Separation of bacterial ubiquinones by reverse-phase high-pressure liquid chromatography

Moss, C W; Guerrant, G O

doi:10.1128/jcm.18.1.15-17.1983

Cited by 33 publications

(22 citation statements)

References 7 publications

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“…Ubiquinone analysis. Ubiquinones were extracted by the method of Moss and Guerrant (12), with the following modifications. Isolates were grown on two BCYE agar plates for 3 to 4 days.…”

Section: Methodsmentioning

confidence: 99%

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Problems associated with identification of Legionella species from the environment and isolation of six possible new species

Wilkinson

Sangster

Ratcliff

et al. 1990

Appl Environ Microbiol

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Following investigation of an outbreak of legionellosis in South Australia, numerous Legionella-like organisms were isolated from water samples. Because of the limited number of commercially available direct fluorescent-antibody reagents and the cross-reactions found with some reagents, non-pneumophila legionellae proved to be difficult to identify and these isolates were stored at-70°C for later study. Latex agglutination reagents for Legionella pneumophila and Legionella anisa developed by the Institute of Medical and Veterinary Science, Adelaide, Australia, were found to be useful as rapid screening aids. Autofluorescence was useful for placing isolates into broad groups. Cellular fatty acid analysis, ubiquinone analysis, and DNA hybridization techniques were necessary to provide definitive identification. The species which were isolated most frequently were L. pneumophila, followed by L. anisa, Legionella jamestowniensis, Legionella quinlivanii, Legionella rubrilucens, Legionella spiritensis, and a single isolate each of Legionella erythra, Legionellajordanis, Legionella birminghamensis, and Legionella cincinnatiensis. In addition, 10 isolates were found by DNA hybridization studies to be unrelated to any of the 26 currently known species, representing what we believe to be 6 possible new species.

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“…Ubiquinone analysis. Ubiquinones were extracted by the method of Moss and Guerrant (12), with the following modifications. Isolates were grown on two BCYE agar plates for 3 to 4 days.…”

Section: Methodsmentioning

confidence: 99%

“…Ubiquinones were detected at 275 nm. Peaks were said to be ubiquinones if the peak height at 275 nm was significantly greater than at 248 nm (12). The chain lengths were determined by a comparison of the retention times with that of commercial standards Q6, Q9, and Q10, supplied by Sigma Chemical Co. DNA relatedness studies.…”

Section: Methodsmentioning

confidence: 99%

Problems associated with identification of Legionella species from the environment and isolation of six possible new species

Wilkinson

Sangster

Ratcliff

et al. 1990

Appl Environ Microbiol

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“…After 48 h of incubation, the cells were scraped from three plates (100 by 15 mm) in sterile, distilled water and placed into a screw-capped test tube (20 by 150 mm) fitted with a Teflon-lined cap. The quinones were extracted after saponification and analyzed by reverse-phase thin-layer chromatography and high-performance liquid chromatography as described previously (10,15).…”

Section: Methodsmentioning

confidence: 99%

Legionella sainthelensi: a new species of Legionella isolated from water near Mt. St. Helens

Campbell¹,

Bibb²,

Lambert³

et al. 1984

Appl Environ Microbiol

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Six strains of a new species, Legionella sainthelensi, were isolated from freshwater in areas affected by the volcanic eruptions of Mt. St. Helens in the state of Washington. Strains of L. sainthelensi are culturally and biochemically similar to other legionellae. They grow on buffered charcoal yeast agar but not on media that lack cysteine. They are gram-negative, nonsporeforming, motile rods that are positive in reactions for catalase, oxidase, gelatin liquefaction, and beta-lactamase. They are negative in reactions for urease, hydrolysis of hippurate, reduction of nitrates, fermentation of glucose, and blue-white autofluorescence. Their cell wall fatty acid composition is qualitatively similar to those of other legioneilae, with 50 to 62% branched-chain fatty acids. They contain the isobranched-chain 14and 16-carbon acids and anteisobranched-chain 15and 17-carbon acids and relatively large amounts of straight-chain 16-carbon acid. All strains of L. sainthelensi contain approximately equal amounts of ubiquinones Q9, Q10, Qil, and Q12, a pattern similar to those of Legionella bozemanii, Legionella dumoffii, and Legionella longbeachae.

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“…Identification of fatty acids and determination of double-bond positions in monounsaturated acids were accomplished by gas-liquid chromatography and gas-liquid chromatography-mass spectrometry (30). Isoprenoid quinones were extracted from 100 mg of lyophilized cells and analyzed by reverse-phase high-performance liquid chromatography (29).…”

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confidence: 99%

Bordetella holmesii sp. nov., a new gram-negative species associated with septicemia

et al. 1995

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CDC nonoxidizer group 2 (NO-2) currently consists of 15 gram-negative, rod-shaped, oxidase-negative, asaccharolytic, brown soluble pigment-producing strains isolated from blood cultures, usually from young adults. On the basis of their cellular fatty acid profiles, NO-2 strains formed a single group that was identical with the profile of Bordetella avium. 16S rRNA sequencing of one NO-2 strain and the type strains of B. pertussis, B. parapertussis, B. bronchiseptica, and B. avium showed a high degree of homology (Ն98% over 1,525 bases). The NO-2 guanine-plus-cytosine content (61.5 to 62.3 mol%) and major ubiquinone analysis (ubiquinone-8) results were both consistent with those for the genus Bordetella. DNA relatedness studies (hydroxyapatite method) confirmed a close relatedness between NO-2 and Bordetella species and demonstrated that NO-2 strains were a single new species. The name B. holmesii sp. nov. is proposed for CDC group NO-2. CDC nonoxidizer group 2 (NO-2) is the vernacular name given to gram-negative, nonoxidizing, brown soluble pigmentproducing rods sent for identification to the Special Bacteriology Reference Laboratory at the Centers for Disease Control and Prevention (CDC). The first of these strains was received in 1983. Since then, 14 additional strains have been received, all of which were isolated from blood cultures obtained, with few exceptions, from young adults. At least four of these strains were isolated from more than one blood culture per patient. Eleven NO-2 strains, including two isolates discovered outside the United States, have been received since 1989, raising the possibility that this group is an emerging pathogen. In this study we characterized the strains of NO-2 biochemically, by cellular fatty acid (CFA) and ubiquinone analysis, guanine-plus-cytosine (GϩC) content, 16S rRNA sequence analysis, and DNA-DNA hybridization. The results indicate that group NO-2 is a single, previously undescribed species whose closest relatives are in the genus Bordetella. We propose the name Bordetella holmesii for NO-2. MATERIALS AND METHODS Bacterial strains and culture conditions. The NO-2 strains and their sources are shown in Table 1. Bordetella pertussis ATCC 9797 T , Bordetella parapertussis ATCC 15311 T , Bordetella avium ATCC 35086 T , and Bordetella bronchiseptica ATCC 19395 T were obtained from the American Type Culture Collection, Rockville, Md. In addition, CFA profiles were determined for B. parapertussis F6288 and F6289, B. bronchiseptica B5821 and G1696, and B. pertussis 042, 083, and 087. The B. parapertussis and B. bronchiseptica strains were submitted to CDC as patient isolates and were identified by the Special Bacteriology Reference Laboratory. The B. pertussis strains were obtained from P. Cassiday, Pertussis Laboratory, Division of Bacterial and Mycotic Diseases, CDC. All strains were suspended in defibrinated rabbit blood and stored in a liquid nitrogen freezer until studied. Unless otherwise indicated, all strains except B. pertussis were grown on heart infusion agar with...

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Separation of bacterial ubiquinones by reverse-phase high-pressure liquid chromatography

Abstract: A procedure was developed for the separation of ubiquinones by high-pressure liquid chromatography on a reverse-phase C18 column. Ubiquinones Q6 through Q14 were resolved in 20 min and were distinguished from menaquinones by comparing UV spectra at 248 and 275 nm.

Cited by 33 publications

References 7 publications

Problems associated with identification of Legionella species from the environment and isolation of six possible new species

Problems associated with identification of Legionella species from the environment and isolation of six possible new species

Legionella sainthelensi: a new species of Legionella isolated from water near Mt. St. Helens

Bordetella holmesii sp. nov., a new gram-negative species associated with septicemia

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