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Background: ESBL-producing bacteria are a clinical problem in the management of diseases caused by these pathogens. Worldwide, systemic infections with BL enzymes are evolving by mutations from classical bla genes in an intensified manner and they continue to be transferred across species. Results: E.cloacae BF1417 isolate and its transconjugants gave positive results with the DDST, suggesting the presence of ESBL. Sequence analysis revealed a blaSHV-ESBL-type gene that differs from the gene encoding SHV-1 by five point mutations resulting in three amino acid substitutions in the coding region: C123R, I282T and L286P. This novel SHV-type enzyme was designated SHV-128. The conjugation tests and plasmid characterization showed that the blaSHV-128 is located on a conjugative plasmid IncFII type. Expression studies demonstrated that the above mutations participated in drug resistance, hydrolysis of extended spectrum β-lactam and the change of the isoelectric point of the protein. Conclusion: These findings underscore the diversity by which antibiotic resistance can arise and the evolutionary potential of the clinically important ESBL enzymes. In addition, this study highlights the need for systematic surveillance of ESBL-mediated resistance as well as in clinical areas and communities. [ABSTRACT FROM AUTHOR]

Klebsiella pneumoniae ML2508 was isolated from a patient at the surgery unit of the Military Hospital (Hôpital Militaire de Tunis), Tunisia. It was identified as a producer of extended-spectrum β-lactamases (ESBLs) by the double-disk synergy test. The β-lactamases produced by the strain were characterized by isoelectric focusing, determination of the specific activities against penicillins and cephalosporins, determination of the inhibitory concentration required to inhibit 50% of enzyme activity (IC), and the inhibition effect of EDTA on putative metallo-β-lactamases. The crude extract of K. pneumoniae ML2508 contains five different β-lactamases with pI 5.5, 7.3, 7.6, 8.1, and 8.6. Only the β-lactamase with pI 8.1 was transferred by transformation and conjugation experiments. Molecular characterization of these genes was performed by PCR and sequencing. The four chromosomal β-lactamases are TEM (pI 5.5), 2 SHV (pI 7.3 and 7.6), and CTX-M-28 (pI 8.6). The β-lactamase with pI 8.1 was encoded by bla gene located on a 50-kb highly conjugative plasmid. The study of the genetic context of bla was realized by PCR-mapping and DNA sequencing. A novel variant of tnp designated IS Ecp1C was detected upstream of the gene. [ABSTRACT FROM AUTHOR]

clinical isolate of Enterobacter cloacae BF280 showing a multidrug resistance phenotype including resistance to β-lactams and quinolones is the subject of this study. This strain was isolated from a patient at the intensive care unit of a military hospital (hôpital Militaire de Tunis). By polymerase chain reaction amplification and sequencing, this isolate was found to produce quinolone resistance determinant QnrS1. In addition, it was found to produce CTX-M-28, OXA-1 and TEM-1. The qnrS, bla, bla and bla genes were located on a highly conjugative plasmid. [ABSTRACT FROM AUTHOR]

Klebsiella pneumoniae ML0306 was isolated from a patient in the intensive care unit of the Military hospital of Tunis in Tunisia. The strain was resistant to β-lactams, aminoglycosides, quinolones, phenicols and tetracyclines. Crude extract from the isolate showed a broad-substrate profile by hydrolyzing benzylpenicillin, ampicillin, cloxacilline, cephalothin, cefotaxime and ceftriaxone but not ceftazidime, cefoxitin, cefpirome, imipinem and aztreonam. The isolate was identified as a producer of pI 7.6 β-lactamase. Sequencing of the β-lactamase gene showed that it coded for a penicillinase SHV-11. This is the first description of this enzyme in Tunisia. The gene encoding this enzyme was located on the chromosome and a 50 kb conjugative plasmid. The overproduction of SHV-11 was involved in the wading of the hydrolytic spectrum of the strain. [ABSTRACT FROM AUTHOR]