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| Amyl octanoate (spicy, clove, chemical, plastic) <ref name="Lucy_2015" /> || Amyl alcohol and caprylic acid || || C<sub>13</sub>H<sub>26</sub>O<sub>2</sub> <ref>[http://www.thegoodscentscompany.com/data/rw1004161.html "Amyl octanoate". The Good Scents Company. Retrieved 02/24/2017.]</ref> || Also known as pentyl octanoate, it is a flavoring agent <ref>[https://pubchem.ncbi.nlm.nih.gov/compound/Pentyl_octanoate "Amyl octanoate". PubChem website. Retrieved 02/24/2017.]</ref>.
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| [[Ethyl acetate]] (fruity, pineapple, pear, solventy, nail polish remover) || [[Acetic Acid]] and ethanol || 33ppm (odor), 100ppm (flavor) || C<sub>4</sub>H<sub>8</sub>O<sub>2</sub> <ref>[http://pubchem.ncbi.nlm.nih.gov/compound/ethyl_acetate PubChem. Ethyl Acetate. Retrieved 08/15/2015.]</ref> || High flavor threshold; pineapple or pear-like in low amounts and nail polish in high amounts. Increases production with higher temperatures and oxygen. Also produced by ''Saccharomyces'' species <ref name="HubbeHubbe_MTF">[https://www.facebook.com/groups/MilkTheFunk/1407620505932826/ Effect of mixed cultures on microbiological development in Berliner Weisse (master thesis). Thomas Hübbe. 2016.]</ref>.
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| Ethyl butyrate (pineapple, mango, tropical fruit <ref>[http://www.flavoractiv.com/products/ethyl-butyrate-beer-flavour-standards/ Ethyl Butyrate Beer Flavour Standard. FlavorActIV. Retrieved 6/20/2015.]</ref>, juicy fruit gum <ref>Private corrospondance with Richard Preiss by Dan pixley. 12/1/2016]</ref>) || [[Butyric Acid]] and ethanol || 0.4ppm (flavor) <ref>[http://www.flavoractiv.com/products/ethyl-butyrate-beer-flavour-standards/ Flavoractiv. Ethyl butyrate. Retrieved 1/18/2015.]</ref> || C<sub>6</sub>H<sub>12</sub>O<sub>2</sub> <ref name="pubchem_ethylbutyrate">[http://pubchem.ncbi.nlm.nih.gov/compound/ethyl_butyrate PubChem. Ethyl Butyrate. Retrieved 08/15/2015.]</ref> || Low levels of production by some species of Brettanomyces; production decreases with higher acidity <ref name="yakobson1">[http://www.brettanomycesproject.com/dissertation/pure-culture-fermentation/pure-culture-fermentation-discussion/ Yakobson, Chad. Pure Culture Fermentation Characteristics of Brettanomyces Yeast Species and Their Use in the Brewing Industry. Pure Culture Fermentation Discussion. 2011.]</ref>. Also known as ethyl butanoate <ref name="pubchem_ethylbutyrate"></ref>.
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| Ethyl caproate (sweet, fruity, pineapple, banana, apple or aniseed) || Caproic acid and ethanol <ref>[https://books.google.com/books?id=1b1CAgAAQBAJ&pg=RA2-PA320&lpg=RA2-PA320&dq=Ethyl+caproate+precursors&source=bl&ots=myHXfoVz9f&sig=fHGkce4UmeJVC4M3Kk4TXUCO-Nc&hl=en&sa=X&ei=ip68VOqjFY-tyASpmoHoCA&ved=0CEQQ6AEwBA#v=onepage&q=Ethyl%20caproate%20precursors&f=false Encyclopedia of Food Microbiology. Batt, Carl A. Academic Press. Sep 28, 1999. Pg 320.]</ref> || 0.2ppm (flavor) <ref>[http://www.aroxa.com/beer/beer-flavour-standard/ethyl-hexanoate/ Aroxa. ethyl hexanoate. Retrieved 1/18/2015.]</ref> || C<sub>8</sub>H<sub>16</sub>O<sub>2</sub> <ref>[http://pubchem.ncbi.nlm.nih.gov/compound/31265 PubChem. Ethyl Caproate. Retrieved 08/15/2015.]</ref> || Also known as Ethyl hexanoate, Ethyl butyl acetate, and butylacetate <ref>[http://www.chemspider.com/Chemical-Structure.29005.html Chemspider. Ethylhexanoat. Retrieved 1/18/2015.]</ref>. Can also be produced by ''Saccharomyces'' species <ref name="HubbeHubbe_MTF"></ref>.
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| Ethyl caprylate (Sweet, waxy, fruity and pineapple with creamy, fatty, mushroom and cognac notes <ref>[http://www.thegoodscentscompany.com/data/rw1056351.html Ethyl Octanoate. The Good Scents Company. Retrieved 5/28/2015.]</ref>) || Caprylic acid (contained in buckwheat; produced by yeast autolysis) and ethanol <ref>[https://www.youtube.com/watch?v=2QgXptgw1ys Chop & Brew - Episode 37: Influence of Mashing on Sour Beer Production by Michael Tonsmeire. NHC 2014 Presentation. At 26 minutes. Retrieved 5/28/2015.]</ref> || 15ppb (flavor) <ref>[http://www.leffingwell.com/esters.htm Esters Detection Tresholds & Molecular Structures. Leffingwell & Associates. Retrieved 5/28/2015.] </ref> || C<sub>10</sub>H<sub>20</sub>O<sub>2</sub> <ref name="pubchem_ethylcaprylate">[http://pubchem.ncbi.nlm.nih.gov/compound/7799 PubChem. Ethyl Caprylate. Retrieved 08/15/2015.]</ref> || Also known as ethyl octanoate <ref name="pubchem_ethylcaprylate"></ref><ref>[https://pubchem.ncbi.nlm.nih.gov/compound/Ethyl_caprylate "Ethyl caprilate". PubChem website. Retrieved 01/30/2017.]</ref>.
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| Ethyl Decanoate/Ethyl Caprate (brandy, fruity, oily, grape) || Decanoic acid (Capric Acid) and ethanol <ref name="wikipedia_ethyldecanoate">[http://en.wikipedia.org/wiki/Ethyl_decanoate Wikipedia. Ethyl Decanoate. Retrieved 1/18/2015.]</ref> || 0.5ppm (flavor in water) <ref>[https://books.google.com/books?id=ax1OvyH8jGoC&pg=PA222&dq=Ethyl+Decanoate+threshold&hl=en&sa=X&ved=0CCsQ6AEwAmoVChMIv865pbSsxwIVEaWICh0OIQVt#v=onepage&q=Ethyl%20Decanoate%20threshold&f=false Flavours and Fragrances: Chemistry, Bioprocessing and Sustainability. Ralf Günter Berger. Springer Science & Business Media, Mar 6, 2007. Pg 222.]</ref> || C<sub>12</sub>H<sub>24</sub>O<sub>2</sub> <ref name="wikipedia_ethyldecanoate"></ref> || Also known as Ethyl caprate, Ethyl caprinate, and Capric acid ethyl ester <ref>[http://www.michiganbrewersguild.org/docs/Gary-Spedding_off-flavor-notes_2014WC.pdf Spedding, Gary. Flavor notes for Michigan Craft Guild Conference. 2014.]</ref>. Can also be produced by ''Saccharomyces'' species <ref name="HubbeHubbe_MTF"></ref>
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| Ethyl hexanoate <ref name="Lucy_2015" /> (Apple or aniseed <ref name="aroxa_ethylhexanoate">[http://www.aroxa.com/beer/beer-flavour-standard/ethyl-hexanoate/ "Ethyl hexanoate". Aroxa Website. Retrieved 01/31/2017.]</ref>) || Hexanoic acid and ethanol <ref>[https://en.wikipedia.org/wiki/Ethyl_hexanoate "Ethyl hexanoate". Wikipedia. Retrieved 01/31/2017.]</ref>. || 0.2ppm (flavor in beer) <ref name="aroxa_ethylhexanoate" /> || C<sub>8</sub>H<sub>16</sub>O<sub>2</sub> || Also produced by both ale and lager yeast; it is a key flavor note in many beers <ref name="aroxa_ethylhexanoate" />. High amounts Produced by two strains out of 9 ''B. bruxellensis'' in one study <ref name="Lucy_2015" />.
| Pentyl formate (artificial fruit, candy, chemical) <ref name="Lucy_2015" /> || Pentanol and formic acid <ref>[https://en.wikipedia.org/wiki/1-Pentanol "1-pentanol". Wikiepedia article. Retrieved 02/24/2017.]</ref><ref>[https://en.wikipedia.org/wiki/Formate "Formate". Wikipedia article. Retrieved 02/24/2017.]</ref> || || C<sub>6</sub>H<sub>12</sub>O<sub>2</sub> || Also known as amyl formate <ref>[http://www.thegoodscentscompany.com/data/rw1015511.html "Amyl format". Good Scents Company website. Retrieved 02/24/2017.]</ref>.
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| Phenethyl acetate (sweet, honey, rose flower like) || Acetyl-CoA and 2-phenylethanol <ref>[http://pubchem.ncbi.nlm.nih.gov/compound/7654 PubChem. Phenethyl Acetate. Retrieved 08/15/2015.]</ref><ref>[http://www.ymdb.ca/compounds/YMDB00574 YMDB. Phenethyl acetate.]</ref> || 3-5ppm (odor), 5-10ppm (flavor) <ref>[https://books.google.com/books?id=A8OyTzGGJhYC&pg=PA1521&lpg=PA1521&dq=Phenethyl+acetate+flavor+threshold&source=bl&ots=Ygvq_JFvhQ&sig=06Lj57fWYemcXizkEHnYzwfOXX8&hl=en&sa=X&ei=Zz-nVOPCKI61oQSFwIGABQ&ved=0CEAQ6AEwBA#v=onepage&q=Phenethyl%20acetate%20flavor%20threshold&f=false Burdock, George A]. Fenaroli's Handbook of Flavor Ingredients, Fifth Edition. CRC Press. 2005. pg 1521. </ref> || C<sub>10</sub>H<sub>12</sub>O<sub>2</sub> || Produced in very small amounts in Lambic <ref name="Spaepen"></ref><ref>[https://vtechworks.lib.vt.edu/bitstream/handle/10919/19203/Witrick_KA_D_2012.pdf?sequence=1 Characterization of aroma and flavor compounds present in lambic (gueuze) beer. Katherine A Thompson Witrick. 2012.]</ref>. Can also be produced by ''Saccharomyces'' species <ref name="HubbeHubbe_MTF"></ref>
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| Phenethyl formate (artificial floral, perfume, wild flower, solvent) <ref name="Lucy_2015" /> || 2-phenylethanol and formic acid || || C<sub>9</sub>H<sub>10</sub>O<sub>2</sub> ||
===Starter Information===
When pitching just ''Brettanomyces'' from a commercial pure or blended culture and no other microbes, it is recommended to make a starter for the culture. If the ''Brettanomyces'' is being pitched into secondary, no starter is necessary unless the brewer suspects that the ''Brettanomyces'' has lost a lot of viability due to age, heat exposure, etc., or prefers higher cell count pitches (current information suggests that there is no significant flavor difference between high and low pitching rates in secondary pitches of ''Brettanomyces''; see [[Brettanomyces secondary fermentation experiment]]). ''Brettanomyces'' growth is inhibited by the concurrent growth of ''S. cerevisiae'' under anaerobic conditions. In aerobic conditions, oxygen appears to help ''Brettanomyces'' out-compete ''S. cerevisiae''. The presence of lactic acid bacteria does not greatly effect ''Brettanomyces'' growth <ref name="Hubbe"></ref><ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1362220513806159/ MTF post by Brian Martyniak on ''Brettanomyces'' growth in competition with ''Saccharomyces'' under aerobic and anaerobic conditions. 07/29/2016.]</ref>. Therefore, when making starters for mixed cultures of ''Brettanomyces'' and ''Saccharomyces'', the brewer can favor ''Saccharomyces'' by limiting oxygen, or favor ''Brettanomyces'' by introducing oxygen during growth. Data from Thomas Hübbe and Mark Trent support that the initial pitching rate doesn't have a great effect on the final cell count in pure ''Brettanomyces'' starters or beer, indicating that ''Brettanomyces'' is fairly forgiving in regards to small initial cell counts <ref name="Hubbe"></ref><ref>[[Brettanomyces_Propagation_Experiment|MTF ''Brettanomyces'' Propagation Experiment]]</ref>.
Just like in other yeast species, temperature has a direct effect on the rate of growth for ''Brettanomyces''. The optimal growth rate temperature range for ''Brettanomyces'' is between 25-32°C (77-90°F). Growth is about half as slow at 20°C (68°F). ''Brettanomyces'' will still grow at temperatures as low as (and maybe lower than) 15°C (59°F) and will be much slower, however one study showed a slightly higher viability during the full-time period of fermentation at 15°C as opposed to the optimal growth temperature range of 20-32°C. At a temperature of 35°C (95°F), both growth and viability over time are greatly inhibited <ref name="Brandam_2008">[http://oatao.univ-toulouse.fr/1595/1/Brandam_1595.pdf Effect of temperature on Brettanomyces bruxellensis: metabolic and kinetic aspects. Brandam C, Castro-Martínez C, Délia ML, Ramón-Portugal F, Strehaiano P. 2008.]</ref>.
====Cell Counting====
The use of methylene blue, although popular in breweries, has been shown to be inaccurate when counting cells of ''Brettanomyces''. Trypan blue staining has been shown to give more accurate cell counting results <ref name="Hubbe"></ref><ref>[https://www.facebook.com/groups/MilkTheFunk/1407620505932826/?comment_id=1407723619255848&comment_tracking=%7B%22tn%22%3A%22R8%22%7D Discussion with Richard Preiss regarding counting Brettanomyces with methylene blue versus trypan blue staining. 09/15/2016.]</ref>.
====Example of a Home Lab Orbital Shaker====
In order to explore Yakobson's anecdotal observations in a more controlled manner, Mark Trent performed an experiment on storing one strain of ''Brettanomyces'' in wort, MYPG, buffered wort (buffered to prevent a drop in pH), and buffered MYPG, and compared storage of the ''Brettanomyces'' in each of the storage solutions at room temperature versus cold temperatures for 100 days. This single ''Brettanomyces'' strain survived best in unbuffered MYPG at room temperature, and second best in unbuffered wort at room temperature, and survived less in cold storage conditions for all media. See the [[Brettanomyces Storage Survival Experiment]] for more details. Therefore, when storing ''Brettanomyces'' for one month or less in wort (or perhaps beer), it should be stored refrigerated. However, if the ''Brettanomyces'' will be stored for more than one month in wort (or perhaps beer), it should be stored at room temperature (until more data improves our understanding). Note that at best these storage techniques will decrease viability greatly (80%+) within 3 months, and a starter should be used to try and revive the culture before use <ref>[[Brettanomyces_Storage_Survival_Experiment]]</ref>.
Occasional feeding has been shown to keep ''Brettanomyces'' alive in beer for brewers who do not have a lab, however many variables may come into play as far as how effective this will be for individual strains and in different environments. Although no research has been done to indicate what the best practices are for feeding ''Brettanomyces'' to keep it alive in beer, we recommend trying this method: every 3-6 months swirl the vessel so as to suspend all of the yeast and then decant 70-90% of the beer and suspended yeast slurry, and replace it with a 1.040 starter wort with yeast nutrients. This method will discard a lot of the old yeast cells, while retaining enough living cells for replication <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1345924532102424/?comment_id=1345979272096950&reply_comment_id=1346020438759500&comment_tracking=%7B%22tn%22%3A%22R3%22%7D Conversation with Mark Trent and Richard Preiss on MTF regarding occasional feeding of ''Brettanomyces'' to keep it alive. 07/07/2016.]</ref>. Some strains may survive extended periods of aging in beer, however their viability and vitality will be greatly reduced over time. Interestingly, ''Brettanomyces'' remains '''more''' viable over time if it was co-fermented with ''S. cerevisiae'' than if it was fermented without the presence of ''S. cerevisiae''; i.e. [[100%25_Brettanomyces_Fermentation|100% ''Brettanomyces'' beers]] or ''Brettanomyces'' and ''Lactobacillus <ref name="Hubbe"></ref>.
Another method for storing ''Brettanomyces'' has reportedly worked for MTF member Justin Amaral. This method involves storing the culture in isotonic sodium chloride. ''Brettanomyces'' cultures have been reported by Amaral to survive at least for 6-7 months. This includes other microbes as well (RVA Orchard Brett, ECY Dirty Dozen, Bright Yeast Labs Brett Chateaux, ''T. delbrueckii'', ''L. plantarum'' isolated from goodbelly, Omega Lacto blend, ''Pediococcus damnosus'', Bootleg Biology Sour Weapon, and Funk Weapon 2 and 3, and a ''Brettanomyces'' isolate from Yeast Bay). For more information on this method, see [https://eurekabrewing.wordpress.com/2012/09/05/yeast-banking-3-isotonic-sodium-chloride/ this Eureka Brewing blog article] <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1631448736883334/ Amaral, Justin. Milk The Funk Facebook group. 03/30/2017.]</ref>.