Journal of Applied Microbiology 2005, 98, 1400–1409
A REVIEW
Microbiology of pressure-treated foods
M.F. Patterson
Department of Agriculture and Rural Development, Northern Ireland and Queen’s University, Belfast, UK
2004/0950: received 16 August 2004, revised and accepted 10 December 2004
1. Summary, 1400
2. Introduction, 1400
3. High-pressure processing equipment, 1401
4. Fundamental effects of pressure on microbial cells, 1401
4.1 Effect of pressure on cell membranes, 1401
4.2 Effect of pressure on cell morphology, 1401
4.3 Effect of pressure on biochemical reactions, 1402
4.4 Effect of pressure on genetic mechanisms, 1402
5. High pressure inactivation of micro-organisms, 1402
5.1 Inactivation kinetics, 1402
5.2 Pressure injury, 1402
1. SUMMARY
High hydrostatic pressure has the potential to produce high
quality foods that are microbiologically safe and with an
extended shelf-life. Micro-organisms vary in their response
to high pressure. Bacterial spores are the most resistant group
and they cannot be significantly inactivated by pressure
alone. Combination treatments using high pressure and heat
have been proposed as a method of producing shelf-stable
low acid foods. Viruses are less resistant than bacterial spores
and their infectivity can be abolished without destroying their
ability to elicit antibodies, leading to the possibility of vaccine
production. Yeasts, moulds and vegetative bacteria vary in
their response to pressure, depending on factors such as
species, strain, processing temperature and substrate, and
these are reviewed in the paper. A knowledge of how these
factors interact is necessary in order to select the optimum
processing conditions for foods. A number of pressure-
treated foods are already commercially available and these are
discussed in the paper.
2. INTRODUCTION
Consumers in the 21st century are demanding high quality
foods that are free from additives, fresh tasting, microbio-
Correspondence to: Margaret Patterson, Agricultural, Food and Environmental
Science Division (Food Microbiology Branch), Agriculture and Food Science Centre,
Newforge Lane, Belfast BT9 5PX, Northern Ireland, UK
(e-mail: margaret.patterson@dardni.gov.uk).
doi:10.1111/j.1365-2672.2005.02564.
5.3 Bacterial endospores, 1402
5.4 Viruses and prions, 1404
5.5 Vegetative bacteria, 1405
5.6 Yeasts and moulds, 1405
6. Extrinsic factors affecting the sensitivity of micro-
organisms to pressure, 1405
6.1 Effect of substrate, 1405
6.2 Effect of temperature, 1406
7. Pressure treatment to improve the microbiological quality
of foods, 1406
8. References, 1407
logically safe and with an extended shelf-life. One food
technology that has the potential to meet these demands is
high pressure processing. High pressure processing, also
known as high hydrostatic pressure or ultra-high pressure
processing, uses pressures up to 900 MPa (c. 9000 atmos-
pheres, c. 135 000 pounds per square inch) to kill many of
the micro-organisms found in foods, even at room tempera-
ture. These pressures are immense. A mid-range food
processing pressure of 500 MPa is equivalent to the weight
of three elephants on a strawberry.
The idea of using high pressure in food processing is not
new. The first report of high pressure being used as a food
preservation method was by Hite (1899). He reported that
milk
kept sweet for longer after a pressure treatment of c.
600 MPa for 1 h at room temperature. Hite et al. (1914)
also reported that while pressure could be used to extend
the shelf-life of fruits, it was less successful with vegeta-
bles. He concluded that fruits and fruit juices responded
well to high pressure because the
yeasts and other
organisms having most to do with decomposition are very
susceptible to pressure. Vegetables, however, he
aban-
doned as hopeless due to the presence of spore-forming
bacteria that survived the pressure treatment and could
grow in the low acid environment. The problem of
pressure resistant spores still remains one of the challenges
for the technology today. Much experimental data has been
produced over the last 100 years but it was not until the
early 1990s that the first commercial food applications
of the technology were seen. There are considerable
ª 2005 The Society for Applied Microbiology

engineering problems involved in repeatedly generating
and containing the immense pressures in a vessel suitable
for food products. However, within the last 25 years a
range of specialist high pressure vessels, based on those
used routinely in the production of polymers, ceramics and
artificial diamonds, became available and reopened the
possibility of commercial production of pressure-treated
foods.
This paper will review how high pressure can be used to
improve the microbiological safety and quality of foods,
including the problem of spore-forming bacteria and
commercial food applications of the technology will also be
discussed.
3. H I G H - P R E S S U R E P R O C E S S I N G
EQ U I P M E N T
A typical pressure treatment system consists of a pressure
vessel, the pressure transmission fluid (usually water) and
one or more pumps to generate the pressure. It is
traditionally a batch process and pressure vessels used
for commercial food production having capacities of 35–
350 l.
Food packages are loaded into the vessel, the top is
closed and the pressure transmission fluid is pumped into
the vessel from the bottom. Once the desired pressure is
reached, pumping is stopped, valves are closed and the
pressure can be maintained without further need for
energy input. The pressure is transmitted rapidly and
uniformly throughout the pressure fluid and the food. The
high pressure is applied in an isostatic manner so that all
parts of the food are subjected to the same pressure at
exactly the same time, unlike heat processing where
temperature gradients are established. As it is equal from
all sides, the pressure does not significantly affect the
product shape. The pressure is released after the desired
treatment time and the food packages can be unloaded. In
the case of liquids, such as fruit juices, the whole vessel
can be filled with the juice, which itself becomes the
pressure transmission fluid. After treatment, the juice can
be transferred to an aseptic filling line, similar to that used
for UHT (ultra-high pressure treatments) liquids. A series
of these vessels can work in sequence, with a vessel filling
with juice, a vessel pressurizing and another emptying, all
operating simultaneously, so the overall system can become
semi-continuous.
High pressure equipment suitable for food use is special-
ized and the capital equipment cost is relatively high,
although running costs are relatively low. Typically a
commercial vessel can cost £500 000 to over £1 million,
depending on its size. It is likely that these costs would
reduce if the use of the technology grows and more vessels
are sold.
ª 2005 The Society for Applied Microbiology, Journal of Applied Microbiology, 98, 1400–1409, doi:10.1111/j.1365-2672.2005.02564.x
PRESSURE TREATMENT OF FOOD 1401
4. FUNDAMENTAL EFFECTS OF PRESSURE
ON MICROBIAL CELLS
There is much published research on the changes induced
by pressure treatment of microbial cells, including altera-
tions in the cell membrane, cell morphology, effects on
proteins, including enzymes and effects on genetic mech-
anisms of micro-organisms (for reviews see Hoover et al.
1989; Smelt et al. 2001). However, despite this effort, the
mechanisms of microbial inactivation are still not fully
understood (Pagán and Mackey 2000).
4.1 Effect of pressure on cell membranes
The cell membrane is generally acknowledged to be a primary
site of pressure damage in micro-organisms (Morita 1975;
Ulmer et al. 2000; Casadei et al. 2002). Evidence of physical
damage to the cell membrane has been demonstrated as leakage
of ATP or UV-absorbing material from bacterial cells subjected
to pressure (Smelt et al. 1994) or increased uptake of fluores-
cent dyes such as propidium iodide that do not normally
penetrate membranes of health cells (Benito et al. 1999).
Stationary phase cells are normally more pressure resistant than
exponential-phase cells. Mañas and Mackey (2004) have
proposed that exponential-phase cells are inactivated under
high pressure by irreversible damage to the cell membrane. In
contrast, stationary-phase cells have a more robust cytoplasmic
membrane that can better withstand pressure treatment. This
proposal was based on the fact that exponential-phase cells
showed changes in their cell envelopes that were not seen in
stationary-phase cells. These changes included physical pert-
urbations of the cell envelope structure, a loss of osmotic
responsiveness and a loss of protein and RNA to the
extracellular medium. Loss of membrane functionality result-
ing from pressure treatment has also been described by
Wouters et al. (1998), who reported that in Lactobacillus
plantarum, pressure treatment at 250 MPa reduced F0F1
ATPase activity. The ability to maintain a DpH was also
reduced and the acid reflux was impaired.
4.2 Effect of pressure on cell morphology
The cell wall is less affected by high pressure than the
membrane and generally no morphological changes can be
observed in prokaryotes and lower eukaryotes by observation
under a light microscope. However, intracellular damage can
be observed using electron microscopy. Ritz et al. (2001),
using scanning electron microscopy (SEM), reported that
bud scars appeared on the cell surface of Listeria mono-
cytogenes after a 10 min pressure treatment at 400 MPa in
citrate buffer. Similarly, Park et al. (2001) studied the effect
of pressure on the ultrastucture of L. viridescens. Nodes on
cell walls of organisms treated at 400 MPa and above for
1402 M . F . P A T T E R S O N
5 min at 25
C were observed using SEM. Transmission
electron micrographs indicated empty cavities between the
cytoplasmic membrane and the cell wall after this treatment.
4.3 Effect of pressure on biochemical reactions
High pressure treatment favours biochemical reactions that
lead to a volume decrease while it can inhibit or retard
reactions that lead to a volume increase. Most biochemical
reactions result in a volume change and are therefore affected
by pressure. Studies carried out on volume changes in
proteins have shown that the main targets of pressure are
hydrophobic and electrostatic interactions while hydrogen
bonding, which stabilizes the a-helical and b-pleated sheet
forms of proteins, is not significantly influenced by pressure
(see Heremans 2001 for detailed review). Enzymes vary
greatly in their ability to withstand pressure. Certain micro-
bial enzymes, such as Bacillus subtilis a-amylase, can withstand
pressures of 500 MPa (Suzuki and Kitamura 1963), while
others, such as L. monocytogenes phosphoglucomutase and
aconitase, are inactivated by 200 MPa (Simpson and Gilmour
1997a). However, in the latter case, L. monocytogenes was little
affected at this pressure treatment, suggesting that the
inactivation of these enzymes was not critical to survival. In
addition, there was no relationship between pressure resist-
ance of the 13 enzymes studies and the pressure resistance of
the three L. monocytogenes strains included in the study.
Covalent bonds are generally unaffected at the pressures used
in food processing. This means that many of the components
responsible for the sensory and nutritional quality of foods, such
as flavour components and vitamins, are not destroyed by high
pressure. This is an important benefit for the food industry.
4.4 Effect of pressure on genetic mechanisms
Nucleic acids are relatively resistant to high pressures and as
the structure of the DNA helix is largely the result of
hydrogen bond formation, it is also stable under pressure.
However, the enzyme-mediated steps involved in DNA rep-
lication and transcription are disrupted. It has been reported
that pressure causes a condensation of nuclear material in
L. monocytogenes, Salmonella Typhimurium (Mackey et al.
1994) and L. plantarum (Wouters et al. 1998). Chilton et al.
(1997) postulated that at elevated pressures, DNA comes into
contact with endonucleases that cleave the DNA.
5. HIGH PRESSURE INACTIVATION OF
MICRO-ORGANISMS
5.1 Inactivation kinetics
High pressure inactivation of micro-organisms is complex,
and plotting the log of surviving numbers against time does
ª 2005 The Society for Applied Microbiology, Journal of Applied Microbiology, 98, 1400–1409, doi:10.1111/j.1365-2672.2005.02564.x
not always form a straight line relationship (first-order
kinetics). Often there is an initial linear decrease in numbers
followed by a decrease in the rate of kill leading to a
pressure-resistant
tail. Studies have shown that when this

tail population is isolated, grown and again exposed to
pressure, there is no significant difference in pressure
resistance between it and the original culture (Metrick et al.
1989). Such tails are also found with heat processing but the
phenomenon seems to be more pronounced with high
pressure processing. The tailing effect is not fully under-
stood. It may be because of inherent phenotypic variation in
pressure resistance in some cells. Experimental conditions,
such as the substrate and growth conditions, may also be a
factor. Tailing phenomena can make calculation of pressure
D-values difficult and have to be taken into account when
doing challenge studies designed to optimize processing
conditions for various foods.
5.2 Pressure injury
A high pressure treatment may not always completely
inactivate micro-organisms but rather may injure a propor-
tion of the population. Recovery of the injured cells will
depend on the conditions after treatment and this has
implications for microbiological enumeration. Compounds
such as sodium chloride added to plating media can act as
selective agents and inhibit growth of injured cells. Using
selective agars can, therefore give inaccurate estimates of
numbers of survivors (Patterson et al. 1995).
5.3 Bacterial endospores
Micro-organisms vary in their response to pressure
(Table 1). Bacterial endospores can be extremely resistant
to high pressure, just as they are resistant to other physical
treatments such as irradiation and heat, and can survive
treatments of more than 1000 MPa (for a review see Smelt
1998). There can be significant variation between spores of
different species and also between strains of the same
species. Clostridium botulinum spores are among the most
pressure resistant, especially nonproteolytic type B (Reddy
et al. 2001). However, relatively low pressures (below
200 MPa) can trigger spore germination (Gould and Sale
1970). This has led to the suggestion that spores could be
killed by applying pressure in two stages. The first pressure
treatment would germinate the spores while the second
treatment, at a higher pressure, would kill the germinated
spores (for a review see Heinz and Knorr 2001). This
process could be repeated several times leading to the idea
that pressure cycling between relatively low and then high
pressures could be one way of overcoming the problem of
spore resistance. However, the extent of the inactivation can
be highly variable (Heinz and Knorr 2001) and a small
 PRESSURE TREATMENT OF FOOD 1403

Table 1 Sensitivity of selected micro-organisms to high hydrostatic pressure

Inactivation
Treatment (log10 units
Micro-organism Substrate conditions of reduction) Comments Reference

Spore-forming bacteria
C. botulinum type Sorensen phosphate 827 MPa/5 min/ 5 Reddy et al. 1999
E spores (Alaska) buffer 50
C
(0Æ067 mol l)1,
pH 7Æ0)
C. sporogenes spores Chicken breast 680 MPa/80
C/ 2 Level of inactivation Crawford et al. 1996
20 min increased if spores
were subsequently
irradiated to 3 kGy
B. stearothermophilus Water 600 MPa/5 min/ 5 Hayakawa et al. 1994
70
C · 6 cycles
C. sporogenes, Meat emulsion 621 MPa/98
C/ >5 (C. sporogenes) Method relies on adiabatic Wilson and Baker
B. subtilis, 5 min >9 (B. subtilis) heating occurring. 2001
B. stearothermophilus >10 (B. stearothermophilus) Product temperature
spores peaked at 29Æ4
C
Viruses
HIV-1 Culture medium 550 MPa/25
C/ Infectivity titre reduced Otake et al. 1997
10 min by 4log10 units
Foot and Mouth Culture medium 250 MPa/ Infectivity titre reduced Treated virus, although no Ishimaru et al. 2004
Disease virus )15
C/1 mol l)1 by >4log10 units longer infective, can still
urea/60 min elicit neutralizing antibody
production in rabbits.
Gives the possibility of
novel method of viral
vaccine production
Feline calicivirus Tissue culture 275 MPa/21
C/ 7 log 10 tissue culture Kingsley et al. 2002
medium 5 min infectious dose for 50%
of the cultures was
completely inactivated
Hepatitis A Tissue culture 450 MPa/21
C/ >6log10 reduction in Kingsley et al. 2002
medium 5 min plaque-forming units
Sea water 450 MPa/21
C/ <2log10 inactivation
5 min
Poliovirus Tissue culture 450 MPa/21
C/ No reduction in Kingsley et al. 2002
medium 5 min plaque-forming units
Prions
Hamster-adapted Hamster brain 700–1000 MPa/ Increase in survival rate Garcı́a et al. 2004
scrapie agent homogenate 60
C/2 h of hamsters following
infection (47%) and
delayed onset of disease
in those that were infected
(from 80 to 153 d)
Vegetative bacteria
Campylobacter jejuni Pork slurry 300 MPa/25
C/ 6 Shigehisa et al. 1991
10 min
Salmonella Strained baby 340 MPa/23
C/ <2 Metrick et al. 1989
Senftenberg food 10 min
775W
Escherichia coli UHT milk 600 MPa/20
C/ <2 Pressure-resistant strain Patterson et al. 1995
O157:H7 Poultry meat 15 min 3
NCTC 12079

ª 2005 The Society for Applied Microbiology, Journal of Applied Microbiology, 98, 1400–1409, doi:10.1111/j.1365-2672.2005.02564.x
1404 M . F . P A T T E R S O N

Table 1 (Continued)

Inactivation
(log10 units
Micro-organism Substrate Treatment conditions of reduction) Comments Reference

Staphylococcus aureus UHT milk 600 MPa/20
C/15 min 2 Patterson et al. 1995
Poultry meat 3
Listeria monocytogenes UHT milk 375 MPa/20
C/15 min <1 Most resistant of three Patterson et al. 1995
Poultry meat 2 strains studied
Vibrio parahaemolyticus Oysters 300 MPa/10
C/3 min 5 Most resistant of Cook 2003
O3:K6 10 strains studied
Lactobacillus helveticus Ewe’s milk 500 MPa/10
C/10 min 3 Gervilla et al. 1997b
Pseudomonas fluorescens Ewe’s milk 450 MPa/10
C/10 min 4 Gervilla et al. 1997b
Yeasts and moulds
Saccharomyces cerevisiae Pork slurry 300 MPa/20
C/10 min 2 Shigehisa et al. 1991
Byssochlamys nivea Grape juice, aw ¼ 0Æ97 700 MPa/70
C/30 min 4 Butz et al. 1996
ascospores Bilberry jam, aw ¼ 0Æ84 <1

proportion of each spore population seems to remain
resistant to pressure-induced germination (Gould 1973).
Another approach to the problem of the pressure
resistance of bacterial spores is to combine high temperature
along with pressure treatment. There have been many
reports indicating that this can be very successful (Heinz
and Knorr 2001). This approach is now being actively
considered for the commercial production of shelf-stable
foods and is the subject of a number of patents designed to
achieve the commercial sterilization of foods that have a pH
greater than 4Æ5. One such patent describes a process
involving two or more cycles of high heat (>70
C) and high
pressure (>530 MPa) with a pause between the cycles
(Mayer 2000). The temperature, pressure level, treatment
time and time interval between the cycles can be varied
depending on the product but are designed to give greater
than the equivalent of a 12D process for C. botulinum. These
treatments use an initial temperature of below 100
C and
rely on the fact that adiabatic heating occurs when the
product is pressure treated. Adiabatic heating results from
the work of compression during pressure treatment leading
to an increase in the temperature of food. The extent of the
temperature increase varies with the composition of the food
but is normally 3–9
C/100 MPa. The overall treatment
conditions are less severe than conventional retorting. This
results in shelf-stable products which are of higher quality in
terms of texture, flavour and retention of nutrients than
those obtained by conventional processing (Master et al.
2004).
5.4 Viruses and prions
There is relatively little information on pressure inactiva-
tion of viruses compared with other micro-organisms but it
ª 2005 The Society for Applied Microbiology, Journal of Applied Microbiology, 98, 1400–1409, doi:10.1111/j.1365-2672.2005.02564.x
does appear that viruses can vary significantly in their
response to treatment (Table 1). Polio virus in tissue
culture medium appears to be relatively resistant with
450 MPa for 5 min at 21
C giving no reduction in plaque-
forming units (PFU). The same treatment conditions with
hepatitis A resulted in a 6log10 PFU ml)1 stock culture
being reduced to undetectable levels (Kingsley et al. 2002).
However, treatment in sea water increased the pressure
resistance of hepatitis A virus, suggesting a protective
effect of the salts. Feline calicivirus, a Norwalk virus
surrogate, (Kingsley et al. 2002) and human rotavirus
(Khadre and Yousef 2002) were more pressure sensitive
than hepatitis A when treated in tissue culture medium.
There are also reports that HIV-1 is relatively sensitive to
pressure with 400–600 MPa for 10 min at 25
C resulting
in 4–5log10 reduction in viable particles when treated in
tissue culture medium (Otake et al. 1997), although
different strains varied in their pressure resistance. Foot
and mouth disease virus (FMDV) is also relatively
sensitive to high pressures. A treatment of 250 MPa at
)15
C and 1 mol l)1 urea for 1 h destroyed FMDV
infectivity but maintained the integrity of capsid structure.
The treated virus could also elicit neutralizing antibody
production in rabbits. These results suggest that high
pressure could be a safe, simple, cheap and reproducible
method of producing viral vaccines (Ishimaru et al. 2004).
Evidence is emerging that high pressure may have some
effect on prions. Garcı́a et al. (2004) used hamster-adapted
scrapie strain 263k to infect hamsters intracerebrally.
Pressure treatment of the prions to >700 MPa at 60
C
for 2 h significantly increased survival rate of the animals.
Further work still needs to be done but the early work
suggests the possibility of producing safe products for
specialized, high added-value markets, such as baby foods.

5.5 Vegetative bacteria
In general terms Gram-positive bacteria tend to be more
resistant to pressure than Gram-negatives and cocci are
more resistant than rod-shaped bacteria (Table 1). It has
been suggested that the cell membrane structure is more
complex in Gram-negative bacteria, making it more sus-
ceptible to environmental changes caused by pressure
(Shigehisa et al. 1991). However, there are many exceptions
to these general rules. Certain strains of Escherichia coli
O157:H7, for example, can be exceptionally pressure
resistant. Benito et al. (1999) reported that an E. coli
O157:H7 strain isolated from a major hamburger patty
outbreak in the US showed less than a 1log10 reduction
when treated in laboratory medium at 500 MPa at <45
C
for 30 min. This strain was also more resistant to heat, acid,
oxidative and osmotic stresses than a pressure-sensitive
strain. However, other studies with pathogens such as
Salmonella, have shown only a weak, or no correlation,
between pressure resistance and resistance to other stresses
(Sherry et al. 2004).
5.6 Yeasts and moulds
Yeasts are generally not associated with food-borne disease
but are important in spoilage, especially in acidic foods.
They are relatively sensitive to pressure (Table 1) and this is
one reason why pressure treatment of fruit products to
extend shelf-life is particularly successful.
There is relatively little information on the pressure
sensitivity of moulds but it has been shown that vegetative
forms are relatively sensitive, while ascospores are more
resistant (Butz et al. 1996; Voldrich et al. 2004). The effect
of pressure on preformed mycotoxins is thought to be
limited as the treatment has little effect on covalent bonds.
However, one study reported that patulin, a mycotoxin
produced by several species of Aspergillus, Penicillium and
Byssochlamys, was found to be degraded by pressure (Brůna
et al. 1997). The patulin content in apple juice decreased by
42, 53 and 62% after 1 h treatment at 300, 500 and
800 MPa, respectively, at 20
C. An explanation for this was
not given.
6. EXTRINSIC FACTORS AFFECTING
THE SENSITIVITY OF MICRO-ORGANISMS
TO HIGH PRESSURE
High pressure is no different from other physical preserva-
tion methods in that its effectiveness against micro-organ-
isms is influenced by a number of factors. These all interact
and contribute to the lethal effect and therefore have to be
considered when designing process conditions to ensure the
microbiological safety and quality of pressure-treated foods.
ª 2005 The Society for Applied Microbiology, Journal of Applied Microbiology, 98, 1400–1409, doi:10.1111/j.1365-2672.2005.02564.x
PRESSURE TREATMENT OF FOOD 1405
6.1 Effect of substrate
The chemical composition of the substrate during treatment
can have a significant effect on the response of micro-
organisms to pressure. Certain food constituents such as
proteins, carbohydrates and lipids can have a protective
effect (Simpson and Gilmour 1997b). Inactivation data
obtained using buffers or laboratory media, therefore,
should not be extrapolated to real food situations where a
more severe pressure treatment may be needed to achieve
the same level of inactivation. For example, a treatment of
375 MPa for 30 min at 20
C in phosphate buffer (pH 7Æ0)
gave a 6log10 inactivation of a pressure-resistant strain of
E. coli O157:H7. However, the same treatment gave a
2Æ5log10 reduction in poultry meat and only 1Æ75log10
reduction in milk (Patterson et al. 1995). Cations, such as
Ca2+, can be baroprotective and this may explain why many
micro-organisms appear more pressure resistant when
treated in certain foods, such as milk (Hauben et al. 1998).
A low water activity protects micro-organisms against the
effects of pressure (Palou et al. 1997). Oxen and Knorr
(1993) reported that reducing aw of the medium from 0Æ98–
1Æ0 down to 0Æ94–0Æ96 resulted in better survival of
Rhodotorula rubra when it was subjected up 200–400 MPa
for 15 min at 25
C. However, the nature of the solute is
important. At the same aw, cells were more pressure
sensitive in glycerol than in monosaccharides and disaccha-
rides. Trehalose is reported to confer most protection (see
review by Smelt 1998).
The pH of acidic solutions decreases as pressure increases
and it has been estimated that in apple juice, there is a pH
drop of 0Æ2 U per 100 MPa (Heremans 1995). To date, the
pH change which occurs during pressure treatment cannot
be measured directly in solid food, but methods have been
developed for in situ pH measurement during pressure
treatment of liquids (Hayert et al. 1999; Stippl et al. 2004).
When the pressure is released, the pH reverts to its original
value but it is not known whether these sudden changes in
pH affect microbial survival in addition to the effect of
pressure. It is known that pH and pressure can act
synergistically leading to increased microbial inactivation.
Linton et al. (1999) has shown that initial pH had a
significant effect on inactivation rates of E. coli O157:H7 in
orange juice. As pH was lowered, the cells were more
susceptible to pressure inactivation and sublethally injured
cells failed to repair and died more rapidly during subse-
quent storage of the juice.
Food additives can have varying effects on microbial
resistance to pressure. Pressure has been used to sensitize
Gram-negative bacteria such as Salmonella, as well as Gram-
positive bacteria such as L. monocytogenes to nisin and
lysozyme (Kalchayanand et al. 1998; Masschalk et al. 2001).
The combination of high pressure and nisin was also used to
1406 M . F . P A T T E R S O N
increase the inactivation of B. cereus spores in cheese
(López-Pedemonte et al. 2003). However, in this case the
pressure treatment conditions were relatively severe. The
treatment (60 MPa at 30
C for 210 min to germinate
the spores followed by 400 MPa at 30
C for 15 min to
kill the vegetative cells) was carried out in the presence of
1Æ56 mg l)1 nisin in the cheese. This resulted in approx.
2Æ4log10 inactivation of the spores.
Pediocin AcH also works synergistically with pressure. A
combination of 345 MPa for 5 min at 50
C in the presence
of 3000 AU ml)1 pediocin AcH gave at least a 7log10
inactivation of a range of bacteria including L. monocyto-
genes, Salm. Typhimurium, Staphylococcus aureus, E. coli
O157:H7 L. sake and Pseudomonas fluorescens. This level of
inactivation could not be achieved by pressure alone
(Kalchayanand et al. 1998). Similarly, the combination of
high pressure with other antimicrobial agents such as
lacticin 3147 (Ross et al. 2000), lactoperoxidase (Garcia-
Graellis et al. 2003) and carvacrol (Karatzas et al. 2001) can
work synergistically to enhance the kill of micro-organisms,
including pathogens.
6.2 Effect of temperature
Temperature during pressure treatment can have a signifi-
cant effect on microbial survival. Increased inactivation is
usually observed at temperatures above or below 20
C
(Takahashi et al. 1992).
High temperatures (>70
C) can be particularly effective in
helping to achieve high pressure sterilization, as discussed
above. The combination of elevated temperatures (<50
C)
with pressure has also been suggested as a practical way to
overcome the problem of pressure resistant strains of
vegetative cells. Patterson and Kilpatrick (1998) reported
approx. a 6log10 inactivation of a pressure-resistant E. coli
O157:H7 in poultry mince and a 5log10 inactivation in milk
using a treatment of 400 MPa at 50
C for 15 min. Neither
heat nor pressure alone could achieve this level of inactivation.
Refrigeration temperatures can also enhance pressure
inactivation. Gervilla et al. (1997a) reported that ewes milk
pressurized at 450 MPa at 2
C for 15 min was more effective
at inactivating L. innocua than the same treatment at 25
C but
less effective than the pressure treatment at 50
C.
7. PRESSURE TREATMENT TO IMPROVE
T H E M I C R O B I O LO G I C A L Q U A L I T Y O F
FOODS
Pressure-treated fruit jams and sauces first became com-
mercially available in Japan in the early 1990s. Treatment of
fruit jams with around 400 MPa for up to 5 min at room
temperature can significantly reduce the number of micro-
organisms, especially yeasts and moulds. Refrigeration of the
ª 2005 The Society for Applied Microbiology, Journal of Applied Microbiology, 98, 1400–1409, doi:10.1111/j.1365-2672.2005.02564.x
jam after processing is necessary due to browning and
flavour changes caused by enzymatic activities. These
products have a shelf-life of around 30 d and have superior
sensory quality compared with those prepared in a conven-
tional manner (Ludikhuyze and Hendrickx 2001).
Fruit juices are normally processed at 400 MPa or greater
for a few minutes at 20
C or less. This can significantly
reduce numbers of yeasts and moulds and so extend shelf-
life for up to 30 d. Pathogens, such as E. coli O157:H7 can
also be destroyed by this treatment (Linton et al. 1999;
Ramaswamy et al. 2003). Pressure-treated orange and
grapefruit juices have been available in France since 1994,
while pressure-treated apple juice is available in Italy.
Pressure treatment of vegetable products is problematic
because of their relatively high pH along with the possibility
of survival and growth of pathogenic spore-forming organ-
isms. However, one of the most successful pressure-treated
foods in the USA is guacamole. Its market share continues
to grow and is reportedly based on the consumer preference
for the
fresher taste of guacamole processed in this manner
compared with heat-treated or frozen products. Treatment
of around 500 MPa for 2 min is sufficient to extend shelf-
life from 7 to 30 d at refrigeration temperatures. Challenge
studies with a variety of pathogens have shown that this
treatment is sufficient to give a 5log10 reduction in numbers
(Parnell 2003) and the process has been approved by the US
Food and Drug Administration.
Sliced cooked ham and other delicatessen meat products,
in flexible pouches, can be successfully treated using
500 MPa for a few minutes. The sensory properties of
ham are preserved, and shelf-life can be extended to 60 d
under chilled storage. Cooked delicatessen products have a
risk of postprocessing contamination from pathogens such as
L. monocytogenes. High pressure treatment as a final
preservation step, after packaging, can give additional
microbiological safety assurance. During challenge studies
a treatment of 500 MPa can cause a 5log10 reduction in L.
monocytogenes in dry-cured ham (Minerich and Krug 2003).
Pressure-treated cooked and vacuum-packaged ham is
available in Spain and in the USA.
Another example of commercially successful pressure-
treated foods available in the USA are oysters. The initial
aim of the pressure treatment was to eliminate Vibrio spp.
from oysters, which are often eaten raw or only lightly
cooked. Vibrio spp. are relatively sensitive to high pressure,
although there can be species variation (Cook 2003). Typical
treatments of 250–350 MPa for 1–3 min at ambient tem-
perature are used commercially without significantly affect-
ing sensory quality. An additional benefit of pressure-treated
oysters is the mechanical shucking effect it causes, releasing
the adductor muscle from the shell (He et al. 2002). For this
reason, a heat shrink plastic band is placed around each
oyster prior to processing so that the shell is kept shut until

the meat is required. The main processor of pressure-treated
oysters in the USA uses a trademark plastic gold band and
these products have achieved several national awards for
quality products. Pressure will successfully shuck other
shellfish, such as mussels, Nephrops and crabs as well as
improving their microbiological quality and it is likely that
the technology will be more widely used for this purpose.
There is also increasing interest in pressure-treating fin fish
to improve microbiological safety and quality as well as
using the technology to produce a range of novel food
products (Lakshmanan and Dalgaard 2004), including surmi
gels (Ohshima et al. 1993; Ashie and Simpson 1996).
Pressure treatment of milk and dairy products to improve
microbial safety and quality has been of interest since the
early work of Hite (1899). However, it is likely that the
technology will only be used commercially for niche
applications, where it can provide a commercial advantage
over existing, usually heat-treated, products. For example,
pressure treatment may be of value in treating milk that is to
be used in the manufacture of raw milk cheese, where it
could reduce the numbers of pathogens such as L.
monocytogenes. However, some pathogens, such as certain
strains of E. coli O157:H7, are known to be extremely
pressure resistant in milk (Patterson et al. 1995). Therefore,
this approach will not solve all the microbiological safety
problems associated with raw milk cheese. High pressure
treatment of yoghurt has also been investigated. Tanaka and
Hatanaka (1992) investigated the effectiveness of using high
pressure to prevent after-acidification of yoghurt during
storage. They concluded that a treatment of 200–300 MPa
for 10 min at room temperature prevented the continued
growth of lactic acid bacteria during storage and so
maintained the yoghurt quality.
The range of products now being considered for high
pressure treatment continues to grow year-on-year. A range of
complete meal kits have recently been launched in the USA.
The kits consist of pressure-treated cooked meat or chicken,
salsa, guacamole, peppers and onion are now also available.
Only the flour tortillas are not pressure treated. The products
have a chilled shelf-life of at least 30 d and only required to be
reheated in a microwave before consumption. It is likely that
the range of added-value, high quality pressure-treated foods
will increase within the next 5 years.
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