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PubMed2005

Plasmid-mediated, inducible AmpC beta-lactamase (DHA-1)-producing Enterobacteriaceae at a Korean hospital: wide dissemination in Klebsiella pneumoniae and Klebsiella oxytoca and emergence in Proteus mirabilis.

The aim of the study was to investigate the phenotypic and genetic characteristics of recently emerging cefoxitin-resistant and induction-positive isolates of Escherichia coli, Klebsiella species, and Proteus mirabilis. Strains of Enterobacteriaceae were isolated at a Korean tertiary care hospital between June and December 2002. Induction was tested using cefoxitin and aztreonam disks, the blaDHA allele was detected by PCR, and pulsed-field gel electrophoresis (PFGE) patterns were also analyzed. Among the cefoxitin-resistant isolates, 2.7% of E. coli, 21.1% of Klebsiella pneumoniae, 32.0% of Klebsiella oxytoca, and 8.3% of P. mirabilis isolates showed induction, and were blaDHA-1 allele positive. To the best of our knowledge, this is the first report of blaDHA-1 in P. mirabilis. The MICs of ceftazidime, cefotaxime, and aztreonam increased significantly by higher inoculum, suggesting that their clinical usefulness is limited. Presence of multiple PFGE patterns and identical patterns in some isolates suggest that the widely disseminated blaDHA-1 in Klebsiella species was because of both horizontal and clonal spread.

Anti-Bacterial Agents↗
PubMed1978

[Klebsiellae isolated from patients and healthy subjects. III. Biochemical types of klebsiellae].

By the difference in fermentation of glucose, lactose, saccharose, inosite, adonite, behaviour in IMViC tests, relation to urine and hydrogen sulfide formation there were revealed 10 biochemical types among 120 strains of klebsiellae. Klebsiellae of biotypes 1 and 2 were the most frequently isolated from the patients; biotypes 3--10 were less frequent. Klebsiella of biotypes 1 and 2 were revealed in healthy persons; biotypes 3--10 were never found. Biochemical reactions on whose basis klebsiellae were referred to the definite biochemical type proved to be stable.

Digestive System↗
PubMed2004

O antigen seroepidemiology of Klebsiella clinical isolates and implications for immunoprophylaxis of Klebsiella infections.

Prevention of Klebsiella infections by passive immunotherapy has received more attention during the last decade. Both K antigen-and O antigen-specific antisera and monoclonal antibodies (mAbs) have been studied with respect to phagocytosis-enhancing and in vivo protective capacities. Our own work has focussed on the generation of O serogroup-specific rabbit antisera and O antigen specific murine antibodies. O-specific rabbit sera were absorbed extensively with heterologous O antigen strains in order to obtain highly specific typing reagents. Using these for typing a collection of 378 clinical strains, we found that 82% of them belonged to one of the four serogroups O1, O2ab, O3 and O5. Phagocytosis experiments using antisera and mAbs showed that O antigen specific antibodies were able to opsonize non-encapsulated strains, while fully encapsulated bacteria were rather resistant against the opsonizing effect. Nevertheless, in vivo experiments demonstrated a prophylactic effect on both Klebsiella septicemia and pneumonia in a mouse model of lethal infection. Given the limited number of O serogroups, O antigen-specific antibodies may be suited to supplement K antigen-specific hyperimmune globulins for passive immunoprophylaxis of Klebsiella infections.

Animals↗
PubMed1998

[Hemolytic activity of Klebsiella pneumoniae and Klebsiella oxytoca].

We demonstrated that Klebsiella pneumoniae and Klebsiella oxytoca possess a selective haemolytic activity on rabbit erythrocytes. Thirty one Klebsiella strains (18 strains of K. pneumoniae and 13 strains of K. oxytoca) were isolated from hospitalized patients. The liquid (Trypcase-soy broth--TSB) and solid (Trypcase-soy agar--TSA) medium, containing the red cells were used for the tests. All the screened strains showed a haemolytic effect on rabbit erythrocytes, provided that the supernatants of the cultures were preincubated with beta-mercaptoethanol or calcium chloride. There was no human and sheep erythrocyte lysis.

Animals↗
PubMed2001

A monoclonal antibody with specificity for the genus Klebsiella binds to a common epitope located in the core region of Klebsiella lipopolysaccharide.

A mouse monoclonal antibody (mAb) which has been obtained after immunization of mice with heat-killed Klebsiella pneumoniae strain R20/O1(-) followed by standard plasmacytoma cell fusion protocols was investigated for its ability to identify various species of the genus Klebsiella. Based on the published observation that the antibody binds to an epitope located in the core region of lipopolysaccharide (LPS) of strain R20/O1(-), we tested whether this epitope is shared and exposed by other species of the genus Klebsiella. The antibody was able to bind to LPS of clinical isolates of K. pneumoniae (n = 77), K. oxytoca (n = 50), K. terrigena (n = 49) and K. planticola (n = 50) in 93%, 98%, 96% and 100%, respectively, but did not bind to LPS of other Gram-negative genera (n = 159) as tested by Western blots and dot blots using proteinase K-digested whole cell lysates as antigens. Western blot analyses indicated that the antibody bound only to those LPS molecules which did not carry an O-antigen and that the antibody is thus different from those already published.

Animals↗
PubMed1984

[Protective activity of a cell-free Klebsiella vaccine in relation to different Klebsiella pneumoniae serovars].

The capacity of dried Klebsiella cell-free vaccine, obtained from strain No. 204 by the disintegration of microbial cells with hydroxylamine, for protecting mice from Klebsiella septic infection caused by the homologous serovar and 9 heterologous serovars of K. pneumoniae was studied. The newly developed preparation was found capable of stimulating immunity not only to the homologous K. pneumoniae serovar, but also to other K. pneumoniae heterologous serovars: K1, K9, K11, K16, K20, K61. The protective capacity of the preparation with respect to these serovars was not inferior to that of the vaccines prepared by the same method from the corresponding homologous strains. The capacity of the vaccine to protect mice from Klebsiella sepsis was manifested irrespective of the virulence of the strains used for challenge.

Animals↗
PubMed1980

Studies concerning the sensitivity of Klebsiella strains to a set of Klebsiella bacteriocins.

Optimal crude preparations of Klebsiella bacteriocins were obtained bu U.V. irradiation of bacteriocin producer strains. Of 308 clinical isolates of Klebsiella 88.9% could be typed on the basis of their sensitivity to different Klebsiella bacteriocins. The method affords further development of bacteriocin typing as a suitable procedure for hospital epidemiological investigation.

Bacteriocins↗
PubMed1989

Clonal analysis of the lac operons from Klebsiella M5al and the Lac plasmid (pRE2) from Klebsiella V9A.

The chromosomal lac region of the coliform bacterium Klebsiella M5al was cloned into the multicopy plasmid pBR322 to give pHE7 and pHE8. pHE8 contains 12.6 kb of M5al DNA, including its complete lac operon, and pHE7 contains 2.5 kb of M5al DNA and includes the complete lac Y gene and a small segment of lacZ. The M5al operon has the same gene order as the Escherichia coli lac operon. The lac genes of the Lac plasmid of Klebsiella V9A were cloned into pBR322 to give pHE1 and pHE2, of approximately 39 and 43 kb. Both plasmids were unstable in an E. coli RecA-strain, in contrast to the stability of pHE8. Polyacrylamide gel electrophoresis tests suggested that the M5al beta-galactosidase monomer is about 5% longer, i.e. has about 50 more amino acids, than that of the E. coli Z gene. Tests made on the enzymes coded by the lac operons of M5al, another Klebsiella strain (V9A) and its resident Lac plasmid, and several Lac+ Enterobacteria, led to the conclusion that only Escherichia coli among the Enterobacteria contains an active lacA gene.

Acetyltransferases↗
PubMed1986

Ankylosing spondylitis, HLA-B27, and Klebsiella: a study of lymphocyte reactivity of anti-Klebsiella sera.

Twenty three anti-Klebsiella antisera were tested for their cytotoxic activity and four for their binding capacity for peripheral blood lymphocytes (PBL) from patients with HLA-B27 positive ankylosing spondylitis (AS+B27+) and from B27 positive (AS-B27+) and B27 negative (AS-B27-) healthy individuals. None of the antisera showed specific activity against PBL from any particular group. The antisera tested included two anti-Klebsiella K43 sera provided by an Australian group, who have reported them to be specifically cytotoxic for AS+B27+ PBL, four antisera raised against a Klebsiella K43 strain provided by this group, and an antiserum from another group, who have reported it as having increased binding capacity for AS+B27+ and AS-B27+ PBL compared with AS-B27- PBL. The results of other workers who have attempted to reproduce the results of either group are reviewed and the possible reasons for the repeated failure to confirm the reported findings are discussed.

Animals↗
PubMed2003

Characterisation, biotyping, antibiogram and klebocin typing of Klebsiella with special reference to Klebsiella oxytoca.

400 strains of Klebsiellae identified by culture characteristics and biochemical reactions were subjected to biotyping, antibiogram and klebocin typing. Based on indole production, pectin and gelatin liquefaction 16.0% of all the isolates were Klebsiella oxytoca. Maximum sensitivity was shown to Amikacin (72%) and maximum resistance to Ampicillin (87.5%). Klebocin typability was 73.5%. So by combining biotyping, antibiogram and Klebocin typing, Klebsiella could be differentiated better than based on any single marker.

Anti-Bacterial Agents↗
PubMed2000

Epidemiology of quinolone resistance of Klebsiella pneumoniae and Klebsiella oxytoca in Europe.

The epidemiology of quinolone resistance and the concomitant resistance to other antibiotic classes was investigated in 445 Klebsiella pneumoniae and 238 Klebsiella oxytoca isolates. Decreased susceptibility to ciprofloxacin was found in 7.2% and 3.4% of these two species, respectively. Ciprofloxacin resistance was significantly linked to ceftazidime resistance, the hallmark of extended-spectrum beta-lactamase production, as well as to resistance to all antibiotic classes tested. Using automated ribotyping, seven intrahospital- and interhospital-transmitted clones of ciprofloxacin-resistant isolates were found. The newer fluoroquinolones sitafloxacin and clinafloxacin may become increasingly valuable, since they proved to be active also against ciprofloxacin-resistant isolates.

Anti-Bacterial Agents↗
PubMed1993

Plasmid DNA analysis, biotyping, and antimicrobic susceptibility as subtyping tests for Klebsiella pneumoniae and Klebsiella oxytoca.

We compared plasmid DNA analysis, biotyping by Vitek, and disk diffusion antimicrobic susceptibility as subtyping tests of Klebsiella pneumoniae and Klebsiella oxytoca. The 92 tested isolates were from alternate, culture-positive patients over 6 months. No outbreak or cluster of infections was recognized during this interval. Plasmid DNA was detected in 85% of the isolates. Each isolate except one had a reproducible absence of plasmid DNA or a reproducible plasmid DNA profile on repetitive testing. Restriction endonuclease enzyme analysis of plasmid DNA was necessary to distinguish differences among some isolates that had only large plasmids. Isolates with only large plasmids represented 18% of the collection. Of the 78 isolates with plasmid DNA, all but two were considered different from one another by plasmid DNA analysis. Biotyping and antimicrobic susceptibility testing were not highly reproducible. In addition, biotyping did not demonstrate a sufficient variety of patterns among the isolates for subtyping purposes. We conclude that plasmid DNA analysis is very useful as a subtyping test for isolates of K. pneumoniae and K. oxytoca. Neither biotyping nor antimicrobial susceptibility as performed in our laboratory had sufficient discriminatory power and reproducibility for subtyping these organisms.

Anti-Bacterial Agents↗
PubMed2000

Screening and confirmatory testing for extended spectrum beta-lactamases (ESBL) in Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca clinical isolates.

Escherichia coli and Klebsiella spp. were screened for ESBL based on routine susceptibility testing results. Isolates with intermediate or resistant susceptibilities for extended spectrum cephalosporins or aztreonam were reported as probable ESBL producers. By using the NCCLS proposed ESBL confirmatory method, we tested 61 screen-positive isolates from 42 patients, 30 randomly selected susceptible isolates, and 12 isolates with previously characterized beta-lactamases. Ceftazidime contributed to 97% of screen-positive isolates, whereas aztreonam added a single patient isolate. An ESBL was confirmed in 86% of K. pneumoniae, 100% of K. oxytoca, and 20% of E. coli screen-positive single patient isolates. None of the susceptible isolates were shown to produce ESBL. Based on these findings a comment regarding the presence of ESBL seems sufficient for Klebsiella spp. but confirmatory testing is indicated for E. coli. 0.25 microg/mL was used to indicate the presence of ESBL, the specificity of the assay increased to 100%. The NCCLS ESBL phenotypic confirmatory method was reproducible and accurate enough to be used in the clinical laboratory.

Anti-Bacterial Agents↗
PubMed1977

Isolation of Klebsiella ozaenae and Klebsiella rhinoscleromatis in a general hospital.

Klebsiella ozaenae and Klebsiella rhinoscleromatis are uncommon organisms usually associated with granulomatous or necrotizing disease of the upper airways. Seven patients are described, non of whom had characteristic clinical manifestations of infection with these species. The organisms were recovered from mixed wound infections or sputum of five patients, and from the blood of three patients. Antibiotic sensitivity patterns were unusual and included susceptibility to both ampicillin and carbenicillin.

Adolescent↗
PubMed2001

CMY-2-producing Salmonella enterica, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis and Escherichia coli strains isolated in Spain (October 1999-December 2000).

CMY-2 plasmid-mediated AmpC beta-lactamase (CMY-2) was detected in 21 isolates from two hospitals located in different geographical regions of Spain between October 1999 and December 2000. The isolates comprised two Salmonella enterica serovars (Mikawasima and Montevideo), 16 Escherichia coli, one Klebsiella pneumoniae, one Klebsiella oxytoca and one Proteus mirabilis. In addition to the expected resistance to beta-lactams, including extended-spectrum cephalosporins and cefoxitin, all isolates showed a broad spectrum of associated resistance. All were resistant to sulfamethoxazole, chloramphenicol, tetracycline and streptomycin, and all but two were also resistant to gentamicin. Five isolates were studied in detail and all transferred CMY-2 and other resistance determinants by conjugation. Genomic DNA restriction pattern analysis of the E. coli isolates excluded the dissemination of a single clone. To the best of our knowledge this is the first time that CMY-2 has been detected in P. mirabilis, K. oxytoca and S. enterica serovars Mikawasima and Montevideo. It is also the first time that CMY-2 has been described in Spain.

Anti-Bacterial Agents↗
PubMed1997

Identification of clinical isolates of indole-positive Klebsiella spp., including Klebsiella planticola, and a genetic and molecular analysis of their beta-lactamases.

In a collection of 43 indole-positive Klebsiella clinical isolates, which were initially identified as Klebsiella oxytoca, there were 18 isolates which exhibited a pattern characteristic of extended-spectrum beta-lactamase (ESBL) resistance. This study aimed to confirm their identity by biochemical tests and by PCR and to determine the genetic basis for their resistance to the beta-lactams and broad-spectrum cephalosporins. Chromosomal beta-lactamase genes were analyzed by PCR, and plasmid-mediated beta-lactamase genes were analyzed by conjugation and transformation. There were 39 isolates which grew on melezitose but failed to grow on 3-hydroxybutyrate, confirming them as K. oxytoca. PCR analysis of their beta-lactamase genes divided these isolates into two groups, the bla(OXY-1) group and the bla(OXY-2) group. Each group had beta-lactamases with different isoelectric points; the bla(OXY-1) group had beta-lactamases with isoelectric points at 7.2, 7.8, 8.2, and 8.8, and the more common bla(OXY-2) group had beta-lactamases with pIs at 5.2, 5.4 (TEM-1), 5.7, 5.9, 6.4, and 6.8. A pI of 5.2 was the most frequently detected and accounted for 59% of all the bla(OXY-2) beta-lactamases. Hyperproduction of clavulanate-inhibited chromosomal beta-lactamases was detected in 17 K. oxytoca isolates, resulting in an ESBL phenotype. K. oxytoca isolates having a plasmid-mediated genetic basis for their ESBL phenotype were not found, confirming that, in K. oxytoca, plasmids are rarely involved in conferring resistance to the newer cephalosporins. Four isolates proved to be isolates of K. planticola in which the beta-lactamase genes failed to react with the primers used in the PCR. One K. planticola isolate contained a transferable plasmid harboring the SHV-5 beta-lactamase gene and showed an ESBL phenotype, while the other non-ESBL K. planticola isolates contained chromosomal beta-lactamases with isoelectric points at 7.2, 7.7, and 7.9 plus 7.2.

3-Hydroxybutyric Acid↗
PubMed1979

Taxonomic position of Klebsiella atlantae and Klebsiella edwardsii.

The taxonomic position of two species of the genus Klebsiella (K. atlantae and K. edwardsii) is being introduced. 211 strains of different origin were studied: 80 strains of K. pneumoniae, 46 strains of K. oxytoca, 50 strains of K. atlantae and 35 strains of K. edwardsii. Sixty biochemical characteristics were determined 40 of these pertaining to carbohydrate metabolism. It was discussed whether both are to be considered species or biotypes of another Klebsiella species, however, by determining citrate as carbon source, by MR test and by tests on malonate, gluconate, methyl-xyloside, 1 (--) sorbose, inulin, amylose, methyl-d-mannoside, glycogen, melezitose, VP test, amygdalin, d-tartrate and gas from glucose, we arrived at the conclusion that both could be considered species of the genus. These conclusions were confirmed by the determination of biotypes of both (according to C. Richard). Later, we expect to study their participation in human infective processes and their sensitivity by antibiotic therapy.

Carbohydrate Metabolism↗
PubMed1977

[Cyclodextrin glucanotransferase from Klebsiella pneumoniae. 1. Formation, purification and properties of the enzyme from Klebsiella pneumoniae M 5 al (author's transl)].

1. The strain M 5 al of Klebsiella pneumoniae grows excellently with starches. We were able to show that besides the pullulanase associated with the external membrane of the cells the bacterium produces an inducible, extracellular cyclodextrin glucanotransferase [1,4-alpha-D-glucan-4-alpha-(1,4-alpha-glucano)-transferase (cyclising) (EC 2.4.1.19)]. Potato starch and cyclohexaamylose or cycloheptaamylose were found to be the best "inducing" carbon sources for the synthesis of the enzyme. When the bacteria are grown batchwise, maltose is a poorly "inducing" carbon source; larger quantities of the enzyme are synthesized by continuous cultivation with maltose as growth limiting factor. 2. For the determination of the cyclodextrin glucanotransferase-activity an assay method wsa worked out. 3. The enzyme could be separated from the culture filtrate and purified to more than 90% in few steps. At a total yield of 61.2% related to the activity of the culture filtrate employed we received an enzyme solution with the specific activity of 26.6 units/mg protein. Some properties of the enzyme are described. 4. The products formed from amylopectin by the enzyme were analyzed. Somewhat more than half the amylopectin was found as cyclodextrins. 29.3% of the cyclodextrin fraction were cycloheptaamylose, 47.2% cyclohexaamylose and 10.7% exo-branched cyclohexaamylose. 12.8% of cyclohexaamylose were obtained from a cyclodextrin glucanotransferase-limit dextrin after debranching by pullulanase and exposing the product to the action of the glucanotransferase again. 5. The importance of the cyclodextrin glucanotransferase for the utilization of starches by this strain of Klebsiella pneumoniae is discussed. After a first characterization the enzyme is compared to the amylase of Bacillus macerans.

Amylopectin↗