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==Procedures==
A strain of ''Brettanomyces'' believed to be the species ''B. bruxellensis '' was isolated by Trent from a bottle of Orval in 2012. This ''Brettanomyces'' strain was subjected to four different propagation treatments: aerobic (stir plate set to high with a foil cover), semi-aerobic (orbital shaker set to 80 RPM's with a foil cover), anaerobic with constant agitation (Co2 purged and then set on an orbital shaker set to 80 RPM's with an airlock), and still (foil cover, no agitation). All treatments were duplicated.
The initial inoculum of ''Brettanomyces'' was grown in 3 steps to 300 mL slurry from a single colony. For each treatment 35 mL of the inoculum ''Brettanomyces'' was added to 230 mL of 10°P wort in 500 mL Erlenmeyer flasks for an initial cell density of 112.87 million cells per mL.