[FOOD SAFETY & QUALITY]
by Neil H. Mermelstein
Testing for Alicyclobacillus,
a quality control technician
draws a sample of a juice
beverage. Photo courtesy of
Cott Beverages Inc.
pg 62
Preventing Juice Spoilage
S
helf-stable juices, juice bever-
ages, and certain other
beverages and concentrates
are susceptible to spoilage by
Alicyclobacillus (ACB), a non-
pathogenic, aerobic, spore-
forming, thermoaciduric bacterium
typically found in soil and on har-
vested fruit. The spoilage occurs
only in shelf-stable products
[Alicyclobacillus] produces heat-resistant spores that can survive pasteurization
and can germinate and grow under acidified or high-acid conditions.
because the spores do not germi-
nate and grow in chilled and frozen
juices and concentrates. It does
not pose a public health risk but
can cause economic damage to
the juice products industry.
Unlike other spoilage
microbes, the heat- and acid-lov-
ing bacterium doesn’t produce
visible signs of spoilage, such as
11.12 • www.ift.org
gas, turbidity, or sediment. Rather,
it produces off-flavors from com-
pounds formed during growth,
primarily guaiacol, which is a
chemical with an unpleasant
smoky and medicinal odor or taste
that is detectable by humans at
leves as low as 1 ppb.
Because of ACB ’s heat-resis-
tant nature, the typical process
temperatures used during pas-
teurization of juice products
increases the likelihood of its sur-
vival in the finished product. Of
several ACB species,
Alicyclobacillus acidoterrestris is
most closely associated with spo-
radic spoilage of shelf-stable juice
products.
Handling the Problem
Sean Leighton, Director of Quality
& Food Safety, Contract Manu-
facturing at The Coca-Cola
Company (the no. 1 global buyer of
fruit for juice consumption), said
that the rate and extent of product
spoilage depend on a number of
factors such as product storage
temperature, package type, char-
acteristics of the growth matrix
(product or ingredient), and the
specific microorganism present.
He added that multiple overlapping
strategies should be used by com-
panies to reduce the impact of
these organisms on their business.
ACB, he said, is still a very rel-
evant challenge for the beverage
and food manufacturing industry.
The primary challenge, he said, is
that ACB produces heat-resistant
spores that can survive pasteuri-
zation and can germinate and
grow under acidified or high-acid
conditions. It’s a novel challenge
for the juice industry. The typical
strategy for the juice industry is to
provide a kill step with a thermal
process because the acidity of the
product can guarantee extended
shelf life. But ACB and other ther-
moaciduric bacteria spores
survive and grow in an acidified
environment, resulting in a greatly
reduced shelf life. The saving
grace is that it can’t grow under
refrigerated conditions.
Susceptible products include
heat-stable juice products, iso-
tonic sports drinks, and
vitamin-enhanced waters.
Randy McNeal, Technical
Director at Cott Beverages Inc.
(the largest private-label beverage
company in the United States),
said that spoilage of juice products
by ACB is not as serious or wide-
spread a problem as it was five to
ten years ago because a lot of
work has been done by a lot of
people. Juice processors have put
a lot of pressure on their ingredi-
ent suppliers, and they have
responded by making changes in
their process. The result is fewer
positive samples on incoming
ingredients. The incidence of
spoilage has gone down over the
years but hasn’t gone away.
 A challenge now, he said, is
to prevent spoilage of newer
products that combine fruit juices
with vegetable juices. Vegetable
root crops such as carrots and
beets are naturally contaminated
with soil-based ACB spores and
are therefore more problematic,
but as the industry matures, sup-
pliers will learn how to clean
them up. Juices that are clear,
such as apple juice, can be fil-
tered to eliminate ACB spores, he
said. Ultrafiltration will remove
all spores, but then the trick is not
to re-contaminate the filtered
juice. Juices containing pulp,
such as citrus juices, can’t be
filtered, so sanitation of the
incoming fruit is very important.
The very slow-growing
microorganism needs heat and
oxygen for sporulation and
growth. It takes about two to four
weeks at 100°F before the unde-
sirable flavor develops in apple
juice, he said. Packaging is also
an important factor. ACB won’t
grow in cans because they con-
tain very little air and the air
reacts with the lining of the can.
It can grow in glass bottles that
have a large headspace and in
PET bottles, which are not per-
fect oxygen barriers.
Cott requires its ingredient
suppliers to provide a certificate
of analysis that includes the
results of ACB testing. The com-
pany does some spot-checking
and sends the samples to third-
party laboratories for analysis.
The company also conducts taste
tests as part of its normal quality
control program. The off-flavor is
quite difficult to detect in a nor-
mal quality control program,
McNeal said, since it takes a
week or two to develop. Even in
inoculated trials with 10–20 bot-
tles, only 20% of the samples
may show spoilage after one
week. If guaiacol is present at a Alicyclobacillus colonies are growing on YSG
low level, people are more likely media after three days of incubation. Photo
courtesy of The Coca-Cola Company.
to taste it than smell it. At a rela-
tively high level, people can smell
it right away, and if tasted, the
flavor may linger and cause sen-
sory fatigue.
Analytical Methodology
ACB was identified as the cause
of juice spoilage (initially apple
juice) in 1984, and methods for
detecting it and preventing it
from causing spoilage have been
worked on over the years by
numerous researchers,
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[FOOD SAFETY & QUALITY]
Preventing Juice Spoilage continued...
Relevant Organizations
The International Society of
Beverage Technologists (ISBT)
(www.bevtech.org) is dedi-
cated to the scientific and
technical aspects of the nonal-
coholic beverage industry. Its
members are engaged in the
science, technology, or pro-
duction of soft drinks or
beverages, including suppliers
engaged in scientific or techni-
cal positions. Its next annual
conference, BevTech ® 2013,
will be held April 29 – May 1,
2013, in Fort Lauderdale, Fla. In
2008, ISBT’s Alicyclo-
bacillus Subcommittee worked
to identify and validate rapid
test methods for the detection
and enumeration of spoilage-
pg 64 11.12 • www.ift.org
causing ACB species.
The International Federation
of Fruit Juice Producers (IFU)
(www.ifu-fruitjuice.com) is an
association of fruit juice pro-
ducers’ organizations in more
than 30 countries. It serves as
a nongovernmental organiza-
tion, acts as an information
and communication facilitator,
harmonizes standards and
practices for juice products
manufacturers, and coordi-
nates scientific activities to
benefit the fruit juice industry.
It issued its revision of its IFU
Method No. 12, “Method on
the Detection of Taint
Producing Alicyclobacillus in
Fruit Juices,” in 2007.
The Technical Committee for
Juice and Juice Products
(www.tcjjp.org) is an interna-
tional nonprofit organization
with members from govern-
ment, industry, and academia.
Among its objectives are to
develop and adopt accurate
methods for the analysis of
juices and juice products; facil-
itate collaborative testing of
new analytical methods; facili-
tate acceptance of methods as
official AOAC International
methods; improve communica-
tion among government,
academic, industrial, and pri-
vate laboratory scientists
working on juice research; rec-
ommend and acquire funding
for development of new analyt-
ical methods; and work with
the AOAC to publish a compen-
dium of official juice and juice
product analytical methods.
The Juice Products
Association (www.juiceprod-
ucts.org) is an international
trade association of major
packers and distributors of
fruit and vegetable juices, juice
beverages, and other fruit
products. Its members include
a majority of the juice and juice
beverage processors and sup-
pliers. The organization
promotes high safety and
quality standards and good manufactur-
ing and inspection practices in the
production and distribution of juice prod-
ucts; provides input regarding technology,
trade, legislative, and regulatory propos-
als; and sponsors research.

The European Fruit Juice Association

(AIJN) (www.aijn.org) is an organization
whose purpose is to work for the best
possible political, regulatory, and eco-
nomic framework at the European Union
level to add value and promote the growth
of the fruit juice industry. In 2008, AIJN
issued its “Alicyclobacillus (ACB) Best
Practice Guideline” for the reduction and
control of ACB in the production, packing,
and distribution of fruit juices, juice con-
centrates, purees, and nectars. The
guideline also references AIJN’s “Guide
of Good Hygiene Practice for the Fruit
Juice Industry,” which encourages estab-
lishment of good manufacturing practice
standards. The guideline recommends
use of IFU Method No. 12 for juice con-
centrates and other raw materials, or
alternative methods such as Japan Fruit
Juice Association’s “Unified Test Method
for Thermo-Acidophilic Bacilli.”

companies, and associations. Since a wide

variety of methods were being used for
detection of ACB in fruit juices, the
International Federation of Fruit Juice
Producers (IFU) took the initiative to develop
an internationally accepted method. The
result, IFU Method No. 12 (IFUMB12),
“Method on the Detection of Taint Producing
Alicyclobacillus in Fruit Juices,” was issued in
2004. Revised in 2007, it provides procedures
for testing (1) raw materials such as fruit con-
centrates, syrups, etc. where ACB is likely to
be present as spores; (2) final products such
as juices, drinks, sauces, and other ready-to-
consume products sampled directly after
heat processing; (3) final products taken from
the market; and (4) process water.
Various media have been developed for
the detection of ACB species. Because con-
ventional culture methods can take up to 12
days to detect ACB and another 24 hr to con-
firm guaiacol production, a number of rapid

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 [FOOD SAFETY & QUALITY]
Preventing Juice Spoilage continued...
The foodproof ® Alicyclobacillus Detection Kit detects Alicyclobacillus species and
A. acidoterrestris in the same reaction. Photo courtesy of Merck Millipore.
methods have been developed.
Flow cytometry requires a pre-
incubation or enrichment step
but cuts the culturing time to
about 10 hr. Real-time PCR also
requires a pre-incubation or
enrichment step but cuts the
culturing time to 48 hr. The food-
proof ® Alicyclobacillus Detection
Kit detects ACB species and A.
acidoterrestris in the same reac-
tion; it is available from Merck
Millipore (www.merckmillipore.
com) and Biotecon Diagnos-
tics (www.bc-diagnostics.com)
and will soon be launched in the
United States under the EMD
Millipore brand (www.emdmilli-
pore.com). Gene probe
technology uses a 48-hr pre-
incubation without the need for
DNA extraction and provides
results within 3 hr, and the VIT-
Alicyclobacillus Detection Test
using VIT ® gene probe technol-
ogy is available from Vermicon
(www.vermicon.com).
Industry Approach
Patricia Faison, Technical
Director at the Juice Products
Association, said that although
work to overcome the economic
spoilage risk of ACB in juice prod-
ucts continues to be done, two
decades of research has given
industry a better understanding
of the ecology of this microorgan-
ism. Using the knowledge gained,
the industry has implemented
multiple processes across the
supply chain to minimize risk of
spoilage from ACB. Among them
are use of Good Agricultural
Practices during harvesting,
Good Hygiene Practices during
raw-fruit washing and process-
ing, modified sanitation practices
across process steps, supply-
chain and distribution
improvements, extensive shelf-
life testing to minimize potential
impacts, and implementation of
testing programs. These hurdle
approaches have been effective
in reducing or limiting the impact
of ACB in juice products.
The industry routinely tests
for ACB across the supply chain
using various methods to validate
and verify the efficacy of their
programs—such as IFU’s
Method No. 12 and “Isolation of
Alicyclobacillus from Fruit Juice
Concentrates, Fruit Juices and
Other Acid Products,” which is pub-
lished in the American Public
Health Association’s Compendium
of Methods for the Microbiological
Examination of Foods. The indus-
try continues to search for novel
risk-mitigation strategies as well
as rapid detection techniques for
ACB, she added. The Juice
Product Association’s Technical
Affairs Committee evaluates
methods to detect ACB to ensure
that the industry is kept abreast
of current technology. FT
Neil H. Mermelstein, a Fellow of
IFT, is Editor Emeritus of
Food Technology
• nhmermelstein@comcast.net
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