Resum

All clinically relevant Enterobacteriaceae strains isolated between 1994 and 1996 without inducible chromosomal b-lactamase, that showed decreased susceptibility to broad-spectrum cephalosporins and/or aztreonam, were selected. The mechanism implicated in the decreased susceptibility was determined by analytical isoelectric focusing, PCR and/or sequenciation. The results obtained showed that the most frequent mechanism implicated in the decreased susceptibility to broad-spectrum cephalosporins and/or aztreonam in E. coli and K. oxytoca was the hyperproduction of the chromosomal b-lactamase, followed by the SHV-1 hyperproduction in E. coli and K. pneumoniae. In our hospital, the incidence of plasmid-mediated extended-spectrum b-lactamases (ESBLs) between 1994 and 1996 was low (0.14%). The b-lactamase that couldn't be identified by the techniques previously described, was characterised by conjugation studies, clonation, and sequenciation experiments, allowing us to describe a new b-lactamase, CTX-M-9, that was the most frequent ESBL detected in our laboratory.

The CTX-M-type b-lactamases has been described in various species of the family Enterobacteriaceae in geographically and temporally widely distant areas, most of them being plasmid-encoded. To know more about the dissemination of these enzymes around the world, we decided to study the environment of blaCTX-M-9. This study suggested us that blaCTX-M-9 is contained in a new complex integron, In60. This integron has the common 5'-CS and 3'-CS conserved sequences of class 1 integron and two gene cassettes encoding for a dihydrofolate reductase (dfrA16) and aminoglycoside-adenylytransferase (aadA2). Downstream of the first sul1 gene is present the orf513. Following this region there is the blaCTX-M-9 and an orf3-like region showing both about 80% identity with blaKLUA-1 and orf3 of K. ascorbata, respectively. Downstream of the orf3-like region, a new insertion sequence designated IS3000, is found. This IS is flanked by two imperfect inverted repeats and its deduced amino acid sequence presents the DD(35)-E motif highly conserved in the transposases. Close to the downstream IS3000 inverted repeat the second 3'-CS, is found. The presence of In60 was evaluated in a total of 37 enterobacteria isolated between 1994 and 1999 carrying a b-lactamase compatible with CTX-M-9. The results obtained showed that 33 strains had the environment compatible with In60 and four strains showed modifications or did not share this environment.

La difusió d'aquesta tesi per mitjà del servei TDX ha estat autoritzada pels titulars dels drets de propietat intel.lectual únicament per a usos privats emmarcats en activitats d'investigació i docència. No s'autoritza la seva reproducció amb finalitats de lucre ni la seva difusió i posada a disposició des d'un lloc aliè al servei TDX. No s'autoritza la presentació del seu contingut en una finestra o marc aliè a TDX (framing).

Aquesta reserva de drets afecta tant al resum de presentació de la tesi com als seus continguts. En la utilització o cita de parts de la tesi és obligat indicar el nom de la persona autora.