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(4) Their isolation requires BCYE or BCYE-derived media, but these aren't routinely used.
For the isolation and counting of Legionella bacteria, 0.1 mL of the homogenized biofilm suspension was inoculated into BCYE agar supplemented with GVPN directly and after treatment with acid and heat as described above.
pneumophila isolates were cultured on BCYE agar, and their growth was confirmed using a serotyping kit.
The authors used a method which consisted of membrane filtration (0.2 [micro]m) with 1 L of potable water followed by aseptically cutting the membrane filter and vigorously shaking the filter homogenate for 15 min in 10 ml of phosphate buffer solution (PBS; pH 7.2) and plated on BCYE. This study uses the same method with the following modifications.
The gram-negative rod grew after 4 days of incubation on buffered-charcoal yeast extract (BCYE) agar.
The samples were decontaminated by acid treatment (0.2 KCl-HCl) followed by plating on buffered charcoal yeast extract agar (BCYE) (Oxoid, UK) and BCYE supplemented with polymyxin(80 u/ml), anisomycin (40 [micro]g/ml) and cefamandole (4 [micro]g/ml) to make BCYE selective for respiratory samples (13).
Strain was revived and maintained in Buffered Charcoal Yeast Extract agar (BCYE) (BD, Becton, Dickenson & Co.
A tracheal aspirate collected on day 14 plus postmortem lung tissue samples subsequently yielded colonies of a Legionella species on buffered-charcoal yeast extract (BCYE) agar (Oxoid Ltd., Basingstoke, England).
At 48 to 72 hours, F tularensis may form small (1 mm to 2 mm) colonies on 5% SBA, chocolate agar (CA), buffered charcoal yeast extract (BCYE) agar, and even Thayer-Martin (TM) agar.
All colonies from plates with [less than or equal to] 10 and 10-20 random colonies were subcultured on buffered charcoal yeast extract (BCYE) agar (with cysteine) and charcoal yeast extract agar (cysteine-free) media (Oxoid) for [greater than or equal to] 2 days.
Since Legionella is spread by aerosolized water, the organism can be recovered from the air environment by concentrating it with the N-6 impactor on culture media composed of buffered-charcoal yeast extract (BCYE) agar and BCYE agar plus polymyxin B, anisomycin, and vancomycin (BCYE/PAV).
When Gimenez-stained bacteria were observed, the culture was subcultured on BCYE agar (BioMerieux, Marcy L'Etoile, France) and sheep blood agar (BioMerieux) and on amoebal microplates without antibiotics or with vancomycin (10 [micro]g/mL) or colistin (500 U/mL).
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