María Inés Mota

Enteropathogenic Escherichia coli Strains Carrying Genes Encoding the PER-2 and TEM-116 Extended-Spectrum β-Lactamases Isolated from Children with Diarrhea in Uruguay

ABSTRACT We studied 13 extended-spectrum β-lactamase (ESBL)-producing enteropathogenic Escherichia coli isolates from children suffering acute diarrhea in Uruguay. ESBL characterization in crude extracts showed a single band at pI 5.4. PCR amplification and sequencing data allowed identification of blaPER-2 and blaTEM-116. Retrospective analysis suggests that these strains were disseminated in the community, even if unnoticed, prior to their access to the hospital environment more than a decade ago.

Bacterial pathogens associated with bloody diarrhea in Uruguayan children

Diarrheal disease continues to be a serious health problem, especially in developing countries. Bloody diarrhea represents approximately 20-30% of all cases and has higher morbidity and mortality. Treatment with antibiotics is beneficial in cases of Shigella, Campylobacter, Yersinia and Salmonella infection, principally in those children with a higher risk of invasive disease. The aims of this study were to detect the bacterial agents associated with bloody diarrhea in children and to determine their antimicrobial susceptibility patterns. Between June 2001 and January 2008, 249 children with bloody diarrhea were studied. Shigella and Shiga toxin-producing Escherichia coli (STEC) were recovered from 48 (19.3%) and 3 (1.2%) of the total of cases, respectively. In 49 out of 249 children, in whom other enteropathogens were investigated, we recovered Campylobacter jejuni from 7 children (14.3%), Salmonella spp. from 2 (4.1%) and Aeromonas spp. from 1 (2%) in addition to Shigella from 7 children (14.3%). Thirty-four (70%) Shigella isolates showed resistance to ampicillin and 13 (27%) to trimethoprim-sulfamethoxazole. All Shigella isolates were susceptible to nalidixic acid, ciprofloxacin and ceftriaxone. Salmonella and STEC isolates were susceptible to all antibiotics assayed. Thus, the use of trimethoprim-sulfamethoxazole or ampicillin would not be appropriate for the empirical treatment of Shigella - associated diarrhea.

New TEM-derived extended-spectrum beta-lactamase and its genomic context in plasmids from Salmonella enterica serovar derby isolates from Uruguay.

ABSTRACT A small (8.2-kb) ColE1 plasmid encoding TEM-144 (a new β-lactamase with a ceftazidimase profile) was sequenced by a gene-walking strategy. The blaTEM allele was carried on a Tn2 element, disrupting a Rom protein gene. TEM-144 differs from TEM-1 by two mutations (R164C and E240K) and from the ceftazidime-hydrolyzing TEM-91 by one mutation (T182M).

Characteristics of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strains isolated from skin and soft-tissue infections in Uruguay.

We analyzed 90 nonduplicates community-associated methicillin-resistant S. aureus (CA-MRSA) strains isolated from skin and soft-tissue infections. All strains were mecA positive. Twenty-four of the 90 strains showed inducible macrolide-lincosamide-streptogramin B resistance. All strains produced α-toxin; 96% and 100% of them displayed positive results for lukS-F and cna genes, respectively. Eigthy-five strains expressed capsular polysaccharide serotype 8. Six different pulsotypes were discriminated by pulsed-field gel electrophoresis (PFGE) and three predominant groups of CA-MRSA strains (1, 2, and 4) were identified, in agreement with phenotypic and genotypic characteristics. Strains of group 1 (pulsotype A, CP8+, and Panton-Valentine leukocidin (PVL)+) were the most frequently recovered and exhibited a PFGE band pattern identical to other CA-MRSA strains previously isolated in Uruguay and Brazil. Three years after the first local CA-MRSA report, these strains are still producing skin and soft-tissue infections demonstrating the stability over time of this community-associated emerging pathogen.

Identification of the first blaCMY-2 gene in Salmonella enterica serovar Typhimurium isolates obtained from cases of paediatric diarrhoea illness detected in South America

The objectives of this study were to investigate clinical isolates of Salmonella enterica serovar Typhimurium resistant to β-lactam antibiotics, to characterise their mechanisms of antibiotic resistance and to evaluate the possible biological cost of expressing resistance genes. Two oxyimino-cephalosporin-resistant Salmonella isolates obtained from children with diarrhoea were characterised. The occurrence of plasmid-encoded blaCMY-2 genes was confirmed by molecular methods and conjugation assays; transcription levels were determined by quantitative real-time PCR (qRT-PCR). The genomic context of the β-lactamases, replicon type and addiction systems were analysed by PCR. Genomic relatedness of both isolates was studied by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) assays. Growth curves, motility and invasiveness assays in Caco-2 cells were performed to analyse the bacterial fitness of both isolates. Both isolates carried a blaCMY-2-like allele in an IncI plasmid and belonged to the same MLST sequence type (ST19); nevertheless, they showed extensive differences in their PFGE profiles and virulotypes. Isolate STM709 appeared to lack the Salmonella virulence plasmid and displayed less motility and invasiveness in cultured cells than isolate STM910. qRT-PCR showed that isolate STM709 had higher blaCMY-2 mRNA levels compared with STM910. Altogether, the results suggest that a plasmid carrying blaCMY-2 could be disseminating among different clones of S. Typhimurium. Different levels of blaCMY-2 mRNA could have an effect on the fitness of this micro-organism, resulting in lower invasiveness and motility.

Prevalence and serotype distribution of Listeria monocytogenes isolated from foods in Montevideo-Uruguay

The aim of this work was to study the prevalence of Listeria monocytogenes in foods obtained in retail shops and food industries located in Montevideo-Uruguay, and to identify the serogroups of the obtained isolates. Three-thousand one-hundred and seventy-five food samples (frozen, deli meats, ready-to-eat and cheese) were analyzed. The obtained isolates were serogrouped by multiplex PCR and serotyped by conventional procedure. Genetic comparisons were performed using pulsed-field gel electrophoresis on a sub-set of isolates belonging to the same serotype successively recovered from the same establishment. L. monocytogenes was isolated from 11.2% of samples. The highest prevalence was observed in frozen foods (38%), followed by cheese (10%). 1/2b and 4b were the most frequently identified serotypes. In six of 236 analyzed establishments we successively recovered L. monocytogenes isolates belonging to the same serotype. Most of them corresponded to serotype 1/2b. Pulsed-field gel electrophoresis profiles suggest that at least 33% of L. monocytogenes 1/2b isolates are genetically related and that may remain viable for prolonged periods. The observed prevalence of L. monocytogenes was lower than reported in neighboring countries. Our findings highlight the role that frozen foods may play in the spread of this pathogen, and the relevance of serotypes 1/2b and 4b.

Risk factors for the acquisition of extended-spectrum beta-lactamase-producing Enterobacteriaceae in hospitalized children

1 Departamento de Bacteriología y Virología, Instituto de Higiene, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay 2 Departamento de Pediatría, Centro Hospitalario Pereira Rossell, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay 3 Departamento de Farmacología y Terapéutica. Hospital de Clínicas “Dr. Manuel Quintela”, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay 4 Laboratorio Central del Hospital Pereira Rossell, Ministerio de Salud Pública, Montevideo, Uruguay

Rhombencephalitis caused by Listeria monocytogenes in a pastured bull

A pastured 2-y-old cross-breed bull developed brainstem encephalitis (rhombencephalitis); Listeria monocytogenes was isolated from the brain. In the brainstem, there was perivascular cuffing, multiple microabscesses, and positive immunostaining for L. monocytogenes. Samples of bovine feces, water, feedstuffs, milking parlor soil, and bulk tank milk were collected from the dairy farm. Seven isolates of the genus Listeria were obtained, 6 of L. innocua and 1 of L. monocytogenes, which was found in the pasture where the bull grazed. Both isolates belonged to serotype 4b and were positive for internalins A, C, and J. According to the DNA fragment patterns of pulsed-field gel electrophoresis, the isolates were closely related. The source of infection was the pasture, implying that listeriosis should not be discounted in cases with compatible clinical signs but the absence of silage feeding.

Bacteriemia por Shigellaflexneri serotipo 6 en pacientes con SIDA: Presentación de dos casos clínicos

Las infecciones por Shigella spp., en general, permanecen localizadas en el tracto digestivo y tienen una evolucion autolimitada. La bacteriemia es una complicacion potencialmente letal que ocurre en pacientes con algun tipo de inmunocompromiso. Presentamos dos casos de bacteriemia causadas por Shigella en dos adultos con SIDA. En ambos pacientes, se recupero Shigella flex-neri en muestras de deposiciones y sangre. Los aislados correspondieron al serotipo 6, fueron resistentes solo a cotrimoxazol y mostraron un perfil de bandas similar por PFGE. Los pacientes recibieron tratamiento antimicrobiano prolongado y la evolucion fue favorable. No se registraron otros casos de diarrea en individuos admitidos en el servicio de emergencia. La hipotesis fue que el paciente 2 adquirio la infeccion en el hospital a partir del paciente 1. Sin embargo, no pudimos establecer el modo de transmision. Aunque poco frecuente, es importante tener presente la ocurrencia de bacteriemia por Shigella spp. o por otros enteropatogenos bacterianos en pacientes inmunocomprometidos con diarrea.