Changes

| [[Craft Cultures]] || CCYL512 || L. brevis || Heterofermentative || || Typically produces more lactic acid than Lactobacillus delbrueckii. Commercial pitches only.

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| [[Wyeast]] || 5335 || L. buchneri || Heterofermentative <ref name="mtf_wiki_shaner"></ref> || 1 liter starter for a 5 gallon batch of beer, 1.020 DME sterile wort, no stir plate, no O2, starter at 90°F if possible 5-7 days || Incubate at 90°F for 5-7 days for greater lactic acid production. Cell count: 1.0 x 10⁸ (100 million) cells/mL (10 billion cells in a 100 mL homebrew pouch) <ref name="wyeast_cellcounts">[https://drive.google.com/folderview?id=0B8CshC9nxYHdZmE4MmoyLXA2WVk&usp=sharing Wyeast Specifications 2015 Retail Products. 2015.]</ref>.

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| [[Omega Yeast Labs]] || OYL-605 || L. brevis, <span style="text-decoration: line-through;">delbrueckii</span>, and plantarum blend || Hetero/Hetero <ref name="mtf_wiki_shaner"></ref> || 1 liter starter for a 5 gallon batch of beer at room temperature for 24-48 hours. No stir plate unless kept anaerobic. || Quick souring. Pitch into 65°F-95°F <ref name="adi_oyl605"></ref>. Holding temperature is not required. No longer contains delbruekii <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1065268213501392/?comment_id=1065669443461269&offset=0&total_comments=18&comment_tracking=%7B%22tn%22%3A%22R%22%7D Conversation with Raymond Wagner of Oso Brewing Co on Milk The Funk. 4/30/2015.]</ref>. Don't use any hops if possible. 2 IBU is a good target if hops must be used <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1092523807442499/?comment_id=1092571350771078&offset=0&total_comments=6&comment_tracking=%7B%22tn%22%3A%22R1%22%7D Conversation with Lance Shaner on MTF in regards to IBU tolerance of OYL-605. 6/15/2015.]</ref>. Contains ~150 billion cells per homebrew pitch <ref name="sbb2.0">[http://sourbeerblog.com/lactobacillus-2-0-advanced-techniques-for-fast-souring-beer/ Lactobacillus 2.0 – Advanced Techniques for Fast Souring Beer. Sour Beer Blog. Matt Miller. 11/18/2015. Retrieved 11/19/2015.]</ref>.

| [[SouthYeast Labs]] || Lactobacillus 2 || Unknown || Homofermentatative || || Source: Prickly pear fruit (South Carolina). Best suits strong sours, and lambic (high acidity).

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| [[Escarpment Laboratories]] || Lactobacillus Blend || L. brevis and L. plantarum || Heterofermentative || || This blend is designed to be usable at a wide range of temperatures, and is especially suited for kettle souring/sour worting. We recommend pre-acidifying wort to 4.5 with lactic acid, then pitching the Lactobacillus blend in a CO2-purged kettle or fermentor at 32-42°C.

| [[Escarpment Laboratories]] || Lactobacillus brevis || L. brevis || Heterofermentative || || This strain is moderately hop-tolerant, and as such it can also be used for long-term souring of <10IBU beers. It also performs well in kettle souring/sour worting where fast and clean lactic acidity is desired. We recommend pre-acidifying wort to 4.5 with lactic acid, then pitching the Lactobacillus blend in a CO2-purged kettle or fermentor at 35-45°C.

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====[[White Labs]] on WLP672====

"It is intended for secondary, so you only need to do a starter if you are doing a primary fermentation with it. DME would be the best substrate... Since its a Lacto species, you don't really want to aerate it. A slow stir-plate would be good, to keep it moving, but not much more than that." - Sarah Neel, White Labs, Sales and Customer Service (quoted with permission).

===General Advice===

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 should still be considered unless the brewer is confident that high growth rate has been achieved in the starter. 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.

Although more experiments and probably needed, agitation is believed to be an important factor for both yeast and bacteria in general. Gentle stirring on a stir plate or orbital shaker, or frequent gentle manual agitation leads to faster growth and a higher number of organisms. Agitation keeps the microbes in solution. It also maximizes the microbes' access to nutrients and disperses waste evenly. In a non-agitated starter, the microbes are limited to the diffusion rate of nutrients, leading to a slower and more stressful growth <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1168024059892473/?comment_id=1174865305875015&reply_comment_id=1176092372418975&total_comments=1&comment_tracking=%7B%22tn%22%3A%22R9%22%7D Conversation with Bryan of Sui Generis Blog about starters and agitation. 11/09/2015.]</ref>. If agitation is not possible for whatever reason, a successful starter can be made without agitation. Sam Aeschlimann reported good success with ''Lactobacillus'' starters that are not agitated <ref name="Sam_starter2"></ref>.

Although 100% apple juice or 100% DME starters will "work" for ''Lactobacillus'' starters, they do not provide optimal growth conditions. [https://eurekabrewing.wordpress.com/2015/05/18/evaluate-starter-media-to-propagate-lactobacillus-sp/ Samuel Aeschlimann from Eureka Brewing Blog] ran a set of experiments that found a DME based recipe for starter wort that produces a very high cell density similar to that of MRS media, which provides optimal growth rates for ''Lactobacillus''.

# Add 100 grams of DME to around 900 mL of water and heat pasteurize/boil as you would normally do for a starter. This should make 1.040 SG (10°P) starter wort.

<blockquote>

Hops contain multiple compounds which are bacteriostatic. Alpha acids are the best understood, but other compounds such as beta acids, a number of polyphenols (e.g. xanthohumol), and even some of the aromatic oils (e.g. humulene) have been found to have some inhibitory effects on ''lactobacilli''. The later compounds (especially the beta acids) are why aged hops retain inhibitory characteristics, despite being nearly devoid of alpha acids. In all cases these compounds appear to inhibit the bacteria in the same way - all of these compounds contain fairly large, flat-ish, hydrophobic regions. These regions do not "like" to be in water, and thus will be driven into the hydrophobic core of the bacterial plasma membrane. This opens minute holes in the membrane which prevents the bacteria from maintaining ion (in particular, proton) gradients, leading to suppression of growth and even death of the bacteria.

Hop resistance is generally due to the induced expression of "multi-drug transport" (MDT) genes, which are "pumps" that recognize the general chemical signature of membrane-disruptive compounds, and then pump them out of the cell. Other mechanisms may also be involved - a few papers have identified changes in the lipid make-up of the plasma membrane, which may increase stability. This change also occurs in response to alcohol (to improve stability), so its not clear if that particular change has anything to do with hop resistance.

</blockquote>

See also:

* [http://www.garshol.priv.no/blog/337.html How hops prevent infection, by Lars Garshol].

* [https://www.youtube.com/watch?v=J2g5P7ZlGn4 Per Buer's Video Demonstration of how dry hopping inhibits ''Lactobacillus''.]

====Storage====

* See [http://basementbrewlab.com/lab/lab-media/sla/ Rogosa SL Agar (Lactobacillus); Basement Brew Lab, by Dustin Metzger] for more information.

== Commercially available Lactobacillus strains and their pH change over time ==

| Enterococcus faecalis <ref name="fao"></ref> || ||

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====100% Lactobacillus Fermentation====

The amount of CO2 produced is very small in heterofermentative species. Lance Shaner of Omega Yeast Labs noted that although ''L. brevis'' is classified as obligatory heterofermentative, the human eye cannot detect any CO2 production in the Omega Yeast Lactobacillus blend (OYL-605). Lance still needs to test this blend to see if it produces any CO2 at all. There have been reliable reports of pure ''Lactobacillus brevis'' cultures producing a layer of bubbles on the surface of wort if roused <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1354678291227048/?comment_id=1354678411227036&reply_comment_id=1355288821165995&notif_t=group_comment_reply&notif_id=1468974761019794# Conversation with Richard Preiss on MTF regarding pure Lactobacillus fermentation. 07/19/2016.]</ref>. It is clear though that any type of ''Lactobacillus'', regardless of whether it is heterofermentative or homofermentative, cannot produce a krausen. Krausens are sometimes seen even with the use of commercially available ''Lactobacillus'' cultures and good sanitation techniques. If a krausen develops in wort when it is the only culture that is pitched, this is indicative of cross contamination of ''Saccharomyces'' or ''Brettanomyces'' in either the wort, or the ''Lactobacillus'' culture itself <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1083842231643990/?comment_id=1084646124896934&offset=0&total_comments=26&comment_tracking=%7B%22tn%22%3A%22R8%22%7D Discussion with Lance Shaner on MTF. 6/7/2015.]</ref>. In addition to this, heterolactic fermentation by ''Lactobacillus'' can only produce 10-20% of the ethanol that Saccharomyces can produce <ref name="PhysioLacto">[http://phdinbeer.com/2015/04/13/physiology-of-flavors-in-beer-lactobacillus-species/ Humbard, Matt. Physiology of Flavors in Beer – Lactobacillus Species. Retrieved 6/14/2015.]</ref>, therefore a high level of attenuation cannot be achieved by ''Lactobacillus'' and is again a sign of cross contamination by yeast.

<youtube>9a-ZpF2LDm8</youtube>

* See also [[100% Lactobacillus Fermentation]].

The type of grain that the ''Lactobacillus'' is fermented in may also play a role in the types and amounts of secondary metabolites that are produced. One study compared volatile acids produced by a probiotic strain of ''L. plantarum'' (NCIMB 8826) when fermented in oats, barley, malted barley, and wheat. In oats, there was slight increase in oleic acid and linoleic acid and a decrease when fermented in wheat, barley, or malted barley. In malted barley, there were small increases in flavor active compounds such as furfural ("almond" flavor), 2-ethoxyethyl acetate and isoamyl alcohol, but little to none detected when fermented in oats, wheat, or unmalted barley. Acetic acid production was higher in barley and malted barley than it was in oats and wheat. Many other organic acids in the oats, wheat, barley, and malted barley were supposedly taken up by the ''L. plantarum'' during fermentation. In barley, there were trace amounts of new acids created that were not already in the barley itself <ref>[http://www.sciencedirect.com/science/article/pii/S0308814609004373 Volatile compounds produced by the probiotic strain Lactobacillus plantarum NCIMB 8826 in cereal-based substrates Ivan Salmeron, Pablo Fuciños, Dimitris Charalampopoulos, Severino S. Pandiella. 2009.]</ref>. Some species of ''Lactobacillus'', including ''L. lactis'' and ''L. plantarum'', produce diacetyl (which can be reduced to acetoin and 2,3-butanediol) as an intermediate metabolite from consuming sugar, citrate, and amino acids. However, citrate levels are rather low in malted barley (but higher in sorghum), and diacetyl production has been observed to be very low in barley and oat based worts <ref name="peyer_review"></ref>.

Aging has a large impact on the aromas and flavors produced by ''Lactobacillus'' fermentation over time and is typically influenced by temperature of the environment, oxygen exposure, and the byproducts of fermentation. Generally, fermentation has a positive effect on preserving some aroma and flavor compounds. Other compounds may change, causing aroma and flavor changes. For example, one study characterized wort freshly fermented with ''L. plantarum'' as "butter" and honey", and when aged as "yogurt" and "sour". In the same study, ''L. reuteri'' was characterized as "sour" when fresh, and "honey" and "pungent" when aged. ''L. brevis'' was characterized as "soy sauce" when fresh, and "yeasty" and "cider" when aged <ref name="Peyer"></ref>.

See the [[Lactobacillus#100.25_Lactobacillus_Fermentation|Elke Arendt video presentation above]] on the referenced study, starting at ~14:45.

===External Resources===

* [https://matthumbard.wordpress.com/2015/04/13/physiology-of-flavors-in-beer-lactobacillus-species/ Physiology of Flavors in Beer – Lactobacillus Species] - Matt Humbard's overview of different species of Lactobacillus physiology, discussion on homofermentative vs heterofermentative physiology, which species can ferment different types of sugars, and secondary metabolites. Extensive data points included.

* [http://phdinbeer.com/2015/08/05/beer-microbiology-lactobacillus-ph-expeirment/ Beer Microbiology – Lactobacillus pH experiment] - Matt Humbard's experiment to determine final pH of individual ''Lactobacillus'' strains used in the graphs on this page.