Occurrence and characterization of class 1 integrons in Escherichia coli from healthy individuals and those with urinary infection.

PURPOSE: Class 1 integrons are among the main vehicles that facilitate the spread of antibiotic-resistance genes, with serious public health consequences. The aim of this cross-sectional study was to investigate the presence of class 1 integrons and to characterize their variable regions, as well as the antimicrobial resistance profiles and phylogenetic groups of a collection of Escherichia coli isolates recovered from healthy subjects (n=42) and those with urinary infection (n=40). METHODOLOGY: The methods used included PCR, sequencing and antimicrobial susceptibility testing. RESULTS: PCR screening for the integrase gene (intI1) revealed a higher incidence of class 1 integrons in uropathogenic E. coli (65 %, UPEC) than in commensal isolates (11.9 %). Eight of 31 intI1-positive isolates, all of them UPEC, harboured empty integrons. The variable regions of the other 23 contained gene cassettes encoding resistance to ß-lactams (blaOXA-1), aminoglycosides (aadA1 and aadA5), trimethoprim (dfrA1 and dfrA17) and an ORF. To our knowledge this is the first report of an ORF identified as a putative phage tail protein associated with a class 1 integron. The aadA1 and dfrA17-addA5 arrays prevailed in commensal E. coli and UPEC, respectively. UPEC isolates were highly resistant to the antimicrobials tested, in contrast to commensal isolates. The E. coli isolates carrying gene cassettes associated with class 1 integrons were found to be unrelated to any phylogroup or multiresistance. CONCLUSION: Co-resistance to clinically relevant fluoroquinolone and trimethoprim-sulfamethazole in all UPEC isolates is a cause for concern. These results expand the current knowledge of gene cassettes in both commensal and pathogenic E. coli.

Identification of new bacteria harboring qnrS and aac(6')-Ib/cr and mutations possibly involved in fluoroquinolone resistance in raw sewage and activated sludge samples from a full-scale WWTP.

Wastewater treatment plants (WWTPs) harbor bacteria and antimicrobial resistance genes, favoring gene exchange events and resistance dissemination. Here, a culture-based and metagenomic survey of qnrA, qnrB, qnrS, and aac(6')-Ib genes from raw sewage (RS) and activated sludge (AS) of a full-scale municipal WWTP was performed. A total of 96 bacterial isolates were recovered from nalidixic acid-enrichment cultures. Bacteria harboring the aac(6')-Ib gene predominated in RS, whereas qnrS-positive isolates were specific to AS. Novel qnrS- and aac(6')-Ib-cr positive species were identified: Morganella morganii, Providencia rettgeri, and Pseudomonas guangdongensis (qnrS), and Alcaligenes faecalis and P. rettgeri (aac(6')-Ib-cr). Analysis of qnrS and aac(6')-Ib sequences from isolates and clone libraries suggested that the diversity of qnrS is wider than that of aac(6')-Ib. A large number of amino acid mutations were observed in the QnrS and AAC(6')-Ib proteins at previously undetected positions, whose structural implications are not clear. An accumulation of mutations at the C72, Q73, L74, A75 and M76 positions of QnrS, and D181 of AAC(6')-Ib might be important for resistance. These findings add significant information on bacteria harboring qnrS and aac(6')-Ib genes, and the presence of novel mutations that may eventually emerge in clinical isolates.

The Microbiota and Abundance of the Class 1 Integron-Integrase Gene in Tropical Sewage Treatment Plant Influent and Activated Sludge.

Bacteria are assumed to efficiently remove organic pollutants from sewage in sewage treatment plants, where antibiotic-resistance genes can move between species via mobile genetic elements known as integrons. Nevertheless, few studies have addressed bacterial diversity and class 1 integron abundance in tropical sewage. Here, we describe the extant microbiota, using V6 tag sequencing, and quantify the class 1 integron-integrase gene (intI1) in raw sewage (RS) and activated sludge (AS). The analysis of 1,174,486 quality-filtered reads obtained from RS and AS samples revealed complex and distinct bacterial diversity in these samples. The RS sample, with 3,074 operational taxonomic units, exhibited the highest alpha-diversity indices. Among the 25 phyla, Proteobacteria, Bacteroidetes and Firmicutes represented 85% (AS) and 92% (RS) of all reads. Increased relative abundance of Micrococcales, Myxococcales, and Sphingobacteriales and reduced pathogen abundance were noted in AS. At the genus level, differences were observed for the dominant genera Simplicispira and Diaphorobacter (AS) as well as for Enhydrobacter (RS). The activated sludge process decreased (55%) the amount of bacteria harboring the intI1 gene in the RS sample. Altogether, our results emphasize the importance of biological treatment for diminishing pathogenic bacteria and those bearing the intI1 gene that arrive at a sewage treatment plant.