Enterotoxins, colonization factors, serotypes and antimicrobial resistance of enterotoxigenic Escherichia coli (ETEC) strains isolated from hospitalized children with diarrhea in Bolivia

Enterotoxigenic Escherichia coli (ETEC) is recognized as the main cause of bacterial diarrhoea among children in Asia, Africa and Latin America but less investigated in Bolivia. OBJECTIVE: To determine the relation between enterotoxins, CFs and serotypes as well as the antimicrobial resistance patterns in a set of ETEC isolates collected from hospitalized children with acute diarrhea. In the present study we characterized 43 ETEC strains isolated from 2002 to 2006 from hospitalized children (0-5 years) with acute diarrhea in Bolivia. The strains were analyzed for heat-labile (LT) and heat-stable (ST) enterotoxins and colonization factor (CF) profiles, as well as for serogroups and antimicrobial resistance using phenotypic (ELISA, dot blot, slide agglutination and disc diffusion) and genotypic (Multiplex PCR) methods. Among the ETEC isolates tested, 30 were positive for LT, 3 for STh and 10 for LT/STh. Sixty-five percent (28/43) of the strains expressed one or more CF. The most common CFs were CS17 (n = 8) and CFA/I (n = 8). The phenotypical and genotypical results for toxins and CFs were congruent except for CS21 that was amplified in 10 of the strains by multiplex PCR, but CS21 pili was only detected phenotypically in four of these strains. The ETEC strains had diverse O and H antigens and the most common types were O8:H9 LT CS17 (n = 6; 14 percent) and O78:HNM LT-ST CFA/I (n = 4; 9 percent). The analysis of antibiotic resistance showed that 67 percent (n = 29/43) of the strains were resistant to one or several of the antimicrobial agents tested. Presence of CFs was associated with antibiotic resistance. CONCLUSION: The most common toxin profile was LT 70 percent, LT/STh 23 percent and STh 7 percent. High antimicrobial resistance to ampicillin among serogroups O6, O8 and O78 were the most common.

From cholera to enterotoxigenic Escherichia coli (ETEC) vaccine development.

It was shown earlier that immune responses against cholera toxin (CT) as well as Vibrio cholerae lipopolysaccharide (LPS) or whole bacterial cells (WC) were protective and that these different antibody specificities co-operated synergistically for protection against experimental cholera. Similarly, antibodies against the heat-labile toxin (LT) and major colonization factors (CFs) of enterotoxingenic Escherichia coli (ETEC) co-operated synergistically for protection against LT-producing ETEC expressing homologous CFs. Studies in humans revealed that repeated oral antigen administration was optimal in inducing intestinal immune responses. Based on these findings oral inactivated vaccines consisting of toxin antigen and whole cells, i.e. the licensed recombinant cholera B subunit (rCTB)-WC cholera vaccine Dukoral®, and candidate ETEC vaccines have been developed. In different trials the rCTB-WC cholera vaccine has provided very high (85-100%) short term protection, which was significantly higher than that induced by the WC component alone, whereas rCTB-WC and WC alone provided comparable (50-60%), long term protection. An oral ETEC vaccine consisting of rCTB and formalin-inactivated E. coli bacteria expressing major CFs was shown to be safe and immunogenic in adults and children in different countries. The vaccine also induced significant protection against non-mild ETEC diarrhoea, i.e. diarrhoea interfering with daily activity in American travellers but not against ETEC diarrhoea in young children in Egypt. Against this background, a modified ETEC vaccine consisting of recombinant E. coli strains overexpressing the major CFs and a more LT like hybrid toxoid (LCTBA) has been developed. This vaccine will be tested soon alone and together with a mucosal adjuvant, i.e. dmLT, in clinical trials.

Relationship between enterotoxigenic Escherichia coli and diarrhea among children in Buenos Aires

La diarrea aguda infecciosa es el problema de salud más importante en los países en desarrollo. Diversos estudios en Latinoamérica han demonstrado que Escherichia coli enterotoxigénica (ETEC) y rotavirus son los patógenos más frecuentemente asociados con la diarrea infantil. Para causar la diarrea, ETC coloniza, se multiplica en el intestino delgado y produce la enterotoxina lábil (LT) y/o la estable (ST) al calor. La colonización está mediada por estructuras fimbriales de superficie llamadas factores de colonización (CFAs). Tres CFAs han sido descriptos y bien caracterizados: CFA/I, CFA/II y CFA/IV. Las cepas de ETEC aisladas de pacientes con diarrea pertenecen a un número reducido de serotipos y se ha sugerido que existe una relación entre el patrón de fermentación de azúcares y el serotipo. En este trabajo se estudió la incidencia de ETEC en 85 niñoscon diarrea aguda internados en el Hospital de Niños "Pedro de Elizalde" de Buenos Aires y en 38 niños sanos. La edad en ambos grupos, estuvo comprendida entre 0 y 4 años y ninguno de los niños recibió antibioticoterapia en los 7 días previos a la toma de muestra. Se aislaron cepas de ETEC en 9 de los 85(10,6%) niños con diarrea. De estos casos positivos, 6 estuvieron asociados con ETEC productores de ST, 1 con LT y 2 con ambas (LT/ST). Dentro del grupo control sólo en 1 caso (2,6%) se detectó ETEC-LT. En 5 de los 9 casos de diarrea asociados a ETEC (55,5%), se asislaron cepas que expresaron algunos de los CFAs investigados: 4 fueron CFA/I y 1 CFA/II. En 23 cepas de E. coli, aisladas de los 10 niños ETEC positivos, se estudiaron los factores de colonización, el biotipo, serotipo y antibiotipo en relación al perfil toxigénico que presentaban. De 12 cepas productoras sólo de ST, 5 (41,7%) expresaron CFA y 2 (16,7%) CFA/II (CS2 + CS3)...