Frecuencia de clostridium difficile en pacientes pediátricos con diarrea asociada al uso de antibióticos. Hospital Vicente Corral Moscoso Cuenca 1997-1998

El presente estudio se realizó en el departamento de Pediatría del Hospital Vicente Corral Moscoso [H.V.C.M.] de la ciudad de Cuenca, el año 1997; en pacientes ambulatorios de las zonas urbana y rural. Sus objetivos fueron; determinar la prevalencia del Clostridiun Difficile en pacientes menores de dos años con diarrea asociada o no al uso de antibióticos, y el grado de asociación entre el uso de estos, la presencia de C. Difficile y aparición o no de diarrea. El método epidemiológico utilizado fue: Descriptivo de corte comparativo el universo estuvo conformado por los niños menores de dos años atendidos en el H.V.C.M., en el año 1997; la muestra quedó constituida por 85 unidades de estudio [según la tabla del College Outline Series for Stadísticians]. El método de diagnóstico de laboratorio empleado fue; Test Premier C. Difficile Toxin A de los Laboratorios Meridian Diagnostics Ind. USA. Los resultados indican que la proporción de prevalencia de punto de diarrea asociada al uso de antibióticos en toda la población de estudio fue del 60 por ciento. La proporción de prevalencia de punto en la asociación uso de antibióticos y presencia de diarrea fue de 67 por ciento. La razón de prevalencia [relación entre proporción de prevalencia de expuestos y no], alcanzó a 1.34 que fueron no significativos estadísticamente. Las autoras encontraron C. Difficile en los grupos que recibieron antibióticos y presentaron diarrea. Al término del estudio se concluyó que la presencia del C. Difficcile, esta relacionado estrechamente con el uso de antibiótico. Sobre estos resultados, se recomienda: profundizar la investigación a través de estudios locales analíticos o de intervención y que las instituciones de salud se preocupen sobre el problema

Predictores de riesgo para desarrollo de infección por Clostridium difficile

Clostridium diffícile (CD) es el agente responsable de la mayoría de los episodios de diarrea de origen nosocomial. El objetivo de este estudio fue identificar los factores de riesgo asociados a la infección por C. difficile (ICD) en los pacientes con diarrea de origen nosocomial en el Hospital Universitario Dr. Manuel Quintela. Para ello se llevó adelante un estudio observacional, analítico, de casos y controles no pareados. Se incluyeron todos los pacientes mayores de 18 años que instalaron diarrea posterior a las 72 horas del ingreso al hospital y en quienes se solicitó investigación de CD durante el período abril a diciembre de 2013. Veintisiete pacientes (18,6%) cumplieron con la definición de caso, 105 (72,4%) cumplieron con la definición de control y 13 (9,0%) se excluyeron por presentar solo la prueba de glutamato deshidrogenasa (GDH) positiva. En relación con la presentación clínica, se destaca la asociación significativa de la ICD con la presencia de fiebre. En el análisis univariado el haber recibido antibióticos por más de 30 días y el uso previo de clindamicina estuvieron significativamente asociados a ICD. En el análisis multivariante solo el uso previo de clindamicina (p = 0,004 OR 5,881; IC95% 1,743-19,841) se mantuvo como factor de riesgo independiente. Este hallazgo pone en evidencia que en situación de endemia la implementación de una política de uso racional de clindamicina es una de las medidas necesarias para disminuir la frecuencia de ICD.

Clostridium difficile ribotypes in humans and animals in Brazil

Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies about C. difficile polymerase chain reaction (PCR) ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficile ribotypes isolated from humans and animals in Brazil. Seventysix C. difficile strains isolated from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf were distributed into 24 different PCR ribotypes. Among toxigenic strains, PCR ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4%) and eight (10.5%) samples, respectively. Fourteen different PCR ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.

Role of phospholipase A(2) and tyrosine kinase in Clostridium difficile toxin A-induced disruption of epithelial integrity, histologic inflammatory damage and intestinal secretion

Clostridium difficile-associated disease causes diarrhea to fulminant colitis and death. We investigated the role of phospholipase A(2) (PLA(2)) inhibitors, aristolochic acid (AA), bromophenacyl bromide BPB and quinacrine (QUIN) on the C. difficile toxin A-induced disruption of epithelial integrity, histologic inflammatory damage and intestinal secretion. Toxin A caused severe hemorrhagic and inflammatory fluid secretion at 6-8 h in rabbit ileal segments, an effect that was significantly inhibited by QUIN (71%, P < 0.01), AA (87%, P < 0.0001) or by BPB (51%, P < 0.01). The secretory effect of toxin A was also inhibited in segments adjacent to those with AA (89%, P < 0.01). Furthermore, QUIN or AA substantially reduced the histologic damage seen after 6-8 h in rabbit ileal segments. The cyclooxygenase inhibitor, indomethacin, also significantly inhibited (96%; n = 6) the secretory effects of toxin A in ligated rabbit intestinal segments. The destruction by toxin A of F-actin at the light junctions of T-84 cell monolayers was not inhibited by AA or BPB. AA or QUIN had no effect on the T-84 cell tissue resistance reduction over 8-24 h after toxin A exposure. All the inhibitors were shown to be effective in the doses administered direct in ileal loops to inhibit PLA(2) activity. The data suggest that PLA(2) is involved in the major pathway of toxin A-induced histologic inflammatory damage and hemorrhagic fluid secretion. Cop. right (C) 2008 John Wiley & Sons, Ltd.

Characteristics of Clostridium difficile infection in patients with discordant diagnostic test results

Background: Clinical features of Clostridium difficile infection (CDI) cases diagnosed by detection of polymerase chain reaction (PCR), with negative toxin enzyme immunoassay results (EIA) have not been fully elucidated. The purpose of this study was to determine the magnitude of CDI patients who had negative EIA toxin determinations but positive PCR tests, and their differences in clinical presentation. Methods: We performed a retrospective study comparing the clinical features of CDI cases detected by EIA (toxins A + B) with cases detected by PCR (toxin negative, PCR positive) over a 16-month period. Only patients with an initial Clostridium difficile infection episode that fulfilled a standardized definition were included. Results: During the study period, 107 episodes of CDI were detected. Seventy-four patients (69%) had positive glutamate dehydrogenase (GDH) antigen and EIA determinations (EIA positive patients). Thirty-three patients (31%) had GDH positive, negative toxin EIA and positive PCR determination (PCR positive patients). PCR positive patients were younger, 57 (27) years (mean [SD]), than EIA positive patients, 71 (16) years, (p < 0.001). Fewer PCR positive patients were receiving proton pump inhibitors (21 patients, 64%) than EIA positive patients (61 patients, 82%, p = 0.034). The clinical presentation was similar in both groups. In the multivariate analysis, lower age was identified as the only independent variable associated with PCR positive patients. Conclusions: One third of Clostridium difficile infection patients present negative toxin EIA and PCR positive tests. Performing PCR determination after the negative EIA test is more relevant in younger patients.

COMPUTATIONAL PREDICTION OF THE SPORULATION NETWORK IN CLOSTRIDIUM THERMOCELLUM

Sporulation is a process in which some bacteria divide asymmetrically to form tough protective endospores, which help them to survive in a hazardous environment for a quite long time. The factors which can trigger this process are diverse. Heat, radiation, chemicals and lacking of nutrition can all lead to the formation of endospores. This phenomenon will lead to low productivity during industrial production. However, the sporulation mechanism in a spore-forming bacterium, Clostridium theromcellum, is still unclear. Therefore, if a regulation network of sporulation can be built, we may figure out ways to inhibit this process. In this study, a computational method is applied to predict the sporulation network in Clostridium theromcellum. A working sporulation network model with 40 new predicted genes and 4 function groups is built by using a network construction program, CINPER. 5 sets of microarray expression data in Clostridium theromcellum under different conditions have been collected. The analysis shows the predicted result is reasonable.

Estabilidade oxidativa e microbiológica em carne de galinha mecanicamente separada e adicionada de antioxidantes durante período de armazenamento a -18 °C

Com o objetivo de acompanhar a estabilidade físico-química e microbiológica da carne mecanicamente separada (CMS) de diferentes origens e estocada durante 99 dias a -18 °C, foi realizada prévia mistura de conservante (nitrito de sódio) e antioxidante (eritorbato de sódio) em CMS obtida de duas linhagens de aves: galinhas matrizes de corte e galinhas poedeiras comerciais brancas. Na CMS de cada linhagem foram realizados três diferentes tratamentos: 1) controle (sem aditivos); 2) adição de 150 ppm de nitrito; e 3) adição de 150 ppm de nitrito e 500 ppm de eritorbato. Os resultados encontrados demonstraram que a adição de nitrito isoladamente não impediu a oxidação lipídica, avaliada através do índice de TBARS, nem a alteração na cor, avaliada em colorímetro. Por outro lado, a adição de nitrito juntamente com eritorbato foi efetiva na redução dos problemas de oxidação lipídica na CMS de galinhas matrizes, e em menor grau, na CMS de poedeiras. A adição de nitrito e eritorbato na CMS também melhorou a preservação da cor vermelha desejável (a*) ao longo do tempo. A avaliação da estabilidade microbiológica da CMS, realizada no primeiro e último dia de estocagem congelada, para microrganismos mesófilos, Escherichia coli, Staphylococcus aureus, Clostridium perfringens e Pseudomonas spp., e quinzenalmente para microrganismos psicrotróficos, indicou que não houve uma variação significativa nas contagens em função do tratamento utilizado (diferentes aditivos adicionados). Não foi detectada Salmonella spp. em nenhuma das amostras analisadas. Em função da melhoria da estabilidade oxidativa, recomenda-se a adição de nitrito (150 ppm) e eritorbato (500 ppm) em CMS de galinhas matrizes a ser estocada congelada por um período prolongado.

Microbiological quality of hamburgers sold in the streets of Cuiabá - MT, Brazil and vendor hygiene-awareness

This study evaluated the microbiological quality of hamburgers and the microbe community on the hands of vendors in Cuiabá, Mato Grosso, Brazil, in relation to vendors´ awareness as to what constitute acceptable food-handling practices as part of a broad-spectrum research programme on street foods in Brazil . Sale of the hamburger known as the 'baguncinha' is common and widespread in urban Cuiabá, Mato Grosso, Brazil. Food inspectors encounter various difficulties in carrying out inspections. One hundred and five hamburgers samples were evaluated using conventional methods including tests for facultative aerobic and/or anaerobic mesophytic bacteria, coliform counts at 45 °C, the coagulase test for Staphylococcus, Gram-staining for the presence of Bacillus cereus, Clostridium sulphite reductase and Salmonella spp. The hamburgers were categorized as unsuitable for human consumption in 31.4% of samples, with those testing positive for coliforms and Staphylococcus at unacceptably high levels by Brazilian standards. High levels of microbiological contamination were detected on the hands of the food handlers and mesophytic bacterial counts reached 1.8 × 10(4) CFU/hand. Interviews were carried out by means of questionnaires to evaluate levels of awareness as to acceptable food handling practices and it was found that 80,1% of vendors had never participated in any kind of training.

Quality of water sources used as drinking water in a Brazilian peri-urban area

The objective of this paper was to assess bacteriological quality of drinking water in a peri-urban area located in the Metropolitan Region of São Paulo, Brazil. A total of 89 water samples were collected from community plastic tanks and 177 water samples from wells were collected bimonthly, from September 2007 to November 2008, for evaluating bacteriological parameters including: Escherichia coli, Enterococcus and heterotrophic plate count (HPC). Clostridium perfringens was investigated in a subsample (40 samples from community plastic tank and 40 from wells). E. coli was present in 5 (5.6%) samples from community plastic tanks (2.0 - 5.1x10(4) MPN/100mL) and in 70 (39.5%) well samples (2.0 - 8.6x10(4) MPN/100mL). Thus, these samples were not in accordance with the Brazilian Regulation. Enterococcus was detected in 20 (22.5%) samples of the community plastic tanks (1 to 79 NC/100mL) and in 142 (80.2%) well samples (1 to >200 NC/100mL). C. perfringens was detected in 5 (12.5%) community plastic tanks samples and in 35 (87.5%) wells samples (2.2 to >16 MPN/100mL). HPC were above 500 CFU/mL in 5 (5.6%) waters from community plastic tanks. In wells samples, the HPC ranged from <1 to 1.6x10(4) CFU/mL. The residual chlorine did not attend the standard established in the drinking water legislation (0.2 mg/L), except in 20 (22.5%) samples. These results confirm the vulnerability of the water supply systems in this peri-urban area what is clearly a public health concern.

Matched case-control study evaluating the frequency of the main agents associated with neonatal diarrhea in piglets

A case-control study was carried out in litters of 1 to 7-day-old piglets to identify the main infectious agents involved with neonatal diarrhea in pigs. Fecal samples (n=276) from piglets were collected on pig farms in the State of Rio Grande do Sul, Brazil, from May to September 2007. Litters with diarrhea were considered cases (n=129) and normal litters (n=147) controls. The samples were examined by latex agglutination test, PAGE, conventional isolating techniques, ELISA, PCR, and microscopic methods in order to detect rotavirus, bacterial pathogens (Escherichia coli, Clostridium perfringens type A and C, and Clostridium difficile), and parasites (Coccidian and Cryptosporidium spp.). Outbreaks of diarrhea were not observed during sampling. At least one agent was detected in fecal samples on 25 out of 28 farms (89.3%) and in 16 farms (57.1%) more than one agent was found. The main agents diagnosed were Coccidia (42.86%) and rotavirus (39.29%). The main agents identified in litters with diarrhea were Clostridium difficile (10.6%), Clostridium perfringens type A (8.8%) and rotavirus (7.5%); in control litters, Clostridium difficile (16.6%) and Coccidian (8.5%). Beta hemolytic Escherichia coli and Clostridium perfringens type C were not detected. When compared with controls, no agent was significantly associated with diarrhea in case litters. These findings stress the need for caution in the interpretation of laboratorial diagnosis of mild diarrhea in neonatal pigs, as the sole detection of an agent does not necessarily indicate that it is the cause of the problem.

Performance evaluation and phylogenetic characterization of anaerobic fluidized bed reactors using ground tire and pet as support materials for biohydrogen production

This study evaluated two different support materials (ground tire and polyethylene terephthalate [PET]) for biohydrogen production in an anaerobic fluidized bed reactor (AFBR) treating synthetic wastewater containing glucose (4000 mg L(-1)). The AFBR, which contained either ground tire (R1) or PET (R2) as support materials, were inoculated with thermally pretreated anaerobic sludge and operated at a temperature of 30 degrees C. The AFBR were operated with a range of hydraulic retention times (HRT) between 1 and 8 h. The reactor R1 operating with a HRT of 2 h showed better performance than reactor R2, reaching a maximum hydrogen yield of 2.25 mol H(2) mol(-1) glucose with 1.3 mg of biomass (as the total volatile solids) attached to each gram of ground tire. Subsequent 16S rRNA gene sequencing and phylogenetic analysis of particle samples revealed that reactor R1 favored the presence of hydrogen-producing bacteria such as Clostridium, Bacillus, and Enterobacter. (C) 2010 Elsevier Ltd. All rights reserved.

Hydrogen production from soft-drink wastewater in an upflow anaerobic packed-bed reactor

The production of hydrogen from soft-drink wastewater in two upflow anaerobic packed-bed reactors was evaluated. The results show that soft-drink wastewater is a good source for hydrogen generation. Data from both reactors indicate that the reactor without medium containing macro- and micronutrients (R2) provided a higher hydrogen yield (3.5 mol H(2) mol(-1) of sucrose) as compared to the reactor (R1) with a nutrient-containing medium (3.3 mol H(2) mol(-1) of sucrose). Reactor R2 continuously produced hydrogen, whereas reactor R1 exhibited a short period of production and produced lower amounts of hydrogen. Better hydrogen production rates and percentages of biogas were also observed for reactor R2, which produced 0.4 L h(-1) L(-1) and 15.8% of H(2), compared to reactor R1, which produced 0.2 L h(-1) L(-1) and 2.6% of H(2). The difference in performance between the reactors was likely due to changes in the metabolic pathway for hydrogen production and decreases in bed porosity as a result of excessive biomass growth in reactor R1. Molecular biological analyses of samples from reactors R1 and R2 indicated the presence of several microorganisms, including Clostridium (91% similarity), Enterobacter (93% similarity) and Klebsiella (97% similarity). Copyright (C) 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.

Fermentative hydrogen production by microbial consortium

Heat pre-treatment of the inoculum associated to the pH control was applied to select hydrogen-producing bacteria and endospores-forming bacteria. The source of inoculum to the heat pre-treatment was from a UASB reactor used in the slaughterhouse waste treatment. The molecular biology analyses indicated that the microbial consortium presented microorganisms affiliated with Enterobacter cloacae (97% and 98%), Clostridium sp. (98%) and Clostridium acetobutyricum (96%), recognized as H, and volatile acids` producers. The following assays were carried out in batch reactors in order to verify the efficiencies of sucrose conversion to H-2 by the microbial consortium: (1) 630.0 mg sucrose/L, (2) 1184.0 mg sucrose/L, (3) 1816.0 mg sucrose/L and (4) 4128.0 mg sucrose/L. The subsequent yields were obtained as follows: 15% (1.2 mol H-2/mol sucrose), 20% (1.6 mol H-2/mol sucrose), 15% (1.2 mol H-2/mol sucrose) and 4% (0.3 mol H-2/mol sucrose), respectively. The intermediary products were acetic acid, butyric acid, methanol and ethanol in all of the anaerobic reactors. (C) 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.

Degradation of detergent (linear alkylbenzene sulfonate) in an anaerobic stirred sequencing-batch reactor containing granular biomass

This study aimed to determine the efficiency of an anaerobic stirred sequencing-batch reactor containing granular biomass for the degradation of linear alkylbenzene sulfonate (LAS), a surfactant present in household detergent. The bioreactor was monitored for LAS concentrations in the influent, effluent and sludge, pH, chemical oxygen demand, bicarbonate alkalinity, total solids, and volatile solids. The degradation of LAS was found to be higher in the absence of co-substrates (53%) than in their presence (24-37%). Using the polymerase chain reaction and denaturing gradient gel electrophoresis (PCR/DGGE), we identified populations of microorganisms from the Bacteria and Archaea domains. Among the bacteria, we identified uncultivated populations of Arcanobacterium spp. (94%) and Opitutus spp. (96%). Among the Archaea, we identified Methanospirillum spp. (90%), Methanosaeta spp. (98%), and Methanobacterium spp. (96%). The presence of methanogenic microorganisms shows that LAS did not inhibit anaerobic digestion. Sampling at the last stage of reactor operation recovered 61 clones belonging to the domain bacteria. These represented a variety of phyla: 34% shared significant homology with Bacteroidetes, 18% with Proteobacteria, 11% with Verrucomicrobia, 8% with Fibrobacteres, 2% with Acidobacteria, 3% with Chlorobi and Firmicutes, and 1% with Acidobacteres and Chloroflexi. A small fraction of the clones (13%) were not related to any phylum. Published by Elsevier Ltd.