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Brettanomyces

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Biofilm
[[Image:Brettanomyces.jpg|thumb|200px|right|Brettanomyces]][[File:Brett-aroma-wheel.jpeg|thumb|200px|right|Brett Aroma Wheel by Dr. Linda Bisson and Lucy Joseph at UC Davis]]
'''''Brettanomyces''''', also referred to by brewers as "Brett" or "Bretta", is Greek for "British Fungus" and is a [https://en.wikipedia.org/wiki/Facultative_anaerobic_organism facultative anaerobic] yeast that was originally thought of as an important yeast for producing the character of some 17th century and prior English ales. Since the wide adoption of pure cultures of ''Saccharomyces cerevisiae'' and ''S. pastorianus'' in the brewing and wine industries starting in the late 1800's, ''Brettanomyces'' has been mostly viewed as a spoilage yeast, except in Belgian lambic, Flanders red/brown beers, and a handful of styles of wine. More recently ''Brettanomyces'' has gained popularity in the United States (and subsequently the brewing industries of other countries) as a yeast that can contribute desirable and novel characteristics to beer and other alcoholic beverages. The genus name ''Dekkera'' is used interchangeably with ''Brettanomyces'', as it describes the teleomorph or spore-forming form of the yeast, although this form is extremely rare or perhaps even non-existent <ref>[http://en.wikipedia.org/wiki/Brettanomyces Wikipedia. Brettanomyces. Retrieved 2/24/2015.]</ref><ref name="Avramova_2018">[https://www.nature.com/articles/s41598-018-22580-7 Brettanomyces bruxellensis population survey reveals a diploid-triploid complex structured according to substrate of isolation and geographical distribution. Marta Avramova, Alice Cibrario, Emilien Peltier, Monika Coton, Emmanuel Coton, Joseph Schacherer, Giuseppe Spano, Vittorio Capozzi, Giuseppe Blaiotta, Franck Salin, Marguerite Dols-Lafargue, Paul Grbin, Chris Curtin, Warren Albertin & Isabelle Masneuf-Pomarede. 2018. doi:10.1038/s41598-018-22580-7.]</ref>. Known for its barnyard, fecal, horsey, metallic or Band-Aid flavors, ''Brettanomyces'' continues to be unwelcome in many breweries and most wineries <ref>[https://www.cambridge.org/core/journals/journal-of-wine-economics/article/brettanomics-i-the-cost-of-brettanomyces-in-california-wine-production/295D206007B358EC1B07CEF4879BE06B Brettanomics I: The Cost of Brettanomyces in California Wine Production. Julian M. Alston, Torey Arvik, Jarrett Hart and James T. Lapsley. 2020. DOI: https://doi.org/10.1017/jwe.2020.20.]</ref>. However, ''Brettanomyces'' also produces high levels of fruity esters that are desirable in some styles like saison, [[lambic]], and American sour beers. ''Brettanomyces'' can also form a [[pellicle]] during fermentation. See ''[[Lactobacillus]]'', ''[[Pediococcus]]'', ''[[Saccharomyces]]'', [[Mixed Cultures]], [[Kveik#Commercial_Availability|Kveik]], and [[Nonconventional Yeasts and Bacteria]] charts for other commercially available cultures.
==Introduction of History, Characteristics, and Taxonomy==
===Environment and Survival===
''Brettanomyces'' has been thought to occur naturally on the skins of fruit such as apples and grapes. However, there are only a handful of reports of ''Brettanomyces'' being identified on the skins of fruit, and in some cases where ''Brettanomyces'' has been found, its abundance is extremely minimal <ref name="Aires_2025">[https://www.mdpi.com/2076-2607/13/3/538 Aires, C.; Maioto, R.; Inês, A.; Dias, A.A.; Rodrigues, P.; Egas, C.; Sampaio, A. Microbiome and Microbiota Within Wineries: A Review. Microorganisms 2025, 13, 538. https://doi.org/10.3390/microorganisms13030538.]</ref><ref>[https://onlinelibrary.wiley.com/doi/full/10.1002/jib.154 Lentz, M., Putzke, T., Hessler, R. and Luman, E. (2014), Genetic and physiological characterization of yeast isolated from ripe fruit and analysis of fermentation and brewing potential, J. Inst. Brew., 120: 559– 564. DOI: 10.1002/jib.154.]</ref><ref name="Comitini">[https://www.frontiersin.org/articles/10.3389/fmicb.2019.00415/abstract Occurrence of Brettanomyces bruxellensis on grape berries and in related winemaking cellar. Francesca Comitini, Lucia Oro, Laura Canonico, Valentina Marinelli, Maurizio Ciani. 2019. DOI: 10.3389/fmicb.2019.00415.]</ref><ref name="Renouf_2007">[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>. In contrast, there are also studies that indicate ''Brettanomyces'' only being found during or after food processing, which indicates that the processing equipment may be the primary source for the ''Brettanomyces''. In addition, ''Brettanomyces'' has been isolated in abundance from the surfaces of equipment/processed materials in wineries and breweries <ref name="Aires_2025" /><ref name="smith_divol_2016" /><ref name="Schifferdecker" /><ref name="Loureiro_2003">[https://www.ncbi.nlm.nih.gov/pubmed/12892920 Spoilage yeasts in the wine industry. Loureiro V, Malfeito-Ferreira M. 2003.]</ref><ref name="Steensels" /><ref name="Barata_2008">[https://www.ncbi.nlm.nih.gov/pubmed/18077036 Survival patterns of Dekkera bruxellensis in wines and inhibitory effect of sulphur dioxide. f Barata A, Caldeira J, Botelheiro R, Pagliara D, Malfeito-Ferreira M, Loureiro V. 2008.]</ref> (Table 1). For example, an ongoing survey of wild yeasts in different regions of United States wilderness areas 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 and Alaska; substrates sampled included leaves, soil, bark, moss, mushrooms, needles, pine cones, twigs/wood, and other plant matter) <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>. It is therefore unclear that ''Brettanomyces'' found on grape skins originated there or from the industrial processing where it is more abundant. It is also thought to disperse via fruit-flies (called "vectors" in the scientific literature), similar to how ''Saccharomyces'' travels, although direct evidence for this has been reported rarely and only on fruit-flies in wineries that are likely to come into contact with equipment/processed material that is already contaminated with ''Brettanomyces'' <ref>[https://youtu.be/G2nhUM5PIrg?t=309 Dr. Bryan Heit. BotB - Where (Do) The Wild Brettanomyces Roam?. ~5 mins in. Retrieved 07/10/2022.]</ref><ref name="Renouf_2007" /><ref name="Steensels">[http://www.sciencedirect.com/science/article/pii/S0168160515001865 Brettanomyces yeasts — From spoilage organisms to valuable contributors to industrial fermentations. Jan Steensels, Luk Daenen, Philippe Malcorps, Guy Derdelinckx, Hubert Verachtert, Kevin J. Verstrepen. International Journal of Food Microbiology Volume 206, 3 August 2015, Pages 24–38.]</ref><ref name="Barata_2008" /><ref name="Loureiro_2003" />. ''Brettanomyces'' is known to be difficult to grow in a lab due to slow growth, specific nutrient requirements, or perhaps because of a "VBNC" state (see [[Wild_Yeast_Isolation#Wild_Brettanomyces|Wild ''Brettanomyces'']] for more information), which may account for the lack of evidence for fruit being the primary natural habitat for ''Brettanomyces''. More recently, techniques have been invented to more easily isolate and grow ''Brettanomyces'' <ref name="Renouf_2007" /><ref name="Comitini" />. There is also significant evidence that the natural habitat of ''Brettanomyces'' might actually be the root systems of certain plants, known as the [https://www.nature.com/scitable/knowledge/library/the-rhizosphere-roots-soil-and-67500617/ "rhizosphere"]. The rhizosphere refers to the complex symbiotic community of microbe populations that live on and around the root system of plants. Wild strains of ''Brettanomyces'' have been found in the root systems of dill, common beans, sunflowers, maize, corn, jute, cassava, and grey mangroves found in the estuaries of Indonesia <ref>[https://onlinelibrary.wiley.com/doi/abs/10.1111/aab.12309 Weisany, W., Raei, Y., Salmasi, S., Sohrabi, Y. and Ghassemi-Golezani, K. (2016), Arbuscular mycorrhizal fungi induced changes in rhizosphere, essential oil and mineral nutrients uptake in dill/common bean intercropping system. Ann Appl Biol, 169: 384-397. https://doi.org/10.1111/aab.12309.]</ref><ref>[https://archive.aessweb.com/index.php/5003/article/view/3333 I.O, S. ., & G.P, O. . (2012). Diversity of Fungal Populations in Soils Cultivated With Cassava Cultivar TMS 98/0505. Journal of Asian Scientific Research, 2(3), 116–123. Retrieved from https://archive.aessweb.com/index.php/5003/article/view/3333.]</ref><ref>[https://www.ajol.info/index.php/swj/article/view/149513 Rhizosphere and non-rhizosphere soil mycoflora of Corchorus olitorius (Jute). G.S. Olahan, I.O. Sule, T Garuba, Y.A. Salawu. Science World Journal. 2016.]</ref><ref>[https://ojs.unud.ac.id/index.php/jbb/article/view/36023 NOERFITRYANI, Noerfitryani; HAMZAH, Hamzah. THE EXISTENCE OF ENTOMOPATHOGENIC FUNGI ON RICE PLANTS RHIZOSPHERE. International Journal of Biosciences and Biotechnology, p. 12-24, dec. 2017. ISSN 2655-9994. doi: https://doi.org/10.24843/IJBB.2017.v05.i01.p02.]</ref><ref>[https://www.sciencedirect.com/science/article/abs/pii/S2452219818300259 Marcela Sarabia, Saila Cazares, Antonio González-Rodríguez, Francisco Mora, Yazmín Carreón-Abud, John Larsen, Plant growth promotion traits of rhizosphere yeasts and their response to soil characteristics and crop cycle in maize agroecosystems, Rhizosphere, Volume 6, 2018, Pages 67-73, ISSN 2452-2198, https://doi.org/10.1016/j.rhisph.2018.04.002.]</ref><ref>[https://www.sciencedirect.com/science/article/abs/pii/S1049964419303238 Nivien A. Nafady, Mohamed Hashem, Elhagag A. Hassan, Hoda A.M. Ahmed, Saad A. Alamri. The combined effect of arbuscular mycorrhizae and plant-growth-promoting yeast improves sunflower defense against Macrophomina phaseolina diseases. Biological Control. Volume 138, 2019, 104049. ISSN 1049-9644, https://doi.org/10.1016/j.biocontrol.2019.104049.]</ref><ref>[http://ejurnal.its.ac.id/index.php/sains_seni/article/view/5613 Isolation and Characterization of Yeast from Rhizosphere Avicennia Marina Wonorejo. Sitatun Zunaidah, Nur Hidayatul Alami. 2014. DOI: 10.12962/j23373520.v3i1.5613.]</ref>. See Dr. Bryan Heit's video [https://www.youtube.com/watch?v=G2nhUM5PIrg "Where (Do) The Wild Brettanomyces Roam?"] and [https://www.facebook.com/groups/MilkTheFunk/posts/5940213029340195 his comments in Milk The Funk], as well as [https://www.youtube.com/watch?v=BrR7G_YyfmA "Philip Poole. Plant Control of the Rhizosphere Microbiome"]. For documented isolation attempts from plant rhizospheres, see [[Wild_Yeast_Isolation#Wild_Brettanomyces|Wild Yeast Isolation]].
The occurrence of ''Brettanomyces'' has been more commonly identified in industrial food processing areas (wine, beer, kombucha, soft drinks, dairy products, tea, sourdough, etc.) <ref name="Crauwels_2016">[https://academic.oup.com/femsyr/article-abstract/17/1/fow105/2670560/Fermentation-assays-reveal-differences-in-sugar?redirectedFrom=fulltext Fermentation assays reveal differences in sugar and (off-) flavor metabolism across different Brettanomyces bruxellensis strains. Fermentation assays reveal differences in sugar and (off-) flavor metabolism across different Brettanomyces bruxellensis strains. Sam Crauwels, Filip Van Opstaele, Barbara Jaskula-Goiris, Jan Steensels, Christel Verreth, Lien Bosmans, Caroline Paulussen, Beatriz Herrera-Malaver, Ronnie de Jonge, Jessika De Clippeleer, Kathleen Marchal, Gorik De Samblanx, Kris A. Willems, Kevin J. Verstrepen, Guido Aerts, and Bart Lievens. 2016]</ref>. For example, ''B bruxelensis'', ''B. anomala'', and ''B. custersianus'' have mostly been isolated from wine or beer production, while ''B. naardenensis'' has mostly been isolated from soda production <ref name="Tiukova_2019">[https://www.mdpi.com/2076-2607/7/11/489 Assembly and Analysis of the Genome Sequence of the Yeast Brettanomyces naardenensis CBS 7540. Ievgeniia A. Tiukova, Huifeng Jiang, Jacques Dainat, Marc P. Hoeppner, Henrik Lantz, Jure Piskur, Mats Sandgren, Jens Nielsen, Zhenglong Gu, and Volkmar Passoth. 2019. DOI: https://doi.org/10.3390/microorganisms7110489.]</ref>. ''Brettanomyces'' is not considered to be airborne; however, one study has demonstrated a very small amount of cells in the air at wineries where wine with ''Brettanomyces'' in it was being handled (most of the yeasts found in the air were ''Aureobasidium'' and ''Cryptococcus'', which aren't considered spoilage organisms in beer and wine). This set of studies also determined that very specific methodology was needed in order capture ''Brettanomyces'' from the air, and indicated that the yeast was "stressed". A [https://oeno-one.eu/article/view/8015 second study] carried out in three wineries in the Bordeaux region of France reported finding ''B. bruxellensis'' in the air from three out of ten samples between two of the breweries; all three of the breweries had ''B. bruxellensis'' found on various surfaces within the wineries. While it is possible for ''Brettanomyces'' to be briefly carried by gusts of air, it only happens in the vicinity where the ''Brettanomyces'' beer or wine is being bottled (more so) or is actively fermenting (less so) <ref>[http://www.sciencedirect.com/science/article/pii/S0956713513002284 Screening of yeast mycoflora in winery air samples and their risk of wine contamination. E. Ocón, P. Garijo, S. Sanz, C. Olarte, R. López, P. Santamaría, A.R. Gutiérrez. Food Control Volume 34, Issue 2, December 2013, Pages 261–267.]</ref><ref name="Montagner_2024">[https://oeno-one.eu/article/view/8015 Le Montagner, P., Etourneau, L., Ballestra, P., Dols-Lafargue, M., Albertin, W., Maupeu, J., … Masneuf-Pomarède, I. (2024). Critical areas for Brettanomyces bruxellensis contamination and biofilm formation in the cellar: on the origin of wine spoilage. OENO One, 58(3). https://doi.org/10.20870/oeno-one.2024.58.3.8015.]</ref>. Good cleaning and sanitation and cold temperatures should be employed to keep ''Brettanomyces'' from contaminating other equipment; however, flying insects could also be a potential cause for ''Brettanomyces'' contamination (although direct evidence for this is lacking).
Montagner et al. (2024) examined the biofilms of several strains of ''B. bruxellensis'' that were found at one of three different wineries in the Bordeaux region of France. They reported that phenol production occurred within the biofilm and hypothesized that it is possible that phenol contamination in wine could be a result of the wine coming into contact with the biofilm rather than solely from ''Brettanomyces'' growing in the wine. They also observed the detachment of ''B. bruxellensis'' cells from the biofilm and into the wine <ref name="Montagner_2024" />.
 
Biofilm formation in some strains of ''S. cerevisiae'' have been linked to genes that code for flocculation, specifically the ''Flo11'' gene. This gene is capable of expressing the so-called ''flor'' trait (a reference to sherry flor), which is the adhesion of cells to surfaces as well as the air-liquid interface of sherry flor biofilm (also refereed to as "[[Pellicle|pellicles]]" by brewers) <ref>[https://www.sciencedirect.com/science/article/abs/pii/S1567135605001005 Jennifer C. Bayly, Lois M. Douglas, Isak S. Pretorius, Florian F. Bauer, Anne M. Dranginis. Characteristics of Flo11-dependent flocculation in Saccharomyces cerevisiae. FEMS Yeast Research, Volume 5, Issue 12, 2005, Pages 1151-1156. ISSN 1567-1356. DOI: https://doi.org/10.1016/j.femsyr.2005.05.004.]</ref>. [https://www.sciencedirect.com/science/article/pii/S2666517425000343#bib0002 Di Canito et al. (2025)] demonstrated that ''Brettanomyces'' strains also have a wide range of ''Flo1'' and ''Flo11'' genes that code for flocculation, with ''B. bruxelensis'' strains being commonly more flocculant than strains of ''S. cerevisiae''. They also demonstrated that the presence of environmental stressors such as ethanol or SO<sub>2</sub> increase the flocculation of flocculant positive strains of ''B. bruxellensis''. This work was the first step in linking the ''Flo11'' gene to biofilm formation in ''B. bruxellensis'' <ref>[https://www.sciencedirect.com/science/article/pii/S2666517425000343#bib0002 Alessandra Di Canito, Roberto Foschino, Ileana Vigentini. Flocculation Mechanisms in Brettanomyces bruxellensis: Influence of ethanol and sulfur dioxide on FLO gene expression. Current Research in Microbial Sciences, Volume 8, 2025, 100372, ISSN 2666-5174. DOI: https://doi.org/10.1016/j.crmicr.2025.100372.]</ref>.
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