13,591
edits
Changes
added EBB
A few different media can be used to isolate ''Brettanomyces'' but DBDM medium is commonly used. WLD with additions of cycloheximide can also be used. A simple Malt agar with cycloheximide has been shown to grow ''Brettanomyces'' that has been adapted to the brewing environment more efficiently than Dekkera medium (recommended by the European Brewing Convention, but not listed here) and [https://foodsafety.neogen.com/pdf/acumedia_pi/7574_pi.pdf universal beer medium] (recommended by the Brewery Convention of Japan; also not listed here) <ref>[https://www.tandfonline.com/doi/abs/10.1094/ASBCJ-2008-0917-01 Effects of Beer Adaptation on Culturability of Beer-Spoilage Dekkera/Brettanomyces Yeasts. Koji Suzuki, Shizuka Asano, Kazumaru Iijima, Tomoo Ogata, Yasushi Kitagawa & Tsunehiro Ikeda. 2018.]</ref>.
Renouf et al. (2007) and Comitini et al. (2019) demonstrated that an "enrichment ''Brettanomyces bruxellensis''" media called EBB is more efficient at first growing up ''Brettanomyces'' before trying to culture it on DBDM. ''Brettanomyces'' was allowed to grow for 80 days in the EBB media, and then streaked onto DBDM for selection for ''Brettanomyces'' (other wild yeast such as ''Hanseniaspora'' and ''Pichia'' grew much more readily than ''Brettanomyces'' that was cultured from wine grapes) <ref>[https://www.sciencedirect.com/science/article/pii/S0944501306000231?via%3Dihub Development of an enrichment medium to detect Dekkera/Brettanomyces bruxellensis, a spoilage wine yeast, on the surface of grape berries. Vincent Renouf, Aline Lonvaud-Funel. 2007. DOI: https://doi.org/10.1016/j.micres.2006.02.006.]</ref><ref>[https://www.frontiersin.org/articles/10.3389/fmicb.2019.00415/abstract Occurrence of Brettanomyces bruxellensis on grape berries and in related winemaking cellar. Francesca Comitini1, Lucia Oro, Laura Canonico, Valentina Marinelli, Maurizio Ciani. 2019. DOI: 10.3389/fmicb.2019.00415.]</ref>.
'''EBB Media Recipe'''
{| class="wikitable"
|-
! Chemical !! Usage Amount
|-
| Red grape juice (possibly replaceable with DME?) || 200 mL/L
|-
| Ethanol || 40 mL/L
|-
| Malt extract || 1.5 g/L
|-
| Yeast extract || 1.5 g/L
|-
| (NH4)2 SO4 (sourced from Sigma-Aldrich) || 0.5 g/L
|-
| MgSO4 || 0.2 g/L
|-
| Tween 80 || 0.5 mL/L
|-
| Biphenyl (limits mold) || 0.2 g/L (w/v)
|-
| Chloramphenicol (limits bacteria) || 0.05 g/L (w/v)
|-
| Adjust pH to 5.0 with sodium hydroxide ||
|-
|}
Notes: gentle agitation during growth (120 rpm).
'''DBDM Media Recipe'''