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Biochemical characterization of a cefotaximehydrolysing β-lactamase encoded by a conjugative plasmid
Abstract
During the nosocomials infections occurring in the intensive care unit of the military hospital of Tunis in 2005, the Enterobacter cloacae BW 1150 strain was isolated from a stool culture. This strain was found to have a high level resistance to broad-spectrum -lactams. Resistance profile against the
various families of antibiotics was determined using the disc diffusion test. The minimal inhibitory concentrations values showed that this strain was resistant to the -lactams such as ampicillin and the extended spectrum cephalosporins (cefotaxime, ceftriaxon and cefpirome). Analysis of this strain by the disk diffusion test revealed synergies between amoxicillin-clavulanate (AMX-CA) and ceftriaxon, ceftazidime and cefotaxime. Cell sonicate of this isolate is very active against cefotaxime and showed a
specific activity (AS) of 7.54 U/mg for the same antibiotic. This activity was inhibited by the sulbactam and the clavulanic acid. Isoelectrofocusing methods revealed that the crude extract of the E. cloacae BW 1150 strain showed 1 - lactamase activity with an isoelectric piont (pI) of about 8. This activity was transferred by conjugation and was highly expressed in the transconjugant.
various families of antibiotics was determined using the disc diffusion test. The minimal inhibitory concentrations values showed that this strain was resistant to the -lactams such as ampicillin and the extended spectrum cephalosporins (cefotaxime, ceftriaxon and cefpirome). Analysis of this strain by the disk diffusion test revealed synergies between amoxicillin-clavulanate (AMX-CA) and ceftriaxon, ceftazidime and cefotaxime. Cell sonicate of this isolate is very active against cefotaxime and showed a
specific activity (AS) of 7.54 U/mg for the same antibiotic. This activity was inhibited by the sulbactam and the clavulanic acid. Isoelectrofocusing methods revealed that the crude extract of the E. cloacae BW 1150 strain showed 1 - lactamase activity with an isoelectric piont (pI) of about 8. This activity was transferred by conjugation and was highly expressed in the transconjugant.
