PubMed Health⌕ Search

Biomedical subjects

C W Moss

Publications and source records attributed to C W Moss.

At least 73 records · Page 4Linked to original sources

Legionella bozemanii serogroup 2: a new etiological agent.

A newly discovered bacterium, Toronto 3, isolated from a lung aspirate of a patient with pneumonia, has been characterized. The isolate was identified as Legionella bozemanii by chemical data from cellular fatty acid and ubiquinone analyses and by DNA relatedness studies. The isolate, however, differs phenotypically from L. bozemanii by its colonial characteristics and strong interspecies serological cross-reactions, which are unique among clinical isolates of legionellae. The name L. bozemanni serogroup 2 is proposed. The reference strain is Toronto 3. The clinical illness caused by L. bozemanii serogroup 2 was not distinguishable from other Legionella infections. L. bozemanii is the third Legionella species with more than one serogroup. Rapid laboratory diagnosis of this strain by direct fluorescent antibody test may be complicated in the absence of culture isolation.

DNA, Bacterial↗

Isoprenoid quinone content and cellular fatty acid composition of Campylobacter species.

A total of 36 strains of Campylobacter species were examined for isoprenoid quinones and cellular fatty acids. The isoprenoid quinone content was determined by reverse-phase high-pressure liquid chromatography, and the fatty acids were determined by capillary gas-liquid chromatography. All Campylobacter species contained menaquinone-6 (2-methyl-3-farnesyl-farnesyl-1,4-naphthoquinone) and a methyl-substituted menaquinone-6 (2,[5 or 8]-dimethyl-3-farnesyl-farnesyl-1,4-napthoquinone) as the major isoprenoid quinones. The latter menaquinone has not been reported in other bacteria and may prove to be a useful chemical marker of Campylobacter species. Campylobacter jejuni and most Campylobacter coli were distinguished from other campylobacteria by the presence of a C19 cyclopropane acid, and Campylobacter sputorum subsp. mucosalis differed from other species by the presence of lauric acid.

Campylobacter↗

Frequency-pulsed electron capture gas-liquid chromatographic analysis of metabolites produced by Clostridium difficile in broth enriched with amino acids.

Clostridium difficile strain CDC A-567 was cultured in Trypticase (BBL Microbiology Systems)-yeast-salt broth supplemented with 0.2% L-leucine, L-norleucine, L-isoleucine, L-tyrosine, or L-tryptophan. Four extractions were done on the spent medium, three at pH 2 and one at pH 10, using CHCL3 or ether. Derivatizations were done with trichloroethanol, heptafluorobutyric anhydride, and heptafluorobutyric anhydride-ethanol. All samples were analyzed with frequency-pulsed electron capture gas-liquid chromatography. A dedicated computer was used to assist in data analysis. C. difficile produced both short-chain and aromatic acids in Trypticase-yeast-salt broth; hydroxy acids were also detected. p-Cresol, indoleacetic acid, 4-methylthio-2-hydroxybutyric acid, and some unidentified alcohols were observed. The basic chloroform extraction contained cadaverine and putrescine. Leucine, norleucine, and isoleucine influenced the production of C5 and C6 acids and alcohols. L-Tyrosine underwent successive degradation to produce p-cresol and aromatic acids as final products. Tryptophan increased the production of indoleacetic, indolepropionic, and indolebutyric acids. Isocaproic acid was produced in relatively high concentrations regardless of medium substitution. The consistent production of iC6 under various substrate conditions indicates that the production of this compound might be consistent enough in vitro to form the basis of a rapid test for detection of C. difficile in stool specimens by frequency-pulsed electron capture gas-liquid chromatography.

Alcohols↗

Rapid differentiation of enterotoxigenic Escherichia coli that produce heat-stable and heat-labile toxins by frequency-pulsed electron capture gas-liquid chromatography analysis of diarrheal stool specimens.

Thirty-three stool specimens from infants in the village of Tamooh near Cairo, Egypt, were studied by frequency-pulsed electron capture gas-liquid chromatography (FPEC-GLC). In 13 of the diarrheal cases, the suspected causative agent isolated was Escherichia coli which produced heat-stable toxin (ST), and in 10 other cases E. coli that produced heat-labile toxin (LT) were isolated. Ten control stool samples, collected from infants from whom no pathogenic organisms were isolated, were analyzed at the same time. Comparisons also were made against healthy control stools from individuals in the United States who had been previously analyzed by FPEC-GLC (Brooks et al., J. Clin. Microbiol. 20:549-560, 1984). The stools were suspended in water and centrifuged, and the supernatant was extracted with organic solvents and derivatized to form electron-capturing derivatives of carboxylic acids, hydroxy acids, alcohols, and amines. Results from the study showed distinct differences among the FPEC-GLC profiles of E. coli ST-positive stools, of E. coli LT-positive stools, and of the control stool samples. An unidentified compound appearing in the ether-soluble hydroxy acid fraction from E. coli ST-positive stools was tentatively identified by mass spectrometry as 6-methoxy-2-hydroxyhexanoic acid. 6-Methoxy-2-hydroxyhexanoic acid was found in all stools that contained E. coli ST but was not present either in stools from which E. coli LT was isolated or in control samples. 6-Methoxy-2-hydroxyhexanoic acid may prove to be an important marker for use in the identification of E. coli ST. In addition to 6-methoxy-2-hydroxyhexanoic acid, the carboxylic acid, alcohol, and amine FPEC-GLC profiles obtained from stools were very different between these two organisms. The data indicate that FPEC-GLC analysis of diarrheal stool specimens might be a rapid way to distinguish diarrhea caused by E. coli ST, E. coli LT, Clostridium difficile, and rotavirus.

Bacterial Toxins↗

Legionella pneumophila serogroup 9: a cause of human pneumonia.

A new serogroup of Legionella pneumophila was isolated from bronchoscopic washings and an open-lung biopsy specimen of a patient from California with pneumonia. A serologically identical isolate was obtained from tap water of a hospital ward in the Netherlands, and a fatal case of pneumonia in a patient from Virginia was shown retrospectively to have been caused by this new organism. The type strain of what is now serogroup 9 of L. pneumophila is IN-23-G1-C2 (American Type Culture Collection no. 35289). Disease caused by L. pneumophila serogroup 9 is not apparently different from that caused by other L. pneumophila serogroups.

Agglutination Tests↗

Cellular fatty acid composition and ubiquinone content of Legionella feeleii sp. nov.

The cellular fatty acid composition of Legionella feeleii was determined by capillary gas-liquid chromatography, and the ubiquinone content was determined by reverse-phase high-pressure liquid chromatography. As in other Legionella species, this new species is characterized by relatively large amounts of branched-chain fatty acids and by major amounts of ubiquinones with more than 10 isoprene units in the side chain.

Chromatography, Gas↗

Cellular fatty acid composition of Pseudomonas marginata and closely associated bacteria.

The cellular fatty acid compositions of the lectotype strain and four clinical isolates of Pseudomonas marginata were determined by gas-liquid chromatography and compared with 11 strains of the Centers for Disease Control Pseudomonas-like group 2, which are similar to P. marginata in a number of conventional biochemical tests. Isolates of P. marginata were readily distinguished from Pseudomonas-like group 2 by the presence of a C17:0 cyclopropane acid and hydroxy acids 3-OH-C14:0, 2-OH-C16:0, 3-OH-C16:0, and 2-OH-C18:1, whereas strains of Pseudomonas-like group 2 contained C16:1 delta 9 as a major acid with small amounts of 3-OH-C12:0 and 2-OH-C14:0 acids. Our data show that cellular fatty acid composition provides useful additional information that can be combined with selected conventional tests to provide a more reliable and rapid identification of P. marginata and related bacteria.

Chromatography, Gas↗

Comparison of the effects of acid and base hydrolyses on hydroxy and cyclopropane fatty acids in bacteria.

The cellular fatty acid compositions of Legionella oakridgensis, Brucella suis, Pseudomonas aeruginosa, and Francisella tularensis were compared after base hydrolysis (saponification), acid hydrolysis, and acid methanolysis procedures were used to release the fatty acids. The branched-chain, unsaturated, saturated, and ester-linked hydroxy acids were released as effectively with saponification at 100 degrees C for 30 min as with acid hydrolysis or acid methanolysis at 85 degrees C for 16 h. Although the amide-linked hydroxy acids were released more effectively by acid hydrolysis or acid methanolysis, these methods degraded the cyclopropane fatty acids, producing a number of new peaks or artifacts in the chromatograms. Cyclopropane fatty acids were not degraded by saponification, and at least 50% of the hydroxy acids were released when the cells were saponified with 15% NaOH in 50% aqueous methanol. Thus, the results show that saponification for 30 min at 100 degrees C with 15% NaOH, followed by methylation is an excellent method for routine fatty acid analysis of bacteria and for screening cultures whose identity and fatty acid composition are unknown.

Bacteria↗

Legionella jordanis: a new species of Legionella isolated from water and sewage.

Legionella jordanis sp. nov., as found in two cultures, is described. One isolate was from river water in Indiana and the other isolate was from sewage in DeKalb County, Ga. The former is the type strain of the species, and is designated BL-540 (ATCC 33623). L. jordanis had a partial relationship to L. bozemanii by direct fluorescent-antibody tests but was unrelated to L. pneumophila, L. dumoffii, L. micdadei, L. gormanii, or L. longbeachae. Legionella phenotypic characteristics, including large amounts of branched-chain cellular fatty acids, were shown by the isolates. Studies of DNA relatedness showed that the two cultures of L. jordanis were only slightly related to the six previously described species of Legionella but were more than 90% related to each other. Indirect fluorescent-antibody tests with human sera suggested that unrecognized human infections with L. jordanis may be occurring.

DNA, Bacterial↗

Isoprenoid quinones of the genus Legionella.

Representative strains of each of the named species of Legionella were examined for isoprenoid quinones by reverse-phase thin-layer chromatography. All strains contained three or more ubiquinones (Q9, Q10, Q11, Q12, Q13) which were useful for placing the species into one of three distinct groups. Group 1 contained L. longbeachae, L. bozemanii, L. dumoffi, and L. gormanii; group 2 contained only L. micdadei; and group 3 contained only L. pneumophila. The identities of the quinones were established by UV spectroscopy and mass spectrometry.

Legionella↗

30 years of campylobacters: biochemical characteristics and a biotyping proposal for Campylobacter jejuni.

Several biochemical test systems were studied for their potential usefulness for the examination of strains of Campylobacter species. Most (81%) of the C. jejuni strains hydrolyzed sodium hippurate, but strains of C. fetus, C. sputorum, and C. fecalis did not. Some (46%) of the C. jejuni strains and all of the C. sputorum subsp. sputorum, C. sputorum subsp. bubulus, and C. fecalis strains hydrolyzed DNA, but the C. fetus and C. sputorum subsp. mucosalis strains did not. Strains of all species of Campylobacter grew on charcoal-yeast extract agar, but 47% of the C. jejuni strains did not. Alkaline phosphatase activity was recorded for some strains of C. jejuni, but all other species were negative for this activity. Aryl sulfatase activity was detected in 7% of the C. jejuni, 15% of the C. fetus subsp. fetus, and all of the C. sputorum subsp. sputorum, C. sputorum subsp. bubulus, and C. fecalis strains, but it was not detected in the C. fetus subsp. venerealis and C. sputorum subsp. mucosalis strains. Most (93%) of the C. jejuni but none of the other Campylobacter strains contained lactobacillic acid when examined for cellular fatty acids. On the basis of results from three of these tests (hippurate hydrolysis, DNA hydrolysis, and growth on charcoal-yeast extract agar), clinical strains of C. jejuni were placed in eight biotypes.

Alkaline Phosphatase↗

Analysis of short-chain acids from anaerobic bacteria by high-performance liquid chromatography.

A standard mixture of 25 short-chain fatty acids was resolved by high-performance liquid chromatography, using an Aminex HPX-87 column. The acids produced in culture media by anaerobic bacteria were analyzed by high-performance liquid chromatography after extraction with ether and reextraction into a small volume of 0.1 N NaOH. The presence of fumaric acid in culture extracts of Peptostreptococcus anaerobius was confirmed by gas chromatography-mass spectrometry analysis of the trapped eluent fractions from the high-performance liquid chromatography column.

Bacteroides fragilis↗

Cellular fatty acids of Capnocytophaga species.

The cellular fatty acid composition of 18 clinical isolates and 4 reference strains of Capnocytophaga species (Capnocytophaga ochracea, Capnocytophaga gingivalis, and Capnocytophaga sputigena) was determined by gas-liquid chromatography. The fatty acid profiles of the 22 cultures were essentially identical and were characterized by major amounts (60% or greater) of a saturated, iso-branched-chain, 15-carbon acid (13-methyl-tetradecanoate) and the presence of two relatively uncommon saturated, iso-branched, 3-hydroxy acids (13-methyl-3-hydroxy-tetradecanoate and 15-methyl-3-hydroxy-hexadecanoate). The presence and relative amounts of these acids distinguish Capnocytophaga spp. from other gliding bacteria.

Capnocytophaga↗

Determination of cellular fatty acid compositions of various yeasts by gas-liquid chromatography.

The cellular fatty acid composition of 51 cultures of various species of yeasts was determined by gas-liquid chromatography. Analysis was done with a fused-silica gas-liquid chromatography capillary column, with resolution of all components including mono-, di-, and tri-unsaturated 18-carbon acids. The cultures were placed into one of four distinct gas-liquid chromatography groups on the basis of large quantitative differences in fatty acids. Group I contained only Saccharomyces species and group II only Torulopsis glabrata. Most Candida species were placed into group III, and group IV contained only basidiomycetous yeasts.

Chromatography, Gas↗

Legionella wadsworthii species nova: a cause of human pneumonia.

A new species of Legionella that caused pneumonia in a patient with chronic lymphocytic leukemia has been characterized. The Legionella wadsworthii species nova is proposed for this bacterium. The type strain is Wadsworth 81-716A (American Type Culture Collection 33877). The new species differs phenotypically from L. pneumophila in that the predominant cellular fatty acid is methyl-12-methyltetradecanoic acid (a-15:0) rather than methyl-14-methylpentadecanoic acid (i-16:0), and in its failure to react with L. pneumophila antiserum. The clinical illness caused by L. wadsworthii was not apparently different from that seen with other legionella infections, except for prolonged fever and slow resolution of pulmonary infiltrates.

Aged↗

Gas-liquid chromatography as an analytical tool in microbiology.

The use of gas-liquid chromatography (GLC) in clinical and diagnostic bacteriology laboratories has increased significantly in recent years. GLC analysis of metabolic products from bacterial growth and chemical components of bacterial cells has provided useful information for rapid detection and identification of several bacterial groups or species. The use of short-chain acid products and cellular fatty acid composition for identifying and classifying Pseudomonas species and other medically important gram-negative non-fermentative bacteria is illustrated. Application of the flexible, fused, silica-glass, capillary column for increased resolution of bacterial fatty acids is also discussed.

Alcaligenes↗