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Wild Yeast Isolation

4,081 bytes added, 28 March
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[[File:Thornton Wild Capture.jpg|thumb|[https://www.facebook.com/groups/MilkTheFunk/permalink/1034954619866085/ Picture of the first step to catching wild yeast by David Thornton of SouthYeast Labs and Carolina Bauernhaus Ales.]]]
'''Wild yeast isolation''', sometimes known as "yeast wrangling" or more formally [https://en.wikipedia.org/wiki/Bioprospecting "bioprospecting"], refers to the process of catching wild, native yeast ([https://www.merriam-webster.com/dictionary/autochthonous autochthonous] yeast), and isolating it with agar plates as a pure culture. This article also contains information on growing up a ''captured'' mixed culture in wort rather than isolating single strain yeast cultures on agar plates. Culturing wild yeast and bacteria should not be confused with [[Spontaneous Fermentation]] because many of the microbes that might make a flavor impact during spontaneous fermentation are killed off during the bioprospecting processes. For general lab techniques, see [[Laboratory Techniques]]. For more information on nonconventional microbes, see [[Nonconventional Yeasts and Bacteria]].
:''Special thanks goes out to Bryan Heit from Sui Generis blog and Jeff Mello from Bootleg Biology for providing the information in this article.''
* [https://drive.google.com/open?id=17wqLEBinSVCx6zKEoXpsUI9hMPN_MAUHgZ22TD552r0 MTF thread with tips on removing bacteria from yeast colonies using selective media and antibiotics.]
* [https://www.facebook.com/groups/MilkTheFunk/permalink/3298177256877132/ Ruth Barry details her process for isolating individual strains in a mixed culture with the goal of characterizing each strain for a more controlled mixed fermentation.]
* [https://www.youtube.com/watch?v=dQzbeE5_ehc Dr. Matt Winans from Imperial Yeast on "Chop n Brew" demonstrates that incubating agar plats at different temperatures can result in widely different growth success rates (~11:00 minutes in).] See also [https://www.facebook.com/groups/MilkTheFunk/posts/6025908170770680/ this MTF thread exploring this approach].
===Identifying===
===Growing and Testing===
* [http://suigenerisbrewing.blogspot.ca/2012/11/yeast-banking-iii-stepping-up-to.html Sui Generis Blog] guide to stepping up yeast to pitchable amounts.
* [https://www.youtube.com/watch?v=CAIS29Pj8AY Dr. Bryan Heit from Sui Generis Brewing blog] discusses and demonstrates his approach to trialing wild yeast isolates.
* [http://braukaiser.com/wiki/index.php?title=Growing_Yeast_from_a_Plate Kai Troester's] guide to growing yeast from an agar plate.
* [https://www.facebook.com/groups/MilkTheFunk/permalink/2089020794459457/?comment_id=2089022064459330&comment_tracking=%7B%22tn%22%3A%22R0%22%7D Some wild yeasts might perform well at lager temperatures.]
==Domestication==
It has been demonstrated that wild yeast can adapt to brewing conditions if they are exposed to a repeated and constant conditions consistent environment in as little as 200 generations. The average beer fermentation produces 3-5 generations, and the average starter for growth produces around 10 generations. For more information, see this [https://www.facebook.com/groups/MilkTheFunk/permalink/4374738789220968/ MTF thread] by Dr. Bryan Heit for a summary of [https://elifesciences.org/articles/63910 "Phenotypic and molecular evolution across 10,000 generations in laboratory budding yeast populations"].
See also:
==Wild ''Brettanomyces''==
''Brettanomyces'' is notoriously difficult to bioprospect from the wild. For example, an ongoing survey of wild yeasts in most of the US which isolated nearly 2,000 isolates with 262 unique species has not yet found a single occurrence of ''Brettanomyces'' in the wild (so far they have only surveyed non-human inhabited wild areas of the US) <ref>[https://www.biorxiv.org/content/10.1101/2021.07.13.452236v1 Substrate, temperature, and geographical patterns among nearly 2,000 natural yeast isolates. William J. Spurley, Kaitlin J. Fisher, Quinn K. Langdon, Kelly V. Buh, Martin Jarzyna, Max A. B. Haase, Kayla Sylvester, Ryan V. Moriarty, Daniel Rodriguez, Angela Sheddan, Sarah Wright, Lisa Sorlie, Amanda Beth Hulfachor, Dana A. Opulente, Chris Todd Hittinger. bioRxiv 2021.07.13.452236; doi: https://doi.org/10.1101/2021.07.13.452236.]</ref>. While it has been alleged by many that the natural environment for ''Brettanomyces'' is the skins of fruit or the bodies of insects, it wasn't until 2007 that science was able to show that ''Brettanomyces'' survives on the skins of grapes, although it does so in a very low population and a possibly [[Quality_Assurance#Viable_But_Nonculturable|"viable but not culturable"]] state. 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).
Using EBB, Comitini et al. (2019) found that after 80 days of incubation in the EBB medium, half of grapes samples from a vineyard with several varieties of grapes had wild ''Brettanomyces'' on their skins. The researchers also tested the air, the trailers, and the winery where the grapes were processed. They found no ''Brettanomyces'' in the air or the trailers, but did find a couple of valves that were used to transfer must have ''Brettanomyces''. Out of the four varieties of grapes sampled, Merlot grapes did not have any ''Brettanomyces'' growing on them, indicating that the variety of the grape might play a role in the ability of ''Brettanomyces'' to survive on grape skins. The researchers found a wide range of strains that matched the strains found in the winery, indicating that the ''Brettanomyces'' strains did originate from the grape skins <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>.  More recent information indicates that the natural habitat of ''Brettanomyces'' might be the root systems of certain plants, known as the [https://www.nature.com/scitable/knowledge/library/the-rhizosphere-roots-soil-and-67500617/ "rhizosphere"]. See [[Brettanomyces#Environment_and_Survival|''Brettanomyces'' Environment and Survival]] and the links below for more information.
See also:
* [https://suigenerisbrewing.com/index.php/2022/10/06/this-is-not-the-yeast-your-are-looking-for/ Dr. Bryan Heit's first attempt to find wild ''Brettanomyces'' in the rhyzosphere of plants.]** [https://suigenerisbrewing.com/index.php/2022/11/02/this-is-the-wild-brettanomyces-your-are-looking-for/ Second attempt which may have been successful (pending genetic testing).] *** A follow up [https://www.facebook.com/groups/MilkTheFunk/posts/6294804440547717/?comment_id=6294865953874899&reply_comment_id=6297362253625269 thread in MTF] reports Dr. Heit finding a strain of ''B. nanus'' and (likely) ''B. custersianus''.*** [https://suigenerisbrewing.com/index.php/2023/03/24/wild-brettanomyces-fermentation-test/ A blog update on the flavor and aroma results from fermenting with rhyzosphere-isolated ''Brettanomyces''.]* [[Laboratory_Techniques#Brettanomyces|EBB recipeAgar recipes for ''Brettanomyces'']].
* [https://www.facebook.com/groups/MilkTheFunk/permalink/1620419974652877/ MTF discussion on the rare but possible isolation of wild ''Brettanomyces''.]
* [[Brettanomyces#Environment_and_Survival|''Brettanomyces'' natural environment.]]
==Misc==
* Escarpment Labs presentation on catching wild yeast:
: <youtube width="300" height="200">hvgyVrMpa4k</youtube>
 
* Two recent studies have found a very high occurrence of ''Lactobacillus acetotolerans'' in many mixed fermentation sour beers (see [https://www.facebook.com/groups/MilkTheFunk/posts/4652629078098603 this associated MTF thread for discussions]:
** [https://www.sciencedirect.com/science/article/pii/S0740002020302471 Characteristics of bacterial and yeast microbiomes in spontaneous and mixed-fermentation beer and cider; Tyakht et al. (2021).]
** [https://www.biorxiv.org/content/10.1101/2021.07.21.453094v1 Mixed culture metagenomics of the microbes making sour beer; Bochman et al. (2021).]
===MTF Wild Capture Swap===
* [http://brouwerij-chugach.com/?p=1171 "Yeast Wrangling Adventure", Brian Hall of Brouwerij Chugach blog shares wild yeast hunting techniques in the Alaskan wilderness.]
* [https://www.youtube.com/channel/UC8_kozGERzHnjVIizhnh43g/videos "Brewing Viking" home yeast lab videos on YouTube.]
* [https://www.mbaa.com/publications/tq/tqPastIssues/2022/Pages/TQ-59-3-1204-01.aspx "Methods for Harvest, Isolation, Selection, and Utilization of Saccharomyces and Non-Saccharomyces Yeast Strains for Commercial Brewing​​," by Tim Faith and Alex Nham, MBAA Technical Quarterly, 2022.] See also their interview on [http://sites.libsyn.com/315398/episode-114-yeast-foraging-harvesting-isolating-and-selection-w-tim-faith-alex-nham Bru Lab podcast].
* [https://surradelupulo.com.br/projeto-manipueira-resultados-cientificos/ "Manipueira, wild beers and scientific results" by Nana Ottoni's podcast; interviewing Brazilian scientists Estela Virgílio and Ana Carolina about microbes identified during spontaneous fermentation in Brazilian beer (English translation available).]
====Books====

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