Determination of the Gram Type Using the Reaction Between Polymyxin B and Lipopolysaccharides of the Outer Cell Wall of Whole Bacteria

Wiegel, Juergen; Quandt, L.

doi:10.1099/00221287-128-10-2261

Cited by 24 publications

(22 citation statements)

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“…Such behavior also is known for some other clostridia (19). Thus, C. thermobutyricum is classified as Gram type positive as defined by Wiegel(l8) and Wiegel and Quandt (19). The organism has an m-Dpm-direct cell wall type (A17 peptidoglycan type) (N. Weiss, personal communication).…”

Section: Resultsmentioning

confidence: 99%

“…The following properties indicate that the new isolate is a member of the genus Clostridium: rods forming endospores, strictly anaerobic growth, absence of dissimilatory sulfate reduction, and positive Gram type (18,19) (including testing of polymyxin B-lipopolysaccharide interaction) of cell walls. The positive Gram type differentiates the organism from the gram type-negative sporeforming genus Sporomusa (13).…”

Section: Resultsmentioning

confidence: 99%

“…Micrographs of ultrathin sections show a slightly atypical grampositive type of cell wall. The polymyxin B-lipopolysaccharide test (19) gives no indication of the presence of lipopolysaccharides. Such behavior also is known for some other clostridia (19).…”

Section: Resultsmentioning

confidence: 99%

“…The polymyxin B-lipopolysaccharide test (19) gives no indication of the presence of lipopolysaccharides. Such behavior also is known for some other clostridia (19). Thus, C. thermobutyricum is classified as Gram type positive as defined by Wiegel(l8) and Wiegel and Quandt (19).…”

Section: Resultsmentioning

confidence: 99%

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Clostridium thermobutyricum sp. nov., a Moderate Thermophile Isolated from a Cellulolytic Culture, That Produces Butyrate as the Major Product

Wiegel

Kuk

Kohring

1989

International Journal of Systematic Bacteriology

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A new moderately thermophilic clostridium, Clostridium thermobutyricum, was isolated from cellulolytic enrichment cultures inoculated with horse manure. This organism forms subterminal spores, and the sporangium is not swollen. From glucose, the organism produces butyrate, CO,, and H, and minor amounts of acetate and lactate. Yeast extract is required for good growth. The temperature range for growth is between 26 and 61.5OC. The pH range is between 5.8 and 9.0. The guanine-plus-cytosine content is 37 mol%. The type strain is C. thermobutyricum JW171K (= DSM 4928).During the enrichment and isolation of cellulolytic thermophilic anaerobes, butyrate frequently is encountered as a fermentation product. For more than two decades, it was thought that butyrate was an end product of the cellulose degrader Clostridium thermocellum (4, 7, 12). However, when pure cultures were finally obtained, it was found that the butyrate was formed by contaminating organisms. Thus, pure cultures of the known cellulolytic thermophilic clostridia, C. thermocellum and Clostridium stercorarium, do not produce butyrate. In 1949, Enobo (5) described another thermophilic cellulose degrader, which he named "Clostridium thermocelulaseum . " From his cellulolytic enrichment culture he also isolated a butyrate-forming glycolytic organism (5). Unfortunately, neither of his cultures is extant. The pH optimum of the cellulolytic strain of Enebo was above pH 8.0. We tried to isolate such an organism by using various soil and compost samples as inocula. Only the sample from horse manure produced an enrichment culture which degraded cellulose quickly and formed butyrate as the major product. After several dilution steps, the enrichment culture contained two organisms. One was a cellulolytic clostridium which produced the drumstick type of sporangium typical of C. thermocellum. The second clostridium formed central to subterminal spores with little or no swelling of the cells. We present here a formal description of this organism, a new Clostridium species that produces butyric acid as the major fermentation product under moderately thermophilic growth conditions. MATERIALS AND METHODSIsolation and culture conditions. Samples (2 to 10 g, wet weight) of horse manure from a manure pile at the University of Georgia horse stable and from private horse stables were inoculated into 50-ml portions of reduced medium in 150-ml serum bottles (Wheaton 400 brand borosilicate glass; Fisher Scientific Co., Pittsburgh, Pa.). These enrichment cultures were incubated at 59 to 60°C. The medium contained (per liter of distilled water) 0.9 g of KH,PO,, 3.6 g of Na,PO, . 7H20, 0.5 g of NH,Cl, 0.5 g of (NH,),SO,, 0.18 g * Corresponding author. of MgCl, 6H,O, 0.05 g of CaCl,, 5 ml of a trace element solution (7), 0.5 ml of a vitamin solution (modified Wolfe solution [7]) 0.2 g of Na,S 9H20, 0.2 g of cysteic acid, 3 g of yeast extract, and either 10 g of filter paper (Kimwipes; Kimberly Clarke C o p , Roswell, Ga.) (during early enrichment) or 5 g of glucose (during purificat...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Clostridium thermobutyricum sp. nov., a Moderate Thermophile Isolated from a Cellulolytic Culture, That Produces Butyrate as the Major Product

Wiegel

Kuk

Kohring

1989

International Journal of Systematic Bacteriology

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“…The motility of T. pallidum was unaffected by concentrations of polymyxin B more than ten times higher than are lethal for "smooth" strains of Salmonella typhirnurium (Teuber, 1974). Moreover, the effects of polymyxin B include induction of morphological changes in the outer membrane (Wiegel and Quandt, 1982). Comparison of the ultrastructure of polymyxin-treated T. pallidurn and S .…”

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confidence: 99%

The Eighth C. L. Oakley Lecture: Pathogenicity and immunobiology of Treponema pallidum

Penn

1987

Journal of Medical Microbiology

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Coding Microbiological Data for Computers

Rogosa

Krichevsky

Colwell

1986

Springer Series in Microbiology

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Determination of the Gram Type Using the Reaction Between Polymyxin B and Lipopolysaccharides of the Outer Cell Wall of Whole Bacteria

Cited by 24 publications

References 40 publications

Clostridium thermobutyricum sp. nov., a Moderate Thermophile Isolated from a Cellulolytic Culture, That Produces Butyrate as the Major Product

Clostridium thermobutyricum sp. nov., a Moderate Thermophile Isolated from a Cellulolytic Culture, That Produces Butyrate as the Major Product

The Eighth C. L. Oakley Lecture: Pathogenicity and immunobiology of Treponema pallidum

Coding Microbiological Data for Computers

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