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updates to Lacto starters
In addition to the starter information given in the [[Lactobacillus#Manufacturer_Tips|Manufacturer Tips]] above, this section includes general advice for ''Lactobacillus'' starters for homebrewers and brewers. For growing ''Lactobacillus'' in a lab environment, or from an initial grouping of cells from a plate/slant or smaller cell count, MRS media is the most efficient growth media. However, for full pitches of ''Lactobacillus'' in beer/wort, brewers probably don't want to add that much MRS media to their beer since MRS media has a distinct odor and smell that would not be desirable in beer <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1152135114814701/?comment_id=1152674674760745&offset=0&total_comments=9&comment_tracking=%7B%22tn%22%3A%22R0%22%7DConversation with Lance Shaner and Nick Impellitteri on Lacto starters on MTF. 09/22/2015.]</ref>. Therefore, growing ''Lactobacillus'' in a wort-based starter media is recommended for building full pitches of ''Lactobacillus''. We recommend using an Erlenmeyer flask dedicated to growing ''Lactobacillus'' in order to lower the potential for yeast contamination. We also recommend using [[Lactobacillus#Samuel_Aeschlimann.27s_Starter_Procedures|Samuel Aeschlimann's Starter Procedures]]. See [[Lactobacillus#External_Resources|External Resources]] for additional starter guides.
A good pitching rate for ''Lactobacillus'' is 100-125 billion cells per 5 gallons of wort, although pitching rates for ''Lactobacillus'' are more forgiving than they are for yeast. Having a manufacturer supply a cell count or doing a cell count manually after growth in a starter, and then pitching an exact cell count is the best approach to achieving the desired pitching rate. However, counting cells of ''Lactobacillus'' under a microscope can be difficult to achieve due to the small size of bacteria cells, so starter volumes are generally used instead when talking about pitching rates for ''Lactobacillus'' <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1068323126529234/?comment_id=1068337639861116&offset=0&total_comments=47&comment_tracking=%7B%22tn%22%3A%22R9%22%7D Conversation on MTF with Bryan of Sui Generis Blog. 5/6/2015.]</ref>. Pitching ~0.5-1 liter per ~20 liters of wort (~0.75-1 gallon per barrel) of ''Lactobacillus'' starter is the general guideline. Note that the exact advisable pitching rates of commercial cultures may differ from manufacture to manufacturer <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1077639885597558/ MTF thread started by Brad Primozic. 5/29/2015.]</ref>, and their recommendations might differ from ours. Achieving exact pitching rates for ''Lactobacillus'' is generally not necessary because over-pitching does not have a negative impact, and under-pitching but a little bit is also more forgiving tthan under-pitching yeast. For these reasons, and the difficulty in estimating cell counts based on starter media type, strain, and other variables, a pitching calculator for ''Lactobacillus'' does not exist currently. However, estimations based on starter volume and freshness should be adequate.
The amount of growth that occurs in a starter is an unknown unless the brewer can do a cell count. Matt Miller of Sour Beer Blog and Richard Preiss of Escarpment Yeast Labs advise that if ideal growth (1-2 billion cells/mL) can be achieved (for example by using [[Lactobacillus#Samuel_Aeschlimann.27s_Starter_Procedures|Samuel Aeschlimann's starter procedure]] or MRS media), then pitching as little as 100-125 mL of fresh ''Lactobacillus'' starter for 5 gallons of beer can achieve desirable acidity within 24 hours <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1180630378631841/?comment_id=1180733761954836&reply_comment_id=1180740128620866&comment_tracking=%7B%22tn%22%3A%22R9%22%7D Conversation with Richard Preiss on MTF regarding Lacto starters. 11/19/2015.]</ref><ref name="sbb2.0"></ref>. Moderately ideal growth in a starter results in around 500 million cells/mL, resulting in a 400 mL starter for 5 gallons, and low growth results in around 100 million cells/mL, resulting in a 2 liter starter for 5 gallons, according to Matt Miller (see [http://sourbeerblog.com/lactobacillus-2-0-advanced-techniques-for-fast-souring-beer/ Matt Miller's article] for more details) <ref name="sbb2.0"></ref>. If the amount of growth cannot be accurately determined by cell counting under a microscope, then pitching 0.5-1 liter of fresh ''Lactobacillus'' starter culture per 5 gallons of wort ensures that an adequate number of cells will be pitched.
Another thing to consider is that achieving a pH of 4 as fast as possible is advisable for preventing off-flavors from contaminating microbes <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1180630378631841/?comment_id=1181674265194119&reply_comment_id=1181715048523374&comment_tracking=%7B%22tn%22%3A%22R7%22%7D Conversation with Bryan of Sui Generis Blog on MTF regarding speed of acid production with Lacto. 11/20/2015.]</ref>. Larger pitch rates tend to achieve a lower pH faster <ref name="bryan_lacto_starters"></ref>. Therefore, unless using [[Lactobacillus#Samuel_Aeschlimann.27s_Starter_Procedures|Samuel Aeschlimann's starter procedure]], pitching 0.5-1 liters of starter for 5 gallons of wort is advisable in general. Even when using [[Lactobacillus#Samuel_Aeschlimann.27s_Starter_Procedures|Samuel Aeschlimann's starter procedure]], over-pitching ''Lactobacillus'' is not a concern (pitching an overly massive starter wort could produce undesirable flavors), so the same pitch rate of 0.5-1 liters for 5 gallons should still be considered unless the brewer is confident that high growth rate has been achieved in the starter, in which case 100-125 mL for 5 gallons of wort/beer should be adequate (again, exact pitching rates for ''Lactobacillus'' are generally not as important as they are for yeast pitching rates). Other factors that might affect the effectiveness of a volume based starter is the species/strain of the ''Lactobacillus'' being used, how much yeast contamination has occurred, and how old the ''Lactobacillus'' starter is. Some species/strains may require a larger volume of starter, as well as if yeast has contaminated the starter or wort (see [[Lactobacillus#100.25_Lactobacillus_Fermentation|100% Lactobacillus Fermentation]]). If a ''Lactobacillus'' culture is older than 1 month, then a fresh starter should be made. Keeping a separate Erlenmeyer flask for ''Lactobacillus'' starters can help to prevent yeast contamination <ref>Private correspondence with Richard Preiss to Dan Pixley. 11/20/2015.]</ref>, as well as using sterilization equipment such as an autoclave or pressure cooker.
Starter mediums that brewers have used include unhopped DME wort starters and apple juice starters. These tend to be adequate for many brewers. However, [https://eurekabrewing.wordpress.com/2015/05/18/evaluate-starter-media-to-propagate-lactobacillus-sp/ Samuel Aeschlimann from Eureka Brewing Blog] showed that using DME with a little bit of apple juice, chalk, and yeast nutrients provides close to optimal cell densities that match MRS media cell densities. See [[Lactobacillus#Samuel_Aeschlimann.27s_Starter_Procedures|Samuel Aeschlimann's Starter Procedures]]. Specific nutrients that will increase growth is thiamine (vitamin B1), or a combination of thiamine and riboflavin (vitamin B2). For example, it has been shown that thiamine is required in order for ''L. brevis'' to efficiently convert pyruvate into lactic acid and ethanol. The addition of these nutrients can help encourage growth <ref>[http://onlinelibrary.wiley.com/doi/10.1002/jib.385/full The influence of thiamine and riboflavin on various spoilage microorganisms commonly found in beer. Barry Hucker, Melinda Christophersen, Frank Vriesekoop. 2017.]</ref>.