RESUMEN

This study analysed the mechanisms of quinolone resistance among enterotoxigenic Escherichia coli (ETEC) in a periurban area of Lima, Peru. The susceptibility to nalidixic acid and ciprofloxacin, the role of Phe-Arg-b-Naphtylamyde inhibitable-(PAbN) efflux pumps, the presence of mutations in gyrA and parC as well as the presence of aac(6')Ib-cr, qepA, qnrA, qnrB, qnrC, qnrD, qnrVC and oqxAB were determined in 31 ETEC from previous case/control studies of children's diarrhoea. Discordances between disk diffusion, with all isolates showing intermediate or fully resistance to nalidixic acid, and minimal inhibitory concentration (MIC), with 7 isolates being below considered resistance breakpoint, were observed. Twenty-one isolates possessed gyrA mutations (19 S83L, 2 S83A). AAC(6') Ib-cr, QnrS, QnrB and QepA were found in 7, 6, 2 and 1 isolates respectively, with 3 isolates presenting 2 transferable mechanisms of quinolone resistance (TMQR) concomitantly. TMQR were more frequent among isolates with MIC to nalidixic acid ranging from 2 to 16 mg/L (p=0.03), while gyrA mutations were more frequent among isolates with nalidixic acid MIC >= 128 mg/L (p=0.0002). In summary, the mechanisms of quinolone resistance present in ETEC isolates in Peru have been described. Differences in the prevalence of underlying mechanisms associated with final MIC levels were observed. The results suggest two different evolutive strategies to survive in the presence of quinolones related to specific bacterial genetic background.

Asunto(s)

Escherichia coli Enterotoxigénica , Quinolonas , Niño , Humanos , Escherichia coli Enterotoxigénica/genética , Ácido Nalidíxico/farmacología , Quinolonas/farmacología , Ciprofloxacina , Estudios de Casos y Controles

RESUMEN

The aim of this study was to determine the prevalence of Shiga toxin-producing Escherichia coli (STEC) in cattle and pigs as a possible STEC reservoir in Lima, Peru. One hundred and fourteen cattle and 112 pigs from 10 and 4 farms, respectively, were studied. Five E. coli colonies per culture were studied by a multiplex real-time PCR to identify Shiga toxin-producing (stx1, stx2, eaeA), enterotoxigenic (lt, st), enteropathogenic (eaeA), enteroinvasive (ipaH), enteroaggregative (aggR), and diffusely adherent E. coli (daaD). Shiga toxin-producing E. coli were isolated from 16 cattle (14%) but none from pigs. stx1 was found in all bovine isolates, 11 of which also carried eaeA genes (69%); only 1 sample had both stx1 and stx2. Thirteen stx-positive strains were classified as Shiga-toxigenic (81%) using an enzymatic immunoassay, 2 STEC strains were from serogroup O157 (13%), and 7 were sorbitol negative (44%). Enteropathogenic E. coli were detected more frequently in cattle (18%, 20/114) than in pigs (5%, 6/112). To our knowledge, this is the first study on the prevalence of STEC in farms animals in Peru using molecular methods. Further studies are needed in a large number of farms to determine the relevance of these findings and its consequences for public health.

Asunto(s)

Enfermedades de los Bovinos/microbiología , Infecciones por Escherichia coli/veterinaria , Escherichia coli Shiga-Toxigénica , Enfermedades de los Porcinos/microbiología , Animales , Bovinos/microbiología , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/epidemiología , Infecciones por Escherichia coli/diagnóstico , Infecciones por Escherichia coli/epidemiología , Perú/epidemiología , Porcinos/microbiología , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/epidemiología

RESUMEN

The aim of this study was to determine the prevalence, virulence factors (stx, eae, ehxA and astA) and phylogenetic relationships [PFGE and multilocus sequence typing (MLST)] of Shiga toxin-producing Escherichia coli (STEC) strains isolated from four previous cohort studies in 2212 Peruvian children aged <36 months. STEC prevalence was 0.4 % (14/3219) in diarrhoeal and 0.6 % (15/2695) in control samples. None of the infected children developed haemolytic uraemic syndrome (HUS) or other complications of STEC. stx1 was present in 83 % of strains, stx2 in 17 %, eae in 72 %, ehxA in 59 % and astA in 14 %. The most common serotype was O26 : H11 (14 %) and the most common seropathotype was B (45 %). The strains belonged mainly to phylogenetic group B1 (52 %). The distinct combinations of alleles across the seven MLST loci were used to define 13 sequence types among 19 STEC strains. PFGE typing of 20 STEC strains resulted in 19 pulsed-field patterns. Comparison of the patterns revealed 11 clusters (I-XI), each usually including strains belonging to different serotypes; one exception was cluster VI, which gathered exclusively seven strains of seropathotype B, clonal group enterohaemorrhagic E. coli (EHEC) 2 and phylogenetic group B1. In summary, STEC prevalence was low in Peruvian children with diarrhoea in the community setting. The strains were phylogenetically diverse and associated with mild infections. However, additional studies are needed in children with bloody diarrhoea and HUS.

Asunto(s)

Infecciones por Escherichia coli/microbiología , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genética , Adhesinas Bacterianas/genética , Secuencia de Bases , Estudios de Casos y Controles , Preescolar , Estudios de Cohortes , ADN Bacteriano/genética , Electroforesis en Gel de Campo Pulsado , Infecciones por Escherichia coli/epidemiología , Proteínas de Escherichia coli/genética , Femenino , Genes Bacterianos , Proteínas Hemolisinas/genética , Humanos , Lactante , Recién Nacido , Masculino , Tipificación de Secuencias Multilocus , Perú/epidemiología , Filogenia , Prevalencia , Serotipificación , Toxina Shiga/genética , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Factores de Virulencia/genética

RESUMEN

Enterotoxigenic Escherichia coli (ETEC) is a major cause of childhood diarrhea. The present study sought to determine the prevalence and distribution of toxin types, colonization factors (CFs), and antimicrobial susceptibility of ETEC strains isolated from Peruvian children. We analyzed ETEC strains isolated from Peruvian children between 2 and 24 months of age in a passive surveillance study. Five E. coli colonies per patient were studied by multiplex real-time PCR to identify ETEC virulence factors. ETEC-associated toxins were confirmed using a GM1-based enzyme-linked immunosorbent assay. Confirmed strains were tested for CFs by dot blot assay using 21 monoclonal antibodies. We analyzed 1,129 samples from children with diarrhea and 744 control children and found ETEC in 5.3% and 4.3%, respectively. ETEC was more frequently isolated from children >12 months of age than from children <12 months of age (P < 0.001). Fifty-two percent of ETEC isolates from children with diarrhea and 72% of isolates from controls were heat-labile enterotoxin (LT) positive and heat-stable enterotoxin (ST) negative; 25% and 19%, respectively, were LT negative and ST positive; and 23% and 9%, respectively, were LT positive and ST positive. CFs were identified in 64% of diarrheal samples and 37% of control samples (P < 0.05). The most common CFs were CS6 (14% and 7%, respectively), CS12 (12% and 4%, respectively), and CS1 (9% and 4%, respectively). ST-producing ETEC strains caused more severe diarrhea than non-ST-producing ETEC strains. The strains were most frequently resistant to ampicillin (71%) and co-trimoxazole (61%). ETEC was thus found to be more prevalent in older infants. LT was the most common toxin type; 64% of strains had an identified CF. These data are relevant in estimating the burden of disease due to ETEC and the potential coverage of children in Peru by investigational vaccines.

Asunto(s)

Escherichia coli Enterotoxigénica/clasificación , Escherichia coli Enterotoxigénica/aislamiento & purificación , Infecciones por Escherichia coli/microbiología , Antibacterianos/farmacología , Anticuerpos Antibacterianos , Anticuerpos Monoclonales , Toxinas Bacterianas/biosíntesis , Toxinas Bacterianas/genética , Estudios de Casos y Controles , Preescolar , Diarrea/microbiología , Escherichia coli Enterotoxigénica/genética , Escherichia coli Enterotoxigénica/metabolismo , Enterotoxinas/biosíntesis , Enterotoxinas/genética , Ensayo de Inmunoadsorción Enzimática/métodos , Proteínas de Escherichia coli/biosíntesis , Proteínas de Escherichia coli/genética , Femenino , Humanos , Lactante , Masculino , Pruebas de Sensibilidad Microbiana , Perú , Reacción en Cadena de la Polimerasa/métodos , Factores de Virulencia/biosíntesis , Factores de Virulencia/genética

RESUMEN

Los pacientes con alteraciones en las relaciones intermaxilares para la intubación y el mantenimiento del tubo endotraqueal durante la cirugía craneomaxilofacial. La fijación del tubo nasotraqueal puede ser seccionado accidentalmente en el momento de las osteomías. Se describe la técnica de pasaje transmilohiodeodel tubo orotraqueal y se detallan sus indicaciones y resultados. Entre Mayo de 1995 y Abril de 1997 se utilizó el método en diez pacientes con resultados favorables. Se indica como alternativa de intubación el uso de la vía orotraqueal, con abocamiento a la piel a través del músculo milohioideo, con el fin de obtener un mejor espacio quirúrgico y evotar los riesgos de obstrucción, extubación accidental y sección de la cánula endotraqueal, sin tener que recurrir a una traqueostomía

Asunto(s)

Humanos , Intubación Intratraqueal , Cirugía Bucal

RESUMEN

Los pacientes con alteraciones en las relaciones intermaxilares para la intubación y el mantenimiento del tubo endotraqueal durante la cirugía craneomaxilofacial. La fijación del tubo nasotraqueal puede ser seccionado accidentalmente en el momento de las osteomías. Se describe la técnica de pasaje transmilohiodeodel tubo orotraqueal y se detallan sus indicaciones y resultados. Entre Mayo de 1995 y Abril de 1997 se utilizó el método en diez pacientes con resultados favorables. Se indica como alternativa de intubación el uso de la vía orotraqueal, con abocamiento a la piel a través del músculo milohioideo, con el fin de obtener un mejor espacio quirúrgico y evotar los riesgos de obstrucción, extubación accidental y sección de la cánula endotraqueal, sin tener que recurrir a una traqueostomía

Asunto(s)

Humanos , Intubación Intratraqueal , Cirugía Bucal