Changes

At 15°C (59°F), none of the ''Brettanomyces'' strains could match the US-05 attenuation, with most of them falling to around 50-25% less final attenuation after 28 days, and one of the wine strains and one of the USA saison strains fermented almost nothing at all. Still, this data showed that some beer strains of ''B. bruxellensis'' can ferment at lower temperatures. Interestingly, one of the wine strains was almost unaffected by the difference in fermentation temperature; it only lagged for a couple of days longer in the colder 15°C (59°F) fermentation temperature versus the warmer 22.5°C (72.5°F) fermentation temperature <ref name="Tyrawa_2019" />.

The effect on phenol production, 4-ethylguaiacol (clove) and 4-ethylphenol (barnyard), was relatively the same and above flavor threshold for both fermentation temperatures for all of the ''B. bruxellensis'' strains tested, although some strains had slightly more or less of these phenols produced at the different fermentation temperatures. The temperature of the fermentation had a larger impact on the amount of esters produced. Ethyl acetate (pineapple/pear) was significantly higher in the warmer fermentation temperature of 22.5°C (72.5°F) than the cooler temperature of 15°C (59°F) for all strains, with one saison strain producing significantly more ethyl acetate and another saison strain producing significantly less ethyl acetate than the other strains. As expected, the US-05 produced higher amounts of 4-vinylguaiacol (clove) and isoamyle acetate (banana) at 22.5°C (72.5°F) and lower amounts at 15°C (59°F). The US-05 produced comparably high amounts of phenethyl alcohol (dried rose), phenethyl acetate (honey/rose pedal), and isoamyl alcohol (banana/oily) at both temperatures. These were generally not produced at more than low levels by the ''Brettanomyces'' strains, except for 4-vinylguaiacol which was produced more at the lower temperature by BSI Drei and one of the saison strains, indicating that the lower fermentation temperature slowed the process of these strains to convert the 4-VG to 4-EG, and they also produced the lowest amount of the ethyl phenols compared to the other ''Brettanomyces'' strains. The ''Brettanomyces'' strains produced other fatty acid esters at significant levels above tastes threshold that the US-05 produced below tastes threshold. These esters include ethyl caproate (pineapple/apple), ethyl caprylate (pineapple), ethyl decanoate (brandy/apple) and ethyl nonanoate (fruity/rose/waxy). In general, a higher amount of these esters were produced at the higher fermentation temperature, although there were exceptions. Several of the saison strains and the lambic strain produced higher amounts of esters than the BSI Drei control, especially when fermented at the warmer temperature, demonstrating the amount of esters produced is highly variable amount different strains of ''B. bruxellensis'', particularly when fermented at 22.5°C (72.5°F) rather than the lower fermentation temperature of 15°C (59°F). Interestingly, the two wine strains of ''Brettanomyces bruxellensis'' did not produce above threshold levels of any of these esters at either fermentation temperature (the wine strains produced the highest levels of decanoic acid, which were elevated at the higher fermentation temperature) <ref name="Tyrawa_2019" />.