Difference between revisions of "Wort Souring"
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==Various Other Concerns==
==Various Other Concerns==
===Not Boiling and Dimethyl Sulphide (DMS)===
===Not Boiling and Dimethyl Sulphide (DMS)===
This is generally not a concern. See [[Dimethyl Sulfide]] for an explanation why.
This is generally not a concern. See [[Dimethyl Sulfide]] for an explanation why.
Revision as of 00:36, 17 September 2021
Wort Souring is the process of Mixed Fermentation where lactic acid bacteria, usually Lactobacillus (although there are also Pediococcus cultures available that work well for this technique), is given a "head start" as it's pitched before the yeast so that it will be able to produce significant amounts of lactic acid before the Saccharomyces completes the main fermentation. While non-sour beer falls in the range of 3.8-4.6 pH, a final pH between 3.0 and 3.7 is the general target range for the soured wort and also the finished sour beer (although Titratable Acidity is more accurate for measuring perceived sourness). This is a broadly defined technique and there are many variations on souring wort. These include souring in the primary fermenter, souring in a secondary vessel, or souring in the boil kettle itself (kettle souring). There are also various methods of inoculating the wort with Lactobacillus. Finally, the brewer has the option of pasteurizing the wort by heating it to kill the Lactobacillus before adding the yeast for the main fermentation.
Although this process generally does not include the use of Brettanomyces, some creative brewers have applied wort souring techniques to longer aged Mixed Fermentation beers and barrel aged beers that do contain Brettanomyces.
Many brewers prefer this process over Sour Mashing because it can be easier to control, and when implemented properly, it produces a clean sour beer in a short amount of time. However, the environment created provides an ideal situation for contaminations and great care should be taken to prevent contamination of both yeast and spoilage microbes (see Contamination Concerns below).
The possibility of pasteurizing the soured wort also makes this a good method for making sour beers with a lot of residual malt sweetness (e.g. sour barley wines), and also makes it an attractive process for brewers who are concerned about infection issues in their cold side equipment (equipment that is used post-boil) . When souring wort, some brewers first lower the pH of the wort to 4.0-4.3 before pitching Lactobacillus. This sometimes helps the head retention of the beer and it sometimes helps to protect the wort from contaminating microorganisms. For more information preventing the loss of head retention in sour beers, see the Lactobacillus page section on Foam Degradation.
Generally, Pediococcus is not used with this method (Pediococcus is generally used in long aged Mixed Fermentation sours with Brettanomyces), however Bootleg Biology has released a blend of Pediococcus strains that are reportedly good for souring wort.
Important note regarding aluminum pots: souring in an aluminum vessel may strip the aluminum of the protective oxide layer. The oxide layer is only stable at a pH of 4.5 - 8.5. Therefore, kettle souring in an aluminum pot is generally not recommended . Stainless steel (304 and 316) vessels are safe for holding acidified wort or beer, as well as PET and HDPE plastics .
"Mixed culture fermentation for sour beers produces one thing, and (kettle souring) produces another thing. If you’re going to make a malty red ale that is kettle soured, don’t call it a Flanders Red. Honor the tradition." - Sean Burke, Kettle Souring Presentation, CBC 2015.
- 1 Strain Selection
- 2 Processes
- 3 Contamination Concerns
- 4 Various Other Concerns
- 5 See Also
- 6 References
The strain of Lactobacillus is very important. As is explained below, the souring process should not take longer than 2 days, and some strains do not perform well within this timeframe. Not all strains or species are efficient to use for wort souring. Lactobacillus plantarum has proven to be the most successful species for kettle souring because it produces acidity very quickly, and it is not as vulnerable to temperature shifts, although using any hops will usually completely prevent this species from producing lactic acid. L. brevis has also been used with some degree of success, depending on the strain. While Pediococcus is generally not used for wort souring, Bootleg Biology's Sour Weapon P, which is P. pentosaceus, performs well at wort souring (see Commercial Pediococcus cultures). The L. delbruekii culture offered by White Labs is meant for long term souring and is not a good choice for wort souring (other strains of L. delbruekii might perform better) . Review the Lactobacillus commercial cultures before selecting one for wort souring, and review the Alternative Bacteria Sources page for tips on using alternative sources of Lactobacillus such as probiotics, grain, etc.
Lactobacillus Nutrient Requirements
Different strains of Lactobacillus are known to have different nutrient requirements, including some metal ions. Lactobacillus requires manganese, magnesium, potassium, and sodium, with manganese and magnesium being crucial for their growth. Brewer's wort has many of these nutrients naturally, although zinc is generally deficient. One study looked at WLP672 (L. brevis) and found that an increase of magnesium resulted in a faster souring phase, while zinc additions slightly slowed the souring phase. This suggests that in at least the case for this strain of L. brevis, zinc additions should be added after the souring phase for yeast health. Excess magnesium could lead to negative flavor contributions to beer, so magnesium additions should not exceed 30 mg/l . The addition of zinc also had several effects on the flavor and aroma compounds produced. Therefore, experimenting with different nutrient additions for the souring phase may result in widely different tasting beer . See this MTF thread for more commentary on this study.
Souring in the Boiler (Kettle Sour)
Also known as kettle souring, souring in the boil kettle is a simple process that is often used if the brewer wants to subsequently heat pasteurize the wort. Pasteurizing the wort has the advantage of allowing the brewer to rack the pasteurized wort into a fermenter and pitch brewer's yeast without fear of an ongoing Lactobacillus infection in their post-boil equipment. This process was originally invented by a German scientist named Otto Francke, and called the "Francke acidification process". It was designed by Francke as a way to shortcut the mixed culture fermentation of Berliner Weisse; however, this process was never really used in Berliner Weisse production because it did not produce a product that resembled Berliner Weisse that was fermented with a mixed culture of yeast and bacteria (including Brettanomyces). See the Berliner Weisse page for more information on the history of Berliner Weisse production methods .
The brewing process is the same for any all-grain batch up until the first wort and sparge runnings are collected in the boil kettle. The temperatures that a typical mash out/sparge are not enough to completely pasteurize the wort . Therefore, the best approach is to heat the wort for a short boil (1-2 minutes) in order to kill a greater degree (2-3 logs more) of thermotolerant microbes . Once all of the wort is collected in the boil kettle (and preferably brought to a boil), the wort is chilled to around 80-115°F (37-46°C), depending on the Lactobacillus culture that is being used (see the Lactobacillus culture charts). Once chilled to the appropriate temperature, the wort in the kettle is inoculated with a culture of Lactobacillus. Hops should not be added at any point before inoculating the wort with a culture of Lactobacillus as most species of Lactobacillus will be inhibited by the presence of even very small amounts hops (1-2 IBU or even just hop material from dry hopping). When using a pure culture of Lactobacillus, it is generally a good idea to create a 500 mL starter for ~5-6 gallons of wort.
There are various ways of inoculating the wort. A reliable method is pitching a pure culture of Lactobacillus or a blend of Lactobacillus cultures. Alternatively, a handful of unmilled malted barley can be added to the kettle for inoculation, similar to how sauergut is made, instead of a pure culture since the husks of grain carry many microorganisms. If unmilled grain is added it is thought that filling the head space of the kettle with CO2 (or another gas like argon or nitrogen; always provide proper ventilation when working with any gasses ) will help decrease off-flavors such as "footiness" from Isovaleric Acid which are produced by aerobic microbes that are naturally present on the grain . Keeping the temperature between 109-115°F (42.8-46°C) will encourage the Lactobacillus resident on the grain and will discourage other bacteria. Temperature consistency is critical during this process . Lowering the pH of the wort to under 4.5 (ideally 4.0 - 4.3 when souring with grain) with lactic acid or phosphoric acid will discourage other unwanted bacteria and wild yeast from thriving in the wort during the incubation period. This will also help with head retention (see Lactobacillus and head retention) . Souring with grains should occur within 1 or 2 days if done correctly . Do not consume wort that has been soured with grains until after it has been fully fermented by yeast because there is a chance that food poisoning pathogens will be present until ethanol is produced. See Blue Owl Brewing's grain inoculation methodology and data for more information on inoculating with grain. Consider Alternative Bacteria Sources for more reliable approaches to using "wild" Lactobacillus, or Lactobacillus from sources other than yeast labs.
If a pure culture of Lactobacillus bacteria is used it has historically been recommended to fill the head space of the fermenter with CO2 gas or another type of gas like argon or nitrogen to purge the oxygen (oxygen does not cause Lactobacillus to produce butyric acid or isovaleric acid, but some brewers have reported that purging oxygen will help reduce sulfur in the finished beer). Purging oxygen could also discourage mold growth. Mold growth during the souring process has been reported from time to time by homebrewers, and if mold grows in contact with the wort then it probably should be discarded (see the Mold page for more information) . However, many homebrewers and some professional brewers have reported having success without purging with CO2 (~20:30 mins in), and a Brulosophy blind triangle test was unable to find a significant difference between a kettle sour that was CO2 purged and one that was soured in an open vessel. Using some CO2 to keep positive pressure in the kettle could help prevent contaminants that create butyric acid and other off-flavors from entering the kettle due to negative pressure, and is often the approach that commercial brewers take . The kettle should be held at the desired temperature for 24-72 hours (in some cases longer, but no longer than 5 days). Depending on the strain of Lactobacillus, and the desired sour level, the time of incubation is ultimately a variable that is up to the brewer (see the Lactobacillus page for suggested temperatures and times for specific strains). The kettle lid should be firmly in place and optionally sealed with plastic wrap so that other microorganisms do not get in. Potential for the formation of Butyric Acid and Isovaleric Acid when using only a pure culture is extremely slight assuming that contamination does not occur. Temperature shifts during kettle souring are not a concern as long as the temperature does not get too high or low for the specific Lactobacillus culture. Some species, such as L. plantarum, create acidity at room temperature so some brewers will pitch this strain at around 90-100°F and let the temperature fall to room temperature during souring. Other species might not perform as well at colder temperatures so maintaining a fairly consistent hot temperature is desirable. If the temperature is allowed to fall, take precautions on not allowing any dust to get sucked into the fermenter since temperature decreases will create a vacuum inside the fermenter (flushing with CO2 is a good way to prevent a vacuum).
In order to address the challenges of creating a sealed environment in a commercial boil kettle, which is often left open to the environment through the boil stack, some commercial brewers find it beneficial to rack the wort from the boil kettle to a mashtun, brite tank, fermenter (often dedicated), or another vessel that seals and purges well to sour in. The mashtun or other vessel is first cleaned, sanitized, and often purged with CO2. Once the wort is in the vessel, and at the correct temperature for the given culture of Lactobacillus, the Lactobacillus is pitched into the vessel and the vessel is sealed off. If the vessel is sealed and air tight temperature shifts from cooling won't suck air into the vessel and a potentially more sanitary souring fermentation will occur. Using a fermenter or brite tank (if there are multiple) has the advantage of not occupying a bottleneck vessel such as the mashtun or boil kettle, but has more risk if the fermenter is not dedicated to kettle souring. Dedicating removable soft parts like gaskets and hoses will minimize the risk of infection originating from the fermenter or brite tank if it is not dedicated to souring .
Once the level of acidity is reached (this can be tested with a reliable pH meter, or in the case of using a pure culture can safely be taste tested), the wort is brought to a boil. The wort may be boiled normally in the case of any style of beer that requires a longer boiling process, or it may be boiled for no more than a minute or two in the case of making a Berliner Weissbier. Technically speaking, the wort doesn't need to be boiled at all (this is called Raw Ale). Heat pasteurization at 180°F (82°C) for 15 minutes will kill even the most heat tolerant Lactobacillus species . Note that boiling a soured wort that has a pH lower than 5.0 will decrease the amount of protein coagulation, potentially resulting in a hazier beer or more sediment in the fermenter . Haze and sediment can be avoided by choosing to boil long enough to hit a hot break before souring the wort (see example MTF post from Chris Prechel). Raw ales are also fairly hazy in nature due to the lack of hot break from boiling. See Haze below for more information.
Deciding whether or not to boil the wort can also depend on whether or not there was a considerable amount of alcohol produced as commonly happens when the wort is contaminated with yeast. Pure cultures of Lactobacillus do not show typical signs of fermentation that we are used to seeing with yeast fermentations, such as forming a krausen, producing a lot of CO2, or fermenting wort more than ~1.005 gravity points (see 100% Lactobacillus Fermentation). See Dealing With Yeast Contamination below.
Once the soured wort is boiled or heat pasteurized, it can be safely added to the primary fermenting vessel without worries of future infections. The wort is aerated as normal, and brewer's yeast, or Brettanomyces yeast, is then pitched into the wort as normal (usually brewer's yeast is used if infection of cold side equipment is a concern). At a pH of 3.4 or lower, the acidity of the wort can reportedly effect the fermentation of some strains of brewer's yeast . For example, a published study showed that growth of US-05 was 82% at a pH of 3.51, and 53% at a pH of 3.17. Fermentation was delayed by 2-4 days (the lower the pH, the longer the start of fermentation was delayed) . It is recommended to pitch a healthy starter of yeast, possibly with a higher cell count than normal. When using dry yeast, re-hydrating as per the manufacturer's recommendations, using a yeast nutrient like Go-Ferm is recommended. Many yeast strains have been successfully used by MTF members to ferment pre-acidified wort: US05, S04, WY1098/WLP007/OYL-006, Belle Saison, Sacch Trois, Bret brux, B. clausenii, B. custersianus, Bret Drie (BSI), WY3711, WY3726, and kveik to name a few (see references) . Michael Tonsmeire has shown results that suggest that English yeast strains might attenuate slightly more and give better flavor results than other strains; Richard Preiss from Escarpment Labs expressed similar observations . Brewers who are having difficulty fully fermenting pre-acidified wort can try growing their yeast in the soured wort (pasteurize the soured wort first if needed) with yeast nutrients (Fermaid K + DAP, for example). This assumes that the wort still has a lot of sugar left over after souring (if not, DME can be added). See Acid Shock Starters for more information on how to acclimate yeast to a highly acidic environment to improve fermentation. Escarpment Labs reported seeing stalled fermentations in kettle soured wort when the selected Lactobacillus produced acetic acid on the higher end, and they suggest using a species like L. plantarum that does not produce high amounts of acetic acid relative to other strains from species like L. brevis, and that perhaps limiting oxygen can help reduce acetic acid production  (~19:50 mins). Some brewers have reported a pH shift, either up or down, by 0.1 to 0.3, or no shift at all from the point of after the second boil and after the final yeast fermentation . This change in pH caused by the yeast fermentation might be due to many variables such as compounds released by the yeast, consumed by the yeast, strains of yeast/bacteria, water profile, malt recipe, dry hopping, etc.
The usual kettle souring process involves fermenting with a normal ale strain of some sort, and lacks Brettanomyces. Without Brettanomyces, there is a lack of Brettanomyces character which is the result of the unique esters, phenols, and fatty acids that this genus of yeast produces (see Brettanomyces secondary metabolites). For example, traditional Berliner Weisse was fermented with a mixed culture containing Brettanomyces, and this was considered the most important aspect of achieving the fruity ester character of that beer style historically (see Benedikt Koch's table comparing esters of traditional Berliner Weisse versus kettle soured Kindl Weisse and Belgian gueuze). Kettle sours can also be racked into oak barrels and have Brettanomyces pitched at that time. This approach is taken by some commercial brewers who don't want to expose their fermentation vessels to living Lactobacillus or Brettanomyces. It is also one method for controlling the level of sourness in the beer since the Lactobacillus is killed off.
- MTF "The Podcast" interview with Adi Hastings of Omega Yeast Labs on kettle souring and episode #007 Lactobacillus microbiology with Dr. Bryan Heit.
- Brewing Network, Brew Strong interview with Fal Allen from Anderson Valley Brewing (author of "Gose: Brewing a Classic German Beer for the Modern Era") on kettle souring in commercial breweries.
Souring in the Primary Fermenter
Wort can be soured in the primary fermenter before adding other yeasts. This is generally a good approach for brewers who aren't concerned with pasteurization and infections of their cold side equipment. This also has the benefit of possibly producing a more complex sour beer overall or at least a sour beer that will evolve over time. It has the advantage over a more traditional Mixed Fermentation in that Lactobacillus is used to guarantee at least a certain level of sourness. This is also a good process to use for making a Berliner Weissbier.
The process is very similar to the kettle souring technique with the exception that the wort is never pasteurized after it is soured. This all-grain brewing process is the same for any all-grain brewing process except that, after the boil, the beer is only chilled to the recommended temperature for the Lactobacillus strain that the brewer is going to use. Using grain husks for souring with this method is inadvisable since the grain will stay in the fermenter during primary fermentation and unwanted microbes on the grain husks would potentially have a longer exposure to the wort. Instead, the brewer should use a pure strain of Lactobacillus. As a result of not using grains to sour the wort there remains less concern of developing Butyric Acid or Isovaleric Acid with this method. Even still, lowering the pH of the wort to under 4.5 (ideally 4.0 - 4.3 when souring with grain) will also discourage contaminating bacteria from thriving in the wort during the incubation period. This will also help with head retention . There is also the option of using a sour yeast cake from another sour beer as the bacteria inoculation.
Once cooled to the desired temperature, usually around 90-115°F or 32.2-46°C (see the Lactobacillus culture charts for desirable temperature ranges for different Lactobacillus cultures), the wort is racked to the primary fermenting vessel. Note that the wort should contain a low amount of IBUs when using this process since IBUs can inhibit many (but not all) species of Lactobacillus. Eliminating hops can be a good approach to getting more acidity, but if hops are required, then using less than 6 IBUs is a good guideline in general (see the Lactobacillus culture charts for tips on hop tolerance for different Lactobacillus cultures). To achieve a low IBU wort consider mash hopping. Mash hopping has been reported to reduce IBUs by ~70% . It is generally a good idea to create a 500 mL starter beforehand for ~5-6 gallons of wort (see Lactobacillus starters). Once the wort is racked to the primary fermenting vessel the Lactobacillus culture is added directly to the fermenter. No other yeasts are added at this time. The Lactobacillus bacteria is allowed to incubate by itself in the wort for 1-3 days with the target temperature maintained throughout the incubation period (some Lactobacillus species/strains may continue to produce acidity under lower temperatures, for example L. plantarum). Small temperature fluctuations should not adversely affect the souring process as long as the temperature stays within the desirable range for that particular strain of Lactobacillus, although the brewer should take care not to allow any dust to get sucked into the fermentation vessel as a result of temperature decreases. During the incubation time, as long as Lactobacillus is the only microbe growing in the wort, the gravity will not drop more than a few points and the fermentation will be calm (see Lactobacillus fermentation. The exact time frame of incubation depends on the species/strain of Lactobacillus, the manufacturer's recommendation, and the brewer's desired acidity level. Acidity can safely be measured with a reliable pH meter throughout this time. If possible, it is advised that the brewer fills the headspace of the fermenter with CO2. Some brewers report that this helps to reduce sulfur production, but if Brettanomyces is added to the beer later on in the process and allowed to age, this shouldn't be a concern.
After reaching the desired acidity level from the incubating Lactobacillus bacteria the brewer can crash cool the fermenter down to the desired temperature for the primary fermenting yeast. Both Saccharomyces and Brettanomyces, or a blend can be used as primary fermenting yeast. Brettanomyces is often chosen because of it's higher tolerance of a low pH environment (3.4- pH ), although many Saccharomyces strains have been successfully used (see the Souring in the Boil Kettle section above). If the chosen yeast requires aeration and the brewer has the ability, the sour wort should be aerated before pitching yeast. Brewers have had good luck using Fermentis dry yeast products in the non-aerated wort after, of course, rehydrating the dry yeast as per the manufacturer's instructions and with using a yeast nutrient such as Go-Ferm is effective . The wort is then fermented out as normal. The brewer can consider other Brewing Methods such as pitching Brettanomyces, a mixed culture, or commercial sour beer dregs into secondary.
Souring with Malted Grains in Another Vessel Before Racking to the Primary Fermenter
This process is very similar to souring the wort in a kettle. This method is ideal for those who wish to use grains to introduce Lactobacillus to the beer, or for breweries that do not want to tie up their boil kettle. If done properly, the formation of Butyric Acid and Isovaleric Acid should be minimal.
The wort is mashed and sparged as normal (and alternatively brought to a short boil) and is then lowered to somewhere between 109-115°F (42.8-46°C). This temperature favors Lactobacillus while discouraging Enterobacteriaceae. Optionally, the mash pH can be lowered to 4.4 with lactic acid or acidulated malt to further discourage Enterobacteriaceae activity. Once the desired temperature (and optionally pH) is reached, the wort is then transferred to a second vessel such as a glass carboy. A pure culture of Lactobacillus can be pitched, or a handful of fresh malted unmilled grain is added. The vessel should be filled to the very top, minimizing the oxygen levels inside the vessel. The vessel should be stored in a heated environment that maintains a temperature between 109-115°F (42.8-46°C) for 1 to 3 days depending on how much acidity the brewer wants (the faster the souring process the better; Jeff Young from Blue Owl achieves the desired acidity in about 18 hours usually) .
Once the 1 to 4 day time period has been reached, the wort is transferred to the boil kettle and boiled as normal. Boiling will kill all of the microorganisms in the wort, and will provide the option for adding hops and other kettle additions. Just as with kettle souring, the wort doesn't have to be boiled, but can be instead heat pasteurized at 140°F (60°C) for 15 minutes  if a yeast contamination produced a significant amount of alcohol (this temperature effectively heat pasteurizes most organisms while not driving off the aromatics and volatiles from the yeast fermentation). Once boiled or pasteurized, the wort can be chilled and handled in the same way as the above methods for wort souring.
An alternative to this method would be to culture Lactobacillus from the grain first and then use it in a kettle souring process. See Culturing From Grains and Blue Owl Brewing's grain inoculation methodology and data for more information.
Tips on Maintaining Heat for Homebrewers
Keeping the temperature as steady as possible for a pure culture Lactobacillus fermentation is not that important. Just try to stay in the range of the temperature best suited for a given species (see the Lactobacillus culture charts). Lactobacillus plantarum strains are known for being effective at sourness all the way down to room temperatures, and so that is a species that might work well for those who cannot maintain the warmer temperature that other species might require. Here are some tips from MTF members on maintaining warm temperatures for wort souring :
- Editor's note: please be cautious when using heat sources to heat plastic fermenters; PET bottles will melt or warp when applying too much heat to them.
- Electric heating blanket or heating pad.
- A room space heater with towels or a t-shirt to insulate.
- If it fits, place the fermenter in an insulated 10 gallon Rubbermaid cooler (like the kind that many people use as mash tuns).
- For metal vessels such as a kettle, keep it on the stove burner and turn the burner on when heat is lost.
- Put the wort in a plastic bucket and seal with lead, then lower that bucket into a ten gallon water cooler and close the lid. Optionally, insulate with blankets.
- Use a glass carboy FermWrap™ heater or another carboy heater attached to a sensor thermowell through the cap and a temperature controller.
- Use a Brew Belt Fermentation Heating Belt and wrap a "hoody" sweater or some other fabric around it to insulate, and a temperature controller.
- Use a Kenley Fermentation Carboy Heater which comes with a thermostat (also marketed as a "Kombucha Heating kit") .
- Use an aquarium heater (Enheim brand recommended), fill a large plastic cooler with water and keep the water warm with the aquarium heater. Set the souring vessel in the cooler. See this AHA article.
- Use an electric kettle with a temperature controller, such as a Grainfather, Braumeister, Brewer's Edge Mash & Boil, or a home build electric kettle.
- Use a "Seedling Heat Mat" that is used to keep plants warm.
- Use an Anova sous vide device - place the vessel in a larger vessel with water, and keep the heat applied to the outside water with the Anova. Works well for maintaining heat for starters too. See this MTF thread for a Q&A on how to use an Anova for various brewing related methods.
- Similar to the Anova, James Spenser of BasicBrewing.com video on using an InstantPot® to maintain "yogurt" temperatures for 1 gallon batches.
- Build an insulation box out of Styrofoam. See this MTF post and this MTF post.
- Sour in a keg, and heat the keg. If the keg is not fitted with a spunding valve or an airlock, be sure to burp the keg in case of an accidental yeast contamination as that would result in a lot of built up pressure. See this MTF post.
- If not using a plastic vessel, keep the vessel in an oven (if it fits) or an electric smoker.
- Use a ZooMed Reptile Heater Cable inside of a fermentation chamber. Line the inside of the chamber walls with the wire and secure with duct tape.
- If maintaining heat is not possible with any of the techniques mentioned, cool the wort to around 100°F/38°C and let the wort free fall down in temperature while souring with a species such as L. plantarum that produces acidity at room temperature. Omega Yeast Labs OYL-605 works well without hot temperatures.
How to Pre-Acidify
- Note: pre-acidifying applies to the wort that will become the batch of sour beer, not Lactobacillus starters. Pre-acidifying a Lactobacillus starter will slow the growth of the Lactobacillus in a starter. The idea of the starter is to grow an effective amount of cells while pre-acidifying the main batch of wort aims to inhibit contaminating microbes and retain foam positive proteins. See Lactobacillus starters for more information.
After the production of the wort, but before pitching the culture of Lactobacillus, some brewers like to slightly lower the pH of the wort with food grade lactic acid (available at homebrew stores) or phosphoric acid before adding the Lactobacillus. Pre-acidifying wort can be applied to any wort souring method in general, and while using any species/source of Lactobacillus including lab cultures, probiotics, yogurt, grains, sauerkraut, etc. Acidifying the wort before pitching Lactobacillus has several benefits, such as discouraging unwanted microbes that may have accidentally been introduced into the wort, and helping to prevent Lactobacillus from degrading foam proteins (although both of these points have been debated on MTF; see this MTF thread as an example). There have also been observations by Jeff Mello of Bootleg Biology and others that pre-acidification results in .1 to .3 pH points lower than if pre-acidification is not applied (this pH difference is not explained by the addition of acid because pH is logarithmic; see this MTF thread). Pre-acidified wort also appears to reach a low pH in less time after the Lactobacillus is pitched for souring.
There currently is no formula for how much lactic acid to add to a volume of wort due to the different buffering capacities of wort . Water chemistry spreadsheets and formulas geared towards mash pH adjustments may not be accurate for wort pH adjustments since wort does not contain grain material; however there has been reports on MTF and recommendations from Martin Brungard (author of Bru'n Water) that Bru'n Water can accurately determine how much lactic acid is needed to lower a wort's pH, or at least provide a starting point . We encourage readers to experiment with water chemistry calculators to see if they can accurately predict wort pH adjustments or get a starting point for how much acid to add .
Another method of finding out how much acid to add would be to pull a measured portion of the wort out, and add acid in measured amounts until the desired pH is reached. The amount of acid added can then be scaled up to the full volume of the wort. A.J. Delange suggests that the buffering capacity of wort might be half that of the mash (based on the kilograms of malt used in the mash) .
Trial and error might be the most practical approach for homebrewers that don't have an abundance of wort to spare for finding out how much acid to add to a sample and scaling that up. Post boil, the wort pH is generally around 5.0 - 5.2. Adjusting the pH of wort before pitching Lactobacillus can then be done fairly easily by taking a trial and error approach. Using 1 mL of 88% lactic acid per .1 shift in pH for 5 gallons of wort is a good starting measurement. As an example, say that 5 gallons of wort has a pH of 5.0 just before pitching the Lactobacillus culture. Begin by adding 5 mL (1 US teaspoon) of food grade lactic acid to the wort for a target of around 4.4 pH (or somewhere between 4.2 and 4.8; target 4.0 - 4.3 if souring with grain or some other non-purified source of bacteria to help inhibit the growth of unwanted bacteria that could produce off-flavors). Stir gently, then take another pH reading. Continue to add 1-2 mL of lactic acid until the wort has the desired pH. Derek Springer has observed that it takes about one tablespoon (15 mL) of 88% lactic acid to reach a pH of 4.2 - 4.8 for 5 gallons of wort ; however a higher amount may be required if the brewer's water is high in bicarbonate (24 mL for 5 gallons of wort to reach a pH of 4.4 was reported by Sean McVeigh for his water which contains 375ppm of bicarbonates ). Once a pH of 4.2 - 4.8 is reached, pitch the Lactobacillus culture. This small amount of lactic acid shouldn't have much of an impact on flavor. If a more precise method for determining the required amount of lactic acid is required, a sample of the wort can be pulled and lactic acid or phosphoric acid can be added to it until the target pH is reached, and then that amount can be scaled up (a micropipette might be required to measure very small amounts of lactic/phosphoric acid).
Maintaining a Lactic Acid Bacteria Culture
It is possible to maintain a pure culture Lactobacillus culture from batch to batch of beer. One method that is common with commercial brewers is to collect an appropriate volume of the soured wort after souring, but before pitching yeast. Commercial brewers will often use a designated corny keg or sanke keg depending on the volume that is required. After souring the wort a small portion will be transferred using a closed system into the keg. The keg is then refrigerated until it is needed for the next batch. Storing the soured wort cold and adding around 20 grams of chalk for 5 gallons is recommended for lengthening the viability of the Lactobacillus culture while also inhibiting the growth of any potential contaminants, however, the Lactobacillus might perform differently based on a number of variables such as how long it is stored (longer storage means less viability; expect 2-4 weeks of maximum storage) . Re-pitching might lead to an increase in acid production as well. We recommend experimenting with this process and finding the right pitching volume/storage time until repeatability can be achieved. For commercial brewers, Breakside Brewery recommends using 5-7% by volume of stored Lactobacillus buchneri (WY5335) soured wort for a new batch, although this might vary based on many factors such as the strain of Lactobacillus, the storage pH, and the storage time .
Homebrewers can use a similar process. For a 5 gallon batch of soured wort, collect around 1 liter of wort in a jar before pitching yeast. Keep the culture cold until the next use. Don't seal the jar completely tight in case refermentation occurs in the jar. If the culture is kept for more than a couple of months, then create a 500 mL starter to ensure that the bacteria culture is still viable.
Yeast contamination is a concern when storing soured wort, especially in a sealed vessel such as a keg. Pure cultures of Lactobacillus cannot effectively ferment most sugars in wort (see 100% Lactobacillus fermentation), therefore there are a lot of sugars available for contaminants to take advantage of when storing wort that has been soured but not fully fermented with yeast. Good sanitation practices are of the utmost importance. Equipment used should be thoroughly clean and ideally sterilized, or at least boiled if possible (or exposed to 82°C/180°F water for 15 minutes). If a yeast contamination goes unnoticed, then the keg can become heavily pressurized and potentially dangerous even when stored cold. We recommend regularly checking a keg of soured wort to ensure that over-pressurization does not occur, or use a spunding valve to ensure that over-pressurization does not occur.
These methods only work with pure cultures of Lactobacillus (or at least cultures that do not contain an unwanted microbe that can take advantage of the residual sugars in the soured wort, such as yeast). Cultures from grains, mixed cultures, or cultures from other fermented foods such as those listed on the Alternative Bacteria Sources wiki page, should be treated as a mixed culture that potentially contains yeast. See Storing a yeast cake or sample of a mixed culture.
When working with lactic acid-producing bacteria, the brewer’s goal is usually to attain clean-tasting sourness, while obtaining desirable flavor contributions from these bacteria, and simultaneously minimizing off-flavors. It should be noted that off flavors span the range from undesired by most, to desirable to some. For example, isovaleric acid is a compound known for its footy aroma that would be considered an off flavor in many beers, yet it gives a highly desired flavor to certain French cheeses and is acceptable by some in small amounts in mixed fermentation beer.
With that said, contamination issues are among the biggest challenges when pre-souring wort with Lactobacillus. This is because Lactobacillus does not fully ferment wort by itself (see 100% Lactobacillus fermentation). When yeast fully ferments wort into beer, alcohol, hops, and a low pH all work together to prevent most spoilage microorganisms from contaminating the beer (although contamination can certainly happen with beer spoilage microbes such as Brettanomyces, Pediococcus, etc.) . When fermenting with Lactobacillus by itself, either no alcohol is produced or not enough alcohol is produced to have an antimicrobial effect. Usually, hops are not used when souring wort with Lactobacillus because even small amounts of hops completely inhibit most commercial strains, but they also inhibit some spoilage microorganisms. The high available sugars, warm temperatures typically used in souring wort, and a lack of alcohol and hops, therefore, increase the chances for contamination during souring wort with Lactobacillus.
Contaminates can include a variety of molds, yeasts, and bacteria. Contaminations can have a variety of potentially unfavorable flavor effects on soured wort depending on the type of microbe(s) that caused the contamination. One common off-flavor in kettle soured beers has been associated with butyric acid, which is in and smells like human vomit. Although the exact source of butyric acid in kettle soured beers has not been identified that we know of, butyric acid is produced by anaerobic contaminates and not when Lactobacillus is exposed to oxygen (see Effects of Oxygen on Lactobacillus and butyric acid). Isovaleric acid is another off-flavor that can be produced by both anaerobic and aerobic contaminates.
Another common contaminate from improper wort souring is brewer's yeast (S. cerevisiae). Brewer's yeast is not inhibited by Lactobacillus. Conversely, Lactobacillus is can be inhibited by the presence of active S. cerevisiae . The warmer temperatures encourage S. cerevisiae to ferment the wort quickly. This often results in a beer that isn't sour because the Lactobacillus are out-competed by the yeast. Signs that yeast has contaminated the wort include the typical signs of yeast fermentation: the presence of a krausen, a gravity shift of more than 1.005 gravity points (or 0.5-1.0° Plato), and looking at a sample of the wort under a microscope. Sources of yeast contamination can come from poor sanitation, but they can also come from the yeast manufacturer themselves. See see 100% Lactobacillus fermentation for more information.
Another source of contamination, which is arguably desired, happens when using a "wild" source of Lactobacillus. For example, when culturing Lactobacillus using non-plating techniques from grains, fruit, or from some other fermented food such as kefir, sauerkraut, etc., yeast and other microbes can carry over from the culturing process (see Alternative Bacteria Sources). The only way to guarantee that only Lactobacillus from a "wild" source such as this ferment the wort, is to isolate the microbe using plating techniques (see Wild Yeast Isolation). Otherwise, there is a chance that wild yeast will also survive the culturing process.
Oxidative species of yeast can also contaminate wort that is being soured. Their fermentation can resemble that of brewer's yeast visually, however, they often do not attenuate wort. See Dr. Bryan Heit's brief biochemistry explanation for "krausen" or "pellicles" caused by oxidative yeast and why they do not correspond with a drop in gravity if they are formed by oxidative yeast species. Oxidative yeast may produce off-flavors, but not always.
Several techniques can be applied to prevent contamination in kettle soured beers. Many of these are covered in the sections below when they apply to specific processes, however, we will cover them all in this section as well.
Pre-acidifying wort to 4.5 pH or lower helps to kill many microbes that are not tolerant of low pH. Flushing the wort with CO2 during the souring process is thought to help prevent aerobic contaminates . Maintaining a temperature between 113-120°F (45-49°C) helps encourage some species of Lactobacillus and inhibits heat intolerant contaminates, however, some species of Lactobacillus do not do well at these warmer temperatures (see the Lactobacillus culture charts). Pre-boiling the wort has been shown to help greatly with preventing contamination. This is likely due to the killing power of boiling temperatures versus lower temperature pasteurization, as well as the heat and steam killing microbes on the sides of the boil kettle, the lid of the boil kettle, and the head space above the wort. Boil kettles are inherently unsanitary vessels compared to fermentation vessels, so great care should be taken to sanitize the vessel and prevent any air from getting into the vessel during the souring process. Air can get in when temperatures cool and a vacuum is created inside the kettle. Wrapping the boil kettle and lid with plastic wrap has been a typical approach for homebrewers, as well as maintaining a constant temperature so as to avoid creating a vacuum inside the kettle from cooling temperatures. Commercial brewers must also prevent air from getting sucked into the boil kettle. Some have used sanitized and inflated sports/beach balls or something similar to clog the boiler stack during kettle souring, for example. Continuous flushing with CO2 can also help prevent a vacuum from sucking in air. Finally, achieving the desired acidity as quickly as possible helps to cut down on the chances for contamination. Achieving the desired acid development within 48 hours is an ideal goal, but achieving it within 12-24 hours is even better.
Yeast contaminations can be difficult to avoid if they are coming from the manufacturer of the Lactobacillus culture. Look at the culture under a microscope and check for yeast cells, which will be much larger and circular in shape compared to the much smaller, rod-shaped bacteria. Reputable yeast companies will usually offer a replacement for any contaminated Lactobacillus cultures.
Contamination of the Lactobacillus strain itself in a production brewery is always a concern, especially if the Lactobacillus is not boiled. Thorough cleaning and sanitation regimes are required for preventing cross contamination. Brewers will often advise performing two cycles of near boiling water with caustic, followed by a sanitizing rinse. If metal parts can be removed from a system, they should be boiled. Soft parts such as hoses and gaskets should be reserved for sour brewing. Using a Lactobacillus strain that has been tested to be intolerant of 1-2 IBU's can also reduce the risk of cross contamination . Check with your QC lab or yeast lab for procedures for avoiding cross contamination.
Dealing With Yeast Contamination
In the case of accidental yeast contamination, the yeast could produce a beer with off-flavors or a beer that is acceptable to drink, depending on the type of yeast and the stress of the hot fermentation. If the fermentation created a lot of off-flavors, it is best to dump the beer since many flavors may not get cleaned up at this point. If the beer tastes fine, the easiest option is to simply allow the beer to ferment out without doing a second boil to kill the Lactobacillus (and yeast). This decision depends on whether or not the brewer is prepared to deal with having living Lactobacillus exposed to their cold side equipment (racking equipment, fermenters, etc.; see Do I Need Separate Equipment FAQ). Making this a Mixed Fermentation sour beer is also an option if the brewer has the fermentation space for it. If the brewer is not willing to expose their cold side equipment to the living Lactobacillus, boiling the fermented/partially fermented beer is another option.
If a yeast contamination produces a significant amount of alcohol during the souring process, then this presents a problem when it comes to boiling. Although 100% pure ethanol boils at 173.1°F/78.4°C , the lower the concentration of ethanol in the wort (technically beer at this point if it has been fully attenuated by yeast), the higher the temperature required for boiling off the ethanol. For example, at 5% ABV it takes approximately 197°F/92°C for the ethanol to boil . Time is also required to boil off the ethanol, so this may not be as big of a concern as it first appears (warning: vaporized ethanol is highly flammable). Another and perhaps more important consideration is that boiling and high heat pasteurization temperatures can have a negative effect on the flavor of fermented beer. Beer already has anti-bacterial properties, such as low pH, presence of alcohol, and hops (although hops may not be present in wort being soured), so higher pasteurization temperatures aren't necessarily required for beer. For these reasons, the beer industry commonly heat pasteurizes beer at 140°F/60°C for 15 minutes, and this is also adequate for pasteurizing soured wort that accidentally fermented do to yeast contamination . Although the yeast will be killed during the pasteurization, so will the Lactobacillus, and the brewer can trust that equipment downstream shouldn't get contaminated. Other options are to simply dump the batch and start again with a pure culture of Lactobacillus, or do not pasteurize and let the Lactobacillus live. If the Lactobacillus is not pasteurized and allowed to live, the soured wort/beer should be treated with care so that it does not contaminate the rest of the brewery (see 'Do I need Separate Equipment?' FAQ). It is possible that the contaminating yeast out-competed the Lactobacillus, which results in not much acid production. If the beer doesn't taste sour, then it is likely to not sour much during aging unless the Lactobacillus strain is very hardy (alternatively, pitch Pediococcus or a hardier strain of Lactobacillus like delbruekii or brevis to try and achieve acidity over time). If the contaminating yeast doesn't fully attenuate the wort, pitch a fresh slurry of yeast to finish the fermentation.
Boiling soured wort that hasn't had an accidental yeast contamination (and thus still has a high specific gravity) probably has less of an effect on the flavor than it does on fully fermented beer. Other than lactic acid, the flavor components that different strains of Lactobacillus produce are not well defined, so it will be difficult to determine if boiling soured wort will have a negative impact. However, brewers who boil kettle soured wort don't often report that the boiling causes flavor issues . Another potential problem with boiling a fermented beer is scorching from excess trub at the bottom of the kettle. Boiling can also cause off-flavors from the lysis (bursting) of yeast and bacteria cells.
If a Lactobacillus starter gets contaminated with yeast, it is best to throw away the starter and make another one, or use probiotics, yogurt, or some other easy to find source of Lactobacillus.
Various Other Concerns
There are instances where wort souring fails for seemingly unknown reasons. Use a species or strain of Lactobacillus that is adapted well to souring wort within 24 hours, such as L. plantarum. The lactic acid bacteria sometimes do not survive well during storage over long periods of time, so viability should be checked for long stored or mishandled cultures. Contamination with yeast can also prevent souring in some instances but not all. The use of hops should be avoided even for hop tolerant species (unless the brewer is experienced enough to predict the souring based on their lactic acid bacteria), and especially avoided if using L. plantarum. Residual iso-alpha acids on the walls of kettle tanks could be enough to inhibit some strains of Lactobacillus, specifically L. plantarum, so kettles that have previously held highly hopped wort might need to be scrubbed clean before being used as a souring tank (~21 mins in).
Not Boiling and Dimethyl Sulphide (DMS)
This is generally not a concern. See Dimethyl Sulfide for an explanation why.
The pH of boiling wort influences how well proteins are precipitated out, with a pH of about 5.2 being ideal for this mechanism. At a pH lower than 5.0, these proteins gain negative charges, and at a higher pH they gain positive charges, making them resistant to settling out during boiling. Boiling a low pH wort that has been acidified by the fermentation of Lactobacillus therefore does not effectively precipitate out haze-forming proteins. However, haze forming polyphenols, which are also required for haze formation, are greatly reduced by an unknown mechanism during kettle souring, which can potentially lead to good clarity in the final beer regardless of the protein level (although other factors might still lead to a hazy beer) . Additionally, some strains lactic acid bacteria are known to cause haze as a side effect of their lactic acid fermentation.
Maillard and various browning reactions lead to darkening of beer in general. These reactions are both temperature and pH related. A low pH will actually prevent these reactions and results in a quantitatively lighter color beer compared to a control beer that is not acidified .
There is no scientific basis for the idea that a small amount of oxygen itself that is dissolved during wort souring has a great effect on the flavors that Lactobacillus produces. See Effects of Oxygen on Lactobacillus and this MTF thread that discusses the anecdotal experience of brewers who do not purge with CO2. The effects of oxygen on the beer could lead to other stability issues with the beer because the exposure to oxygen during wort production is thought to play a role in beer staling (see Aging and Storage), although hot side aeration is also thought to be a minor problem if at all , and the impact of hot side aeration on sour beer has not been studied. Some brewers believe that purging all oxygen out of the system will help to prevent contaminating microbes that are aerobic (microbes that require oxygen) from creating isobutyric acid. Oxygen does not play a role in the production of Butyric Acid in wort soured beers because this compound is produced by anaerobic bacteria (bacteria that requires the absence of oxygen). Air/dust can carry contaminating microbes, which is a potential problem (see Contamination Concerns above), however, if the air is filtered somehow before entering the fermenting vessel then the oxygen itself shouldn't pose a problem to souring wort. Follow the links in this paragraph for more information.
It is generally recommended to not re-use yeast that has fermented a soured wort. This has a stressful impact on the yeast. Some brewers have reported trying this, and not having good results . However, some commercial brewers have claimed that repitching WY3711 as many as 100 (in one case) to 2000 (in a second case) generations in soured wort conditions has not posed a problem for them. If this is attempted, repitching the yeast from the middle of the yeast cake will select for the more acid tolerant cells. Some yeast strains may not be able to adapt to these conditions as well as others (more data is needed), however, it has been shown that S. cerevisiae can adapt to acidic conditions and become more tolerant of low pH conditions from generation to generation. See terminal acid shock for more information.
Additional Articles on MTF Wiki
- Sour Mashing
- Alternative Bacteria Sources
- Butyric Acid
- Isovaleric Acid
- Mixed Fermentation
- Kettle souring; video presentation by Escarpment Labs.
- "How I Sour Mash & A Recipe"; Sui Generis Blog (includes Wort Souring and microbiology information).
- "Year of the Sour Mash" by Derek Springer. This series of articles were written for his NHC 2015 presentation.
- "Lactobacillus 2.0 - Advanced Techniques for Fast Souring Beer", by Matt Miller of Sour Beer Blog.
- "Overnight Acidification" by Michael Tonesmeire, article for Brew Your Own Magazine on Jan/Feb 2016.
- "Boil vs. No-Boil Berliner Weisse | exBEERiment Results!" on Brulosophy blog.
- Catharina Sour – A Brazilian kettle-soured fruit beer. See also thoughts on this style making it to the BJCP Style Guidelines on MTF.
- Miller, Matt. Dec 20, 2014. "Fast Souring with Lactobacillus – Best Practices, Sensory, & Science". Sour Beer Blog.
- Aluminum Surface Finishing Corrosion Causes and Troubleshooting. W. John Fullen, Boeing Research and Technology & Jennifer Deheck, Boeing, Seattle, Washington, USA. 10/17/2014.
- Chemical Resistance Chart. Plastics International. Retrieved 01/03/2017.
- Conversation with Andrew Addkison on MTF. 01/12/2016.
- "Water Knowledge." Martin Brungard. Bru'n Water website. Retrieved 12/02/2020.
- Chemical Composition of Sour Beer Resulting from Supplementation the Fermentation Medium with Magnesium and Zinc Ions. Aneta Ciosek, Katarzyna Fulara, Olga Hrabia, Paweł Satora, and Aleksander Poreda. 2020. DOI: https://doi.org/10.3390/biom10121599.
- Kurt Marshall. CBC 2012 Presentation.
- Samuel Aeschlimann. Eureka Brewing Blog. "#44 Traditional Berliner Weisse". 03/10/2012. Retrieved 09/02/2017.
- Shut up about Barclay Perkins. "Kettle Souring". Ron Pattinson. 09/17/2020.
- Enhancing the Microbiological Stability of Malt and Beer – A Review. Anne Vaughan, Tadhg O’Sullivan and Douwe van Sinderen. 2005.
- Mapping microbial ecosystems and spoilage-gene flow in breweries highlights patterns of contamination and resistance. Nicholas A Bokulich, Jordyn Bergsveinson, Barry Ziola, David A Mills. 2015.
- The Microbiology of Malting and Brewing. Nicholas A. Bokulich and Charles W. Bamforth. 2013. DOI: 10.1128/MMBR.00060-12.
- Conversation with Bryan of Sui Generis Blog regarding boiling versus lower temperature pasteurization. 11/18/2015.
- Lactobacillus 2.0 – Advanced Techniques for Fast Souring Beer. 11/18/2015. Retrieved 11/19/2015.
- "District Michigan MBAA Technical Meeting Grand Ledge, MI". MBAA Presentation. 2011.
- Allen, Fal. "Gose: Brewing a Classic German Beer For the Modern Era". Brewers Publications. 2018. Pg 126.
- Nick R Jones. "The Oxford Companion to Beer definition of nitrogen". Craft beer & Brewing Magazine website. Retrieved 10/21/2018.
- Personal correspondence with Khristopher Johnson of Green Bench Brewing Co. and Dan Pixley. 05/24/2016.
- Conversation with Jeff Young from Blue Owl Brewing Co on souring from grains. 07/21/2016.
- Pavel Dunn. Milk The Funk Facebook group thread on mold growth during kettle souring. 06/23/2018.
- Various members of Milk The Funk. Milk The Funk Facebook thread on purging kettle sours with CO2. 08/03/2018.
- Adi Hastings. Milk the Funk "The Podcast"; Episode #004. 04/02/2018. Retrieved 04/16/2018.
- Personal correspondence with Steph Cope of CraftHaus Brewing Co. 02/06/2016.
- Various professional brewers. Milk The Funk Facebook post about souring in a mashtun, brite tank, or other vessel for commercial brewers. 08/02/2017.
- Lactobacillus wikipage; Tolerance of Extreme Temperature
- Chris Colby. "Easy way to Hit The Proper Boil pH". Beer and Wine Journal blog. 07/01/2013. Retrieved 03/02/2018.
- Michael Tonsmeire on HBT
- "Quick Sour, then what? Acid Tolerance of Brewer’s Yeast." The Mad Fermentationist blog. Michael Tonsmeire. 12/13/2016. Retrieved 12/14/2016.
- Sour Brewing: Impact of Lactobacillus amylovorus FST2.11 on Technological and Quality Attributes of Acid Beers. Lorenzo C. Peyer, Martin Zarnkow, Fritz Jacob, David P. Schutter, Elke K. Arendt. 2017.
- Conversation on MTF about using specific yeast strains in acidic wort. 7/6/2015.
- Multiple MTF members. Milk The Funk Facebook group thread about fermenting kettle sours with kveik. 05/15/2019.
- MTF conversation with Richard Preiss about English ale strains and pH tolerance. 12/13/2016.
- Nate Ferguson. "Quick Souring with Lactobacillus 101". Escarpment Labs video presentation. 06/09/2020.
- Survey of brewers on pH shift between post-souring and yeast fermentation. 01/03/2019.
- Putting Some Numbers on First Wort and Mash Hop Additions. David Curtis NHC 2014 Presentation.
- "GoFerm". Scott Labs website. Retrieved 09/19/2018.
- Spencer, James. December 15, 2014. Beer and Wine Journal.
- Tips from many MTF members on maintaining heat for wort souring. 04/07/2016.
- Tips from many MTF members on maintaining heat for wort souring 2. 10/07/2016.
- Tips from many MTF members on maintaining heat for wort souring 3.
- Chris Hunter. Milk The Funk Facebook group post on heating kettles for wort souring and kveik. 08/29/2019.
- A Study in the Practical Use of Lactic Acid Bacteria. Greg Doss from Wyeast Laboratories Inc. 2014.
- A.J. Delange. Homebrewtalk Thread. 07/19/2016.
- MTF Thread with Landon Ortiz. 03/03/2016.
- Conversation with Adam Boura and Mark Trent on MTF regarding using Bru'n Water for adjusting wort pH. 07/18/2016.
- Conversation with Derek Springer on MTF regarding acidifying wort. 04/24/2016.
- Conversation with Sean McVeigh on MTF on acidifying wort. 04/23/2016.
- Lance Shaner. Milk The Funk Facebook group thread on storing Lactobacillus. 11/08/2018.
- Roy Ventullo. "Lacto experiment" dataset, tab labelled "Experiment V (condensed)". Uploaded to MTF on 09/23/2016 by Dan Pixley.
- Neil Caron. Milk The Funk Facebook group thread on how much volume of stored soured wort to pitch for a new batch. 04/24/2018.
- "Fact or Fiction? Can Pathogens Survive in Beer?" Sui Generis Blog. 02/18/2014. Retrieved 11/10/2016.
- Effect of mixed cultures on microbiological development in Berliner Weisse (master thesis). Thomas Hübbe. 2016.
- Private correspondence with Khristopher Johnson from Green Bench Brewing Co by Dan Pixley. 05/04/2016.
- Sour Brewing: Impact of Lactobacillus amylovorus FST2.11 on Technological and Quality Attributes of Acid Beers. Lorenzo C. Peyer, Martin Zarnkow, Fritz Jacob, David P. De Schutter, Elke K. Arendt. 2017.
- Boiling temperature of ethanol
- Making Moonshine: Still Temperature. Retrieved 01/11/2016.
- Conversation with Russell Carpenter on MTF. 01/11/2016.
- A suitable model of microbial survival curves for beer pasteurization. Sencer Buzrul. 2006.
- Conversation with Bryan from Sui Generis blog on MTF regarding boiling soured wort. 11/10/2016.
- Alex Flores. Craft Beer & Brewing Podcast. Episode 199: Alex Flores of Urban South Kettle-Sours the Hard Way. Retrieved 09/16/2021.
- Charlie Bamforth. Interview on BeerSmith about Hot Side Aeration. 01/31/2014. Retrieved 08/18/2017.
- Ryan Sandlin. Milk The Funk Facebook group thread on re-using kettle sour yeast. 06/06/2018.