Comparison of there anaerobic culture techniques and media for viable recovery of subgingival plaque bacteria

Abstract

– Various methods are available for viable recovery of bacteria from subgingival plaque, but theire relative efficicency is not clear. In the first experiment, with 10 patients, three anaerobic techniques (Brewer jars, chamber and roll tubes) and three agar media (trypicase soy with 5% sheep blood (TS), brucella agar with 5% blood and 0.5 ug/ml of menadione (B), and modified medium 10 with 3% blood (MM 10)), were compared. In the second experiment, with anaother 10 patients, Brewer jars were compared with a glove box, using TS. Subgingival plaque was sampled with a gas flushed broach passed through a syringe. Significance of differences in viale recoveries was determined by ANOVA. Brewer jars and chamber (aerobic plating) were comparable in efficiency, but excelled roll tubes (anaerobic inoculation). TS was better than B, but equivalent to MM 10. TS in Brewer jars (aerobic plating) matched TS in the glove box (anaerobic plating), but perforemed better than all the other procedures tested. Recoveries did not differ on pre‐and non‐prereduced plates and on commercial and laboratory, freshly made platels. Non‐evacuated jars sealed inside the box (anaerobic plating) provided lower recoveries than jars evacuated, filled and sealed outside (aerobic plating). Roll tubes benefited most by extending incubation from 7 to 14d. Commercial, non‐prereduced TS agar plates incubated in Brewer jars or a glove box are highly recommendable methods for processing of subgingival plaque.

Rapid identification of important periodontal microorganisms by cultivation

“…A nutnber of investigators have shown that strict anaerobic incubation systems, such as roll tubes and anaerotiic chambers, yielded higher recovery rates of oral bacteria than conventional anaerobic jars (1,5,18,24,34). Roll tubes and anaerobic chambers seem to possess similar capacity for recovering oral bacteria (15,20 VMG II>RTF>SBL VMG II>RTF>RSS ' After 10 days of storage in VMG II at refrigeration temperature, some streptococcal species were reeovered in higher numbers than at initial culturing. number of samples and some reveal odd samples which show the higher cultivable counts in the overall least efficient culture system.…”

“…The primary incubation time is also a determinant of total colony counts. Slots (24) and Olsen and Socransky (20) showed that after an incubation period of 5-7 days, which is common (Table 1), as many as 7% of the oral isolates present after 14 days were not detected.…”

Porphyromonas gingivalis Lipid A Phosphatase Activity Is Critical for Colonization and Increasing the Commensal Load in the Rabbit Ligature Model

“…gingivalis wild type (WT). P. gingivalis (strain A7436 [14]) was grown using standard procedures as described previously (15)(16)(17)(18). Briefly, bacteria were cultured on agar plates containing Trypticase soy agar supplemented with 0.5% (wt/vol) yeast extract (Invitrogen Life Technologies), 5% defibrinated sheep red blood cells, 5 g of hemin, and 1 g of vitamin K (Sigma-Aldrich)/ml.…”