An epidemiological study of Serratia marcescens isolates from nosocomial infections by enzyme electrophoresis.

Serratia marcescens isolates from 164 patients with suspected nosocomial infection in several hospitals in the greater Paris region were investigated by analysis of the electrophoretically demonstrable allelic variations of gene loci coding for five esterases and five other enzymes. All the loci were polymorphic and the mean number of alleles per locus was 6.1.

Identification and typing of pyogenic streptococci by enzyme electrophoretic polymorphism.

Polyacrylamide-agarose gel electrophoresis was used to study polymorphism of lactate dehydrogenase (LDH), nucleoside phosphorylase (NSP), phosphoglucose isomerase (PGI), hydroxybutyrate dehydrogenase (HBD), adenylate kinase (ADK) and esterases of 44 strains of Streptococcus pyogenes, 25 group G streptococcal strains, 11 "S. equisimilis" strains, seven S. dysgalactiae strains, four S.

Epidemiology of Staphylococcus aureus in patients with cystic fibrosis.

Seven hundred and thirty-four isolates of Staphylococcus aureus, recovered from the sputum of 238 cystic fibrosis patients in six French hospitals, were characterized by esterase electrophoretic typing, capsular polysaccharide serotyping and phage typing and tested against 14 antibiotics for sensitivity. Thirty-four esterase electrophoretic types were found with a genotypic diversity coefficient of 0.91.

Clonal relationships among Escherichia coli serogroup O78 isolates from human and animal infections.

We investigated the clonal relationships among 63 Escherichia coli strains of antigen serogroup O78 isolated from infections in humans, cattle, sheep, pigs, and chickens. Both septicemic and enterotoxigenic isolates were included in the study. A main group of 55 E.

Genetic heterogeneity of Pseudomonas aeruginosa clinical isolates revealed by esterase electrophoretic polymorphism and restriction fragment length polymorphism of the ribosomal RNA gene region.

The intra-species differentiation of Pseudomonas aeruginosa was analysed by comparing the polymorphism of esterases by conventional polyacrylamide-agarose gel electrophoresis, the physicochemical properties of the variants of the major esterase P3 and the restriction fragment length polymorphism of ribosomal RNA gene regions (ribotyping) to O-serotyping for several panels of strains selected from among a series of 257 clinical isolates and two references strains, (ATCC nos. 10145 and 27853). The electrophoretic variation of four main kinds of esterase (P1-P4) and 11 additional esterases distinguished by their spectra of hydrolytic activity with synthetic substrates and by their sensitivity to di-isopropylfluorophosphate, allowed the discrimination of 67 zymotypes.

Correlation between esterase electrophoretic polymorphism and virulence-associated traits in extra-intestinal invasive strains of Escherichia coli.

The electrophoretic variations of carboxylesterase B and of esterases A, C and I, the presence of mannose resistant haemagglutinin, alpha-haemolysin, cytotoxic necrotizing factor type 1 (CNF1) and certain O antigens were compared in 150 strains of Escherichia coli responsible for extra-intestinal infections. Electrophoretic mobilities of outer membrane proteins (OMP) were also studied for strains belonging to O4, O6, O7, O8 and O75 serogroups. Fast migrating allozymes of carboxylesterase B (pattern B1) were correlated with slow migrating allozymes of esterase C, serogroups O7 and O8, lack of virulence factor, and particular OMP patterns, whereas slow migrating allozymes of carboxylesterase B (pattern B2) were correlated with fast migrating allozymes of esterase C, serogroups O2, O4, O6, O18 and O75, virulence factor production, and distinct OMP patterns.

O, K, and H antigens predict virulence factors, carboxylesterase B pattern, antimicrobial resistance, and host compromise among Escherichia coli strains causing urosepsis.

The O:K:H serotypes of 75 Escherichia coli blood isolates from patients with urosepsis were compared for the presence and expression of determinants for P fimbriae, hemolysin, and aerobactin; antimicrobial resistance; the carboxylesterase B phenotype; and associated compromising host conditions. O groups, K types, and O:K:H serotypes previously associated with urovirulence accounted for 69%, 60%, and 31% of the population, respectively. Chromosomal determinants for P fimbriae, hemolysin, and aerobactin were present in combination more commonly among strains belonging to urovirulence-associated O groups, K types, and O:K:H serotypes.

Genetic structures of the B2 and B1 Escherichia coli strains responsible for extra-intestinal infections.

Escherichia coli strains causing human extra-intestinal infections may be divided into two groups, B1 and B2 according to the electrophoretic patterns of carboxylesterase B. This study compares the restriction fragment length polymorphism (RFLP) of ribosomal DNA (rDNA) for 45 B1 strains and 45 B2 strains to examine the genetic structure of B2 strains and to distinguish them from B1 strains. The isolates were chosen for diversity in their allozymes of esterases, B, A, C and I, their production of virulence factors (alpha-haemolysin, mannose resistant haemagglutinin and cytotoxic necrotizing factor) and certain O antigens, and their pathological and geographical origins.

Isolation and characterization of carboxylesterase E3 from Salmonella enterica.

Three esterases (Est-) hydrolysing alpha-naphthyl acetate: Est-E1, Est-E3 and Est-E4 produced by Salmonella enterica serovar Typhimurium, strain LT2 were separated by DEAE chromatography and gel filtration. Est-E3, the major component of this set of enzymes, clearly differed from the two other esterases by its apparent molecular weight, titration curve, substrate specificity and inactivation. Immunoglobulins raised against Est-E3 completely neutralized the activity of Est-E3 but did not react with Est-E1 or Est-E4; it showed no cross reaction with carboxylesterase B of Escherichia coli or with carboxylesterases from other enterobacteria.

Characterization of Escherichia hermannii by ribosomal DNA restriction fragment length polymorphism.

Ribosomal DNA polymorphism was used to characterize strains of Escherichia hermannii and to differentiate them from E. coli. DNA from 11 E.

Purification and characterization of three carboxylesterases from Enterobacteriaceae.

The carboxylesterases from Proteus vulgaris, Salmonella enterica and Citrobacter amalonaticus were purified 104-, 95- and 120-fold, respectively by chromatography. The enzymes had similar catalytic activities but differed considerably in their inactivation by heat, di-isopropyl fluorophosphate and Cd2+, Zn2+, Hg2+ and Cu2+. Quantitative neutralization of hydrolytic activity with specific immunoglobulins indicated that the three enzymes were antigenically distinct.

Distinctive electrophoretic pattern of esterases produced by Alcaligenes species.

The esterases produced by 34 strains of Alcaligenes faecalis, 16 strains of A. denitrificans subsp. xylosoxydans, 5 strains of A.

Esterase electrophoretic polymorphism of human and animal strains of Clostridium perfringens.

Esterase electrophoretic polymorphism in human and animal strains of Clostridium perfringens was studied by using polyacrylamide-agarose gel electrophoresis. Five types of esterases, designated E-I to E-V and defined by their hydrolytic specificities toward five synthetic substrates, were found in protein extracts of bacteria grown without glucose (glucose-containing media allowed only the expression of esterase E-I). Mobility variants of esterase E-I, which hydrolyzes alpha- and beta-naphthyl acetates and butyrates, were used as a basis for the distribution of strains into 11 zymogroups.

Identification of a clone of Escherichia coli O103:H2 as a potential agent of hemolytic-uremic syndrome in France.

In a French multicenter study, six verocytotoxin-producing Escherichia coli strains were isolated from the stools of 6 of 69 children suffering from hemolytic-uremic syndrome. All strains belonged to serotype O103:H2, a serotype commonly associated with diarrhea in weaned rabbits in France. To determine whether the strains from humans and rabbits were genetically related, they were compared by analyzing their esterase electropherotypes and the restriction fragment length polymorphisms of the ribosomal DNA regions.

Typing of Staphylococcus aureus by pulsed-field gel electrophoresis, zymotyping, capsular typing, and phage typing: resolution of clonal relationships.

Sixty-nine Staphylococcus aureus isolates from two epidemiologically unrelated sources were typed by pulsed-field gel electrophoresis after SmaI digestion of chromosomal DNA (genome typing), and the results were compared with those obtained by other typing methods: phage typing with the international set of phages, capsular serotyping with monoclonal antibodies against capsular polysaccharides type 5 and 8, and zymotyping by polyacrylamide agarose electrophoresis for esterase polymorphism. A good correlation of S. aureus types was found by these four typing methods.

Clonal relationships among bovine pathogenic Escherichia coli producing surface antigen CS31A.

Forty-two Escherichia coli strains producing surface antigen CS31A isolated from bovine infections were characterized with respect to OKH serotypes, outer membrane protein (OMP) electrophoretic patterns, allozymes for esterases A, B, C, I and biotypes. A large majority of the strains could be clustered in a limited number of groups of clonally related strains with diverse O serogroups. CS31A producing Escherichia coli strains thus appear to have a common genetic background and are representative of an important part of bovine pathogenic Escherichia coli.

Molecular epidemiology unravels the complexity of neonatal Escherichia coli acquisition in twins.

Combined analysis of restriction fragment length polymorphism of regions of genes coding for rRNA (ribotyping) and esterase electrophoretic typing was used to document neonatal acquisition of Escherichia coli in twins. Our study shows vertical mother-to-infant transmission of one strain of E. coli to one twin and the development of neonatal septicemia with a distinct nonvirulent carboxylesterase type B1 E.

Enzyme electrophoretic polymorphism differentiates invasive from non-invasive Chlamydia psittaci ruminant isolates.

A group of 24 Chlamydia psittaci strains isolated from ruminants, belonging to serotype 1 and previously classified as invasive in a mouse model of virulence, was compared to a group of 10 non-invasive strains belonging to serotype 2 by using determination of glucose-6-phosphate and L-malate dehydrogenase zymotypes resulting of the infection of cells by these strains. The serotype 1 or invasive isolates represent a homogeneous group by sharing a unique zymotype which was not observed in the non-invasive strains. On the contrary, the serotype 2 or non-invasive isolates constitute a heterogeneous group in generating 2 different zymotypes.

[Characterization of Alcaligenes species using analysis of esterase electrophoretic polymorphism and analysis of antibiotic resistance profiles].

The species of an Alcaligenes bacterial strain may be difficult to determine on the basis of conventional phenotype features. Esterase pattern analysis using acrylamide-agar gel electrophoresis and determination of the antimicrobial resistance profile (agar diffusion method) were performed for A. faecalis (34 strains).

Molecular epidemiological analysis of Pseudomonas aeruginosa strains causing failure of antibiotic therapy in cystic fibrosis patients.

A combination of esterase electrophoretic typing and analysis of the restriction fragment length polymorphism of ribosomal DNA regions (ribotyping) was used to compare 27 Pseudomonas aeruginosa strains isolated before and after two-week courses of anti-pseudomonal treatment in seven cystic fibrosis patients. A total of 12 courses of therapy were studied in which ciprofloxacin, ceftazidime, azlocillin or imipenem were used alone or in combination with tobramycin. Isolates at a count of greater than or equal to 10(6) cfu/ml of sputum were collected when there was evidence of therapeutic failure on the basis of persistence of isolates whether or not they were resistant to the antibiotic used for therapy.

Correlation between DNA polymorphism and enzyme polymorphism argues in favour of the delineation of two species within Providencia alcalifaciens.

Ribosomal DNA (rDNA) polymorphism was compared to enzyme polymorphism and DNA/DNA hybridization data for the intraspecies differentiation of Providencia alcalifaciens. DNA from 27 strains previously classified into two zymotypes A1 and A2 and discriminated by two levels of DNA/DNA hybridization (delta Tm values of 0 to 1 degree C and 6 to 10 degrees C, respectively) were analysed by Southern blotting for rDNA polymorphism. The ribotypes fell into two ribogroups A1 and A2, which correlated with the corresponding zymotypes.

Characterization of highly virulent Escherichia coli strains by ribosomal DNA restriction fragment length polymorphism.

Patterns of ribosomal DNA polymorphism were examined to compare carboxylesterase B type B1 strains and B2 strains of Escherichia coli isolated from extra-intestinal infections. DNA from 14 type B2 strains showing the presence of alpha-haemolysin and mannose-resistant haemagglutinin and lethality to mice and 14 type B1 strains lacking these characteristics, was digested with HindIII, EcoRI, BamHI or BglII restriction enzymes and analysed by Southern blotting. The obtained ribotypes clearly differentiated the B2 strains from the B1 strains.

Association of carboxylesterase B electrophoretic pattern with presence and expression of urovirulence factor determinants and antimicrobial resistance among strains of Escherichia coli that cause urosepsis.

We determined the carboxylesterase B electrophoretic profiles of 74 blood isolates of Escherichia coli from patients with urosepsis. Most strains (64%) exhibited the B2 electrophoretic pattern. P fimbrial and hemolysin genetic determinants were present and expressed significantly more often among strains with the B2 than with the B1 electrophoretic pattern.

Complexity of Pseudomonas aeruginosa infection in cystic fibrosis: combined results from esterase electrophoresis and rDNA restriction fragment length polymorphism analysis.

Esterase electrophoretic typing and restriction fragment length polymorphism of ribosomal DNA regions (ribotyping) were used to differentiate 102 Pseudomonas aeruginosa clinical isolates obtained from chronic lung infection in 23 patients with cystic fibrosis (CF) and two reference strains (including the type strain ATCC 10145). Twenty-five zymotypes were obtained with the former method and 16 ribotypes with the latter. Combination of the two typing systems led to the finding of 30 different types.