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competing flora.6 BCYE w/ GPCV is also recommended by Centers for Disease Control and Prevention (CDC) for testing water samples.7 PRINCIPLE BCYE w/ GPCV contains charcoal and yeast extract to enhance the growth of Legionella. Charcoal also serves to absorb toxic metabolic products and modify the surface tension of the medium.
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1. Introduction. Legionella is a water-born pathogen widely spread in man-made water systems, responsible for a severe pneumonia and a flu-like illness, named Legionnaires' disease (LD) and the Pontiac fever, respectively. Overall, at the present time, 62 Legionella spp. have been identified and less than a half were pathogenic, however, Legionella pneumophila was surely the most frequently.
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Limitations of Buffered Charcoal Yeast Extract (BCYE) Agar It is recommended that biochemical, immunological, molecular, or mass spectrometry testing be performed on colonies from pure culture for complete identification.; It is also recommended that more than one type of medium is used for isolating Legionella species and that non-selective and selective BCYE Agar plates be inoculated in.
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BD BBL™ BCYE (Buffered Charcoal Yeast Extract) Agar is used for the isolation and cultivation of Legionella species. II PERFORMANCE TEST PROCEDURE 1. Inoculate representative samples with the cultures listed below. a. Add 0.1 mL of a culture containing 30-300 CFU/0.1 mL to each plate and spread-inoculate using a sterile glass spreader.
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BCYE Agar with GPVC (Cat. no. W169) contains glycine, vancomycin, polymyxin and cycloheximide for the greater inhibition of secondary microflora and further selective isolation of Legionella spp. BCYE Agar with CCVC (Cat. no. G170) contains cephalothin, colistin, vancomycin, and cycloheximide. This is based on the formulation of Bopp et al.(14.
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Later on, Pasculle et al. supplemented the CYE medium with N-(2-Acetamido)-2-aminoethanesulfonic acid (ACES) buffer, thereby obtaining buffered charcoal yeast extract agar (BCYE) which, under aerobic conditions, enabled a better recovery of Legionella , and Edelstein further increased this medium's sensitivity by adding α-ketoglutarate (i.e.
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Colony Characteristics. After 3-4 days of incubation, small white colonies of Legionella appear in BCYE agar which becomes grey in a few days. The colonies are round and convex with entire edges. The isolate may show a typical ground glass (or iridescent) colony appearance.. Colonies may be examined under long-wave UV light (360 nm ± 20nm) to reveal yellow pigment on BCYE or auto-fluorescence.
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Buffered charcoal yeast extract (BCYE) agar is a selective growth medium used to culture or grow certain types of bacteria, particularly the Gram-negative species Legionella pneumophila. It has also been used for the laboratory diagnosis of Acanthamoeba keratitis, Francisella and Nocardia spp.It contains L-cysteine amino acid and ferric pyrophosphate that assist in the growth of Legionnaire's.
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Pour Tube: Melt the agar in a boiling water bath and cool to 45-50°C. Add BCYE Supplement (R45005) and mix thoroughly. The following selective supplements may also be added to the cooled base: PAC (R45006) or PAV (R45007) Supplement. Dispense melted agar into a sterile Petri dish, allow it to harden, and proceed with the procedure above.