BackgroundExtended-spectrum β-lactamases (ESBLs), including the AmpC type, are important mechanisms of resistance among Enterobacteriaeceae. CTX-M type extended-spectrum β- lactamases, of which there are now over 90 variants, are distributed globally, yet appear to vary in regional distribution. AmpC β-lactamases hydrolyze third generation cephalosporins, but are resistant to inhibition by clavulanate or other β-lactamase inhibitors in vitro. Fecal carriage and rates of colonization by bacteria harboring these resistance mechanisms have been reported in patients with community-acquired infections and in healthy members of their households. Expression of these ESBLs compromises the efficacy of current antibacterial therapies, potentially increasing the seriousness of hospital- and community-acquired Escherichia coli (E. coli) infections.To investigate the occurrence of ESBL-producing E. coli in human fecal flora isolated from two pediatric populations residing in the Libyan cities Zleiten and Abou El Khoms. Isolates were further studied to characterize genes encoding β-lactam resistance, and establish genetic relationships.MethodsAntibiotic resistance profiles of phenotypically characterized E. coli isolates recovered from the stools of 243 Libyan children during two surveillance periods in 2001 and 2007 were determined by the disk diffusion method. ESBL-screening was performed using the cephalosporin/clavulanate double synergy disc method, and the AmpC-phenotype was confirmed by the aminophenyl-boronic acid test. ESBL genes were molecularly characterized. Phylogenetic group and multilocus sequence typing (MLST) were determined for ESBL-producing isolates and PFGE was performed to compare banding profiles of some dominant strains.ResultsESBLs were identified in 13.4% (18/134) of E. coli isolates, and nine isolates (6.7%) demonstrated AmpC activity; all 18 isolates contained a CTX-M gene. Three CTX-M gene families (CTX-M-1, n = 9; CTX-M-15, n = 8 and CTX-M-3, n = 1) were distributed in diverse E. coli backgrounds (phylogenetic group D, 39%; B2, 28%; B1, 22% and A, 11%). MLST analysis revealed 14 sequence type (ST) with six new sequence types. The gene encoding the CMY-2 enzyme was detected in five AmpC-positive E. coli.ConclusionsThese results identified heterogeneous clones of CTX-M-producing E. coli in the fecal isolates, indicating that the intestinal tract acts as a reservoir for ESBL-producing organisms, and a trafficker of antibiotic resistance genes.
Abstract. Diarrheagenic Escherichia coli (DEC) are important enteric pathogens that cause a wide variety of gastrointestinal diseases, particularly in children. Escherichia coli isolates cultured from 243 diarrheal stool samples obtained from Libyan children and 50 water samples were screened by polymerase chain reaction (PCR) for genes characteristic of enteroaggregative E. coli (EAEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enterohemorrhagic E. coli (EHEC), and enteroinvasive E. coli (EIEC). The DEC were detected in 21 (8.6%) children with diarrhea; 10 (4.1%) cases were identified as EAEC, 3 (1.2%) as EPEC, and 8 (3.3%) were ETEC; EHEC, and EIEC were not detected. All DEC were grouped phylogenetically by PCR with the majority ( 70%) identified as phylogenetic groups A and B1. The EAEC isolates were also tested for eight genes associated with virulence using PCR. Multi-virulence ( 3 virulence factors) was found in 50% of EAEC isolates. Isolated EAEC possessed different virulence traits and belonged to different phylogenetic groups indicating their heterogeneity.
Resumo -Bactérias fixadoras de nitrogênio podem contribuir para o crescimento vegetal pela produção de auxinas. Os objetivos deste trabalho foram quantificar a produção de hormônios de crescimento por estirpes de bactérias fixadoras de nitrogênio e avaliar o efeito da inoculação destas estirpes em plântulas de milho e trigo. Todas as estirpes avaliadas começaram a produzir indóis no final da fase logarítmica. Houve efeito da adição de diferentes concentrações de triptofano ao meio de cultivo no aumento da produção de indóis até o nível de 200 µM. Nas estirpes de Azospirillum, as formas de N: NH 4 Cl, (NH 4 ) 2 SO 4 , NH 4 H 2 PO 4 e NH 4 NO 3 estimularam a produção de indóis em níveis baixos. A adição de KNO 3 , NaNO 3 e KNO 2 inibiu a produção de indóis em todas as bactérias testadas e no crescimento de células de Azospirillum. Os efeitos da inoculação foram também comparados com concentrações crescentes do ácido 3-indolacético, KNO 3 e triptofano. Em condições axênicas, a elevada produção de indóis reduz o comprimento das raízes e colmos na presença de triptofano, especialmente quando submetidas à inoculação de Azospirillum.Termos para indexação: bactérias diazotróficas, ácido 3-indolacético, fitormônios, fixação biológica de nitrogê-nio. Effect of inoculation with Azospirillum and Herbaspirillum on production of indolic compounds and growth of wheat and rice seedlingsAbstract -Nitrogen fixing bacteria can contribute to plant growth by the production of auxin. The objectives of this work were to quantify the production of plant growth hormones by different nitrogen-fixing bacteria and to evaluate the effect of this inoculation on growth of wheat and rice seedlings. All strains tested produced indoles during the late logarithmic phase. Increasing the amount of tryptophan in the medium enhanced the production of indoles by the bacteria up to the level of 200 µM. For the Azospirillum strains studied, the nitrogen sources: NH 4 Cl, (NH 4 ) 2 SO 4 , NH 4 H 2 PO 4 and NH 4 NO 3 stimulated indoles production in low levels. The addition of KNO 3 , NaNO 3 and KNO 2 inhibited the production of indole compounds for all bacteria tested as well as the Azospirillum growth. The effects of inoculation were also compared to those caused by increased concentrations of IAA, KNO 3 and tryptophan alone. In vitro conditions, the high amounts of indoles produced by the bacteria reduce the length of roots and stems in the presence of tryptophan, especially in the case of Azospirillum strains.Index terms: diazotrophic bacteria, indole-3-acetic acid, phytohormones, biological nitrogen fixation. IntroduçãoDurante a última década, várias bactérias capazes de reduzir N 2 foram descritas, sendo o gênero Azospirillum o mais estudado. Atualmente, este grupo engloba seis espécies diazotróficas, como o A. brasilense e o A. lipoferum. Além disso, algumas espécies foram descritas como endófitas porque não são capazes de sobreviver por longos períodos no solo (Baldani et al., 1997).A primeira espécie endófita a ser identificada foi o Herbaspirill...
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