Allodemic distribution of plasmids co-harbouring blaCTX-M-15/aac(6')-Ib-cr/qnrB in Klebsiella pneumoniae is the main source of extended-spectrum ß-lactamases in Uruguay's paediatric hospital.

OBJECTIVES: This study aimed to describe the characteristics of clinical isolates of extended-spectrum ß-lactamase (ESBL)-producing enterobacteria (EPE) in Uruguay's paediatric hospital. METHODS: ESBLs, qnr alleles and aac(6')-Ib-cr were sought and characterised in EPE isolated between March 2010 and March 2012. Transfer of resistance determinants was assessed by conjugation. Incompatibility (Inc) groups, plasmid toxin-antitoxin systems (TAS) and plasmid size were determined in transconjugants. Clonality was analysed by pulsed-field gel electrophoresis. Multilocus sequence typing was done for ESBL-producing Klebsiella pneumoniae. RESULTS: A total of 77 EPE isolates were characterised, comprising 43% K. pneumoniae, 19.5% Serratia marcescens, 19.5% Escherichia coli, 17% Enterobacter cloacae and 1% Klebsiella oxytoca. ESBLs belonged mainly to the blaCTX-M family (69.6%) [blaCTX-M-15 (45%) and blaCTX-M-2 (31%)]. The aac(6')-Ib-cr/qnrB duplex was the most frequently detected plasmid-mediated quinolone resistance mechanism; this association was detected in K. pneumoniae harbouring blaCTX-M-15. Transconjugants were obtained for 71% of the EPE. Amongst transconjugants, certain combinations were found between ESBLs and Inc group, e.g. IncA/C-blaCTX-M-2, IncHI1/HI2-blaCTX-M-9 and IncHI1/HI2-blaSHV-12. In addition, the combination ccdAB-blaCTX-M-15 was also found. K. pneumoniae isolates harbouring blaCTX-M-15/aac(6')-Ib-cr/qnrB showed allodemic behaviour, with a predominance of ST14, ST45 and ST48. CONCLUSIONS: In this study, epidemiological changes in ESBL distribution could be explained by the spread of K. pneumoniae harbouring blaCTX-M-15/aac(6')-Ib-cr/qnrB, encoded mainly on conjugative plasmids featuring ccdAB TAS. Since reports of TAS in K. pneumoniae plasmids are scarce, new strategies are needed to combat intrinsic selection pressure exerted by the association, in conjugative plasmids, of resistance mechanisms with TAS.

Do paranoid delusions exist on a continuum with subclinical paranoia? A multi-method taxometric study.

BACKGROUND: There is widespread interest in whether psychosis exists on a continuum with healthy functioning. Previous research has implied that paranoia, a common symptom of psychosis, exists on a continuum but this has not been investigated using samples including both patients and non-patients and up-to-date taxometric methods. AIM: To assess the latent structure of paranoia in a diverse sample using taxometric methods. METHOD: We obtained data from 2836 participants, including the general population as well as at-risk mental state and psychotic patients using the P-scale of the Paranoia and Deservedness Scale. Data were analysed using three taxometric procedures, MAMBAC, MAXEIG and L-MODE (Ruscio, 2016), and two sets of paranoia indicators (subscales and selected items from the P scale), including and excluding the patient groups. RESULTS: Eleven of the twelve analyses supported a dimensional model. Using the full sample and subscales as indicators, the MAMBAC analysis was ambiguous. Overall, the findings converged on a dimensional latent structure. CONCLUSIONS: A dimensional latent structure of paranoia implies that the processes involved in sub-clinical paranoia may be similar to those in clinical paranoia.

A rural worker infected with a bovine-prevalent genotype of Campylobacter fetus subsp. fetus supports zoonotic transmission and inconsistency of MLST and whole-genome typing.

Whole-genome characterisation in clinical microbiology enables to detect trends in infection dynamics and disease transmission. Here, we report a case of bacteraemia due to Campylobacter fetus subsp. fetus in a rural worker under cancer treatment that was diagnosed with cellulitis; the patient was treated with antibiotics and recovered. The routine typing methods were not able to identify the microorganism causing the infection, so it was further analysed by molecular methods and whole-genome sequencing. The multi-locus sequence typing (MLST) revealed the presence of the bovine-associated ST-4 genotype. Whole-genome comparisons with other C. fetus strains revealed an inconsistent phylogenetic position based on the core genome, discordant with previous ST-4 strains. To the best of our knowledge, this is the first C. fetus subsp. fetus carrying the ST-4 isolated from humans and represents a probable case of zoonotic transmission from cattle.

First report of the ceftazidimase CTX-M-19 in South America.

We report the first detection of bla CTX-M-19 in South America, harboured in an Escherichia coli isolate obtained from a urine sample; such an isolate belonged to phylogenetic group A, ST603, and showed a ceftazidimase profile. bla CTX-M-19 was encoded in an approximately 100 kb IncI1/IncF conjugative plasmid, featuring pndAC and hok/sok addiction systems; the ß-lactamase gene was flanked upstream by three tandem-like transposons (IS26, IS10 and ISEcp1), inserted one inside the other, and downstream by IS903.

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.

Prevalence of Salmonella enterica in poultry and eggs in Uruguay during an epidemic due to Salmonella enterica serovar Enteritidis.

Salmonella enterica serovar Enteritidis (S. Enteritidis) is frequently associated with food-borne disease worldwide. Poultry-derived products are a major source. An epidemic of human infection with S. Enteritidis occurred in Uruguay, and to evaluate the extent of poultry contamination, we conducted a nationwide survey over 2 years that included the analysis of sera from 5,751 birds and 12,400 eggs. Serological evidence of infection with Salmonella group O:9 was found in 24.4% of the birds. All positive sera were retested with a gm flagellum-based enzyme-linked immunosorbent assay, and based on these results, the national prevalence of S. Enteritidis infection was estimated to be 6.3%. Salmonellae were recovered from 58 of 620 pools made up of 20 eggs each, demonstrating a prevalence of at least 1 in every 214 eggs. Surprisingly, the majority of the isolates were not S. Enteritidis. Thirty-nine isolates were typed as S. Derby, 9 as S. Gallinarum, 8 as S. Enteritidis, and 2 as S. Panama. Despite the highest prevalence in eggs, S. Derby was not isolated from humans in the period of analysis, suggesting a low capacity to infect humans. Microarray-based comparative genomic hybridization analysis of S. Derby and S. Enteritidis revealed more than 350 genetic differences. S. Derby lacked pathogenicity islands 13 and 14, the fimbrial lpf operon, and other regions encoding metabolic functions. Several of these regions are present not only in serovar Enteritidis but also in all sequenced strains of S. Typhimurium, suggesting that these regions might be related to the capacity of Salmonella to cause food-borne disease.

[Detection and characterization of Shiga toxin-producing Escherichia coli from clinical cases and food in Uruguay]. / Detección y caracterización de Escherichia coli productor de toxina Shiga a partir de casos clínicos y de alimentos en Uruguay.

We have assessed the frequency of Shiga toxin-producing Escherichia coil (STEC) in clinical and food samples as well as studied the genotypic and phenotypic characteristics of the recovered strains. One hundred ninety eight fecal samples from children with bloody diarrhea (BD), 14 from children with hemolytic uremic syndrome (HUS), 220 ground beef samples and 4 STEC isolates from other beef-derived products were analyzed. The STEC strains were isolated from 3 (1.5%) children with bloody diarrhea, 1 (7%) from a child with HUS and 4 (1.8%) from ground beef samples. All strains were eae and ehxA positive. The serotypes found were: O157:H7 (9 strains), O26:H11 (2), O111: NM (1) and O145:HNT (1). All O157:H7 STEC strains harbored the eae subtype gamma1, O26:H11 and O145:HNT strains, subtype beta1 and O111:NM strain, subtype gamma2/theta. The STEC strains of the same serogroup showed high genetic diversity. In Uruguay, STEC is not frequently isolated from cases of bloody diarrhea in children. However, all the recovered STEC strains carried the genes associated with severe disease and 2 out of 3 children infected with STEC developed HUS. Ground beef and other food products might be important vehicles for O157:H7 strains.

Detección y caracterización de Escherichia coli productor de toxina Shiga a partir de casos clínicos y de alimentos en Uruguay / Detection and characterization of Shiga toxin - producing Escherichia coli from clinical cases and food in Uruguay

Establecimos la frecuencia de aislamiento de Escherichia coli productor de toxina Shiga (STEC) a partir de muestras clínicas y de alimentos, así como las características fenotípicas y genotípicas de las cepas recuperadas. Se analizaron 198 muestras fecales de niños con diarrea sanguinolenta (DS), 14 muestras fecales de niños con síndrome urémico hemolítico (SUH) y 220 muestras de carne picada. También se estudiaron 4 cepas STEC aisladas de alimentos embutidos. Se recuperó STEC de 3 (1,5%) de los niños con DS, de 1 (7%) niño con SUH y de 4 (1,8%) de las muestras de carne picada. Todas las cepas fueron eae y ehxA positivas. Los serotipos detectados fueron: O157:H7 (9 cepas), O26:H11 (2 cepas), O111:NM (1 cepa) y O145:HNT (1 cepa). Todas las cepas O157:H7 portaron el subtipo eae-g1; las cepas O26:H11 y O145:HNT portaron el subtipo eae-b1 y la cepa O111:NM portó el subtipo eae-g2/q. Las cepas STEC del mismo serogrupo mostraron alta diversidad genética. En Uruguay STEC no sería agente frecuente de diarrea con sangre en niños. Sin embargo, las cepas recuperadas presentaron los genes asociados con enfermedad severa y 2 de los 3 niños infectados con STEC evolucionaron a SUH. La carne picada y otros alimentos serían vehículos importantes de O157:H7.

We have assessed the frequency of Shiga toxin-producing Escherichia coli (STEC) in clinical and food samples as well as studied the genotypic and phenotypic characteristics of the recovered strains. One hundred ninety eight fecal samples from children with bloody diarrhea (BD), 14 from children with hemolytic uremic syndrome (HUS), 220 ground beef samples and 4 STEC isolates from other beef-derived products were analyzed. The STEC strains were isolated from 3 (1.5%) children with bloody diarrhea, 1 (7%) from a child with HUS and 4 (1.8%) from ground beef samples. All strains were eae and ehxA positive. The serotypes found were: O157:H7 (9 strains), O26:H11 (2), O111: NM (1) and O145:HNT (1). All O157:H7 STEC strains harbored the eae subtype g1, O26:H11 and O145:HNT strains, subtype b1 and O111:NM strain, subtype g2/q. The STEC strains of the same serogroup showed high genetic diversity. In Uruguay, STEC is not frequently isolated from cases of bloody diarrhea in children. However, all the recovered STEC strains carried the genes associated with severe disease and 2 out of 3 children infected with STEC developed HUS. Ground beef and other food products might be important vehicles for O157:H7 strains.

Vigilancia del uso de antibióticos en el Hospital Pediátrico del Centro Hospitalario Pereira Rossell: susceptibilidad antimicrobiana; gasto y consumo de antibióticos / Vigilance of antibiotic use in a pediatric hospital (Hospital Pediátrico del Centro Hospitalario Pereira Rossell): antimicrobial susceptibility; cost and consumption antibiotics

Introducción: el uso de guías para el tratamiento antibiótico empírico constituye una estrategia fundamental para contener el aumento sostenido en la resistencia bacteriana y en los gastos en antibióticos. En 1998, en el Hospital Pediátrico del Centro Hospitalario Pereira Rossell se adecuaron las guías de antibioticoterapia empírica par alas infecciones más frecuentes asistidas en salas generales y en emergencia, en base a la epidemiología local. Desde esa fecha se comenzó la vigilancia deel uso de antibióticos. Objetivos: analizar los resultados de esta vigilancia sobre el gasto y el consumo de antibióticos y sobre la susceptibilidad antimicrobiana. Metodología: se determinó el gasto generado por los antibióticos recomendados en las guías de atención hospitalaria, en salas generales y en emergencia (penicilina, aminopenicilinas, cefalosporinas, macrólidos). Para estimar el consumo en pacientes hospitalizados se calculó la dosis diaria definida (DDD)/100 camas día para cada uno de los antibióticos. Se compararon los resultados de gasto y consumo 2001-2. Se comparó la susceptibilidad antimicrobiana de los gérmenes más frecuentes del período 2001-2 con la previa a la adecuación de las guías terapéuticas. Resultados: los antibióticos recomendados representaron enel año 2001, 57 por ciento del gasto en antibióticos (2.206.652,57 pesos) y en 2002, 54 por ciento (1.441.280 pesos). Los antibióticos más usados fueron las animopenicilinas. El consumo de cefuroxime intravenoso disminuyó 60 por ciento (de 13.1 a 5.36 DDD/100 camas-día). El consumo de ceftriaxona intravenosa y de amoxicilina oral aumentó 38 por ciento y 16 por ciento, respectivamente. Para el resto de los antibióticos el consumo se mantuvo constante. Las tasas de susceptibilidad antimicrobiana de los gérmenes más frecuentes (S. pneumoniae, S. aureus, E, coli, Shigella spp) a los antibióticos recomendados en las guías terapéuticas se mantuvieron por encima de 80 por ciento. Conclusiones: estos son los primeros resultados de la vigilancia del uso ded antibióticos en este hospital y demuestran la vigencia y la aceptación de las guías de antibióticoterapia empírica propuestas. Es difícil interpretar la disminución del gasto en antibióticos debido a la variación en el precio de estos medicamentos en este período.

Standard case management of pneumonia in hospitalized children in Uruguay, 1997 to 1998.

OBJECTIVE: To report the results of the use of antimicrobial guidelines for the management of children with community-acquired bacterial pneumonia. METHODS: Admittance and discharge criteria and algorithms for diagnosis and treatment were established. The decision to treat with antibiotics was based on radiologic findings in pneumonia with pulmonary consolidation and left to the attending physician's criteria in the remaining cases. The use of antibiotics was limited to penicillin and derivatives (ampicillin, amoxicillin) and macrolides. RESULTS: Of the 1163 children treated as bacterial pneumonia, hospitalized in public and private health facilities in Montevideo from September, 1997, through September, 1998, standard case management was applied in 1082 (93%). Age distribution was: <1 month, 1%; between 1 and 11 months, 29%; between 1 and 5 years, 50%; >5 years, 20%. Chest radiography showed evidence of pulmonary consolidation in 843 children (73%). Bacteria were detected in blood culture and/or pleural fluid of 57 children (5%). In 51 the identified microorganism was Streptococcus pneumoniae, susceptible to penicillin in 30, intermediate in 6 and resistant in 5 (maximum MIC, 4 microg/ml); in 10 cases etiologic diagnosis was made by antigen detection. Empyema was present in 62 children (5.3%); 38 (3.27%) required treatment in an intensive care unit; and 5 (0.4%) died. CONCLUSIONS: Compliance with standard case management was highly satisfactory. Outcome of children treated with penicillin and derivatives was good, including children with empyema and pneumatocele and two patients with penicillin-resistant S. pneumoniae. At the present time S. pneumoniae resistant to penicillin is not an important problem in children with pneumonia in Uruguay. Surveillance of identified microorganisms and their antimicrobial susceptibility must continue.

Capsular type distribution and susceptibility to antibiotics of Streptococcus pneumoniae clinical strains isolated from Uruguayan children with systemic infections. Pneumococcus Study Group.

Children under 24 months of age are at high risk for serious infection with Streptococcus pneumoniae but they do not elicit effective immune responses to the currently available capsular polysaccharide vaccines. A polysaccharide protein conjugated vaccine involving the most frequent types has become an urgent need. To produce such a vaccine for Latin America, information on type distribution is required. Recently, Uruguay was 1 of the 6 countries in Latin America where surveillance for invasive pneumococcal infections in children under the age of 5 years was carried out. Seventy percent of the 182 invasive S. pneumoniae isolates were recovered from patients under 24 months of age, and 19% were recovered from infants under 6 months. The 7 most frequent types were 14, 5, 1, 6B, 3, 7F, and 19A; representing 80% of invasive isolates. Twenty-one types were identified, 16 in pneumonia and 14 in meningitis. Resistance to penicillin increased during the study period, from 29% in 1994, to 40% in 1995-1996, mainly because of the spread of type 14 strains resistant to penicillin and trimethoprim/sulphamethoxazol (89% of resistant isolates). The high proportion of systemic pneumococcal infections recorded in patients under 24 months of age and the increasing resistance of these agents to first-choice antibiotics point to an urgent need for a capsular polysaccharide protein conjugated vaccine.

Cutaneous botryomycosis. Report of the two first Uruguayan cases.

The first two Uruguayan cases of botryomycosis were diagnosed in white adult male patients with cutaneous lesions. Staphylococcus aureus was the etiologic agent in both cases. In one of them, however, Pseudomonas aeruginosa was also identified during relapse. General comments on diagnosis and pathogenesis of the disease are made.

Antigenic and immunogenic activity of flagella and fimbriae preparations from uropathogenic Proteus mirabilis.

The antigenic and immunogenic activities of fimbriae and flagella from three uropathogenic strains of Proteus mirabilis were compared. Flagella were obtained by mechanical treatment and fimbriae were isolated from cells by heat shock, ammonium sulfate precipitation, sodium deoxycholate and urea treatment, and gel filtration. Both preparations inoculated to mice demonstrated high antigenicity. Titers up to 1:80,000 were determined by an enzyme-linked immunosorbent assay either against the homologous or heterologous strains. When immunized mice were challenged with homologous or heterologous hematogenous infecting doses, a good cross protection was achieved only when fimbriae were used as antigens. Cross-reactivity found between the three fimbriae antisera, and the presence of common proteins in sodium dodecyl sulfate polyacrylamide gel electrophoresis patterns of fimbriae, should validate the study of these proteins to determine the existence of a shared adhesin.

[Profiles of enzymatic activities of the genera Agrobacterium, Alcaligenes, Alteromonas, Flavobacterium and Pseudomonas]. / Profils d'activités enzymatiques des genres Agrobacterium, Alcaligenes, Alteromonas, Flavobacterium et Pseudomonas.

Profiles of enzymatic activities were studied using 19 chromogenic substrates for 22 species (211 strains) belonging to the genera Agrobacterium, Alcaligenes, Alteromonas, Flavobacterium and Pseudomonas. The observed patterns of reactions may be useful as an aid in identification of these species and for epidemiological studies.