ABSTRACT
The increasing use of broader-spectrum cephalosporins in the first half of the 1990s has become one of the major factors responsible for the high rate of selection of extended-spectrum beta-lactamase (ESBL)-producing microorganisms in Polish hospitals. Thirty-five isolates of seven different species of the family Enterobacteriaceae were identified as ESBL producers, over a 4 month period, in one of Warsaw's hospitals between the end of 1996 and the beginning of 1997. Sixteen per cent of all Klebsiella pneumoniae isolates, 16% of Citrobacter freundii isolates and 32% of Serratia marcescens isolates collected by the hospital microbiology laboratory at that time were expressing these enzymes. The majority of these (27 isolates) were found to express CTX-M-type ESBLs (pI 8.4). This outbreak was due to both plasmid dissemination among unrelated strains and clonal spread of some strains in several wards of the hospital. The remaining isolates produced ESBLs (pI 8.2) belonging to the SHV family of beta-lactamases and demonstrated a high degree of genetic diversity.
Subject(s)
Enterobacteriaceae/enzymology , beta-Lactamases/biosynthesis , Bacterial Typing Techniques , Conjugation, Genetic , DNA Fingerprinting , Enterobacteriaceae/drug effects , Enterobacteriaceae/genetics , Isoelectric Focusing , Microbial Sensitivity Tests , Plasmids , Polymerase Chain ReactionABSTRACT
Twelve SHV-type extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli lac mutant isolates were recovered in October 1997 from 11 patients of the neonatal ward in a Warsaw hospital. The outbreak was clonal; however, some of the isolates expressed a much higher level of resistance to several beta-lactam antibiotics, including expanded-spectrum cephalosporins. This phenotype has been attributed to beta-lactamase hyperproduction correlating with the multiplication of ESBL gene copies, as was demonstrated for representative isolates.
Subject(s)
Cephalosporin Resistance/genetics , Cephalosporins/pharmacology , Cross Infection/microbiology , Disease Outbreaks , Escherichia coli Infections/epidemiology , Escherichia coli/drug effects , beta-Lactamases/biosynthesis , Cross Infection/epidemiology , Escherichia coli/enzymology , Escherichia coli/genetics , Escherichia coli Infections/microbiology , Gene Dosage , Humans , Microbial Sensitivity Tests , Phenotype , Poland/epidemiology , beta-Lactamases/geneticsABSTRACT
A group of cefotaxime-resistant Citrobacter freundii and Escherichia coli isolates were collected by a clinical laboratory in a hospital in Warsaw, Poland, in July 1996. Detailed analysis has shown that all of these produced a beta-lactamase (pI, 8.4) belonging to the CTX-M family, one of the minor extended-spectrum beta-lactamase families with a strong cefotaxime-hydrolyzing activity. Sequencing has revealed that C. freundii isolates produced a new CTX-M-3 enzyme which is very closely related to the CTX-M-1/MEN-1 beta-lactamase, sporadically identified in Europe over a period of 6 years. Amino acid sequences of these two beta-lactamases differ at four positions: Val77Ala, Asp114Asn, Ser140Ala, and Asn288Asp (the first amino acid of each pair refers to CTX-M-1/MEN-1 and second refers to CTX-M-3). The partial sequence of the E. coli CTX-M gene was identical to the corresponding region of bla(CTX-M-3), but a transconjugant of the E. coli isolate expressed higher levels of resistance to beta-lactams than did C. freundii transconjugants. These resistance differences correlated with differences in plasmid DNA restriction patterns. Our results suggest that CTX-M genes have been spread among different species of the family Enterobacteriaceae in the hospital and that the CTX-M-3-expressing C. freundii strain causing routine urinary tract infections has been maintained for a relatively long time in the hospital environment.
