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The Legionella BCYE Agar Base and its supplements have been shown to be optimal for Legionella culture with shorter incubation periods from environmental and clinical samples. Feeley et al. described a modification of F-G Agar in which acid hydrolised casein was replaced by yeast extract as the source of protein, and starch was replaced by activated charcoal. This medium, which they named CYE Agar has been further supplemented with ACES Buffer and a-ketoglutarate and is described in the literature as BCYE-a Medium. BCYE-a Medium has been shown to yield optimal recovery of Legionellacea in a shorter incubation period from environmental samples and clinical specimens. Yeast extract provides vitamins, particularly of the B-group, and other growth co-factors. L-Cysteine provides the required nutrional source. Activated charcoal is a protective agent neutralizing and absorbing toxic metabolites produced by bacterial growth. It decomposes hydrogen peroxide, a toxic metabolic product, and can also collect CO2 and modify surface tension. ISO 11731 recommends the following procedure for the isolation of Legionella and its enumeration in water samples. The samples are concentrated by membrane filtration, diluted or inoculated directly on the plate depending on the origin and characteristics of the sample. Independent fractions of the diluted sample should be subjected to heat or acid treatments in case of a high concentration of Legionella and other bacteria. These samples are transferred to the plates with the selective culture medium chosen for Legionella.
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