested by enzyme immunoassay (EIA), and these strains were investigated in the present study. Of these, 50 strains yielded an RNA electropherotype, most of which (80.0%) had long profiles and 20.0% of which had short profiles. Genotyping of 84 positive samples indicated that 67.9% of the strains could be typed. G1 (57.9%), was the most predominant VP7 genotype, followed by G3 (21.1%), G9 (15.8%) and G2 (5.3%). Among the VP4 genotypes, P[4] (49.1%) was the most prevalent, followed by P[6] 36.4% and P[8] (14.5%). Neither G4 nor G8 nor P[9] types were detected. The most common G-P combinations were G3 P[4] (8.8%) and G9 P[6] (7.0%), followed by G1 P[4] and G1 P[8] (5.3% each). All G1 P[8] strains showed long RNA profiles, whereas G3 P[4] and G9 P[6] displayed both long and short patterns. Mixed infections involved 21.0% of strains. There was a marked diversity among strains collected, and novel strains, including G9, as well as other atypical combinations of G and P genotypes, such as G9 P[6] and G3 P[4], were found (AU)

El artículo describe un estudio de la prevalencia de los genotipos G y P de rotavirus en 253 muestras fecales de niños menores de 3 años afectados de diarrea aguda que vivían en la costa norte de Colombia. Un estudio anterior reveló la presencia de rotavirus A en 90 (36.5%) de las 246muestras sometidas a inmuno ensayo y dichas muestras fueron las usadas en este estudio. En 50 de las 90 muestras usadas se obtuvo un electroferotipo, un 80.0% de perfil de RNA largo y un 20.0% de perfil de RNA corto. De 84muestras sometidas a genotipificación, se logró tipificar 67.9%. El genotipo G1(57.9%) fue predominante entre los asociados a VP7, seguido de G3(21.1%), G9 (15.8%) y G2 (5.3%). Entre los genotipos VP4, el P[4] (49.1%)fue el de mayor prevalencia, seguido de P[6] 36.4% y de P[8] (14.5%). No se detectaron los genotipos G4, G8 o P[9], y las combinaciones G-P más frecuentes fueron G3 P[4] ( 8.8%) y G9 P[6] (7.0%), seguidas de G1 P[4] y G1P[8], con 5.3% cada una. Todas las cepas G1 P[8] tenían perfil de RNA largo, mientras que G3 P[4] y G9 P[6] los tenían tanto largos como cortos. Se detectaron infecciones mixtas en el 21.0% de las cepas. Se observó una marcada diversidad en las cepas recolectadas y se identificaron cepas nuevas, como G9, y también combinaciones atípicas, como G9 P[6] y G3P[4] (AU)