Biomedical subjects
Publications and source records attributed to C W Moss.
The cellular fatty acid composition of 36 strains of the Legionnaires disease bacterium was determined by gas chromatography after growth on different media. The fatty acid profile of each strain was essentially identical on each medium and was characterized by large amounts (greater than 68%) of branched-chain acids.
The Legionnaires' disease (LD) bacterium is a gram-negative organism whose "endotoxicity" appears to differ in several respects from the classic endotoxicity generally associated with gram-negative bacteria. Discrepancies were noted between the high activity of LD bacteria in gelating limulus lysate in vitro and their low pyrogenicity in rabbits. Further in-vivo biologic tests indicated that LD bacteria were relatively weak in "endotoxicity". Analysis of LD bacterial cells and in their cellular fractions by gas-liquid chromatography indicated that LD bacteria did not contain hydroxy fatty acids commonly associated with lipid A of endotoxin. The branched-chain fatty acids that were characteristic of LD bacteria were associated with the cell envelope, and were readily extracted into organic solvents without prior saponification. The presence of 2-keto-3-deoxyoctonate in LD bacteria and cell extracts was detected by a microassay method but remains to be confirmed with gas-liquid chromatography and mass spectrometry. The active principle "endotoxicity" in LD bacteria may be a new type of bacterial lipopolysaccharide.
A bacterium with growth characteristics similar to, but genetically distinct from, either Legionella pneumophila or WIGA (a "rickettsia-like agent") was obtained from a postmortem lung specimen of a patient with fatal atypical pneumonia at the M. D. Anderson Hospital and Tumor Institute in Houston, Texas. This bacterium and WIGA have essentially the same cellular fatty acid composition, which is distinct from that of L. pneumophila. Deoxyribonucleic acid-reletadness studies show that the isolate from Texas is only about 10% related to both L. pneumophila and WIGA and there fore may represent a new species. This new bacterium should be considered in selecting laboratory procedures in the diagnosis of atypical pneumonia.
The cellular fatty acid composition and metabolic products of 12 reference strains of Achromobacter sp. and A. xylosoxidans were determined by gas-liquid chromatography (GLC). Results showed that the two Achromobacter groups are strikingly different and can be readily distinguished on the basis of cellular fatty acids and the short-chain acids produced by Achromobacter sp. The major cellular fatty acids of Achromobacter sp. were octadecenoic (18:1) and a 19-carbon cyclopropanoic (19:0 delta) acid, whereas hexadecanoic (16:0) and a 17-carbon cyclopropanoic (17:0 delta) acid were principal components of the lipids of A. xylosoxidans. Hydroxy acids were not found in strains of Achromobacter sp. but comprised approximately 20% of the cellular fatty acids of A. xylosoxidans. In addition, Achromobacter sp. produced relatively large amounts of 2-ketoisocaproic acid, which was detected in only trace amounts from strains of A. xylosoxidans. The data show that GLC tests provide additional criteria for differentiating groups which are very closely related when evaluated with conventional tests. The GLC tests can be readily adapted in the clinical laboratory because they are rapid, highly reproducible, relatively inexpensive, and simple to perform.
The cellular fatty acid profiles of Flavobacterium meningosepticum and Flavobacterium species group IIb were markedly different from those of related bacteria. The profiles were characterized by the presence of 13-methyl-tetradecanoate and three uncommon acids: 2-hydroxy-13-methyl-tetradecanoate, 15-methyl-hexadecanoate, and 3-hydroxy-15-methyl-hexadecanoate.
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Cellular fatty acids of Peptococcus variables and Peptostreptococcus anaerobius were identified by gas chromatography, mass spectrometry, and associated analytical techniques. Iso- and anteiso-branched-chain acids were major components in both species.
The cellular fatty acids of four isolates from Legionnaires disease and two antigenically related isolates were identified by gas chromatography, mass spectrometry, and associated techniques. The six isolates had essentially the same fatty acid composition, which was characterized by large amounts (greater than 80%) of branched-chain acids.
Two isolates of Neisseria gonorrhoeae from two patients who did not respond to repeated treatment with penicillin were found to contain an active beta-lactamase. This enzyme has not been previously found in other gonococci isolated in the United States. Compared to other gonococci, these isolates had higher penicillin minimal inhibitory concentrations, and gave very small or no zones of inhibition in the disc agar diffusion test. The enzyme was demonstrated with three different rapid tests for beta-lactamase, by disc diffusion assay methods, and by gas-liquid chromatography-mass spectrometry determination of penicilloic acid--the enzymatic end product from penicillin.
The cellular fatty acid compositions of Pseudomonas diminuta UC 501 and Streptococcus mutans OMZ-61 were compared in samples processed by a saponification-methylation procedure (method S) and in samples processed by a transesterification procedure (method T). All samples were heated in an inert atmosphere of nitrogen. The major acids found in samples of P. diminuta treated by method S included 16:0 (palmitic), 18:1 (octadecenoic) and 19 cyc (19 carbon cyclopropane acid). Those found in samples of S. mutans treated by the same method included 16:0, 18:1. 18:0 (stearic), 20:1 (eicosenoic), 20:0 (eicosanoic), 19 cyc and 21 cyc. When method T was used to process samples of both cultures, the cyc acids were degraded and artifacts were produced. Transesterification with boron trihalide reagents is therefore not recommended for routine analysis of bacterial fatty acids.
Mass spectra of n-butyl esters of selected biologically important short chain fatty acids were obtained by using the technique of gas-liquid chromatography-mass spectrometry. The results indicate that simple cleavage is responsible for the primary fragmentation of the molecules. The mass spectral data are considered to be advantageous for the identification of unknown short chain acids.
The cellular fatty acid composition of 112 reference strains and clinical isolates of Pseudomonas species was determined by gas-liquid chromatography (GLC). The presence and relative amounts of cyclopropane, hydroxy, and branched-chain fatty acids were distinguishing features of these strains. Determination of short-chain fatty acids extracted from spent growth media provided an additional means for identifying some strains. Our results show that clinical isolates of pseudomonads can be divided into eight distinct GLC groups. The procedures were especially useful for distinguishing glucose-nonoxidizing pseudomonads, which are difficult to identify by conventional criteria. Since the GLC procedures are simple, rapid, and highly reproducible, they are useful in diagnostic laboratories that process large numbers of cultures. Coupled with selected conventional tests, the analysis of short-chain and cellular fatty acids can be very useful for rapid screening of clinical isolates of Pseudomonas species.
The cellular fatty acid composition of 18 strains of Streptococcus mutans and 17 isolates of related streptococci were examined by gas-liquid chromatography. The cultures could be divided into two distinct groups on the basis of their fatty acids. The first group, which included S mutans and S salivarius, contained palmitic (16:0), octadecenoic (18:1), stearic (18:0), eicosenoic (20:1), and eicosanoic (20:0) acids. The presence of the two 20-carbon fatty acids distinguished S mutans and S salivarius from all of the other species. Two of the S mutans cultures were further distinguished by the presence of 19-carbon and 21-carbon cyclopropane acids. The second group included S sanguis, S mitis, S uberis, and a culture of Lancefield group C streptococci. The percentages of the major acids (16:0, 18:1, and 18:0) found in these species were essentially identical. A culture of Lancefield group E streptococci contained large amounts of 16:1 and 18:1 and small amounts of two unidentified acids which were not present in any of the other cultures.
The cellular fatty acid compositions of strains of Pseudomonas aeruginosa, P. cepacia, and P. maltophilia were determined by gas-liquid chromatography. A variety of acids was detected in these organisms, including branched and straight-chain acids, cyclopropane, and hydroxy acids. Comparison of the presence and relative amounts of these acids was useful in distinguishing the three species. Three branched-chain hydroxy acids not found in other bacteria were present in P. maltophilia and were identified by gas chromatography, mass spectrometry, and infrared spectroscopy.
The cultural characteristics, biochemical activity, and cellular fatty acid composition of Pseudomonas diminuta and Pseudomonas vesiculare provided means for differentiation of these closely related species. Broth cultures of P. diminuta showed turbid growth and a distinct surface pellicle after 24 h at 35 C. P. vesiculare had no pellicle, and only light, diffuse growth was observed. All strains of P. vesiculare oxidized maltose and hydrolyzed esculin to varying degrees; P. diminuta was negative in these tests. The cellular fatty acids of these two species were similar, except that P. diminuta possessed a C19 cyclopropane acid which was not detected in P. vesiculare.
The cellular fatty acid composition of 25 clinical isolates of Alcaligenes and Pseudomonas was determined by gas-liquid chromatography (GLC). The GLC fatty acid profiles of three species of Pseudomonas were markedly different from those of Alcaligenes. The most significant differences were the presence and relative amounts of hydroxy, branched-chain, and cyclopropane fatty acids. One of the major fatty acids in A. faecalis was a 17-carbon cyclopropane (17 delta) acid, whereas a 15-carbon branched-chain acid (13-methyl tetradecanoate) characterized isolates of P. putrefaciens. The determination of these fatty acids by GLC provides a rapid and specific means of distinguishing clinical isolates of Pseudomonas and Alcaligenes.
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A gas-liquid chromatographic procedure was used to determine short-chain acids produced by Pseudomonas diminuta and P. vesiculare after growth on Trypticase soy agar. Each of nine strains of P. diminuta produced glutaric acid, whereas none of the strains of P. vesiculare produced this acid.