Susceptibility to first choice antimicrobial treatment for urinary tract infections to Escherichia coli isolates from women urine samples in community South Brazil

ABSTRACT E. coli is the main pathogen of UTI. It is important to be aware the local epidemiological data for an appropriate initial treatment. Resistance to antimicrobial agents has increased, especially to first-choice antibiotics in the treatment of cystitis. There are few studies on the sensivity profile of community uropathogen in our region. Objective: To characterize antimicrobials the sensitivity profile to E. coli isolated from urocultures of women treated at Basic Health Units and Emergency Care Units of Londrina-Paraná- Brazil during a period of 12 months (June 1, 2016 to June 1, 2017). Methodology: A cross-sectional study was carried out from June 2016 to June 2017. All urine samples collected in the Basic Health Units and Emergency Departments in the city of Londrina (Paraná State, Brazil) were sent to a Central Laboratory where the identification and antimicrobial susceptibility testing were performed. Clinical Laboratory Standards Institute (CLSI) breakpoints were used for the interpretation of susceptibility testing results. Results: 56,555 urine cultures were performed in the period, of which 8,832 were positive, of which 5,377 were women. Of these samples, 4.7% were enterobacteria producing extended-spectrum beta-lactamases (ESBL) and 15.5% resistant to quinolones. TMP- SMX was resistant in more than 30% of the samples in all age groups. Among quinolone-resistant isolates, resistance to cephalothin, ampicillin and sulfamethoxazole-trimethoprim was greater than 60%. Nitrofurantoin was the only antimicrobial that showed 90% of sensitivity. Conclusion: The antimicrobials sensitivity profile was similar to that reported in the literature, with TMP- SMX resistance greater than 30% in the studied samples. Nitrofurantoin maintains high sensitivity rates greater than 90%. Resistance to quinolones increases proportionally with age, as well ESBL.

Antimicrobial activity of oregan and clove essential oils against some foodborne pathogens / Atividade antimicrobiona dos óleos essenciais de orégano e cravo frente a patógenos alimentares

The tendency to replace synthetic antimicrobials for natural ones in food industry and an increase in bacterial resistance to antibiotics resulted in a necessity to find new alternatives, and essential oils are emerging as promising substitutes for synthetic chemicals in food preservation. The objective of this work was to test the antimicrobial activity of oregano (OEO) and clove (CEO) essential oils over a range of bacteria, molds and yeast of importance as pathogens or food spoilage. The antimicrobial activity of oregano and clove essential oils were analyzed by disk diffusion method and broth microdilution test (MIC) of OEO and CEO were determined for each tested microorganism. OEO and CEO were evaluated in natura (IN) and after thermal processing (TP) at 120 o C for 5 min. Both OEO and CEO presented the same inhibition zones for IN and TP samples, for all tested microorganisms, indicating that these oils can be thermally processed maintaining their antimicrobial activity. For OEO and CEO, the more sensitive microorganisms were the fungi (Aspergillus niger, Penicillium citrinum and Candida albicans), followed by Staphylococcus aureus, Bacillus cereus and Methicillin - resistant Staphylococcus aureus (MRSA); the lowest antimicrobial activities were observed against Streptococcus mutans and Enterococcus faecalis. In general, OEO resulted in higher inhibition zones and lower MIC values for all tested microorganisms, suggesting that it was more effective as an antimicrobial agent than CEO (AU)

A preferência mundial para alimentos mais saudáveis e livres de aditivos químicos pelos consumidores, associada ao aumento da resistência bacteriana, resultaram na necessidade de medidas alternativas no setor de alimentos. Os óleos correspondem a antimicrobianos naturais e constituem uma classe emergente como substitutos dos produtos químicos sintéticos na conservação de alimentos. O objetivo deste trabalho foi avaliar a atividade antimicrobiana de óleos essenciais de orégano (OEO) e cravo (CEO ) frente a bactérias, fungos e leveduras de importância no setor de alimentos. OEO e CEO foram avaliados in natura (IN) e após processamento térmico (TP) a 120 o C por 5 minutos. Para avaliar a atividade antimicrobiana frente a cada microrganismo empregou-se o método de discodifusão e o teste de microdiluição em caldo (MIC). Tanto o OEO quanto o CEO apresentaram zonas de inibição semelhantes para amostras IN e TP, indicando que a atividade antimicrobiana desses óleos são resistentes a altas temperaturas. Os microrganismos mais sensíveis para ambos os óleos essenciais foram os fungos (Aspergillus niger, Penicillium citrinum e Candida albicans), seguidos por Staphylococcus aureus, Bacillus cereus e Staphylococcus aureus resistente à meticilina (MRSA). Já as cepas Streptococcus mutans e Enterococcus faecalis apresentaram uma maior resistência frente à atividade antimicrobiana dos óleos essenciais. Em geral, os maiores halos de inibição e menores valores de MIC foram obtidos quando empregado o OEO, sugerindo uma maior atividade microbiana do mesmo quando comparado ao CEO. (AU)

Clonagem, sequenciamento, expressão e caracterização do gene tsh de uma cepa de Escherichia coli patogênica de aves / Cloning, sequencing, expression, and characterization of the tsh gene from an avian pathogenic Escherichia coli strain

A hemaglutinina temperatura sensível (Tsh) pertence à família das serino-proteases autotransporte de Enterobacteriacea (SPATE), as quais são capazes de clivar diferentes substratos. Nós isolamos e caracterizamos o gene de Escherichia coli patogênica aviária (APEC) amostra APEC 13, sorotipo O2:H9, clonado em pET 101. A região de 4.2 kb do DNA clonado codificou uma proteína de aproximadamente 140 kDa (r-Tsh). O plasmídio recombinante pET 101-tsh conferiu um fenótipo de hemaglutinação positivo para a linhagem BL21 (tsh) para eritrócitos de galinha. A proteína r-Tsh foi purificada em coluna de níquel e utilizada na produção de anticorpos anti-Tsh. Um fragmento de 1.6 kb foi amplificado e subclonado em pCR4, e a seqüência parcial mostrou alta homologia com outras seqüências analisadas. O anti-Tsh reagiu com as proteínas r-Tsh e Tsh nativa da amostra APEC13, como demonstrado pela técnica de Western blot, mostrando que a r-Tsh tem epitopos conservados e que sua antigenicidade foi preservada. O anti-Tsh também inibiu a atividade hemaglutinante das amostras APEC 13 e BL12/pET 101-tsh

The temperature-sensitive hemagglutinin (Tsh) belongs to a family of high-molecular-weight serineprotease autotransporters of Enterobacteriaceae (SPATEs), which can cleave different substrates. Weisolated and characterised the tsh gene from an avian pathogenic Escherichia coli (APEC) strain, APEC13serotype O2:H9, which was cloned in pET101. The 4.2 kb region of cloned DNA coded one protein ofapproximately 140 kDa (r-Tsh). The recombinant plasmid pET101-tsh conferred to E. coli BL21 strain(tsh) the hemagglutination-positive phenotype against chicken erythrocytes. The r-Tsh was purified byNi-NTA column and used to produce antibody anti-Tsh. A 1.6 kb fragment of the tsh sequence was alsoamplified and cloned in pCR4, and a partial sequence showed high homology with other sequenceanalysed. The anti-Tsh reacted with the protein r-Tsh and native Tsh of APEC13, as demonstrated byWestern blot, showing that r-Tsh has conserved epitopes and that its antigenicity was preserved. Theanti-Tsh also inhibited the hemagglutinating activity of strains APEC13 and BL21/pET101-tsh

Molecular identification of enteropathogenic Escherichia coli (EPEC) associated with infant diarrhea in Londrina, Parana, Brazil

In this work, the prevalence of enteropathogenic Escherichia coli (EPEC) in children in Londrina-PR, Brazil, was evaluated by means of digoxigenin-labelled DNA probes which identify the plasmid responsible for EPEC adherence factor (EAF), and virulence genes for EPEC as bundle-forming pilus (bfp) and E. coli attaching-effacing factor (eae). In addition, the isolated strains were serotyped and tested for adherence to HEp-2 cells. From 102 children with diarrhoea, 19 strains hybridized with at least one probe, and eleven of them were identified as typical EPEC because they hybridized with the three probes used, showed a localized adherence (LA) pattern, and presented no genes for enterotoxins (ST and LT) or invasion as detected by PCR. Six of the typical EPEC strains belonged to the classical serotype O119:H6 43(per cent); in four strains O antigens could not be determined using antisera against O1 to O173, they were all ONT:H7 29(per cent); one strain belonged to O111:H6. Three strains were classified as atypical EPEC: O26H-, O111:H9 and O119:HNT. Strains O26H- and O111:H9 hybridized with the eae probe only and showed localized adherence like (LAL) pattern; strain O119:HNT hybridized with the bfp and eae probes, and showed a localized adherence/diffuse adherence (LA/DA) pattern after 6 h. A DA pattern was observed in two strains isolated from children with diarrhoea (ONT:H11 and O142:H34), which hybridized with the eae probe. From 46 controls, five strains hybridized with one or two probes, but none hybridized with all probes or presented the LA pattern. Three strains with the DA pattern hybridized with the eae probe. No EPEC strain belonging to classical EPEC serotypes was isolated from faeces of control children.