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Brewery Contaminants, Challenges and Remedies-A Review

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 Brewery Contaminants, Challenges and Remedies - A Review.
Obi, C. N.
Department of Microbiology, College of Natural Sciences,
Michael Okpara University of Agriculture,Umudike, P.M.B. 7267,
Umuahia, Abia State, Nigeria.
E-mail: Telephone: +2348063614241

Abstract: Brewery contaminants are microorganisms that cause spoilage in brewery products. One of the major
challenges that breweries face is the production and maintenance of beer quality. The beers produced by breweries
are very susceptible to attack by spoilage micro-organisms that cause the deterioration of beer quality characteristics.
The microbiological stability of the final product can be compromised from a very early point in its production, with
spoilage organisms able to access the brewing process at every stage, even from start to dispense. Brewing raw
materials, such as malt, hops and occasionally brewing water may be infected by micro-organisms and these have to
be killed during the brewing process to prevent wort and beer spoilage. Microorganisms that cause spoilage in beer
include; bacteria such as lactic acid bacteria, acetic acid, Enterobacteriacea, Pectinatus, Megasphaera, Zymomonas
and wild yeast. Good quality assurance procedures are necessary for ensuring a high level of cleanliness, especially
when there is no sterile filtration of beer or bottle/can pasteurization. An increasing amount of microbiologically
spoiled beers are caused by secondary contaminations even though overall brewery hygiene has greatly improved in
recent years. Sampling for cleanliness and increasing the awareness of hygienic conditions can enable the brewer to
produce beer with longer shelf life, fewer customer complaints, higher production rates and lower rejection rates.

Key words: Bacteria, brewery contaminants, challenges, remedies, wild yeasts.

Introduction Brewing is a complex process and critical

B rewery contaminants are microorganisms that

cause spoilage in brewery products. Due to the
inherent exposure to the environment during the
filling process, bottling and canning lines in breweries
often are considered open production equipment and
steps during brewing, cellaring and packaging need to
be monitored for microbial contamination. If the
equipment, raw materials or yeast are
microbiologically contaminated, the beer will not meet
the brewer’s expectations (Bamforth, 2002). Most of the
have the ability to become contaminated from outside organisms of concern to the brewer will cause off-
sources including the environment (Priest and Stewart, flavours or turbidity in the beer and these are beer
2006). One of the major challenges that breweries face quality related problems (Sakamoto and Konings,
is the production and maintenance of beer quality. The 2003). One of the most important parts of the brewing
beers produced by breweries are very susceptible to process is the proper control of unwanted
attack by spoilage microorganisms that cause the microorganisms (Stewart and Russell, 1998).
deterioration of beer quality and organoleptics. The
microbiological stability of the final product can be Modern Brewery
compromised from a very early point in its production Breweries today are made predominantly of
with spoilage organisms able to access the brewing stainless steel, although vessels often have a decorative
process at every stage. An increasing amount of copper cladding for a nostalgic look. Stainless steel has
microbiologically spoiled beers are caused by many favourable characteristics that make it a well-
secondary contaminations even though overall brewery suited material for brewing equipment. It imparts no
hygiene has greatly improved in recent years flavour in beer, it reacts with very few chemicals which
(Henriksson and Haikara, 1991). Microbes are in means almost any cleaning solution can be used on it
constant movement in the bottling and canning areas (concentrated chlorine [bleach] being a notable
because they settle from the air to surfaces and then are exception) and it is very sturdy. Sturdiness is important
re-circulated into the environment again by the as most tanks in the brewery have positive pressure
turbulence of the air (Henriksson and Haikara, 1991). applied to them as a matter of course, and it is not
Microbiological analysis of brewery environmental air unusual that a vacuum will be formed incidentally
can be used to determine if there are indicator or beer- during cleaning (Miller, 2012). Heating in the brew
spoiling organisms present (Henriksson and Haikara, house usually is achieved through pressurized steam,
1991). Environmental factors, such as humidity, although direct-fire systems are not unusual in small
temperature, and air speed can have a direct impact on breweries. Likewise, cooling in other areas of the
the growth and survival of airborne microorganisms brewery is typically done by cooling jackets on tanks,
(Crozier-Dodson and Fung, 2002). which allow the brewer to control precisely the
*Corresponding author: temperature on each tank individually, although whole-
[email protected];Obi, C. N. room cooling is also common.
Copyright © 2017 Nigerian Society for Microbiology

Nigerian Journal of Microbiology 2017, 31(1): 3926-3940

Published online at www.nsmjournal.org
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Today, modern brewing plants perform myriad conical vessels (CCVs), which have a conical bottom
analyses on their beers for quality control purposes. and a cylindrical top (Hornsey, 2004). The cone's
Shipments of ingredients are analyzed to correct for aperture is typically around 70°, an angle that will allow
variations. Samples are pulled at almost every step and the yeast to flow smoothly out through the cone's apex
tested for oxygen content, microbial infections and at the end of fermentation, but is not so steep as to take
other beer-aging compounds. A representative sample up too much vertical space. CCVs can handle both
of the finished product often is stored for months for fermenting and conditioning in the same tank. At the
comparison when complaints are received. end of fermentation, the yeast and other solids have
fallen to the cone's apex can be simply flushed out
Brewing Process through a port at the apex. Open fermentation vessels
Brewing is typically divided into nine steps: are also used, often for show in brewpubs and in Europe
milling, malting, mashing, lautering, boiling, in wheat beer fermentation. These vessels have no tops,
fermenting, conditioning, filtering, and filling. making it easy to harvest top-fermenting yeasts. The
open tops of the vessels increase the risk of
Mashing contamination, but proper cleaning procedures help to
This is the process of mixing milled, usually control the risk (Loeffler. 2006).
malted grain with water and heating it with rests at Fermentation tanks are typically made of
certain temperatures to allow enzymes in the malt to stainless steel. Simple cylindrical tanks with bevelled
break down the starches in the grain into sugars, ends are arranged vertically and conditioning tanks are
especially maltose. usually laid out horizontally. A very few breweries still
use wooden vats for fermentation but wood is difficult
Lautering to keep clean and infection-free and must be re-pitched
This is the separation of the extracts formed often, perhaps yearly. After high kräusen, the point at
during mashing from the spent grain to create wort. It is which fermentation is most active and copious foam is
achieved in either a lauter tun: a wide vessel with a produced, a valve known in German as the
false bottom, or a mash filter: a plate-and-frame filter “spundapparat” may be put on the tanks to allow the
designed for this kind of separation. Lautering has two carbon dioxide produced by the yeast to naturally
stages: wort run-off, during which the extract is carbonate the beer. This bung device can regulate the
separated in an undiluted state from the spent grains and pressure to produce different types of beer; greater
then sparging in which extract that remains with the pressure produces a more carbonated beer (McGovern,
grains is rinsed off with hot water. 2004).

Wort Boiling Conditioning

Wort boilingensures wort’s sterility, thus When the sugars in the fermenting beer have
helping to prevent contamination with undesirable been almost completely digested, the fermentation
microbes. During the boil, hops are added which process slows and the yeast cells begin to die and settle
contribute aroma and flavour compounds to the beer at the bottom of the tank. At this stage, especially if the
especially their characteristic bitterness. Along with the beer is cooled to around freezing, most of the remaining
heat of the boil, they cause proteins in the wort to live yeast cells will quickly become dormant and settle,
coagulate and cause the pH of the wort to fall and they along with the heavier protein chains, due simply to
inhibit the later growth of certain bacteria. Finally, the gravity and molecular dehydration. Conditioning can
vapours produced during the boil volatilize off-flavours, occur in fermentation tanks with cooling jackets. If the
including dimethyl sulfide precursors. The boil must be whole fermentation cellar is cooled, conditioning must
conducted so that it is even and intense. The boil lasts be done in separate tanks in a separate cellar. Some
between 60 and 120 minutes depending on its intensity, beers are conditioned only lightly, or not at all. An
the hop addition schedule and volume of wort the active yeast culture from an ongoing batch may be
brewer expects to evaporate (Gilliland, 1967). added to the next boil after a slight chilling in order to
produce fresh and highly palatable beer in mass
Fermentation quantity (Miller, 2012).
Fermentation begins as soon as yeast is added
to the cooled wort. This is also the point at which the Filtering
product is first called beer. It is during this stage that Filtering the beer stabilizes flavour and gives it
fermentable sugars won from the malt (maltose, a polished, shiny look. It is an optional process. Many
maltotriose, glucose, fructose and sucrose) are craft brewers simply remove the coagulated and settled
metabolized into alcohol and carbon dioxide. solids and forgo active filtration. In localities where a
Fermentation tanks come in many shapes and sizes, tax assessment is collected by government pursuant to
from enormous cylindro-conical vessels that can look local laws, any additional filtration may be done using
like storage silos, to five-gallon glass carboys used by an active filtering system (Hornsey, 2004). The filtered
home brewers. Most breweries today use cylindro- product finally passes into a calibrated vessel for
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measurement just after any cold conditioning and prior

to final packaging where the beer is put into the
containers for shipment or sale. The container may be a
bottle, can, of keg, cask or bulk tank. Filters come in
many types. Many use pre-made filtration media such
as sheets or candles. Kieselguhr, a fine powder of
diatomaceous earth, can be introduced into the beer and
circulated through screens to form a filtration bed.
Filtration ratings are divided into rough, fine, and
sterile. Rough filters remove yeasts and other solids,
leaving some cloudiness, while finer filters can remove
body and colour. Sterile filters remove almost all
microorganisms.

Sources of Contamination
The microbiological stability of the final
product can be compromised from a very early point in
its production, with spoilage organisms able to access
the brewing process at every stage, even from to
dispense (Hill and Bamforth, 2009). Beer may contain
microbial contaminants originating from a variety of
sources. Primary contaminants originate from the raw
materials and the brew house vessels and secondary
contaminants are introduced to the beer during bottling,
canning or kegging. While approximately half of the
documented microbiological problems can be attributed
to secondary contaminations, the consequences of
primary contaminations may be more catastrophic, with
the potential loss of a complete brew (Vaughan et al.,
2005).
Most potential contaminants of beer originate
from raw materials and/or unclean brewing equipment.
Brewing raw materials, such as malt, hops and
occasionally brewing water, may be infected by
microorganisms and these have to be killed during the
brewing process to prevent wort and beer spoilage (Hill
and Bamforth, 2009).

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Fig 1: How microorganisms spread in the brewery (Back, 2005)

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Malt contamination of pitching yeast compromises product

The effects on brewing and beer of quality and taste and can have a significant effect on the
contamination of growing, stored or malted barley are final beer (Hill and Bamforth, 2009). To prevent
varied. The best known effect of the micro biota of both microbial contamination during the production process,
barley and malt is that of reduced gas stability or the microbiological purity of brewing yeast starters is a
gushing (spontaneous ejection of beer from its necessary condition to maintain high product quality.
container). A variety of different fungi have been Many strains are available on the market and their
associated with gushing, the most notable are Fusarium characterization is necessary for quality control in dry
graminearum and Fusarium moniliforme. Small fungal yeast production (Manzano et al., 2005). The
proteins, hydrophobins, present in fungal cell walls contamination of production strains with wild yeasts
have been isolated from strains of the genera Fusarium (non-Saccharomyces or Saccharomyces) and bacteria
, Nigrospora and Trichoderma and shown to act as may contribute negatively or positively to beer
gushing factors in beer (Sarlin et al., 2005 ). A second properties and characteristics.
consequence of fungal infection of barley and malt is
the potential for release of mycotoxins, compounds Hops
toxic to man or animals. Aflatoxin B1, ochratoxin A, Hops are known for their antiseptic properties.
zearalone, deoxynivalenol (DON) and fumosins B1 and As described, the majority of Gram positive bacteria are
B2 are mycotoxins that may be transmitted from inhibited by hops, although Gram negative bacteria are
contaminated grains into beer. In addition to the unaffected. Whole hops are dried following harvesting
potential harm to humans, mycotoxins may affects (Hill and Bamforth, 2009). This process reduces the
fermentation due to their influence on yeast activity. chances of subsequent microbial contamination, and for
There is also an apparent relationship between the brewers who do use whole hops no beer spoilage effects
ability of strains to produce the mycotoxin zearalone attributable to infected hops have been reported (Hill
and gushing (Hill and Bamforth, 2009). and Bamforth, 2009). Similarly, no beer spoilage
organisms have been reported to have been introduced
Water by other herbs or plant-derived products used in
Breweries and good water have long had a brewing (Hill and Bamforth, 2009).
close association and water quality is generally taken
for granted (Hill and Bamforth, 2009). Water used for Sugars
brewing must be fit for human consumption (potable). Free flowing sugar, syrups or honey are
As such it must be free from contaminating organisms. commonly used adjuncts and are generally added
However, what is fit to drink is not necessarily fit for during wort boiling. The main concern in brewing
brewing use (Hill and Bamforth, 2009). Water for involves transfer of bacterial spores, principally from
brewing is boiled during the process. From a Bacillus spp which can withstand heat treatment
microbiological point of view the main concern is the including boiling and may persist into the finished beer
introduction of spoilage organisms from water (Hill and Bamforth, 2009).
introduced after fermentation, for example during
dilution of beer following high gravity brewing or from Brewery contaminants and challenges
vessels rinsed with contaminated water (Hill and Lactic acid bacteria (LAB)
Bamforth, 2009). Lactic acid bacteria (LAB) are known as
predominant beer spoilers and it has been reported that
60-90% of the microbiological incidents are cause by
Air LAB (Back 1994, 1997). Lactic acid bacteria are
Air currents can carry microbial contamination universally considered useful microorganisms in the
to the filling area from numerous sources including food industry and are used in wide range in fermented
pasteurisers, floor drains, personnel, forklifts, products. At the same time, however, can be harmful
packaging material, and the external environment and spoilage in many foods and beverages (Suzuki,
(Dirksen, 2005). Bacteria of the genera Acetobacter, 2011). The development of spoilage lactic acid bacteria
Bacillus, Lactobacillus, Micrococcus, Pectinatus, has been associated with increased use of hops from
Pediococcus, Pseudomonas, Shigella and Streptococcus 1400BC. Hop compounds added to confer bitter flavour
and various yeasts and moulds have been isolated from on beer are reported to exert an antibacterial effect by
the air around the filling line of several breweries acting as proton ionospheres and dissipate the trans
(Henriksson and Haikara, 1991). membrane pH gradient which prevent Gram-positive
bacteria including most LAB from growing in beer.
Pitching yeast Hop resistance ability has been known as a
The most common source of bacterial distinguishing character of beer spoilage LAB strains.
contamination in the brewery is probably from pitching Two genes, HorA and HorC have been demonstrated to
yeast which can transfer contaminants from confer hop resistance ability on LAB. HorA acts as an
fermentation to fermentation. Any microbial ATP-dependent multidrug transporter and extrudes
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toxic hop compounds out of bacterial cells. HorC, on be considered "foreign", but may, after a latent phase,
the other hand confers multidrug resistance to various adapt themselves to the brewery environment and thus
drugs including hop compound, acting as a proton become harmful (Back, 2005). Occasionally, other
motive force (PFM)-dependent multidrug transporter lactic acid bacteria are also said to be involved in this
(Sakamoto and Konings, 2003). fermentation process besides L. brevis (e.g. L. buchneri,
Resistance to hop compounds is considered a L. lindneri, L. Casei, L. coryniformis, L. plantarum)
discriminatory nature of LAB that could adversely (Back, 2005).
affect the beer. These resistance mechanisms have Differences in cell morphology among the
provided selective advantages for their development in hetero fermentative, closely related species L. brevis, L.
beer compared to other organisms (Fernandez and brevis (frigidus) and L. brevisimilis, scarcely exist. The
Simpson, 1993). The LAB share considerable genetic cells of L. brevis (figidus) are on average a bit shorter
and molecular homology and have long been and appear apart from one another under the
recognized as a natural group. However, a definitive microscope due to the production of slimy capsules. L.
description of the LAB cannot be agreed upon and with brevisimilis is noted for its somewhat slimmer cells,
current developments in the taxonomy of these bacteria which may form short chains or, in some strains, be
is probably more distant now than any previous time. curved (Back, 2005). L. brevis (frigidus) differs from L.
The typical LAB is a Gram-positive, non-sporulating brevis mainly in the production of capsules and in the
rod or coccus. It lacks the enzyme catalase and is fermentation of melezitose. In beer, this species is rare,
strictly fermentative producing either a mixture of lactic but quite dangerous (severe hazes, sediments and
acid, CO2, acetic acid and/or ethanol or almost entirely ropiness, increasing of viscosity). In the worst case, the
lactic acid (homo-fermentation) as the major metabolic latter may be so thick as to cause the beer to appear
end-product from sugar catabolism (Priest, 1996; 2006). viscous at dispensing. Because of the production of
Since both types produce lactic acid, these bacteria have capsules, this species belongs to the beer spoilers being
been adapted to grow in an acidic environment such as most resistant to sanitizing agents, and may tolerate up
that found in fermented beverages. Spoilage LAB to 25 pasteurization units. The taste of the beer turns to
produce in beer excessive acidification, haze and off- acidic, while no other sensory changes are detectable. L.
flavours (Back, 2005). brevis (frigidus) contaminates preferentially the
Lactobacilli unfiltered beer in the cellars. It is capable of surviving
Lactobacilli are the most common beer- for years at those low temperatures. Conditions may
spoilage bacteria, regardless of beer type (Thelen et al., arise, under which it spreads through the filter and
2006). The genus Lactobacillus is the largest among the proliferates in beer-lines, gaskets, valves and other
lactic acid bacteria and includes many species, but only similar receptacles (Back, 2005).
some of them can deteriorate the beer (Rainbow 1981;
Jespersen and Jakobsen 1996). Different species vary in Pediococcus
their ability to grow in beer and in their tolerance to hop Pediococci are important in food technology in
bittering compounds. both a negative and positive sense. P. damnosus is a
major spoilage organism in beer manufacture, since
Lactobacillus brevis growth may lead to diacetyl/acetoin formation, resulting
Lactobacillus brevis is the most commonly in a buttery taste (Salminen et al., 2004). The most
found bacteria in beer (50% of cases) (Back, 2009; important coccus-shaped lactic acid bacterium in the
Vaughan et al., 2005). This obligate hetero- brewery is Pediococcus damnosus. This species mostly
fermentative bacterium is generally tolerant towards referred to as "Beer-sarcina" by practical brewers is
hops and grows optimally at 30°C and pH 4-5 (Priest much feared because of its formation of diacetyl
and Campbell, 1996). Lactobacillus brevis is (unpleasant cheesy flavour). It is a typical, brewery-
physiologically versatile and can also cause various specific organism, which may also be found in
problems in beer such as super attenuation due to its wineries, but not elsewhere, neither in other food
ability to ferment starch and dextrins, haze and industries nor in nature. In contaminated beer bottles, a
acidification (Back, 2005). Lactobacillus brevis is diacetyl flavour develops and sediment deposits on the
widespread in the food industry and in nature. In the bottom. They also can occur as punctiform or "ray-
brewery, it occurs in different types with differing cell patterned" colonies. As the cells tend to settle very
and colony morphology and physiological quickly, hazes are rarely observed. Despite being one of
characteristics. The beer spoiling activity varies the most frequent contaminants of the yeast and of the
considerably depending upon the strain and the origin. sections before the filter (Suzuki, 2011), it results in
Some strains develop spontaneously in all kinds of beer relatively few problems in the packaged beer, as it
causing haze, sediment and acidification, but no accumulates in the yeast dregs on the bottom of the
diacetyl flavour. Other strains are potentially harmful storage tank and is therefore almost quantitatively
and are at best able to grow only in weakly hopped eliminated before filtration. Recently, however, sarcina
beers. Occasionally, some strains also find their way infections have been more frequently noticed in
(through water and air) into the brewery, which could
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Bavarian wheat beers to which trace contaminated yeast and Priest, 1996). Approximately ten species of
has been added for secondary fermentation. Acetobacter have been recognized, whereas
The development of pediococci may also be Gluconobacteroxydans is the most important
responsible for higher than normal contents of Gluconobacter in brewing microbiology (Priest and
histamine and for poor foam stability (as a result of Campbell, 1996); Gluconobacteroxydans in beers with
their enzymatic activity against foam proprieties). A high oxygen content cause hazes, formation of acids,
feature of this species is the formation of tetrads. Cocci and off-tastes, their optimum temperature lies around
in singles or in pairs as well as short chains may also 25-27°C, some strains grow at 34°C, none at 37°C. For
occur. The diameter of the cells is between 0.7 and 1.0 most strains the pH optimum is at 5.5-6.0, some grow
μm depending upon strain, age and culture conditions. even at a pH of 3.6 (Back, 2005).
The bacteria are microaerophilic and grow In the brewery environment, the acetic acid
preferentially in a more or less completely pure CO2 bacteria are ubiquitous: in the bottling hall, on malt, in
environment. Several strains are very demanding as far wort and brewing liquor, and also in culture yeast and
as nutrients and growth components are concerned. As beer. During fermentation and after packaging, the lack
they often require some specific beer-components, they of contact with atmospheric oxygen prevents them from
are unable to grow in the usual media. The optimum growing, as they are strictly aerobic. But they reproduce
temperature lies around 22-25°C, the maximum for themselves actively wherever small residues of beer due
many strains is 35°C. Cellobiose, fructose, galactose, to poor sanitation come in contact with air in valves, in
glucose, maltose, mannose, melezitose and trehalose are cocks, in traps, in the beer-lines, underneath the gaskets
fermented. and so forth (Back, 2005).

Acetic acid bacteria Enterobacteriacea

Acetification of beer was studied by pioneers The family Enterobacteriacea comprises
of microbiology such as Pasteur, Hansen, Henneberg numerous genera of free living and sometimes
and Beijernck. The acetic acid bacteria are Gram pathogenic bacteria. Fortunately, none of the
negative, catalase-positive, oxidase negative, non- pathogenic types, such as Salmonella or Shigella
sporing, motile or non-motile short or coccoid rods, species have been found in beer. The Enterobacteria are
exhibiting strictly aerobic metabolism (Back, 2005). A facultative anaerobes able to grow in the presence or
common characteristic of all the representatives of this absence of air, but they are inhibited by ethanol and low
family is the oxidation of ethanol to acetic acid in acid pH so are only responsible for beer spoilage in low
or neutral media (Back, 2005). This characteristic is alcohol products (< 2% by vol) with a relatively high
used commercially for the production of vinegar, but pH (>4.2) (Priest and Stewart, 2006).
needless to say is very detrimental to the brewer. Enterobacteriaceae are facultative aerobic, Gram
Because beer is should be stored with limited access of negative, catalase-positive and oxidase-negative. Their
air, spoilage by these ubiquitous bacteria should not morphology is slightly variable and they exhibit a
occur. marked tendency toward pleomorphic forms. Usually,
However, bacteria of the genus Acetobacter they form short rods (0.6-1.5 x 1.5-3.0 μm) with weakly
are ubiquitous and can cause problems in public houses illuminating cell walls. The cells are often bellied,
dispensing cask-conditioned beer in which the ale beer bulky or sausage shaped, with round, pointed or
is displaced by air (Priest and Stewart, 2006). Their spindle-shaped ends. Brewery-specific strains are
optimum temperature lies between 25 and 30°C, and the usually non-motile but occasionally they carry
optimum pH at 5 - 6, whereas most strains are able to peritrichous cilia.
develop at pH-values as low as 3.6-3.8. The maximum Among enteric bacteria particularly the two
temperature they can withstand is usually below 37°C. species Obesumbacterium proteus and Enterobacter
The family includes the two genera Acetobacter and agglomerans show up as microorganisms are indirectly
Gluconobacter, marked by their differences in the so- harmful to beer. Enterobacter cloacae, E. aerogenes, E.
called "over-oxidation" and in the cilia. Acetobacter sakazakii and Klebsiella pneumoniae are more seldom.
species oxidise acetic and lactic acid to CO2 and H2O The optimum temperature of O. proteus and of
and carry peritrichous or lateral E.agglomerans is 25 - 28°C, the maximum 32 - 37°C
cilia. Gluconobacter lack the "over-oxidation" (Back, 2005). In the past, these species were
capability and are ciliated at the poles. The genus collectively considered "wort bacteria" or included in
Acetobacter consists of four species: A. aceti, A. the coliform germs. At least for O. proteus and
hansenii, A. liquefaciens and A. pasteiuianus, while E.agglomerans, as these bacteria exhibit a very low
only one species of Gluconobacter is described namely temperature maximum (often below 35°C) and grow,
G. oxydans (Back, 2005). unlike typical coliform bacteria, at temperatures as low
Acetobacter and Gluconobacter spp have been as approx. 3°C. They scarcely occur in the wort
isolated from breweries, there are resistant to the environment, but frequently contaminate the yeast and
bacteriostatic activity of hops, acid and ethanol and are are detected in the whole unfiltered beer area. In the
therefore capable of growing and spoiling in beer (Van culture yeast, they are able at strong contamination
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levels and at temperatures of approximately 8°C, to the most important carbon source by the strictly
induce heavy off-tastes through formation of acetoine, anaerobic beer spoilers, Pectinatus and Megasphaera
dimethylsulphide (DMS) and dimethyldisulphide (Back, 2005).
("celery-taste"). The impact of these metabolites goes
right through to the packaged beer in which they induce Pectinatus spp.
more or less strong off-flavours, depending upon the Pectinatus spp are now recognized as one of
degree of contamination. the most dangerous beer spoilage bacteria. They play a
Moreover, these bacteria secrete peptidases major role in 20 to 30% of bacterial incidents, mainly in
and proteinases into the substrate: at high contamination non-pasteurized beer rather than in pasteurized beer
rates, this may impair the foam stability of the beer (Back, 1994). Pectinatus species were long thought to
(Back, 2005). Hafnia protea, formerly be Zymomonas spp because of their phenotypical
Obesumbacterium proteus, and Rahnella aquatilis, similarities. Pectinatus strains were initially isolated
formerly Enterobacter agglomerans, have been from spoiled beer in the late seventies and assigned to a
detected in pitching yeasts but never in finished beer. new genus and species Pectinatus cerevisiiphilus. Later
They can retard the fermentation process. Beer a new species Pectinatus frisingensis has been
produced with yeasts contaminated with H. protea has a established among the initial isolates (Schleifer et al.,
parsnip-like or fruity odour and flavour (Van, 1996; 1990); the type strain of P. frisingens is a Finnish
Sakamoto and Konings, 2003). During the isolate first identified as P.cerevisiiphilus in 1981
fermentation process O. proteus grows rapidly causing (Helander et al., 2004). The first isolate was obtained
the rate of fermentation to decrease and can lead to an from breweries in 1971 and so were all subsequent
inferior product of high specific gravity and high pH. isolates (Haikara, 1984). The natural habitat of the
The bacterium can cause serious problems to the Pectinatus species is still unknown (Haikara et al.,
fermentation. In addition, growth of O. proteus during 2009). Two species are found in this genus: P.
early fermentation is known to produce organo-sulphur cerevisiiphilus and P. frisingensis (Sakamoto and
compounds, various alcohol sand diacetyl which are Konings, 2003).
thought to contribute to the parsnip-like smell of O. Pectinatus spp are non-spore-forming motile
proteus-contaminated beer (Maugueretand and Walker, rods with lateral flagella attached to the concave side of
2002). the cell body (Sakamoto and Konings, 2003). Both
Significantly the growth of O. proteus can Pectinatus species are Gram-negative, strictly
contribute to the formation of apparent total N-nitroso anaerobic, absolutely harmful beer spoilers. The cells
compounds (ATNC) during fermentation (Fernandez are slim, with parallel walls, slightly arched or snake or
and Simpson, 1993). ATNCs are formed from the helicoidally shaped. On average, their diameter is 0.8
reduction of nitrates, naturally present in brewing raw μm, and their length 4 μm and they have round or
materials, to nitrite during the growth of O. proteus. spindle shaped ends. Sometimes thread-like and bowed
The nitrites formed can be subsequently converted to cells also show up but mostly the rods occur singly or in
nitrosamines by reacting with the amines present in the pairs. The bacteria are ciliated laterally (comb-like) and
wort or beer (Maugueretand and Walker, 2002). flutter rapidly when they are young, but upon growing
ATNCs represent a possible risk to health and older, they slow down and are reminiscent of snakes
consequently their concentration is strictly monitored (Back, 2005). Old cultures are mostly non-motile.
and limited to 20 μg/l (Maugueretand and Walker, Pectinatus grows at temperatures between 15 and 40°C,
2002). Abnormally high levels of diacetyl and dimethyl its optimum being 30-32°C. It ferments several sugars,
sulfide were detected in beer produced from wort sugar alcohols and organic acids. In the presence of
contaminated by R. aquatilis (Van, 1996; Sakamoto and fermentable sugars, the medium is acidified, but when
Konings, 2003). Similar problems arise in unfiltered lactate or pyruvate is fermented, often a slight pH-
wheat beers, if the yeast added to the beer before increase can be noticed. The main metabolites are
packaging is contaminated by these enteric bacteria propionic, acetic and succinic acid, and acetoine (Back,
(Back, 2005). They are classified as indirect beer 2005).
spoilers because the damage they induce is generated Pectinatus spp can be identified based on
during the brewing cycle, whereas they are unable to production of large quantities of propionic acid and
grow in the finished beer, at pH-values below 4.8 hydrogen sulphide in beer (Haikara and Henriksson,
(Back, 2005). 1992). Pectinatus bacteria appear to be common
inhabitants in brewery bottling hall deposits. In biofilms
Obligate anaerobic bacteria on bottling machines, Pectinatus species were regarded
Occasionally beer is contaminated by mixed as occasional invaders flourishing on favourable niches
populations consisting of lactobacilli, pediococci, rather than permanent biofilm members.
Pectinatus and Megasphaera. In such cases, the Characterization of environmental isolates has indicated
spoilage usually occurs in two phases. The lactic acid that several sources may exist in a single brewery
bacteria develop first, scavenging residual oxygen and (Sakamoto and Konings, 2003). Moreover, Pectinatus
producing mainly lactic acid, which is later utilised as bacteria have supposedly been detected in pitching
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yeast and in malt steeping water (Sakamoto and addition, hydrogen sulphide is generated (Engelmann
Konings, 2003). The bacteria grow in any beer, and Weiss, 1985). Their potential for beer spoilage is
provided the pH value is higher than 4.4 and the oxygen restricted by their sensitivity to ethanol (>2.8% v/v) and
content very low (< 0.3 mg/1). Higher values are acid pH (Haikara and Lounatmaa, 1987). They are
tolerated only in cases of very severe contaminations. common spoilage bacteria of unpasteurized packaged
Pectinatus is also capable of reproduction in beers and beers. Signs of spoilage include turbidity and off-
utilizes glycerol or pyruvate as a carbon source. A flavours from the synthesis of organic acid sand
combined contamination with lactic acid bacteria is not sulphuric compounds (Haikara and Helander, 2002).
uncommon: as Pectinatus utilizes the lactate produced: Nevertheless, several weeks may be required before
the spoilage occurs then in two steps. Spoilage of beer turbidity becomes evident (Briggs, 2004). M. cerevisiae
by Pectinatus results in the formation of high is a typical secondary contaminant in bottling-halls, its
concentrations of hydrogen sulphide with its putrid behaviour being very similar to that of Pectinatus.
odour and development of turbidity. Various fatty acids, Mixed contaminations of this species with
especially propionic and acetic, together with some Lactobacillus brevis or L. casei are also frequent. With
acetoin are also produced. the latter, two-stage growth occurs, M. cerevisiae
Pectinatus can be considered as a secondary further utilizing the lactic acid formed (Back, 2005).
contaminant only for bottled beer. The germs are
probably introduced into the breweries by air currents Other bacterial contaminants
or dirty empties and hide in various dead and difficult Bacteria of the genus Zymomonas have a
to clean corners in the filler and crowner area. Frequent unique mode of catabolism among the bacteria in that
sources of contamination are also the conveyors (and they conduct an ethanolic fermentation. This is so
their lubricants), the sewers and loose tiles of bad floors efficient that it has been seriously considered for the
(Back, 2005). Pectinatus cerevisiiphilus is difficult to production of fuel ethanol, but it is not used for potable
detect in the brewery indicated that the most common alcohol. Nevertheless, the bacterium is tolerant of
source of contamination was bottling and capping ethanol (up to about 10% by volume) and has been
equipment in summer. A major concern was the slow associated with spoilage of primed conditioning ale
rate of the microbes reproduction in media a great deal (Priest and Stewart, 2006). The potential beer spoiling
of harm might be done to the pre-shelf life, fewer species Zymomonasmobilis is Gram negative, catalase-
customer complaints, higher production rates and lower positive, oxydase-negative, not spore forming and
rejection rate before detection (Chelack and Ingledew, usually immobile. The bacteria are facultatively or
1987). New species of Pectinatus was first isolated only absolutely anaerobic and form short or long rods (1.0 -
a few years ago from samples collected in breweries 1.5μm x 2.0 - 6.0μm), with rounded ends, usually
and has been comprehensively characterized by occurring singly or in pairs: but bigger agglomerates are
Juvonen (2009). Pectinatus haikarae differs markedly occasionally also formed (Back, 2005).
from both of the other primary representatives of this Zymomonasmobilis tolerates oxygen but grows under
genus, P. cerevisiiphilus and P. Frisingensis, due to its anaerobic conditions. It ferments glucose and fructose
positive catalase reaction. but not maltose. Unlike most of the Enterobacteriaceae,
it tolerates ethanol and reportedly survives high-gravity
Megasphaera cerevisiae fermentations in which 12 - 13 % v/v ethanol are
Megasphaera has emerged in breweries along formed. It has a relatively high optimum growth
with Pectinatus and is responsible for 3 to 7% of temperature of 25 ± 3°C. For this reason, it tends to be a
bacterial beer incidents (Sakamoto and Konings, 2003). more common spoilage bacterium in ale breweries as
Megasphaera cerevisiae is also a dangerous, Gram- opposed to those fermenting lager worts at lower
negative, strictly anaerobic, absolutely harmful beer temperatures. Infected worts develop a characteristic
spoiling organism. It is different from Pectinatus, rotten apple odour due to the formation of acetaldehyde.
however, as it forms oval or round cells, preferably as In addition, ethanol, acetic acid, lactic acid, acetoin and
pairs or in chains of four. The cocci are rather large and glycerol are formed (Van and Jespersen, 1998).
attain diameters of 1.2 - 1.6 μm. The temperature range The pH value of the medium is reduced only
for this species is 15 - 37°C, with an optimum of 28 - slightly or not at all. As the sugars glucose, fructose and
30°C. Fructose, lactate and pyruvate are fermented, but sucrose are essential for growth, Z. mobilis is irrelevant
growth also takes place if no sugars are present. The for highly fermented continental beers, but may cause
main metabolites are butyric, caproic, acetic, propionic problems in British ales, to which sugars are added after
and valeric acid as well as CO2 and H2S is also primary fermentation. In contaminated beers, so-called
produced (Back, 2005). "boiling fermentations" may be triggered. An undesired,
M. cerevisiae forms only slight hazes in beer fruity flavour evolves in the beers, probably related
and almost unnoticeable sediments but causes severe mainly to the metabolites hydrogen sulphide and
off-smells and off-tastes (Back, 2005). Megasphaera acetaldehyde. In addition, strong hazes and sediments
strains produce several organic and fatty acids, notably are also formed, and the beers fail to meet consumer
butyric acid and some acetic, isovaleric and valeric. In expectations (Back, 2005).
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Wild Yeast as causing haze and turbidity (Priest and Stewart,

Wild yeasts are generally defined as those 2006).
yeasts not deliberately used and not under full control Aerobic wild yeast can cause problems in beer
(Gilliland, 1967). The definition of wild yeasts is -dispensing equipment than in the brewing process or in
diffuse and for convenience, is traditionally divided into packaged beer. This is due to the higher oxygen levels
Saccharomyces and non-Saccharomyces (Boulton and and higher temperatures at certain points in the
Quain, 2001). This definition includes brewing strains dispensing system. These conditions favour
that are used for a different style of beer and may have contamination by microorganisms such as aerobic wild
been cross-contaminated in the brewery, as well as non yeast in addition to the oxygen tolerant beer spoilage
brewing yeasts that have gained access from the air or organisms found in the brewery environment (Storgårds
raw materials. It is important to emphasize that there et al., 2006; Storgårds, 2000). Wild yeasts may be
are many genera and species of yeast with diverse isolated from all stages of the brewing process, from
physiologies; the only unifying feature is that the raw materials to packaged beer, and from bar dispense
organisms are predominantly unicellular. However, equipment. However, they typically contaminate
many types of yeast have a semi filamentous lifestyle pitching yeast, and increase in number over successive
and may form mycelia under various environmental re-pitching. Spoilage may occur in the finished product,
conditions (Priest and Stewart, 2006). Although boiling during conditioning, or (to a lesser extent) during
of wort kills most microorganisms and thence is fermentation (Fleet, 1992).
inoculated by pitching yeast, other kinds of unwanted The growth of wild yeasts may cause the
yeasts can get into beer during fermentation; these production of off-flavours particularly phenolic
yeasts are collectively known as wild yeasts (Deàk, compounds. The presence of these volatile phenolic
2008). Wild yeasts were detected in 41 % of pitching compounds is considered undesirable when present in
yeasts investigated (Van and Jespersen, 1998). excessive concentration in bottom fermented pilsner
A contamination with wild yeast results in a beers. Hence the term ‘‘phenolic off-flavour” (POF).
phenolic off - flavour in lager beers. Most brewing Despite being historically catalogued as an off-flavour,
strains are unable to utilize dextrins and these persist in these compounds are known to be essential flavour
beer where they contribute to fullness and mouthfeel. contributors to the characteristic aroma of Belgian
Some strains originally classified as S. diastaticus but white beers (made with un-malted wheat), German
now placed with S. cerevisiae, possess glucoamylase weizen beers (made with malted wheat) and rauch
and in consequence can utilize dextrins. Contamination beers. However, in many other top-fermented blond and
of fermentations with diastatic yeasts leads to super- dark specialty beers the phenolic flavour is essential for
attenuation of the wort and beers with abnormally low the overall flavour perception (Vanbeneden et al, 2008).
present gravity. Occasionally, diastatic yeasts have been Saccharomyces wild yeasts produce phenolic off-
used to produce so called `light' beers. Contamination flavours (such as 4-vinyl guaiacol) by decarboxylating
of unpasteurized bottled beer with diastatic yeast is various phenolic acids, due to the presence of the
potentially hazardous, since abnormally high phenolic off-flavour gene (POF) (Ryder et al., 1978;
concentrations of carbon dioxide can develop with the Thurston, 1986).
consequent risk of bottle explosions.
Pichiamembranefaciens is the most common Remedies to brewery contamination
contaminant of beer and wine in this category. The Hygiene management in production facilities
acetic acid-forming Brettanomyces and Dekkera The role of cleaning and disinfection for both
species, although fermentative, do not usually cause a small and large breweries has grown immensely due to
threat to the brewing process because they cannot production of non-pasteurized products (Kretsch, 1994)
flourish under anaerobic conditions (Priest and Stewart, and due to new products low in alcohol and bitterness
2006). (Storgårds, 2000). In the 1990 edition of the “Society of
The aerobic yeasts such as Debbaromyces, Dairy Technology manual CIP”, Cleaning in Place
Pichia, and Williopsis produce yeasty or estery flavours (CIP) was defined as the cleaning of complete items of
that are most unwelcome (Priest and Stewart, 2006). plant or pipeline circuits without dismantling or
The fermentative yeasts such as Kluyveromyces, opening of the equipment and with little or no manual
Saccharomyces, Torulaspora, and Zygosaccharomyces involvement on the part of the operator. The process
on the other hand can cause serious problems in the involves the jetting or spraying of surfaces or
fermentation. They are potentially able to compete with circulation of cleaning solutions through the plant under
the culture yeast and although they cannot generally kill conditions of increased turbulence and flow velocity
it, if they grow just a little faster than the culture yeast (Tamine, 2008).
they will displace the brewing yeast over successive Cleaning out of place is essentially the
generations. As these wild yeasts neither flocculate well opposite of CIP and refers to most manual cleaning
nor interact with finings, they generally pass into applications. Either the equipment must be broken
conditioning where they can have deleterious down into pieces or major modifications must be
organoleptic effects on post fermentation beers, as well performed before the cleaning can take place. Some
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equipment that is normally cleaned via CIP should be - mechanics (pressure, volume flow, flow speed);
cleaned periodically in a COP mode, such as heat- - chemical (type and concentration of the cleaning
exchangers. Therefore, COP cannot and should not be agent) (Back, 2005).
completely avoided. Some of the drawbacks and
benefits of COP are: Improve the management of cleaning and sanitizing
- Cleaning results may vary with operator/employee; procedures
- Exposure of employee to cleaning solutions; With the use of these reagents discussed
- Time-consuming process; below, the challenges of contamination in breweries
- Easily verifiable through visual inspection; will be reduced.
- May expose residuals left by CIP cleaning
(Loeffler, 2006). Alkaline cleaning agents
Soil adheres to surfaces in very complex ways. It can be The composition of alkaline components in a
trapped mechanically in pores, cracks or other cleaner determines its alkalinity. Acidic product
inclusions, which explains the choice of hard-surface residues as well as carbon dioxide can partially
materials such as finished stainless steel. We also see neutralize and reduce the original free alkalinity.
electrostatic binding forces, both between the surfaces Sodium hydroxide provides the most alkalinity, which
and the soil as well as between different types of soils is one of the key factors in removing organic soil in
such as protein and mineral salts. The sum of all these brewery cleaning (Loeffler, 2006). Sodium hydroxide
binding forces combined can be expressed as the exhibits an excellent emulsifying capacity for protein.
adhesion energy, which is the energy that has to be Accordingly, it is used in a wide variety of applications
achieved during the cleaning process to remove the soil. in breweries. Caustic potash has an even greater
During cleaning, the adhesion energy is derived by capacity for breaking up soiling than sodium hydroxide.
combining the energy from chemicals, mechanics, and It is only used in limited applications; however, due to
temperature, whereas the energy from these three the fact that is several times more expensive.
components is interchangeable within certain limits
(Loeffler, 2006). Acidic Cleaning agents
The process of soil removal can be divided into four Beer-stone and mineral deposits are primarily
major steps: based on mineral components. These types of deposits
1. Transport of the cleaning solution to the soil are virtually impossible to remove with alkaline
with complete wetting of the soil. products alone. However, acids will take water-
2. Chemical reactions and physical processes insoluble salts and chemically transform them into a
during the cleaning process: soluble, rinseable form (Loeffler, 2006).
- reaction of the cleaning solution with hard
water constituents and/or suspended soil; Mineral acids: The corrosiveness as well as the
- convective and diffusive transport of the incompatibility of most mineral acids with other active
cleaning agents from the cleaning solution to components commonly used in cleaning products limits
the soil; their use to primarily phosphoric acid and nitric acid.
- wetting or transport of the cleaning agents Sulfuric acid may be used at temperatures not
within the soil itself; exceeding 30°C. Hydrochloric acid should be avoided
- cleaning reaction with the soil, both at all cost (Loeffler, 2006).
chemically and physically - diffusive transport
of soil particles removed during the cleaning Organic acids: The most important criteria for organic
process. acids are their odour (short- chained carbonic acids),
3. Removal of the soil from the surface and solubility and strength. Commonly found products from
transfer into the cleaning solution via this group are formic acid, oxalic acid, citric acid and
dispersion and/or emulsification. lactic acid (Loeffler, 2006). The most suitable acidic
4. Prevention of re-depositing removed soil cleaners are products with phosphoric acid. Phosphoric
through stabilization in the cleaning solution acid far exceeds nitric acid and sulfuric acid in its
and transport of removed soil away from the cleaning power (Storgards, 2000).
surface (Loeffler, 2006).
Disinfecting
Cleaning is the removal of contamination or The single most important precondition for
undesired residues from hard surfaces with the aid of successful sanitizing is an effective cleaning program
chemical and/or physical cleaning methods and agents. (Loeffler, 2006). Disinfecting agents used in the food
industry are tasked with making production equipment
Factors for successful cleaning include: free of microorganisms after use and subsequent
- temperature (hot cleaning, cold cleaning); cleaning. Microorganisms can be killed physically and
- cleaning time (the longer the cleaning time, the chemically. Physical elimination involves heat
greater the cleaning success); treatment, UV and X –rays and other methods.
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Disinfecting with chemicals is possible via a host of are represented in the following table.
disinfecting agents. The most important disinfectants

Table 1: Disinfecting substances.

Active ingredient Remarks Use
Peroxacetic acid Acidic disinfecting agent with oxidizing effect (destroys cell Bottle cleaning,
membrane); conditionally stackable due to loss effectiveness; CIP cleaning.
automatic dosing via inorganic conductive acids only; sealing
materials may be harmed with extended contact; very broad
range of effectiveness.
Hydrogen peroxide Neutral disinfecting agent with oxidizing effect; very CIP cleaning;
environmentally and waste water friendly, since it decomposes spray
with organic material in water and oxygen; high usage disinfecting
concentration; very broad range of effectiveness.
Active chlorine Alkaline disinfecting agent with oxidizing effects; danger of Bottle cleaning;CIP
(sodium hypoclorite) chlorophenol formation (negatively effects taste of the product); cleaning;drinking –
very broad range of effectiveness; ATTENTION: water disinfecting
when mixing with acidic solutions, chlorine gas is released!
Chlorine dioxide Disinfecting agent with oxidizing effect; two – component Bottle cleaning;CIP
system that is mixed on - site when used; economical operating cleaning; drinking –
costs, but high investment costs; very broad range of water disinfecting
effectiveness.
Quaternary neutral disinfecting agent (surfactants); destroys the cell Static disinfecting;
ammonium membrane; heavily foaming (not suitable for CIP); surface - spray disinfecting
compounds active; relatively difficult to rinse out due to the surface activity
(adheres well to the surface)
Source: (Loeffler, 2006).
- intermediate rinse;
Formulation based on peracetic acid and - additional detergent circulation (optional);
hydrogen peroxide is frequently used for post-cleaning - additional intermediate rinse (optional);
disinfection. Peracetic acid (PAA) penetrates the cell - disinfectant rinse (optional);
and oxidises enzymes and other proteins irreversibly. - drain.
PAA has been shown to be effective against biofilm. The pre-rinse cycle removes the loose soil: the more of
The agents quickly lose their activity in a basic this that can be removed with a simple rinse, the less
environment, making careful rinsing after alkaline the need for the chemical, mechanical and thermal
cleaning essential (Storgårds, 2000). Peracetic acid and energy needed later. Rinsing efficiency can be
hydrogen peroxide-based disinfectants also perform improved through ‘burst rinsing’ of vessels, and
well in the presence of organic soil, but they are through ensuring that draining surfaces are on a slope.
markedly less effective when the temperature is The detergent circulation is usually the key stage in
decreased from ambient (20°C) to 4°C. At low removing residual soil. Circulation time is typically 10–
temperatures, such as in the fermentation cellar, higher 30 min, but the choice of detergent very much depends
concentrations are needed to obtain a good result al on the particular cleaning situation. Examples of
(Storgårds, 2000). detergent/temperature solutions are shown in Table 2.
An intermediate rinse is then required if a further
CIP programmes detergent circulation is going to be used. This is
A typical CIP sequence will comprise the following particularly the case if an acid treatment is necessary,
cycles: usually when scale removal is required (Tamime,
- pre-rinse; 2008).
- detergent circulation;

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Table 2: Typical CIP programmes used in the brewery.

Action Temperature Duration

Prerinsing Cold or hot 5-10 min
Alkali cleaning; sodium hydoxide Cold or hot (60-85°C) 10-16 min
(1.4-4%)
Intermediate rinsing Cold or hot 10-30 min
Acid cleaning; phosphoric, nitric or Cold 10-30 min
sulphuric acid (1-2%)
Intermediate rinsing Cold or hot 10-30 min
Disinfection
- disinfectant solution Cold 10-30 min
- hot water 85-90°C 45-60 min
Final rinsing if necessary
-may contain a disinfectant Cold 5-10 min
at low concentration
Source: (Storgårds, 2000).

Conclusion Back, W. (2005). Brewery. In: Colour Atlas and

The application of inhibitory components Handbook of Beverage Biology. W. Back,
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Published by Nigerian Society for Microbiology 3940

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