The genetic diversity of ''Brettanomyces'' is particularly wide. For example, one study that analyzed the whole genomes of 53 strains of ''B. bruxellensis'' found that the overall genetic diversity between different strains of ''B. bruxellensis'' was higher than strains of ''S. cerevisiae'' (however, the entire gene set, known as the ''pangenome'', of all the genes among all of the strains of '''B. bruxellensis'' is much smaller than the entire gene set of ''S. cerevisiae'') <ref name="Gounot_2019" />. Some studies have indicated that strains of ''B. bruxellensis'' have adapted to specific environments. For example, one study found that strains of ''B. bruxellensis'' isolated from wine had 20 genes involved in the metabolism of carbon and nitrogen, whereas strains isolated from beer did not. This indicated that ''B. bruxellensis'' strains living in wine have adapted to the harsher environment of wine <ref name="smith_divol_2016"></ref>. Another study found that one out of the two strains tested that were isolated from soda could not ferment maltose, and only strains isolated from wine were able to grow in wine and the beer/soda strains did not. The wine strains were also more resistant to sulfites, which are commonly used in the wine industry to prevent microbial contamination <ref name="Crauwels_2016" />. The whole genome sequencing of one strain of ''B. naardenensis'' found that it was missing the genes associated with nitrate utilization, indicating that it is not well adapted to survive in beer where nitrates are abundant due to hops <ref name="Tiukova_2019" />.
A study by Cibrario et al. (2019) looked at the genome The addition of ''B. bruxellensis'' strains in bottles of wine going back as far as the year 1909 revealed that the SO<sup>2</sup> tolerant triploid strains only started appearing after the year 1990, which corresponds to when the wine industry started using SO<sup>2</sup> in most wine production (although it vitamins can also mean that the triploid strains are not as good at surviving in bottles of wine long-term compared to the diploid strains that have been isolated from much older bottles of wine; this will be determined to be the case or not in the future as bottles of wine from the 1990s continue to age). They also identified dozens of examples of wineries throughout France and Italy where the same strain of a positive impact on ''B. bruxellensis'' was found in multiple vintages of bottles of wine going back many decades, indicating that individual Brettanomyces''B. bruxellensis'' strains become long-term residents in wineriesgrowth. Some identical strains have been found in different regions and even different continentsFor example, indicating that some strains have traveled not just due to traditional vectors such as insects or birds, but also probably due to human transportation such as wine bottle imports/exports or exchanges between industrial processes. This also indicates that while ''B. bruxellensisBrettanomyces'' becomes rather sedentary and a constant resident in wineries, it can also adapt to the different winemaking conditions in different regions once it's been transported does not need riboflavin (vitamin B2) or thiamine (different grapes, different climates, different fermentation temperatures, etc.vitamin B1), including adapting in order to improved modern hygienic practices such as higher SO<sup>2</sup> treatment. Overallgrow, the results from this study suggest that presence of either or both of these two vitamins encourages ''Brettanomyces'' is able to adapt to living alongside certain human industries and has done so for at least a couple of centuries growth <ref name="Cibrario_2019">[httpshttp://wwwonlinelibrary.biorxivwiley.orgcom/contentdoi/10.11011002/jib.385/763441v1 Brettanomyces bruxellensis wine isolates show high geographical dispersal full The influence of thiamine and long remanence riboflavin on various spoilage microorganisms commonly found in cellars. Alice Cibrario, Marta Avramova, Maria Dimopoulou, Maura Magani, Cécile Miot-Sertier, Albert Mas, Maria Cbeer. Portillo, Patricia Ballestra, View ORCID ProfileWarren AlbertinBarry Hucker, Isabelle Masneuf-PomaredeMelinda Christophersen, Marguerite Dols-Lafargue. 2019Frank Vriesekoop. DOI: https://doi.org/10.1101/7634412017.]</ref>. The genetic differences between the fermentation substrates Other vitamins such as p-aminobenzoic acid (beerPABA), winefolic acid (vitamin B9), etc.nicotinic acid (vitamin B3) were lower but still significant, and this was explained by the frequent cross-over pantothenic acid (vitamin B5) are also not required for most strains of equipment such as wine barrels being used for beer fermentation''Brettanomyces'' to grow. When comparing The presence of alcohol can increase the geographic differences, they found geography contributed only 5% dependence on vitamins for some strains of genetic differences''Brettanomyces'' to grow. For example, while geography explained more than 50% of genetic differences in nonMyo-wine strains, suggesting that beer, kombucha, inositol (vitamin B8) and tequila strains are more localized genetically than wine strains and that humans probably helped the wine thiamine (vitamin B1) were required by two strains travel across the globe. They also found that although one study reported spore-forming versions of ''B. bruxellensis'' when grown in 10% ethanol but not in 0% ethanol. Biotin (referred to as ''Dekkera bruxellensis''vitamin B7)is the one exception, and it was found that the genetic makeup lack of biotin inhibited the analyzed growth of some strains determined their ability to sporulate to be non-existent or rare (only one study that we know of by [https://link.springer.com/article/10.1007%2FBF02539015 Walt and Kerken in 1960] has reported sporulation in ''BrettanomycesB. bruxellensis'' only on specific agar types with vitamins added. Other studies contradict these previously mentioned findings, indicating showing that sporulation in thiamine was not required by the strains of ''BrettanomycesB. bruxellensis'' is extremely rare) tested, that pyridoxine was required, and biotin was not required. These discrepancies between scientific studies are probably due to the genetic differences between the strains selected, the growth media chosen by the scientists, and/or the growth conditions <ref name="Avramova_2018" />. See also [httpshttp://www.facebookmdpi.com/groups2311-5637/MilkTheFunk/permalink2/20228016810813693/ Richard Preiss's discussion 17 Use of this study on MTFNutritional Requirements for Brettanomyces bruxellensis to Limit Infections in Wine. Nicolas H. von Cosmos and Charles G. Edwards. 2016.]</ref>. ====Sulfur Tolerance====
Sulfite and SO<sub>2</sub> inhibits the growth of ''Brettanomyces'', and is often used in the wine industry to prevent the growth of ''Brettanomyces'' (although ''Brettanomyces'' is usually seen is a contaminant in wine, some wineries have identified small amounts of flavors from ''Brettanomyces'' as being beneficial to certain wine styles, and is said to increase the complexity and impart an aged character in young wines <ref name="smith_divol_2016"></ref>) <ref>[http://onlinelibrary.wiley.com/doi/10.1111/j.1745-4549.2012.00702.x/abstract Removal of Brettanomyces Bruxellensis from Red Wine Using Membrane Filtration. Umiker, Descenzo, Lee, and Edwards. 04/24/2012.]</ref>. However, it has been shown that wine strains of ''B. bruxellensis'' could survive dosages of up to 1 mg/L of molecular SO<sub>2</sub>, and the very high dosage of 2.1 mg/L was needed to kill ''Brettanomyces'' in wine <ref name="Agnolucci_2017" />. This dosage of molecular SO<sub>2</sub> requires a total amount of SO<sub>2</sub> that is beyond legal limits (350 mg/l <ref>[https://grapesandwine.cals.cornell.edu/sites/grapesandwine.cals.cornell.edu/files/shared/documents/Research-Focus-2011-3.pdf Sulphur Dioxide Content of Wines: the Role of Winemaking and Carbonyl Compounds. Nick Jackowetz, Erhu Li, and Ramón Mira de Orduña. 2011.]</ref>; see this [https://grapesandwine.cals.cornell.edu/newsletters/appellation-cornell/2012-newsletters/issue-12/article-contains-sulfites/ Cornell University] blog post that explains the difference between ''free'' and ''molecular'' SO<sub>2</sub>) and has negative effects on wine. One study found that out of 145 strains of ''B. bruxellensis'', 107 of which were wine strains with the rest being from beer, tequila, kombucha, etc., 36% of them were either tolerant (lagged growth, but achieved full growth eventually) or resistant (no lagged growth, and achieved full growth) to 0.6 mg/L of molecular SO<sub>2</sub>. 46 of the 52 resistant/tolerant strains were wine strains, thus demonstrating that wine strains of ''B. bruxellensis'' are generally more tolerant of SO<sub>2</sub> than strains of ''B. bruxellensis'' that are found in other types of beverages. It is thought that the wine strains have adapted to the conditions of winemakers adding SO<sub>2</sub> to wine <ref>[https://www.frontiersin.org/articles/10.3389/fmicb.2018.01260/full Molecular Diagnosis of Brettanomyces bruxellensis’ Sulfur Dioxide Sensitivity Through Genotype Specific Method. Avramova M, Vallet-Courbin A, Maupeu J, Masneuf-Pomarède I, Albertin W. 2018. DOI: 10.3389/fmicb.2018.01260.]</ref>. ''Brettanomyces'' strains isolated from sweet wine tend to be more tolerant of sulfur dioxide than strains isolated from dry wine <ref>[https://www.sciencedirect.com/science/article/pii/S0740002018303988 Sulfur dioxide response of Brettanomyces bruxellensis strains isolated from Greek wine. Maria Dimopoulou, Magdalini Hatzikamari, Isabelle Masneuf-Pomarede, Warren Albertin. 2018. DOI: https://doi.org/10.1016/j.fm.2018.10.013.]</ref>. In addition, it has been proposed that SO<sub>2</sub> can induce a so-called "viable but nonculturable" (VBNC) state in ''Brettanomyces'', which means that ''Brettanomyces'' cells in this state cannot grow or be cultured on traditional media but can remain viable and create a low amount of phenol character (see [[Quality_Assurance#VBNC_In_Yeast|VBNC in Yeast]]). Some strains of ''Candida pyralidae'', ''Wickerhamomyces anomalus'', ''Kluyveromyces wickeramii'', ''Torulaspora delbrueckii'' and ''Pichia membranifaciens'' have been found to produce toxin that inhibits ''Brettanomyces'', and these toxins have been proposed as an alternative to SO<sub>2</sub> as a way to kill ''Brettanomyces'' (killer wine strains of ''Saccharomyces cerevisiae'' do not kill ''Brettanomyces''; see [[Saccharomyces#Killer_Wine_Yeast|Killer Wine Yeast]] for more information). [http://www.laboratoriosenosan.com/en/effectiveness-of-kaolin-silver-complex/ Kaolin silver complex (KAgC)] has been found to inhibit ''Brettanomyces'' and acetic acid bacteria in wine when used in legal dosages, and has been proposed as a replacement for SO<sub>2</sub> or to minimize the use of SO<sub>2</sub> <ref>[https://www.ncbi.nlm.nih.gov/pubmed/29666535?dopt=Abstract Effect of kaolin silver complex on the control of populations of Brettanomyces and acetic acid bacteria in wine. Izquierdo-Cañas PM, López-Martín R, García-Romero E, González-Arenzana L, Mínguez-Sanz S, Chatonnet P, Palacios-García A, Puig-Pujol A. 2018. DOI: 10.1007/s13197-018-3097-y.]</ref>. Other proposed replacements for SO<sub>2</sub> as a way to inhibit ''Brettanomyces'' in wine include [https://en.wikipedia.org/wiki/Pascalization high pressure processing] and [https://www.sciencedirect.com/science/article/abs/pii/S0255270106001929 pulsed electric fields] <ref>[https://www.sciencedirect.com/science/article/pii/S1466856418302972 SO2, high pressure processing and pulsed electric field treatments of red wine: Effect on sensory, Brettanomyces inactivation and other quality parameters during one year storage. Sanelle Van Wyk, Mohammed M. Farid, Filipa V.M. Silva. 2018. DOI: https://doi.org/10.1016/j.ifset.2018.06.016.]</ref><ref>[https://www.sciencedirect.com/science/article/pii/S1466856418307409 Pulsed electric field treatment of red wine: Inactivation of Brettanomyces and potential hazard caused by metal ion dissolution. Sanellevan Wyk, Filipa V.M. Silva, Mohammed M.Farid. 2018. DOI: https://doi.org/10.1016/j.ifset.2018.11.001.]</ref>.
The addition A study by Cibrario et al. (2019) looked at the genome of ''B. bruxellensis'' strains in bottles of vitamins wine going back as far as the year 1909 revealed that the SO<sup>2</sup> tolerant triploid strains only started appearing after the year 1990, which corresponds to when the wine industry started using SO<sup>2</sup> in most wine production (although it can also mean that the triploid strains are not as good at surviving in bottles of wine long-term compared to the diploid strains that have a positive impact on been isolated from much older bottles of wine; this will be determined to be the case or not in the future as bottles of wine from the 1990s continue to age). They also identified dozens of examples of wineries throughout France and Italy where the same strain of ''B. bruxellensis'' was found in multiple vintages of bottles of wine going back many decades, indicating that individual ''BrettanomycesB. bruxellensis'' growthstrains become long-term residents in wineries. For exampleSome identical strains have been found in different regions and even different continents, indicating that some strains have traveled not just due to traditional vectors such as insects or birds, but also probably due to human transportation such as wine bottle imports/exports or exchanges between industrial processes. This also indicates that while ''BrettanomycesB. bruxellensis'' does not need riboflavin becomes rather sedentary and a constant resident in wineries, it can also adapt to the different winemaking conditions in different regions once it's been transported (vitamin B2different grapes, different climates, different fermentation temperatures, etc.) or thiamine (vitamin B1) in order , including adapting to growimproved modern hygienic practices such as higher SO<sup>2</sup> treatment. Overall, the presence of either or both of these two vitamins encourages results from this study suggest that ''Brettanomyces'' growth is able to adapt to living alongside certain human industries and has done so for at least a couple of centuries <refname="Cibrario_2019">[httphttps://onlinelibrarywww.wileybiorxiv.comorg/doicontent/10.10021101/jib.385/full The influence of thiamine 763441v1 Brettanomyces bruxellensis wine isolates show high geographical dispersal and riboflavin on various spoilage microorganisms commonly found long remanence in beercellars. Alice Cibrario, Marta Avramova, Maria Dimopoulou, Maura Magani, Cécile Miot-Sertier, Albert Mas, Maria C. Portillo, Patricia Ballestra, Barry HuckerView ORCID ProfileWarren Albertin, Melinda ChristophersenIsabelle Masneuf-Pomarede, Frank VriesekoopMarguerite Dols-Lafargue. 20172019. DOI: https://doi.org/10.1101/763441.]</ref>. Other vitamins such as p-aminobenzoic acid The genetic differences between the fermentation substrates (PABA)beer, folic acid (vitamin B9)wine, nicotinic acid (vitamin B3etc.)were lower but still significant, pantothenic acid (vitamin B5) are also not required and this was explained by the frequent cross-over of equipment such as wine barrels being used for most strains of ''Brettanomyces'' to growbeer fermentation. The presence When comparing the geographic differences, they found geography contributed only 5% of genetic differences, while geography explained more than 50% of alcohol can increase genetic differences in non-wine strains, suggesting that beer, kombucha, and tequila strains are more localized genetically than wine strains and that humans probably helped the dependence on vitamins for some wine strains of ''Brettanomyces'' to growtravel across the globe. For example, MyoThey also found that although one study reported spore-inositol (vitamin B8) and thiamine (vitamin B1) were required by two strains forming versions of ''B. bruxellensis'' when grown in 10% ethanol but not in 0% ethanol. Biotin (vitamin B7referred to as ''Dekkera bruxellensis'') is the one exception, and it was found that the lack genetic makeup of biotin inhibited the growth of some analyzed strains determined their ability to sporulate to be non-existent or rare (only one study that we know of by [https://link.springer.com/article/10.1007%2FBF02539015 Walt and Kerken in 1960] has reported sporulation in ''B. bruxellensisBrettanomyces''. Other studies contradict these previously mentioned findingsonly on specific agar types with vitamins added, showing indicating that thiamine was not required by the strains of sporulation in ''B. bruxellensisBrettanomyces'' tested, that pyridoxine was required, and biotin was not required. These discrepancies between scientific studies are probably due to the genetic differences between the strains selected, the growth media chosen by the scientists, and/or the growth conditions is extremely rare) <refname="Avramova_2018" />. See also [httphttps://www.mdpifacebook.com/2311-5637groups/MilkTheFunk/2permalink/32022801681081369/17 Use Richard Preiss's discussion of Nutritional Requirements for Brettanomyces bruxellensis to Limit Infections in Wine. Nicolas H. von Cosmos and Charles G. Edwards. 2016.this study on MTF]</ref>.
See also: