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PubMed2004

Comparative activities of doripenem versus isolates, mutants, and transconjugants of Enterobacteriaceae and Acinetobacter spp. with characterized beta-lactamases.

Doripenem (S-4661), a new parenteral carbapenem, was tested against over 250 clinical isolates, mutants, and transconjugants of Enterobacteriaceae and Acinetobacter spp., selected or derived for their beta-lactamase expression characteristics. Imipenem, meropenem, and ertapenem were tested as comparators, along with cephalosporins and piperacillin-tazobactam, by using National Committee for Clinical Laboratory Standards agar dilution methodology. Doripenem MICs were from 0.03 to 0.25 microg/ml for Klebsiella isolates, irrespective of the presence of extended-spectrum beta-lactamases (ESBLs) or plasmid-mediated AmpC or hyperproduced K1 beta-lactamase. Similarly, MICs of doripenem for both AmpC-inducible and -derepressed Enterobacter isolates were 0.06 to 0.5 microg/ml. ESBL production did not raise the MICs of doripenem for Escherichia coli transconjugants, and studies with known expression mutants confirmed that neither inducible nor depressed AmpC beta-lactamase expression was protective in Enterobacter cloacae, Citrobacter freundii, Serratia marcescens, or Morganella morganii. In all of these respects, doripenem resembled meropenem and imipenem, whereas the MICs of ertapenem were raised (but still < or =1 microg/ml) for many ESBL-producing klebsiellas and AmpC-derepressed E. cloacae and C. freundii strains. Resistance to all carbapenems, including doripenem (MICs of mostly 16 to 64 microg/ml, compared with 0.25 to 1 microg/ml for typical strains), was seen in Acinetobacter isolates with metallo-beta-lactamases or OXA-carbapenemases. Isolates of Klebsiella and Serratia spp. with IMP, KPC, and SME beta-lactamases also were resistant to doripenem (MICs, 8 to >64 microg/ml) and to other carbapenems, although the continued apparent susceptibility (MICs, < or =0.5 microg/ml) of E. coli derivatives with cloned IMP-1 and NMC-A beta-lactamases suggested that carbapenem resistance might require other factors besides the enzymes.

Acinetobacter↗
PubMed2006

Emergence of Enterobacteriaceae isolates producing CTX-M extended-spectrum beta-lactamase in Austria.

Among 149 extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae isolates collected from patients in southeast Austria from 1998 to 2004, 38 Escherichia coli isolates and 11 Klebsiella spp. were CTX-M producers. The proportion of CTX-M-producers among all ESBL producers rose from 0% in 1998 to 58% in 2004. In general, CTX-M-producers had heterogeneous pulsed-field gel electrophoresis patterns, but one E. coli isolate was identical to a United Kingdom epidemic CTX-M-15-producing strain, although no epidemiological link with the United Kingdom was apparent.

Electrophoresis, Gel, Pulsed-Field↗
PubMed2006

Increasing prevalence and diversity of metallo-beta-lactamases in Pseudomonas spp., Acinetobacter spp., and Enterobacteriaceae from Korea.

Among imipenem-nonsusceptible isolates, acquired metallo-beta-lactamase genes were detected in 36 of 581 (6.2%) Pseudomonas aeruginosa isolates, 42 of 44 (95.4%) other Pseudomonas species, and 136 of 513 (26.5%) Acinetobacter species from 2003 to 2004 at a Korean hospital. Overall, bla(VIM-2)-like genes were the most prevalent and were also detected in Enterobacteriaceae, including Klebsiella pneumoniae.

Acinetobacter↗
PubMed1974

Evaluation of two standardized disk methods for testing antimicrobial susceptibility of Pseudomonas aeruginosa and of the Enterobacteriaceae.

Two standardized disk diffusion susceptibility testing methods were compared using Pseudomonas aeruginosa and members of the Enterobacteriaceae. The standard Bauer-Kirby method and the agar overlay method of Barry et al. were compared using carbenicillin (50- and 100-mug disks), gentamicin, polymyxin B, and colistin sulfate. Both methods gave nearly comparable zone sizes with the enteric bacilli. However, with P. aeruginosa the agar overlay method gave zones 1 to 2 mm smaller than the Bauer-Kirby method with all drugs except gentamicin. In spite of these small differences, further examination of minimal inhibitory concentration-zone size correlations indicated that the same interpretive zone standards can be applied to either diffusion technique. For testing carbenicillin against Pseudomonas sp., 50-mug disks were unsatisfactory, especially with the agar overlay method; 100-mug disks were far superior with both methods.

Carbenicillin↗
PubMed1975

Differences in susceptibility of Enterobacteriaceae and penicillin-resistant Staphylococcus aureus to tetracycline and minocycline.

Two hundred strains of penicillin-resistant Staphylococcus aureus and 311 isolates of Enterobacteriaceae were compared for their susceptibility to tetracycline and minocycline. Thirteen and one-half percent of the staphylococcal isolates were resistant to tetracycline but susceptible to minocycline. Similarly, 24% of the enterobacterial isolates were found to be tetracycline resistant but susceptible to minocycline. Of a total of 511 recent clinical isolates, 14.5% were susceptible to minocycline but were tetracycline resistant.

Enterobacteriaceae↗
PubMed1975

Susceptibility of clinical isolates of Enterobacteriaceae to BL-S640, a new oral cephalosporin.

The in vitro activity of BL-S640, a 7-(2-aryl-2-aminoacetamido)-3-(heterocyclic-thiomethyl) cephalosporin, was evaluated against 338 clinical isolates of Enterobacteriaceae in comparison with ampicillin, cephalothin, cefazolin, and cephalexin. Against Escherichia coli, BL-S640 was as active as cefazolin and more active than ampicillin, cephalothin, and cephalexin. BL-S640 was as effective as the other cephalosporins tested and far more active than ampicillin against Klebsiella and was more active than cephalexin against Proteus mirabilis and the indole-positive Proteus. The majority of Enterobacter, Serratia, and Citrobacter were resistant to ampicillin and all the cephalosporins tested. With rare exceptions, the zone of inhibition by the BL-S640 30-mug disk was either larger or the same as the zone obtained by the cephalothin 30-mug disk in the Kirby-Bauer disk susceptibility test.

Cephalosporins↗
PubMed1978

Comparison of miniaturized multitest systems with conventional methodology for identification of Enterobacteriaceae from foods.

Four miniaturized multiple test systems were compared with tube methodology used to identify Enterobacteriaceae encountered in foods. Identification aids supplied with each system were used to assign names to isolates at the species level. For the 129 strains tested, the Minitek system demonstrated a 96.9 percent agreement with reactions in tubed media. The Inolex, Analytab, and PathoTec test systems exhibited 94.3, 93.8, and 92.7 percent agreement, respectively. Analytab identified 96.1 percent of the isolates to the species level, whereas the Minitek, PathoTec, and Inolex systems were able to identify 78.3, 32.6, and 27.1 percent, respectively. The results indicate that the Analytab and Minitek systems are acceptable substitutes for the tube methodology routinely employed in identifying enterics from foods. Although the PathoTec system might be used to screen isolates for their identity, neither the presently available PathoTec nor the Inolex systems should be substituted for current methodology when definitive identification of foodborne organisms is required.

Bacteriological Techniques↗
PubMed1979

Enterobacteriaceae isolated from iguanid lizards of west-central Texas.

The prevalence of members of the family Enterobacteriaceae in the intestines of seven species of iguanid lizards native to west-central Texas was determined. Of the 67 lizard specimens examined, 48.7% were infected with Salmonella and 9% were infected with Salmonella arizonae. Two lizard species (Sceloporus olivaceus and Crotaphytus collaris) were shown to have a 100% prevalence of Salmonella.

Animals↗
PubMed1983

Glycine betaine, an osmotic effector in Klebsiella pneumoniae and other members of the Enterobacteriaceae.

Osmoregulation was examined in members of the Enterobacteriaceae. Exogenous glycine betaine at a concentration as low as 1 mM was found to stimulate the growth rate of Escherichia coli, Salmonella typhimurium, and Klebsiella pneumoniae in media of inhibitory osmotic strength. The stimulation was shown to be independent of any specific solutes, electrolytes, or nonelectrolytes. Therefore, the stimulatory effect of glycine betaine was a consequence of high osmotic potential. This effect was found to be far greater than the proline effect previously observed in S. typhimurium. Whereas nitrogen fixation by K. pneumoniae is completely inhibited under conditions of osmotic stress, nitrogenase activity could be partially restored by the addition of exogenous glycine betaine to the culture medium. Furthermore, glycine betaine in combination with proline, especially proline produced internally at a high level because of regulatory mutations affecting proline biosynthesis, strongly stimulated nitrogen fixation activity during osmotic stress. Glycine betaine was accumulated by the cells, and the amount taken up was correlated with the osmolarity of the medium. These findings are discussed in relation to the possible mechanisms by which glycine betaine might cause enhanced osmotolerance.

Amino Acids↗
PubMed1992

Rapid detection of members of the family Enterobacteriaceae by a monoclonal antibody.

Six monoclonal antibodies directed against enterobacteria were produced and characterized. The specificity of one of these antibodies (CX9/15; immunoglobulin G2a) was studied by indirect immunofluorescence against 259 enterobacterial strains and 125 other gram-negative bacteria. All of the enterobacteria were specifically recognized, the only exception being Erwinia chrysanthemi (one strain tested). Bacteria not belonging to members of the family Enterobacteriaceae were not detected, except for Plesiomonas shigelloides (two strains tested), Aeromonas hydrophila (five strains tested), and Aeromonas sobria (one strain tested). This recognition spectrum strongly suggested that CX9/15 recognized the enterobacterial common antigen. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot (immunoblot) experiments, the six antienterobacteria antibodies presented similar specificities; they all revealed only one band with an apparent molecular weight of about 20,000 from the crude extract of an enterobacterium. The six monoclonal antibodies, and especially CX9/15, can be used to develop new tests for rapid and specific detection of enterobacteria.

Antibodies, Monoclonal↗
PubMed1992

Rapid determination of members of the family Enterobacteriaceae in drinking water by an immunological assay using a monoclonal antibody against enterobacterial common antigen.

An immunological method for the detection of members of the family Enterobacteriaceae in drinking water was developed. The method was based on a sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibody immunoglobulin G2a 898 against enterobacterial common antigen. The enterobacterial common antigen sandwich ELISA combined with selective preenrichment culture could be performed in only 24 h. Six hundred sixty-eight water samples from a variety of German public water supplies were screened to verify the effectiveness of the new method. Ninety-eight percent of the results obtained by the immunological method could be confirmed by conventional microbiological methods. The immunological method proved to be considerably faster and more specific and sensitive than the standard method specified by the German drinking water regulations.

Antibodies, Bacterial↗
PubMed1993

Comparison of chemical assay, bioassay, enzyme-linked immunosorbent assay, and dot blot hybridization for detection of aerobactin in members of the family Enterobacteriaceae.

In order to determine the best strategy for detection of aerobactin in members of the family Enterobacteriaceae, we compared the results of three phenotypic assays, including a chemical assay, a cross-feeding bioassay, and an enzyme-linked immunosorbent assay (ELISA), with the results of a dot blot hybridization assay using a specific probe for the aerobactin genes. The sensitivity and specificity of the ELISA were better than those of the chemical and cross-feeding assays, but the results of dot blot hybridization were the most reproducible. However, none of the Serratia and Enterobacter cloacae strains which produced aerobactin hybridized with the probe. We concluded that the best strategy for aerobactin detection is a two-step procedure that combines screening by dot blot hybridization with an ELISA for negative strains.

Biological Assay↗
PubMed1994

Isolation of three hexahydro-1,3,5-trinitro-1,3,5-triazine-degrading species of the family Enterobacteriaceae from nitramine explosive-contaminated soil.

Three species of the family Enterobacteriaceae that biochemically reduced hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) were isolated from nitramine explosive-contaminated soil. Two isolates, identified as Morganella morganii and Providencia rettgeri, completely transformed both RDX and the nitroso-RDX reduction intermediates. The third isolate, identified as Citrobacter freundii, partially transformed RDX and generated high concentrations of nitroso-RDX intermediates. All three isolates produced 14CO2 from labeled RDX under O2-depleted culture conditions. While all three isolates transformed HMX, only M. morganii transformed HMX in the presence of RDX.

Aniline Compounds↗
PubMed2002

Rapid and sensitive enumeration of viable diluted cells of members of the family enterobacteriaceae in freshwater and drinking water.

Water quality assessment involves the specific, sensitive, and rapid detection of bacterial indicators and pathogens in water samples, including viable but nonculturable (VBNC) cells. This work evaluates the specificity and sensitivity of a new method which combines a fluorescent in situ hybridization (FISH) approach with a physiological assay (direct viable count [DVC]) for the direct enumeration, at the single-cell level, of highly diluted viable cells of members of the family Enterobacteriaceae in freshwater and drinking water after membrane filtration. The approach (DVC-FISH) uses a new direct detection device, the laser scanning cytometer (Scan RDI). Combining the DVC-FISH method on a membrane with Scan RDI detection makes it possible to detect as few as one targeted cell in approximately 10(8) nontargeted cells spread over the membrane. The ability of this new approach to detect and enumerate VBNC enterobacterial cells in freshwater and drinking water distribution systems was investigated and is discussed.

Bacteriological Techniques↗
PubMed2005

Diversity of carotenoid synthesis gene clusters from environmental Enterobacteriaceae strains.

Eight Enterobacteriaceae strains that produce zeaxanthin and derivatives of this compound were isolated from a variety of environmental samples. Phylogenetic analysis showed that these strains grouped with different clusters of Erwinia type strains. Four strains representing the phylogenetic diversity were chosen for further characterization, which revealed their genetic diversity as well as their biochemical diversity. The carotenoid synthesis gene clusters cloned from the four strains had three different gene organizations. Two of the gene clusters, those from strains DC416 and DC260, had the classical organization crtEXYIBZ; the gene cluster from DC413 had the rare organization crtE-idi-XYIBZ; and the gene cluster from DC404 had the unique organization crtE-idi-YIBZ. Besides the diversity in genetic organization, these genes also exhibited considerable sequence diversity. On average, they exhibited 60 to 70% identity with each other, as well as with the corresponding genes of the Pantoea type strains. The four different clusters were individually expressed in Escherichia coli, and the two idi-containing clusters gave more than fivefold-higher carotenoid titers than the two clusters lacking idi. Expression of the crtEYIB genes with and without idi confirmed the effect of increasing carotenoid titer by the type II idi gene linked with the carotenoid synthesis gene clusters.

Carotenoids↗
PubMed2005

Evolutionary relationships of three new species of Enterobacteriaceae living as symbionts of aphids and other insects.

Ecological studies on three bacterial lineages symbiotic in aphids have shown that they impose a variety of effects on their hosts, including resistance to parasitoids and tolerance to heat stress. Phylogenetic analyses of partial sequences of gyrB and recA are consistent with previous analyses limited to 16S rRNA gene sequences and yield improved confidence of the evolutionary relationships of these symbionts. All three symbionts are in the Enterobacteriaceae. One of the symbionts, here given the provisional designation "Candidatus Serratia symbiotica," is a Serratia species that has acquired a symbiotic lifestyle. The other two symbionts, here designated "Candidatus Hamiltonella defensa" and "Candidatus Regiella insecticola," are sister groups to one another and together show a relationship to species of Photorhabdus.

Animals↗
PubMed1966

Biochemical differentiation of the Enterobacteriaceae with the aid of lysine-iron-agar.

A procedure is described for identifying members of the family Enterobacteriaceae isolated from clinical specimens. The methods are based on primary differentiation of the various groups of bacteria by the use of Kligler Iron Agar and lysine-iron-agar. For identification of Salmonella, Shigella, and Arizona group organisms from stools, Triple Sugar Iron Agar and lysine-iron-agar are employed. The usefulness of this schema for diagnostic bacteriology laboratories is discussed. It is not intended to replace methods used in reference or research laboratories.

Culture Media↗