Grain

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For a general overview of grain in brewing, see Homebrew Talk Wiki page on Grain.

Uses Specific to Funky/Sour Brewing

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Microbial Populations on Barley

Malting Process

Microbial communities found on the outside surface of barley are very diverse and change significantly throughout the malting process, with more diversity occurring further along as the malting process continues. On barley in fields, Gram-negative bacteria (especially Erwinia herbicola) are abundant. Climate is said to have the most impact on which species grow in barley fields. For example Fusarium species are encouraged by high humidity, and are associated with mycotoxins and gushing problems in beer. During dry storage of barley, spore forming bacteria tend to survive. Xerophilic fungi (fungi that doesn't need much water) also survive on dry barley during storage .

Microbial communities on the husks of barley change quite a bit during the malting process. It is thought that the factors affecting this are the initial microbe populations, interactions between species, the variances in the malted barley characteristics/processes, and additives. Different malting houses are likely to form "in-house" microflora populations. The first step in malting, which is steeping, sees the first large change in microbial populations. The steeping process favors lactic acid bacteria (LAB), which are seen only in very small numbers before steeping. Particularly these are Leuconostoc species. In the case of yeasts, Basidiomycota fungi grow, as well as Fusarium .

The second step of malting, which is germination, sees starches converted to sugars, which causes microbial populations to increase once again. Where Leuconostoc species of bacteria were dominate during steeping, the germination process is where Lactobacillus species begin to dominate the microbial community. A greater diversity of species also occurs during this step in malting. As far as fungi, Ascomycetous yeasts begin to dominate, where as Alternaria and Cladosporium decline .

During kilning, microbial populations are reduced by a factor of 10-100, but are still higher than that of field barley. The high degree of variability is dependent on the kilning temperature and procedure. Heat resistant microbes, perhaps by forming biofilms, can survive the hot temperatures. These include molds such as Rhizopus and Mucor species .

Malted Barley

Most studies on microbe populations on malt have been done using traditional lab media such as agar plates, which are thought to not be as effective at analyzing the overall population as recent DNA sequencing approaches, such as 454 amplicon pyrosequencing. A recent study took this approach to measuring bacterial diversity on barley and malt (fungi was not analysed). This study looked at two malting houses during two different years (2010 and 2011). It found that on malt, the largest communities of bacteria were found to include Enterobacter, Sphingobacterium, Weissella, Lactobacillus, Lactococcus, Streptococcus, Acinetobacter, and Strenotrophomonas, Leuconostoc, Pseudomonas, Wautersiella, Cryseobacterium, Curtobacterium, and Propionobacterium .

Enterobacter spp specifically have been identified as beer spoilage agents, which produce high amounts of 2,3-Butanediol (buttery taste ), as well as acetoin (also buttery taste , lactic acid, acetic acid, succinic acid, high alcohols such as n-propanol, iso-butanol, D-amyl-alcohol, isoamyl alcohol, and sulfur compounds such as dimethyl sulfide (DMS), all under both aerobic and semi-aerobic conditions . Enterobacter is not inhibited by hops, but is inhibited by ethanol and a pH of 4.4. Acinetobacter has also been shown to produce DMS in wort, however they are strictly aerobic and encountered in much smaller amounts than Enterobacter in brewing .

Interestingly, the study also found that the dominate genera of microbes was different between the two years sampled. In 2010, there were more Firmicutes (which includes LAB) and more Actinobacteria, and less Bacteroidetes. In 2011, the reverse was found with fewer Firmicutes and Actinobacteria, and more Bacteroidetes (~15 mins in).

The study examined LAB populations specifically due to their potential to help with malt stability and quality . Streptococcus was the most abundant genus of bacteria in all samples. In 2010, Lactobacillus was more abundant than it was in 2011. Lactococcus and Weisella were more abundant in 2011 than in 2010. Maltster 1 had a greater abundance of both Lactococcus and Weisella in 2011 over Maltster 2. In 2010, Lactobacillus was more abundant for Maltster 2 than it was for Maltster 1. This showed that microbial populations differ not only between malt houses, but also between barley harvest years (~17 mins in). Sometimes culturing from malted grains turns out bad perhaps not because of the brewer, but because of the dominate cultures on the malted grains.

Another study focused on microbial populations in an American coolship brewery detected only Pediococcus on grain samples, but the lack of detection of other microbes was attributed to the methods used .

• See this study for a list of microbes that were found on malt using DNA methods.
• See Table 2 of this study for a list of microbes that were found using traditional plating methods.
• See this video presentation by Bart Lievens at the 2015 Belgian Brewers Conference:

Mash and Wort

During mashing, the population of microbes diminishes greatly due to near pasteurization temperatures. However, thermotolerant microbes do survive. These are usually homofermentative LAB . These microbes can have both positive and negative impact on wort production. Mash acidification by thermotolerant L. amylovorus has shown to improve enzymatic conversion of starches to sugars, increased extract and fermentability, increased TSN and FAN, and even improved head retention and increased shelf stability . Bacterial growth during mashing can also have a negative impact. For example, the thermotolerant Gram-positive and facultative anaerobe Bacillus coagulans can cause the mash to go sour and has been shown to form nitrosamines in wort that was supplemented with nitrate. B. coagulans forms nitrosamines without oxygen between the temperatures of 86°F/30°C and 154°F/68°C, and can withstand a pH of 4.0 or higher . Clostridium, which probably does not necessarily originate from the malt itself (so far studies have shown very little to no Clostridium is present on malt), can create butyric acid off flavors during the mash or during kettle souring . High bacterial growth can cause lautering issues, probably due to the production of dextrans by the bacteria .

See Also

Additional Articles on MTF Wiki

• Culturing Lacto From Grains
• Lactobacillus
• Pediococcus

External Resources

• Homebrew Talk Wiki page on Grain.
• Barley World, Oregon State University barley research website.
• Phenol-Explorer site that discusses phenols of some adjunct grains.