A Co-operative Numerical Analysis of Nonscoto- and Nonphotochromogenic Slowly Growing Mycobacteria

Meißner, Gertrud; Schröder, K. H.; G, Amadio; Anz, W.; Chaparas, Sotiros D.; Engel, H. W. B.; Jenkins, P. A.; Käppler, W; Kleeberg, H. H.; Kubala, E; Kubín, M; Lauterbach, D; Lind, A; Magnusson, M.; Miková, Z; Pattyn, S. R.; Schaefer, Werner B.; Stanford, J. L.; Tsukamura, M; Wayne, Lawrence G.; Willers, I.; Wolinsky, Emanuel

doi:10.1099/00221287-83-2-207

Cited by 77 publications

(35 citation statements)

References 18 publications

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“…Wayne (47) showed subsequently that M. avium and the Battey bacillus were not separable from each other. Meissner et al (18) made the same observation. Marks et al (16) showed the similarity of the lipid composition of M. avium, M. intracellulare, and M. scrofulaceum.…”

Section: Growth On Medium Containing 02\ P-emrnosalicylatesupporting

confidence: 69%

Numerical Classification of 280 Strains of Slowly Growing Mycobacteria

Tsukamura

1983

Microbiology and Immunology

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. nonchromogenicum, M. terrae, "M. novum," and M. triviale. Each of these clusters appeared to be reduced to a single species, M. avium Chester 1901 (7) and M. nonchromogenicum Tsukamura 1965 (33), respectively which have priority in each group. However, considering the wide use of the names of the species of these groups, Tsukamura (40) proposed that the names the M. avium complex and the M. nonchromogenicum complex be used for these clusters. Tsukamura et al (43,44) observed also that M. tuberculosis, 315

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Section: Growth On Medium Containing 02\ P-emrnosalicylatesupporting

confidence: 69%

Numerical Classification of 280 Strains of Slowly Growing Mycobacteria

Tsukamura

1983

Microbiology and Immunology

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“…nonchromogenicum and M . terrae were considered to be closely related but of two different species (18). The results of our study suggest that these are differentiable by their patterns, although the patterns are of considerable similarity.…”

Section: Discussionmentioning

confidence: 41%

“…nouum was considered to be a synonym of M . terrae in an IWGMT study (18), these two showed slightly different patterns (Fig. 5).…”

Section: Discussionmentioning

confidence: 99%

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Differentiation Among Mycobacterial Species by Thin-Layer Chromatography

Tsukamura

Matsumoto

1975

International Journal of Systematic Bacteriology

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Thin-layer chromatography of the ethyl ether-ethanol-soluble fraction after incubation with ["S Imethionine is useful for differentiation among mycobacterial species. Scanning of the radioactivity on chromatograms showed various distribution patterns of radioactive spots, and the patterns were, in most cases, characteristic for the species. bouis showed significantly lower activity than the other mycobacteria. The present study was designed to develop the previous method into a more reliable one. MATERIALS AND METHODSThe strains used are listed in Table 1. Most of the strains were identified through cooperative studies of the International Working Group on Mycobacterial Taxonomy (IWGMT) (11,18,35). Some strains were identified in this laboratory by methods described previously (29, 30). The organisms were grown in Ogawa egg medium a t 37 C for 5 days (rapidly growing mycobacteria, M . phlei through M . flauescens in Table 1) or 14 days (slowly growing mycobacteria, M . kansasii through M . gordonae in Table 1). The cells were washed three times in saline and then weighed; 100-mg amounts (wet weight) of the cells were suspended in 2.0 ml of 0.067 M phosphate buffer solution (pH 7.1) containing 5 pCi of ~-[~~S ] m e t h i onine and 10 pg of sodium acetate per ml. The tubes containing the mixture were stdppered with cotton plugs and incubated a t 37 C for 20 h. After incubation, the cells were centrifuged a t 500 x g for 15 min and washed twice with 5.0 ml of chilled water. The cells were extracted twice with 2.0 ml of 10% trichloroacetic acid, each for 10 min. The acid extracts were separated from the cells by centrifugation and discarded. The cells were then extracted twice with 3.0 ml of ethyl ether-ethanol (l:l, vol/vol) for 10 min each. During the first 5 min of the extraction, the cells were mixed continuously with a glass stick a t room temperature (25 + 3 C); during the second 5 min, the tubes were allowed to stand in an incubator a t 37 C, during which time the cells sedimented. The extracts could then be separated easily by means of a pipette. The combined ethyl ether-ethanol-soluble fractions were labeled as "lipid fraction." The residue was dissolved in 2.0 ml of a 1% NaOH solution by heating a t 100 C in a water bath for 5 min.~-[~~S ] m e t h i o n i n e was a product of New England Nuclear Corp., Boston, Mass. (specific activity, 264Ci/mmol; radiochemical purity, > 93%; containing < 7.0% methionine sulfoxide, <0.2% methionine sulfone, and <0.2% homocysteic acid). Ethyl ether was a product of Katayama Chemical Company, Osaka, Japan (specific gravity, below 0.720; fraction [34 to 35 C], minimum 95% [by volume]; water, maximum 0.3%; nonvolatile, maximum 0.002%; acidity [CH,COOH], maximum 0.003%). Ethyl alcohol also was a product of Katayama Chemical Company, Osaka, Japan (bp, 78 C; specific gravity, below 0.797; nonvolatile, maximum 0.003%; acidity [CH,COOH], maximum 0.003%; heavy metals p b ] , maximum O.O002%; ketones [CH, -CO .CH,], maximum 0.002%; assay by specific gravity, minimum 99.46% [by volume...

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“…In neither study did such tests meet our criteria for inclusion in the published reports. Nevertheless, tests for the ability of an organism to grow a t 25 and 45°C appear to provide useful information (4). As indicated in Results, the poor reproducibility observed in some laboratories appeared to reflect inadequate control of temperatures.…”

Section: Discussionmentioning

confidence: 91%