the use of 16s ribosomal rna analyses to investigate the phylogeny of the family legionellaceae.the 16s ribosomal rna sequences of legionella pneumophila, l. erythra, l. hackeliae, l. spiritensis, l. longbeachae, l. bozemanii (fluoribacter bozemanae) and l. micdadei (tatlockia micdadei) were determined using reverse transcriptase. the sequences were compared with published sequences for gram-negative bacteria and phylogenetic trees were constructed. the data confirm previous work which showed that the family legionellaceae forms a monophyletic subgroup within the gamma subdivision of the p ...19911713950
chemotaxonomic differentiation of legionellae by detection and characterization of aminodideoxyhexoses and other unique sugars using gas chromatography-mass spectrometry.legionellae have been differentiated previously by analyzing their carbohydrate contents by gas chromatography with flame ionization detection. in the present study, total ion mode gas chromatography-mass spectrometry (gc-ms) was used to detect a number of unusual sugars, including one that is structurally related to o-methyldideoxyheptoses. increased sensitivity and selectivity for carbohydrate detection was achieved by selected ion-monitoring gc-ms. two of the uncommon sugars previously discov ...19902324276
application of numerical systematics to the phenotypic differentiation of legionellae.a total of 163 strains, including 106 strains of legionella pneumophila, 28 strains of tatlockia micdadei, and 29 strains of other legionellae (including members of the proposed genus fluoribacter), were studied. ten tests which together could distinguish the genera previously proposed were identified. these tests included catalase-peroxidase, gelatinase, hippurate hydrolysis, starch hydrolysis, medium browning, acetoin production, oxidase, medium fluorescence, colony fluorescence, and the bromc ...19892550514
prevalence of legionnaires' disease in pneumonia patients admitted to a danish department of infectious diseases.during a 14-month study period, 92 patients admitted to the university clinic for infectious diseases with pneumonia were investigated to determine the prevalence and severity of legionnaires' disease (ld). the diagnosis of ld was based on positive serology. antibodies to 10 different legionella antigens--legionella pneumophila serogroups 1-6, fluoribacter (legionella) bozemanae, f. dumoffii, f. gormanii, and tatlockia (legionella) micdadei--were measured by the microagglutination (ma) and indir ...19863532303
agglutinating antibody titers to members of the family legionellaceae in cystic fibrosis patients as a result of cross-reacting antibodies to pseudomonas aeruginosa.the objective of this study was to evaluate the prevalence and significance of antibody titers to organisms in the family legionellaceae in 128 serum samples collected from cystic fibrosis patients at routine examinations. antibody titers were determined for 10 antigenic types of legionellaceae; legionella pneumophila serogroups 1 to 6, fluoribacter (legionella) bozemanae, fluoribacter (legionella) dumoffii, fluoribacter (legionella) gormanii, and tatlockia (legionella) micdadei. the method of a ...19846206086
cross-reactions between legionella pneumophila (serogroup 1) and twenty-eight other bacterial species, including other members of the family legionellaceae.cross-reactions between legionella pneumophila serogroup 1 and 28 other bacterial species were studied by various quantitative immunoelectrophoretic techniques. a sonicated l. pneumophila antigen and purified homologous rabbit antibody were used as a reference system. few antigens (0 to 6) cross-reacted with non-legionellaceae, but two were found in nearly all gram-negative bacteria tested (antigens no. 1 and 66). antigen no. 66 of the l. pneumophila reference system was shown to be antigenicall ...19836404825
plasmid transfer into members of the family legionellaceae.the plasmids rp4 and pph1ji::mu::tn5 were transferred with moderate frequency from escherichia coli to legionella pneumophila (strain bloomington 2) and fluoribacter (legionella) bozemanae (strain wiga). the frequency of transfer to tatlockia (legionella) micdadei (strain tatlock) was much lower. in these transconjugant strains the plasmids were maintained following at least 10 nonselective passages. however, the introduced plasmids could not be transferred from these strains to several other st ...19846491366
clinical laboratory differentiation of legionellaceae family members with pigment production and fluorescence on media supplemented with aromatic substrates.a systematic study of pigment production (browning) and fluorescence (extracellular yellow-green and intracellular blue-white) by nine legionellaceae species was performed. a total of 56 strains representing tatlockia micdadei (pittsburgh pneumonia agent), legionella pneumophila, legionella jordanis, legionella longbeachae, legionella oakridgensis, legionella wadsworthii, fluoribacter bozemanae, fluoribacter gormanii, and fluoribacter dumoffii could be separated on media supplemented with tyrosi ...19846547453
capillary gas chromatographic analysis of carbohydrates of legionella pneumophila and other members of the family legionellaceae.legionella pneumophila, the causative agent of legionnaires disease, and related organisms have previously been characterized primarily by conventional bacteriological methods, dna-dna hybridization, antigenic analysis, and fatty acid analysis. by capillary gas chromatographic analysis for carbohydrates, we have shown that muramic acid and glucosamine, characteristic markers of bacterial cell walls, were present in samples of l. pneumophila and a group of legionella-like organisms. some bacteria ...19846715510
electrophoretic characterization of soluble protein extracts of legionella pneumophila and other members of the family legionellaceae.the soluble peptides of strains of legionella pneumophila, tatlockia micdadei, fluoribacter bozemanae, fluoribacter dumoffii, and fluoribacter gormanii were studied by polyacrylamide gel electrophoresis. characteristic patterns were seen for legionella and tatlockia strains, whereas the patterns for the fluoribacter strains were variable as would be expected for this genetically heterogeneous group. grouping by peptide pattern was consistent with proposed taxons based on dna-dna homology. by usi ...19836874904
dye-containing buffered charcoal-yeast extract medium for differentiation of members of the family legionellaceae.the addition of 0.001% bromocresol purple and 0.001% bromothymol blue to buffered charcoal-yeast extract agar allowed differentiation between members of the family legionellaceae. on this medium, legionella pneumophila grew as relatively flat, pale green colonies, whereas tatlockia micdadei (gen. nov., comb. nov., pittsburgh pneumonia agent) produced blue-gray colonies. fluoribacter spp. (gen. nov., atypical legionella-like organisms) developed glistening colonies which were brighter green than ...19817009642
the legionella (fluoribacter) gormanii metallo-beta-lactamase: a new member of the highly divergent lineage of molecular-subclass b3 beta-lactamases.a metallo-beta-lactamase determinant was cloned from a genomic library of legionella (fluoribacter) gormanii atcc 33297(t) constructed in the plasmid vector pacyc184 and transformed into escherichia coli dh5alpha, by screening for clones showing a reduced susceptibility to imipenem. the product of the cloned determinant, named fez-1, contains a 30-kda polypeptide and exhibits an isoelectric ph of 7.6. sequencing revealed that fez-1 is a molecular-class b beta-lactamase which shares the closest s ...200010817705
bacterial populations colonizing and degrading rice straw in anoxic paddy soil.rice straw is a major substrate for the production of methane, a greenhouse gas, in flooded rice fields. the bacterial community degrading rice straw under anoxic conditions was investigated with molecular methods. rice straw was incubated in paddy soil anaerobically for 71 days. denaturing gradient gel electrophoresis (dgge) of the amplified bacterial 16s rrna genes showed that the composition of the bacterial community changed during the first 15 days but then was stable until the end of incub ...200111229927
biochemical characterization of the fez-1 metallo-beta-lactamase of legionella gormanii atcc 33297t produced in escherichia coli.the bla(fez-1) gene coding for the metallo-beta-lactamase of legionella (fluoribacter) gormanii atcc 33297t was overexpressed via a t7 expression system in escherichia coli bl21(de3)(plyss). the product was purified to homogeneity in two steps with a yield of 53%. the fez-1 metallo-beta-lactamase exhibited a broad-spectrum activity profile, with a preference for cephalosporins such as cephalothin, cefuroxime, and cefotaxime. monobactams were not hydrolyzed. the beta-lactamase was inhibited by me ...200111257043
characterization of oxa-29 from legionella (fluoribacter) gormanii: molecular class d beta-lactamase with unusual properties.a class d beta-lactamase determinant was isolated from the genome of legionella (fluoribacter) gormanii atcc 33297(t). the enzyme, named oxa-29, is quite divergent from other class d beta-lactamases, being more similar (33 to 43% amino acid identity) to those of groups iii (oxa-1) and iv (oxa-9, oxa-12, oxa-18, and oxa-22) than to other class d enzymes (21 to 24% sequence identity). phylogenetic analysis confirmed the closer ancestry of oxa-29 with members of the former groups. the oxa-29 enzyme ...200111709332
probing the specificity of the subclass b3 fez-1 metallo-beta-lactamase by site-directed mutagenesis.the subclass b3 fez-1 beta-lactamase produced by fluoribacter (legionella) gormanii is a zn(ii)-containing enzyme that hydrolyzes the beta-lactam bond in penicillins, cephalosporins, and carbapenems. fez-1 has been extensively studied using kinetic, computational modeling and x-ray crystallography. in an effort to probe residues potentially involved in substrate binding and zinc binding, five site-directed mutants of fez-1 (h121a, y156a, s221a, n225a, and y228a) were prepared and characterized u ...200415159411
legioliulin, a new isocoumarin compound responsible for blue-white autofluorescence in legionella (fluoribacter) dumoffii under long-wavelength uv light.legionella dumoffii is one of the causative agents of legionnaires' disease. there are 50 species in the genus legionella, of which 10 species including l. dumoffii are known to exhibit uncharacterized blue-white autofluorescence. we constructed an l. dumoffii strain that exhibited a high intensity blue-white fluorescence, isolated a fluorescent pigment from the strain, and determined its molecular formula to be c(19)h(14)o(3) by high-resolution mass spectrometry. an nmr analysis revealed that t ...200415381093
conjugative plasmid pld-tex-kl promotes growth of host bacterium legionella dumoffii at low temperatures.legionella (fluoribacter) dumoffii is a resident of various aquatic environments and occasionally causes pneumonia in humans. we found that l. dumoffii strain tex-kl carries a 66-kb circular plasmid. as predicted by the presence of tra genes similar to those of other transferable plasmids, we showed that pld-tex-kl was actually capable of transferring itself to a plasmid-cured derivative of the original strain. unexpectedly, this plasmid-free derivative turned out to be partially defective in te ...200919415237
three-dimensional structure of fez-1, a monomeric subclass b3 metallo-beta-lactamase from fluoribacter gormanii, in native form and in complex with d-captopril.the beta-lactamases are involved in bacterial resistance to penicillin and related compounds. members of the metallo-enzyme class are now found in many pathogenic bacteria and are thus becoming of major clinical importance. the structures of the zn-beta-lactamase from fluoribacter gormanii (fez-1) in the native and in the complex form are reported here. fez-1 is a monomeric enzyme, which possesses two zinc-binding sites. these structures are discussed in comparison with those of the tetrameric l ...200312507470
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