Professional Documents
Culture Documents
Brewery.
Gary Spedding, BDAS, LLC (From a 2000 Seminar).
Abstract.
The brewery microbiologist, expert and novice alike, has been presented
with a very wide array of growth media within the past 50 years. The
choice of which media to choose for a particular purpose (yeast, wild
yeast, mould or bacterial strain selection, detection and identification)
has not always been an easy one. This paper will provide a listing of the
properties of a select few media useful for the brewing microbiologist.
The topic will be handled from the perspective of the novice
microbiologist in the craft brewery environment. In consequence of
this, the considerations for the selection and use of such media will be
discussed. Furthermore, as a prelude to the main topic, a discussion as
to the types of, and the properties of, various microbial contaminants
most commonly found in the brewery will be entertained.
Introduction.
The ideal situation for the lager and ale brewer is to have only one
species of organism ever present in the beer; namely the culture yeast
pitched into the hopped wort. However, by the very nature of the sheer
abundance of microorganisms present in the environment this ideal
situation is never realized. There are many bacterial species and wild
yeasts (also moulds) in the environment or in the brewers raw
materials, which can infect beer; some cause damage while others do
not. Every brewer must therefore be aware of the types of organisms,
the kinds of spoilage they can render on the final product, and how to
effectively remove them as potential contaminants. The detection of
contaminating bacteria and wild yeast strains is often very much easier
than their identification; wild yeast strains being much more difficult to
identify than individual bacterial strains. The purpose, therefore, of
this paper is not to describe in any detail how to detect and recognize
many individual species. Instead, the intention is to illustrate a few of
the most useful kinds of media available which will enable you as a
microbiologist to determine if you do actually have contamination by
wild yeast or beer spoiling bacteria. The mere presence of contaminants
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is sufficient news to send most brewers back into the brewery in order
to locate and clean up the source of contamination.
Some History and Recent Developments.
In 1950 two microbiologists, Green and Grey, introduced the antibiotic
cycloheximide (actidione) as a very powerful inhibitor of culture yeast.
Through the use of cycloheximide the main organism found in
fermenting wort and beer (the culture yeast!) could be suppressed. This
suppression allowed most other contaminating organisms to grow and
to be visualized on/in the culture media. Cycloheximide was thus
introduced into many selective media from the 1950s through the 1980s.
In addition, many other types of growth media were also developed
during this period. Nine media are described in this paper (some
incorporating cycloheximide into the formula and some not) which can
be used to determine bacterial and wild yeast contamination. One
major problem has now, however, arisen for the microbiologist in that
the antibiotic cycloheximide is no longer available. Brewers will need to
be aware of this as they now select the kinds of media that they will use
in their own breweries. More importantly microbiologists now need to
go and find other antibiotics and inhibitors of culture yeast which will
prove as useful as cycloheximide.
Beer Spoilage.
The potential sources of beer spoilage organisms in the brewery include
air, soil, water, raw materials, grain/malt dust, pitching yeast,
processing aids, vermin, human skin, brewery equipment, plant and
machinery. The types of spoilage caused by microorganisms include the
following; turbidity, haze formation, rope (slime) formation, overattenuation, gushing, souring of wort and beer and the production of
varied off-flavors. Some of the key organisms (yeast and bacteria) that
are responsible for beer spoilage are illustrated in Table 1. As a brewer,
a comprehensive knowledge of the types of organisms that can result in
beer spoilage is most important. An understanding of the sources of
infection, together with a knowledge of the kinds of damage (including
the aromas and flavors) caused by microorganisms, can provide a useful
baseline which can be used to guide the type of microbiology testing
needed when a contamination issue arises. Presented in Table 1 is a
listing of bacteria and wild yeast strains, along with key characteristics
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Properties
Bacteria
Lactobacillus
Pediococcus
Enterobacteriaceae
Gluconobacter
Zymomonas
Pectinatus
Yeast
Saccharomyces
Brettanomyces
Dekkera
Kloeckera
Pichia*
Candida*
Hansenula*
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MacConkey Agar
A medium used for the detection, isolation and enumeration of
Enterobacteriaceae. This differential medium, for the detection of
most coliforms, has lactose, crystal violet and a neutral red
indicator as part of the formula. The crystal violet inhibits the
growth of Gram positive organisms.
The differential nature of this medium is also based upon the
production of acids from lactose fermentation by various
organisms (especially coliforms). Acid is produced and the
neutral red is absorbed to give rise to brick-red colonies.
Incubation (for brewery work) is typically allowed for four days
at 30 oC prior to examination of the plates. Coliforms develop as
dark pink to red colonies. Other Gram negative species form
translucent almost colorless colonies. Gram positives are
suppressed. Note, some yeast strains will grow on this medium!
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been the only real choice for suppressing the growth of culture yeast in
media used for the detection of bacteria (see the section on yeastselective media below for alternative strategies for culture yeast
suppression). Microbiologists must now very quickly find a
replacement for this very useful (but dangerous) antibiotic if HLP and
related media are to continue to be a useful tool for the brewery
microbiologist.
A Discussion of Brewery Yeast Strains.
Wild yeast strains come in two groups, Saccharomyces wild yeast and
Non-Saccharomyces wild yeast. Wild yeast in fermenting and primary
aging tanks usually comes from infected pitching yeast. Wild yeast in
bottled product is also a major issue. High wild yeast numbers in a beer
is usually a result of poor sanitizing in the bottling/storage area. Wild
yeast if found to be growing in the product will lead to major off-flavor
issues and spoilage. Again, as for bacteria, a number of types of yeast
culture media have been developed especially within the past 50 years in
order to detect and enumerate spoilage yeast strains. Three types of
selective media are advocated for the suppression of brewing yeast
strains (ale and lager) and the subsequent detection of wild yeasts.
These are Lysine Media, Lins Wild Yeast Medium and Lins Copper
Sulfate Medium. A discussion of each is presented below. These media
were originally developed to test the quality of brewers yeast but can
also be used to test wort and beer. In addition, Wallerstein
Laboratories Nutrient Medium is also useful for evaluation of yeast
strains; a brief discussion follows.
Media Specific or Useful for Yeast.
1.
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4.
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Smith, C. E., Casey, G. P., and Ingledew. W. M. (1987) The Use and
Understanding of Media Used in Brewing Microbiology. Update 1987.
Brewers Digest (October); 12-16, 43.
Van Keer, C., Van Melkebeke, L., Vertriest, W., Hoozee, G. and
Schoonenberghe, E. Van. (1983) Growth of Lactobacillus Species on
Different Media. J. Inst. Brew. 89; 361-363.
Walters, L. S. and Thiselton, M. R. (1953) Utilization of Lysine By
Yeasts. J. Inst. Brew. 59; 401-404.
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These tests and the Gram staining test are useful sources of information
together with that obtained by the culturing of organisms. The
Gram staining test is discussed in the BSI manual and in many other
microbiology methods manuals.
If an organism proves to be Gram positive go to the catalase test.
If catalase negative, the organism is likely a lactic acid bacterium.
If catalase positive, the organism is likely a non-beer spoiler.
If an organism is Gram negative go to the oxidase test.
If oxidase negative, the organism is likely a wort spoiler.
If oxidase positive, the organism is likely a non-beer spoiler.
End.
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