Cultivation on solid media of spirochetes in subgingival plaque from advanced marginal periodontitis in humans

Fiehn, Nils‐Erik; Westergaard, Jytte

doi:10.1111/j.1600-0722.1984.tb00912.x

Cited by 7 publications

(7 citation statements)

References 21 publications

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“…Earlier cultivation studies have revealed great diversity in the nutrient requirements for various types of spirochetes (20). Today media promoting growth of most of the known species of oral spirochetes exist (5), but only a low recovery has been obtained. However, no single medium, which can be used to isolate all the different types of oral spirochetes, is available.…”

Section: Discussionmentioning

confidence: 99%

“…samples small-sized spirochetes with one to three endoflagella were among the predominating kinds (5).…”

mentioning

confidence: 99%

“…solid media were: BHIA-SC and MGM-1, both with 5% horse blood and with the following antibiotics: Rifampicin (Sigma) 2 Mg/ml and polymyxin B (Sigma) 800 U/ml (5).…”

mentioning

confidence: 99%

“…The purpose of this study is to elucidate in detail the effect on growth of small-sized spirochetes of 1) the oxygen tension in the atmosphere, 2) the pH in the medium, 3) the concentration of rabbit serum in the medium, 4) substitution of rabbit serum in the medium with bovine serum, human serum, different serum components, or filtrates from various bacterial cultures. solid media were: BHIA-SC and MGM-1, both with 5% horse blood and with the following antibiotics: Rifampicin (Sigma) 2 Mg/ml and polymyxin B (Sigma) 800 U/ml (5).…”

mentioning

confidence: 99%

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Nutrient and environmental growth factors for eight small‐sized oral spirochetes

Fiehn

Westergaard

1986

European J Oral Sciences

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Fiehn N-E, Westergaard J. Nutrient and environmental factors for eight small-sized oral spirochetes. Scand J Dent Res 1986; 94: 208-18. Abstract -The present investigation was carried out in order to obtain better information about the growth requirements for small-sized oral spirochetes containing two endoflagella from each cell-end. Eight strains of such spirochetes were isolated from subgingival plaque in patients suffering from advanced marginal periodontitis. The strains were maintained under anaerobic conditions in a fluid basal BHI medium with 15% inactivated rabhit serum, 0.07% Noble Agar and 5 |lg/ml cocarboxylase. Firstly, the effect of trace amounts of oxygen in the atmosphere and pH in the medium on growth of the spirochete strains were examined. Secondly, the effect of different sera incorporated in the medium was examined, and thirdly, the effect of itnportant growth factors in serum was studied hy adding different serum components to the fluid basal medium instead of rabbit serum. Growth was always determined after 4 days' incubation at 35°C, either by counting numbers of spirochete cells in a Petroff-Hauser counting chamber or by measuring the turbidity of the culture spectrophotometrically at 600 nm. There was no difference in growth by tising an atmosphere containing 1 % oxygen or an anaerobic atmosphere. It was found that serum (rabhit or human) was an essential growth component, and no single growth factor could replace rabhit serum. Only a long chain fatty acid mixture and an amino acid solution could, to a minor extent, stimulate growth compared to the basal medium without rabbit serum. Sodium bicarbonate inhibited growth of all strains. Finally, none of the strains fermented a series of low molecular weight carbohydrates, but all strains produced HjS and indole.Normally spirochetes make up a substantial to some extent contradictory (1,3,4). Recent part of subgingival plaque at diseased sites observations indicate that the distribution of in individtjals with adult marginal periodon-different types of spirochetes in subgingival titis (1,2). The results from different investi-plaque is extremely variable and cliaractergationsregarding the types of spirochetes are istic of the specific sample site (5). In most

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Section: Discussionmentioning

confidence: 99%

“…samples small-sized spirochetes with one to three endoflagella were among the predominating kinds (5).…”

mentioning

confidence: 99%

“…solid media were: BHIA-SC and MGM-1, both with 5% horse blood and with the following antibiotics: Rifampicin (Sigma) 2 Mg/ml and polymyxin B (Sigma) 800 U/ml (5).…”

mentioning

confidence: 99%

mentioning

confidence: 99%

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Nutrient and environmental growth factors for eight small‐sized oral spirochetes

Fiehn

Westergaard

1986

European J Oral Sciences

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“…Another advantage of this method was that spirochetes could be cultured without using antibiotics. Leschine and Canale-Parola (8) firstly used rifampicin to select for oral spirochetes, and many studies used media containing antibiotics such as rifam-picin and polymyxin B (6,15,23) to select for spirochetes. Such selective agents clearly reduced the numbers of culturable spirochetes (Table 3).…”

Section: Discussionmentioning

confidence: 99%

Improved detection of oral spirochetes with an anaerobic culture method

Umeda¹,

Ishikawa²,

Benno³

et al. 1990

Oral Microbiology and Immunology

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A method for routine culturing of oral spirochetes was studied in periodontitis patients and subjects with healthy gingivae. Subgingival plaque bacteria, including oral spirochetes were cultured anaerobically using the steel wool jar method, the glove-box procedure and the plate-in-bottle method. They were also observed directly with a phase-contrast microscope. The number of colonies of recovered spirochetes was significantly higher with the plate-in-bottle method when Medium 10 (M10) or M10 supplemented with 10% rumen fluid was used. In almost all cases, spirochetes were detected with this culture method. The ratios of spirochetes from patient samples, cultured by the plate-in-bottle method, and of other microorganisms, cultured in Eggerth-Gagnon (EG) medium by the steel wool jar method, were almost identical of those observed by phase-contrast microscopy. These results indicate that it can be just as possible to culture certain oral spirochetes as other anaerobic bacteria.

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