Indol and Skatol Determination in Bacterial Cultures

Fellers, C. R.; Clough, Ray W.

doi:10.1128/jb.10.2.105-133.1925

Cited by 31 publications

(13 citation statements)

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“…However, only a few strains have been reported to produce 3-MI from tryptophan. Recently, two strains of ruminal bacteria have been isolated which produce 3-MI in the absence of IAA (Attwood et al 2006) in rich medium, however, Clostridium scatologenes ATCC 25775 is the best studied bacterial strain known to be capable of 3-MI production from tryptophan (Fellers and Clough 1925;Elsden et al 1976;Hilton 1978, 1979;Jensen et al 1995;Kusel et al 2000;Liou et al 2005). It is an objective of this laboratory to elucidate regulatory factors which influence 3-MI production in axenic bacterial cultures as this information could lead to a greater understanding of 3-MI production in animal wastes.…”

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confidence: 99%

3-Methylindole production is regulated inClostridium scatologenesATCC 25775

Doerner

Cook

Mason

2009

Letters in Applied Microbiology

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Aims: 3‐Methylindole (3‐MI) is a degradation product of l‐tryptophan and is both an animal waste malodorant and threat to ruminant health. Culture conditions influencing 3‐MI production in Clostridium scatologenes ATCC 25775 were investigated. Methods and Results: Extracellular 3‐MI levels in cells cultured in brain heart infusion (BHI) medium (pH 7·0) at 33°C and 37°C for 72 h were 907 ± 38 and 834 ± 121 μmol l−1, respectively. Cells cultured in tryptone‐yeast (TY) extract medium at 37°C for 48 h produced 104 ± 86 μmol l−1 3‐MI; however, addition of 1 mmol l−1 l‐tryptophan failed to increase extracellular levels (113 ± 50 μmol l−1 3‐MI). Specific activity of indole acetic acid decarboxylase measured in BHI, TY and TY plus 1 mmol l−1 tryptophan‐grown cells displayed 35‐, 33‐ and 76‐fold higher levels than in semi‐defined medium‐grown cells. Conclusions: When cultured in rich medium, at 33°C or 37°C and pH 7·0, Cl. scatologenes ATCC 25775 optimally produced 3‐MI. Addition of l‐tryptophan to medium did not lead to significant increases in extracellular 3‐MI levels. Whole cell assays indicate growth in rich medium significantly up‐regulated 3‐MI production. Significance and Impact of the Study: Information presented here may prove useful in understanding what factors influence 3‐MI production in malodorous animal wastes.

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confidence: 99%

3-Methylindole production is regulated inClostridium scatologenesATCC 25775

Doerner

Cook

Mason

2009

Letters in Applied Microbiology

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“…Since several named taxa and unnamed biovars of flavobacteria, but not of other nonfermenters, are indole positive, a positive test can promote early identification of a clinical isolate. Numerous procedures have been described for the indole test (7,19,20,23,26,36,37). Several of these were examined.…”

Section: Resultsmentioning

confidence: 99%

Methods for identification of flavobacteria

Pickett

1989

J Clin Microbiol

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reports disagree on the best features for detecting and distinguishing between Flavobacterium meningosepticum (biovar HIa) and Flavobacterium species CDC group IIb (biovar IIb; Flavobacterium indologenes). This report discloses that at least some of these disagreements may reflect the methods used. To detect production of indole, a modified Kovacs reagent (not Ehrlich) and a buffered tryptophan medium were optimal, but not all strains of these two biovars produced indole. To distinguish the two biovars, hydrolysis of corn starch was preferable to that of soluble potato starch. Both biovars may hydrolyze DNA; the differentiation achieved varied with the methods used. Both biovars presented pigmented growth; only IIb, however, was obviously pigmented on a 2-day blood agar plate. Acidification of D-arabinose definitively distinguished these two biovars; several additional features were useful but not definitive. Several reports on glucose-and indole-positive biovars of flavobacteria appear to disagree on their features (5, 10, 14, 27-29, 31, 41). Hence, several methods for determining these features, particularly pigmentation, formation of indole, and hydrolysis of DNA, starch, and urea, were examined. Optimal methods for discriminating between Flavobacterium meningosepticum (biovar IIa) and the Centers for Disease Control Flavobacterium species group IIb (biovar IIb) were then used to characterize several flavobacteriumlike isolates. MATERIALS AND METHODS Except as noted otherwise, all incubations were at 35°C, all biochemical test media were dispensed in 3-ml amounts into screw-cap tubes (13 by 100 mm), and all heat sterilizations were at 121°C for 15 min. Bacterial strains examined were obtained from B. Holmes (Central Public Health Laboratory, London, England), G. L. Gilardi (North General Hospital, New York, N.Y.), R. E. Weaver (Centers for Disease Control, Atlanta, Ga.), and local hospitals. Colony morphology, hemolysis, and pigmented growth were recorded from 24-and 48-h blood agar plates (Trypticase soy agar basal medium with 5% sheep blood; BBL Microbiology Systems, Cockeysville, Md.). Pigmentation was also recorded after an additional 48 h at room temperature. Both soluble and insoluble pigments were noted with modified King medium B (21) incubated at 25°C, the modification being supplementation of the medium with 0.2% KNO3 and 0.1% L-tryptophan (KBNT). Tests for the flexirubin type of pigment were made by adding 1 drop of 3% KOH to a bit of cell paste; a change from yellow-orange to red was recorded as positive (41). Except as noted otherwise, all tests for acidification or alkalinization of substrates were made with aerobic low-peptone (ALP) basal media (12). These basal media contained 0.1% (NH4)2HP04, 0.02% KCl, 0.02% MgSO4. 7H20, 0.05% yeast extract (Difco Laboratories, Detroit, Mich.), 0.05% Casitone (Difco), 0.002% phenol red, and 1.5% agar. Basal medium for sugars (carbohydrates, alcohols, and other acidogenic substrates) was adjusted to pH 7.8. Basal medium for salts (salts of organic acids, gelat...

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“…Qualitative tests for indole were made on cultures after 1, 2, 5 and 10 days' incubation with Fellers and Clough's (1925) modification of Ehrlich's reagent.…”

Section: Indole Formationmentioning

confidence: 99%

Nitrate, Nitrite and Indole Reactions of Gas Gangrene Anaerobes

Reed

1942

J Bacteriol

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In an attempt to work out a scheme for the rapid identification of the gas gangrene organisms (Reed and Orr, 1941), many of the published reports of nitrate reduction and indole formation by this group of organisms were found to be contradictory. The nature of the contradictions suggested that this was largely the result of faulty interpretation of results so clearly described for the nitrate reduction of bacteria in general by Conn (1936). NITRATE REDUCTION Woods (1938) found that washed suspensions of Clostridium welchii catalyzed the reduction of NOs, NO2 and NH20H to NH3 by molecular hydrogen. Moreover, during the reduction of NO3 to NH3 he demonstrated the appearance and disappearance of NO2. This made it probable that tests for NO2 alone in growing cultures of this and related species would give little indication of the nitratereducing action of the organisms. A somewhat more detailed examination has therefore been made. Cultures were tested for nitrite production in the following medium, Reed and Orr (1941):

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Indol and Skatol Determination in Bacterial Cultures

Cited by 31 publications

References 9 publications

3-Methylindole production is regulated inClostridium scatologenesATCC 25775

3-Methylindole production is regulated inClostridium scatologenesATCC 25775

Methods for identification of flavobacteria

Nitrate, Nitrite and Indole Reactions of Gas Gangrene Anaerobes

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