Available online at www.sciencedirect.com
International Journal of Food Microbiology 120 (2007) 51 – 70
www.elsevier.com/locate/ijfoodmicro
Bacteriocin-based strategies for food biopreservation
Antonio Gálvez ⁎, Hikmate Abriouel, Rosario Lucas López, Nabil Ben Omar
Área de Microbiología, Facultad de Ciencias Experimentales, Universidad de Jaén, Spain
Abstract
Bacteriocins are ribosomally-synthesized peptides or proteins with antimicrobial activity, produced by different groups of bacteria. Many lactic
acid bacteria (LAB) produce bacteriocins with rather broad spectra of inhibition. Several LAB bacteriocins offer potential applications in food
preservation, and the use of bacteriocins in the food industry can help to reduce the addition of chemical preservatives as well as the intensity of heat
treatments, resulting in foods which are more naturally preserved and richer in organoleptic and nutritional properties. This can be an alternative to
satisfy the increasing consumers demands for safe, fresh-tasting, ready-to-eat, minimally-processed foods and also to develop “novel” food products
(e.g. less acidic, or with a lower salt content). In addition to the available commercial preparations of nisin and pediocin PA-1/AcH, other bacteriocins
(like for example lacticin 3147, enterocin AS-48 or variacin) also offer promising perspectives. Broad-spectrum bacteriocins present potential wider
uses, while narrow-spectrum bacteriocins can be used more specifically to selectively inhibit certain high-risk bacteria in foods like Listeria
monocytogenes without affecting harmless microbiota. Bacteriocins can be added to foods in the form of concentrated preparations as food
preservatives, shelf-life extenders, additives or ingredients, or they can be produced in situ by bacteriocinogenic starters, adjunct or protective
cultures. Immobilized bacteriocins can also find application for development of bioactive food packaging. In recent years, application of bacteriocins
as part of hurdle technology has gained great attention. Several bacteriocins show additive or synergistic effects when used in combination with other
antimicrobial agents, including chemical preservatives, natural phenolic compounds, as well as other antimicrobial proteins. This, as well as the
combined use of different bacteriocins may also be an attractive approach to avoid development of resistant strains. The combination of bacteriocins
and physical treatments like high pressure processing or pulsed electric fields also offer good opportunities for more effective preservation of foods,
providing an additional barrier to more refractile forms like bacterial endospores as well. The effectiveness of bacteriocins is often dictated by
environmental factors like pH, temperature, food composition and structure, as well as the food microbiota. Foods must be considered as complex
ecosystems in which microbial interactions may have a great influence on the microbial balance and proliferation of beneficial or harmful bacteria.
Recent developments in molecular microbial ecology can help to better understand the global effects of bacteriocins in food ecosystems, and the study
of bacterial genomes may reveal new sources of bacteriocins.
© 2007 Elsevier B.V. All rights reserved.
Keywords: Bacteriocin; Biopreservation; Hurdle technology; Lactic acid bacteria; Food
1. Introduction
In spite of modern advances in technology, the preservation
of foods is still a debated issue, not only for developing countries
(where implementation of food preservation technologies are
clearly needed) but also for the industrialized world. Amelioration of economic losses due to food spoilage, lowering the
food processing costs and avoiding transmission of microbial
pathogens through the food chain while satisfying the growing
⁎ Corresponding author. Área de Microbiología, Departamento de Ciencias de la
Salud, Facultad de Ciencias Experimentales, Universidad de Jaén, Campus Las
Lagunillas s/n. 23071-Jaén, Spain. Tel.: +34 953 212160; fax: +34 953 212943.
E-mail address: [email protected] (A. Gálvez).
0168-1605/$ - see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.ijfoodmicro.2007.06.001
consumers demands for foods that are ready to eat, fresh-tasting,
nutrient and vitamin rich, and minimally-processed and preserved are major challenges for the current food industry. The
extent of microbiological problems in food safety was clearly
reflected in the WHO food strategic planning meeting (WHO,
2002): (i) the emergence of new pathogens and pathogens not
previously associated with food consumption is a major concern;
(ii) microorganisms have the ability to adapt and change, and
changing modes of food production, preservation and packaging
have therefore resulted in altered food safety hazards.
The empirical use of microorganisms and/or their natural
products for the preservation of foods (biopreservation) has been
a common practice in the history of mankind (Ross et al., 2002).
The lactic acid bacteria (LAB) produce an array of antimicrobial
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A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
substances (such as organic acids, diacetyl, acetoin, hydrogen
peroxide, reuterin, reutericyclin, antifungal peptides, and
bacteriocins (Holzapfel et al., 1995; El-Ziney et al., 2000;
Holtzel et al., 2000; Magnusson and Schnürer, 2001). Bacteriocins are ribosomally synthesized antimicrobial peptides or
proteins (Jack et al., 1995). LAB produce a variety of
bacteriocins, most of which can be grouped in one of the classes
proposed by Klaenhammer (1993). The structure, biosynthesis,
genetics and food application of LAB bacteriocins have been
reviewed recently (Cleveland et al., 2001; O'Sullivan et al.,
2002; Chen and Hoover, 2003; Cotter et al., 2005; Fimland et al.,
2005; Deegan et al., 2006; Drider et al., 2006).
The bacteriocins produced by LAB offer several desirable
properties that make them suitable for food preservation: (i) are
generally recognised as safe substances, (ii) are not active and nontoxic on eukaryotic cells, (iii) become inactivated by digestive
proteases, having little influence on the gut microbiota, (iv) are
usually pH and heat-tolerant, (v) they have a relatively broad
antimicrobial spectrum, against many food-borne pathogenic and
spoilage bacteria, (vi) they show a bactericidal mode of action,
usually acting on the bacterial cytoplasmic membrane: no cross
resistance with antibiotics, and (vii) their genetic determinants are
usually plasmid-encoded, facilitating genetic manipulation.
The accumulation of studies carried out in recent years clearly
indicate that the application of bacteriocins in food preservation
can offer several benefits (Thomas et al., 2000): (i), an extended
shelf life of foods, (ii) provide extra protection during temperature abuse conditions, (iii) decrease the risk for transmission
of foodborne pathogens through the food chain, (iv) ameliorate
the economic losses due to food spoilage, (v) reduce the
application of chemical preservatives, (vi) permit the application
of less severe heat treatments without compromising food safety:
better preservation of food nutrients and vitamins, as well as
organoleptic properties of foods, (vii), permit the marketing of
“novel” foods (less acidic, with a lower salt content, and with a
higher water content), and (viii) they may serve to satisfy
industrial and consumers demands. In this respect some of the
trends of the food industry in Europe, such as the need to
eliminate the use of artificial ingredients and additives, the
demands for minimally-processed and fresher foods, as well as
for ready-to-eat food or the request for functional foods and
nutraceuticals (Robertson et al., 2004) could be satisfied, at least
in part, by application of bacteriocins. The present review will
address different aspects related to food preservation by bacteriocins including factors influencing bacteriocin activity in
food systems, hurdle technology, and the impact of recent
advances in molecular biology and the analysis of bacterial
genomes on bacteriocin studies and application.
2. Bacteriocins and food systems
2.1. Supplementing foods with bacteriocins
Foods can be supplemented with ex situ produced bacteriocin
preparations, or by inoculation with the bacteriocin-producer
strain under conditions that favour production of the bacteriocin
in situ (Schillinger et al., 1996; Stiles, 1996). In the first case,
bacteriocin preparations obtained by cultivation of the producer
strain in a fermentor at industrial scale followed by adequate
recovery and processing can be added as partially purified or
purified concentrates, which would require specific approval as
preservatives from the legislative point of view. So far, nisin is
the only bacteriocin licensed as a food preservative (E234).
Many preliminary studies on the activity of bacteriocins in vitro
or in food systems are carried out with partially-purified preparations obtained from cultured broths. In most cases, a low
concentration of bacteriocin is often recovered, which limits the
efficacy of such preliminary tests.
Ex situ produced bacteriocins can also be added in the form
of raw concentrates obtained by cultivation of the producer
strain in a food-grade substrate (such as milk or whey). The
resulting preparations may be regarded as food additives or
ingredients from the legal point of view, since some of their
components may play a recognised function in the food (such as
increase in protein content, or thickening). They also contain the
cell-derived antimicrobial metabolites (such as lactic acid) and
bacteriocins, affording an additional bioprotectant function. In
addition to already-marketed concentrates such as ALTA™
2341 or Microgard™, other milk-based preparations have been
described recently such as lacticin 3147 (Morgan et al., 1999;
Guinane et al., 2005) and variacin (O'Mahony et al., 2001).
Ex situ produced bacteriocins can also be applied in the form of
immobilized preparations, in which the partially-purified bacteriocin or the concentrated cultured broth is bound to a carrier. The
carrier acts as a reservoir and diffuser of the concentrated bacteriocin molecules to the food ensuring a gradient-dependent
continuous supply of bacteriocin. The carrier may also protect the
bacteriocin from inactivation by interaction with food components and enzymatic inactivation. Moreover, the precise localized
application of bacteriocin molecules on the food surface requires
much lower amounts of bacteriocin (compared to application
in the whole food volume), decreasing the processing costs. A
variety of methods have been proposed for bacteriocin immobilization, including adsorption to the producer cells (Yang et al.,
1992; Mattila et al., 2003; Ghalfi et al., 2006), silica particles or
corn starch powder (Coventry et al., 1996; Dawson et al., 2005),
liposome encapsulation (Degnan and Luchansky, 1992), and
incorporation on gel coatings and films of different materials such
as calcium alginate, gelatin, cellulose, soy protein, corn zein,
collagen casings, polysaccharide based films, cellophane, silicon
coatings, polyethylene, nylon or other polymer plastic films
(Daeschel et al., 1992; Cutter and Siragusa, 1995b; Ming et al.,
1997; Siragusa et al., 1999, Natrajan and Sheldon, 2000; Gill and
Holley, 2003; Scannell et al., 2000a; Ko et al., 2001; Dawson
et al., 2002; Franklin et al., 2004; Luchansky and Call, 2004;
Guerra et al., 2005; Lungu and Johnson, 2005). In most cases,
immobilized bacteriocin preparations are applied on the surface of
the processed food to avoid post-process contamination and
surface proliferation of unwanted bacteria. A recent advance in
this field is the use of immobilized bacteriocins in the development of antimicrobial packaging. A polyethylene film containing immobilized bacteriocin 32Y from L. curvatus reduced viable
counts of L. monocytogenes during storage in the packaged pork
steak and ground beef as well as in frankfurters (Mauriello et al.,
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
2004; Ercolini et al., 2006). Similarly, a nisin-containing cellophane coating reduced viable counts of total aerobic bacteria in
fresh veal meat stored at 8 °C (Guerra et al., 2005), and an active
package obtained from nisin-treated film reduced viable counts of
M. luteus ATCC 10,240 cells in broth as well as in raw milk and
pasteurized milk during storage (Mauriello et al., 2005).
Therefore, the use of antimicrobial films containing bacteriocins
can improve the quality and safety and prolong the shelf-life of
food products.
In situ bacteriocin production offers several advantages
compared to ex situ production regarding both legal aspects and
costs. Lowering the costs of biopreservation processes may be
highly attractive, especially for small economies and developing
countries, where food safety may be seriously compromised
(Holzapfel, 2002). Many studies have focused on the selection
and development of bacteriocinogenic cultures for food application (Ross et al., 2000; Työppönen et al., 2003; Peláez and
Requena, 2005; Foulquié Moreno et al., 2006; Leroy et al.,
2006). The use of bacteriocinogenic cultures requires careful
selection of strains that are well-adapted to the particular food
environment in which they will be used and able to grow under
the food processing and/or storage conditions and to produce
enough bacteriocin amounts as to inhibit the target pathogenic or
spoilage bacteria. Therefore, it is necessary to implement the
right experimental approaches to select bacteriocin-producing
strains that are suitable for use in food production. The strain
properties and the amount of bacteriocin produced could be
improved by heterologous expression of bacteriocin genes
(Rodríguez et al., 2003; Zhou et al., 2006), and the precise
moment of bacteriocin production could also be tailored by
using inducible production systems (Zhou et al., 2006).
Bacteriocinogenic strains can be used either directly as starter
cultures, as adjunct or co-cultures in combination with a starter
culture, or as protective cultures (especially in the case of nonfermented foods). When used as a starter culture, the bacteriocinogenic strain must be able to carry out the desired fermentation process optimally besides being able to produce
enough bacteriocin amounts to afford protection. In some cases,
bacteriocin production may also serve to increase the implantation capacity, competitiveness and stability of the starter
(Todorov et al., 1999). Adjunct cultures do not need to contribute
to the fermentation, but they must not interfere with the primary
function of the starter culture. For this reason, bacteriocin resistance of the starter culture may be a key factor. This may be
achieved by selection of natural resistant mutants, by adaptation
through repeated subcultivation with increasing bacteriocin
concentrations, or by genetic modification. Nevertheless, sometimes this may not be necessary as the bacteriocin may just not be
active on the starter culture (as may be the case of many of the
bacteriocins that predominantly show antilisterial activity) or this
may be much more tolerant to the bacteriocin than the target
bacteria in the food system. Differences in inoculum density, a
faster growth rate of the starter or a delayed bacteriocin production
may also permit the starter to grow without interference from the
bacteriocinogenic adjunct culture. As an example, inoculation of
milk with an enterocin AS-48 producer enterococcal strain as
adjunct culture in combination with a commercial starter culture
53
for cheese manufacture had no effect on growth of the starter or
the physicochemical properties of the produced cheese. At the
same time, enough bacteriocin was produced in the cheese to
ensure inhibition of Bacillus cereus (Muñoz et al., 2004).
Bacteriocinogenic protective cultures can be used to inhibit
spoilage and pathogenic bacteria during the shelf life period of
non-fermented foods. A protective culture may grow and produce bacteriocin during refrigeration storage of the food, and/or
during temperature abuse conditions. In the first case, growth
of the protective cultures must have a neutral impact on the
physicochemical and organoleptic properties of the food, while
under temperature abuse conditions the protective culture may
even act as the predominant spoiler, ensuring that pathogenic
bacteria do not grow and that the spoiled food is not consumed
(Holzapfel et al., 1995).
2.2. Factors influencing the efficacy of bacteriocins in food
systems
Comparisons of data obtained in culture media with those
obtained in food systems reveal that the efficacy of bacteriocins is
often much lower in the later (Schillinger et al., 1996). Sometime,
at least ten-fold higher bacteriocin concentrations must be added
to foods in order to achieve an equivalent inhibitory effect. The
efficacy of bacteriocins in foods will greatly depend on a number
of food-related factors (Table 1) that in most cases involve
interaction with food components, precipitation, inactivation, or
uneven distribution of bacteriocin molecules in the food matrix.
As an illustrative example, application of nisin in meat products
faces several limitation derived from its interaction with
phospholipids emulsifiers and other food constituents (Henning
et al., 1986; Jung et al., 1992; Aasen et al., 2003), poor solubility
at pH above 6.0, and inactivation by formation of a nisinglutathione adduct (Rose et al., 2003). However, inactivation is
Table 1
Bacteriocin efficacy in foods: limiting factors
Food-related factors
–Food processing conditions
–Food storage temperature
–Food pH, and bacteriocin unstability to pH changes
–Inactivation by food enzymes
–Interaction with food additives/ingredients
–Bacteriocin adsorption to food components
–Low solubility and uneven distribution in the food matrix
–Limited stability of bacteriocin during food shelf life
The food microbiota
–Microbial load
–Microbial diversity
–Bacteriocin sensitivity
–Microbial interactions in the food system
The target bacteria
–Microbial load
–Bacteriocin sensitivity (Gram-type, genus, species, strains)
–Physiological stage (growing, resting, starving or viable but non-culturable
cells, stressed or sub-lethally injured cells, endospores...)
–Protection by physico-chemical barriers (microcolonies, biofilms, slime)
–Development of resistance/adaptation
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A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
lower in cooked meats due to the loss of free sulphydryl groups
during cooking as a result of the reaction of glutathione with
proteins (Stergiou et al., 2006).
Foods are complex ecosystems with a range of microbial
compositions. These vary from (commercially) sterile foods to
raw or fermented foods. In commercially sterile foods, microorganisms from post-process contamination may easily proliferate because of the lack of competitors. Under these conditions,
the efficacy of the bacteriocin will depend more directly on the
microbial load of the contaminant, and a higher bacteriocin
concentration will always be required in order to inactivate a
higher number of target cells. In raw foods, the autochthonous
microbiota may counteract development of potential pathogens.
However, a complex food microbiota may frequently cause a
lower efficacy of bacteriocins due to the presence of resistant
bacteria (such as Gram-negatives) and/or the higher chances for
production of inactivating enzymes (such as proteases). In
addition, the bacteriocin may not always target the desired
bacterial group (for example, bacteriocin inactivation of the
LAB responsible for food fermentation would be clearly undesirable). In processed foods, microorganisms can also be
found in a variety of physiological stages, which may greatly
influence bacteriocin sensitivity. As an example, cells that are
not actively growing may be more resistant to bacteriocins, and
changes of the target organisms in response to environmental
stress factors may also result in decreased bacteriocin sensitivity.
Inactive bacterial forms (endospores) may also be resistant to
bacteriocins, although processing treatments may trigger spore
germination and outgrowth, increasing bacteriocin sensitivity.
Therefore, the efficacy of bacteriocins in food will greatly depend on the food microbial composition and microbial physiological stage (Table 1). It should also be considered that
microorganisms are seldom distributed homogeneously as
planktonic cells in the food matrix. Most probably, they will
tend to form microcolonies in a solid or particulate food, or grow
in the form of slime-covered biofilms on food surfaces. The
protective effect of biofilms against antimicrobial substances in
the food industry is well documented (Kumar and Anand, 1998),
and may clearly limit the efficacy of bacteriocins as well.
Variations in strain sensitivities and development of strain
resistance/adaptation are also of major concern for application of
bacteriocins. For example, not all strains of L. monocytogenes
show the same degree of sensitivity to antilisterial bacteriocins
(Martínez et al., 2005). Bacteriocin-resistant L. monocytogenes
have been reported to appear at frequencies of 10− 3 to 10− 9 for
nisin, depending on the strain (Ming and Daeschel, 1993; Davies
and Adams, 1994; Mazzotta and Montville, 1997; Martínez et
al., 2005), of 10− 4 to 10− 6 for the class IIa bacteriocins leucocins
A, B, E and sakacin A (Dykes and Hastings, 1998), and of 10− 3
to 10− 4 for mesenterocin 52, curvaticin 13 and plantaricin C19
(Rekhif et al., 1994). The fitness costs of bacteriocin resistance
may vary greatly depending on the strain, the bacteriocin and the
environmental conditions (Gravesen et al., 2002a). Most worrying, the developed resistance to one bacteriocin may also
afford protection against other bacteriocins, and the observed
cross-resistance between pediocin-like bacteriocins has been
attributed to a general mechanism of resistance (Gravesen et al.,
2002b). The needs to find novel bacteriocins showing no cross
resistance with existing ones are clearly patent.
In addition to the factors influencing the effectiveness of
bacteriocins as antimicrobials in food systems, factors influencing bacteriocin production are of most importance when using
bacteriocinogenic cultures (Fig. 1). The use of bacteriocinogenic
cultures faces several limitations directly related to the producer
strain such as (i) the spontaneous loss of the bacteriocinogenic
trait, (ii) the susceptibility of the producer strain to infection by
bacteriophages, (iii) antagonism of other bacteria towards the
producer strain, (iv) inadequacy of producer strain as a starter,
(v) difficulties for genetic manipulation and transfer of bacteriocinogenic trait to suitable starters, and, the most important,
(vi) a low capacity for bacteriocin production in the food system
(which will depend greatly on the food storage conditions, the
capacity of the producer strain for implantation and proliferation,
or the induction of bacteriocin production). In situ bacteriocin
production may also depend greatly on physicochemical factors
(such as pH, temperature, aw, CO2, O2, redox potential, time
of incubation…) as well as food-related factors (such as the
food structure — fluidity, particulate matter, emulsions…, —
Fig. 1. Influence of different factors on the efficacy of in situ bacteriocin production for biopreservation.
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
buffering capacity, composition — available nutrients, additives, antimicrobials… — and processing conditions — freezing
and thawing, pressure homogeneization or other procedures that
may indirectly damage bacterial cells, as well as thermal treatments and other treatments intended to reduce the microbial
load). Since the bacteriocinogenic strain will thrive within a
(more or less complex) microbial population in the food ecosystem, bacteriocin production may also depend greatly on
microbial-related factors such as the microbial load and
diversity, the microbial interactions occurring in the food during
storage (such as the competition for nutrients or the production
of other antagonistic substances), and the physiological state of
the bacteria (Holzapfel et al., 1995; Rodríguez et al., 2003;
Devlieghere et al., 2004). All these factors will be sensed by the
bacteriocinogenic strain which, in turn, will also have a direct
influence on the food environment through the consumption of
and competition for nutrients, production of metabolites and
modification of the food microbial balance through the produced
bacteriocin. Therefore, bacteriocin production in food must be
understood as a dynamic process where the different interactions
change over time, dictating the ultimate result as far as food
preservation (Fig. 1).
Several illustrative examples may explain how bacteriocin
production can change dramatically by altering environmental
conditions, and how optimum production may require a certain
combination of influencing factors (Leal-Sánchez et al., 2002).
A suboptimal temperature (22 °C) and a moderate NaCl stress
(0.65 M) stimulated bacteriocin production by Lactobacillus
pentosus B96 (Delgado et al., 2005). Amylovorin production by
Lactobacillus amylovorus DCE 471 was stimulated by NaCl
(Neysens et al., 2003) as well as carbon dioxide (Neysens and De
Vuyst, 2005). Several other stress factors (such as ethanol,
oxygen, competing microbiota, etc.) may also stimulate bacteriocin production.
Bacteriocin production is often an inducible trait that depends
on cell density and concentration of the inducer (which may be
the bacteriocin itself). Therefore, a minimum concentration of
inoculum is often required. The interactions of the induction
factor with the food matrix (such as adsorption or inactivation)
may have a great influence on bacteriocin production. On the
other hand, the food matrix may also facilitate the concentration
of the induction factor around cells or microcolonies formed by
the bacteriocinogenic culture in the food. The presence of competing microorganisms can be an environmental factor stimulating production of LAB bacteriocins such as lactacin B
(Barefoot et al., 1994) or divercin (Sip et al., 1998). Similarly,
specific Gram-positive bacteria activate both plantaricin NC8
(PLNC8) production by Lactobacillus plantarum NC8 and the
PLNC8-autoinducing activity (Maldonado et al., 2004).
Since bacteriocin production is linked to cell growth, it may
also depend on factors affecting this parameter (such as inhibitory substances like salt or nitrite) or the lack of available
nutrients (such as manganese in the case of many LAB). As an
example, the production of enterocins A and B by Enterococcus
faecium CTC492 was significantly inhibited by sausage ingredients and additives, with the exception of nitrate (Aymerich
et al., 2000). The addition of sodium chloride and pepper de-
55
creased production by 16-fold. The temperature and pH
influenced enterocin production, with optima between 25 and
35 °C, and from 6.0 to 7.5 of initial pH (Aymerich et al., 2000).
Bavaricin A production was negatively affected at NaCl
concentrations of 3% or higher, while cells were still growing
(Larsen et al., 1993). Growth and sakacin production in meat
sausages by Lactobacillus sakei CTC 494 was affected negatively by salting and curing with nitrite as well as manganese
limitation (Leroy and De Vuyst, 1999, 2005). Similarly, growth
and curvacin A production by Lactobacillus curvatus LTH 1174
were negatively affected by nitrite (even by a concentration as
low as 10 ppm), and also by sodium chloride (Verluyten et al.,
2003, 2004a). The effect on bacteriocin production was even
detected at low NaCl concentrations where there was no growth
inhibition, and NaCl was shown to interfere with bacteriocin
induction (Verluyten et al., 2004a). Growth and curvacin A
production were also negatively affected by some species (especially by nutmeg) in a meat system (Verluyten et al., 2004b). A
model was proposed to illustrate the influence of environmental
factors on sakacin production in a meat system (Leroy and De
Vuyst, 2003, 2005).
3. Bacteriocins and hurdle technology
The concept of hurdle technology began to apply in the food
industry in a rational way after the observation that survival of
microorganisms greatly decreased when they were confronted
with multiple antimicrobial factors (Leistner, 1978; Leistner and
Gorris, 1995; Leistner, 2000). After exposure of a bacterial
population to a single antimicrobial factor there is often a heterogeneous response, depending on the intensity of treatment as
well as many other factors. A fraction of the population may
receive a lethal dose of the antimicrobial factor, leading to cell
death. The remaining fraction may survive due to several reasons: (i) receiving a sub-lethal dose; (ii) showing an increased
resistance because of its physiological state (e.g. stationary
phase cells, or cells already stressed in response to other unfavourable environmental conditions), and (iii) cells naturally
resistant to the antimicrobial agent. Sub-lethally injured cells as
well as cells with increased resistance may repair the damage
caused by the antimicrobial agent and survive, having a good
opportunity to develop mechanisms of resistance or adaptation
that would make them immune to further challenges with the
particular antimicrobial factor. By contrast, when cells are exposed to a combination of antimicrobial factors, the intensity of
damage may be higher since some of the antimicrobial factors
may act on the same cellular target. The repair of multiple
damages may also require much higher energy costs, leading to
energy exhaustion and cell death. Therefore, the probabilities for
survival and proliferation for cells confronted with multiple
hurdles are very low. In addition, the synergy between different
antimicrobial factors may allow the use of lower doses compared
to their individual application.
Over 60 potential hurdles have been described to improve
food stability and/or quality (Leistner, 1999). The application of
bacteriocins as part of hurdle technology has received great
attention in recent years (Chen and Hoover, 2003; Ross et al.,
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A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
2003; Deegan et al., 2006), since bacteriocins can be used
purposely in combination with selected hurdles in order to
increase microbial inactivation (Fig. 2). The combination of
hurdles to be applied will depend greatly on the type of food and
its microbial composition. This must be carefully considered,
since different hurdles usually have different effects on
the members of a microbial community. As an example, food
acidification may select for aciduric bacteria while heat treatment may select for endospore formers. Also, elimination of
some members of the population may provide a more favourable environment for others, due to the lack of competition. The interaction of different antimicrobial factors may also
modify their individual antimicrobial spectra. For example,
Gram-negative bacteria may become sensitized to bacteriocins
and other molecules upon exposure to hurdles that destabilise
the bacterial outer membrane, as will be discussed further on.
3.1. Combination of bacteriocins with chemical substances and
natural antimicrobials
Previous studies have shown that the presence of NaCl
enhanced the antimicrobial action of bacteriocins such as nisin,
leucocin F10, enterocin AS-48 and others (Harris et al., 1991;
Thomas and Wimpenny, 1996; Mazzotta et al., 1997; Parente
et al., 1998; Ananou et al., 2004). However, nisin activity was
also antagonized by low concentration of NaCl (Bouttefroy
et al., 2000). Sodium chloride also decreased the antilisterial
activity of acidocin CH5 (at 1–2%; Chumchalová et al., 1998),
lactocin 705 (at 5–7%; Vignolo et al., 1998), leucocins 4010 (at
2.5% NaCl; Hornbæk et al., 2004), pediocin (at 6.5% NaCl;
Jydegaard et al., 2000), curvacin (Verluyten et al., 2002) and
Carnobacterium piscicola A9b bacteriocin (at 2–4% NaCl;
Himmelbloom et al., 2001). The protective effect of sodium
chloride may be due to interference with ionic interactions
between bacteriocin molecules and charged groups involved in
bacteriocin binding to target cells (Bhunia et al., 1991). Sodium
chloride may also induce conformational changes of bacter-
iocins (Lee et al., 1993) or changes in the cell envelope of the
target organisms (Jydegaard et al., 2000).
Reduction of nitrite content by addition of bacteriocins may
be beneficial in the food industry. The combinations of nisin and
nitrite delayed botulinal toxin formation in meat systems and
showed increased activity on clostridial endospores outgrowth
(Rayman et al., 1981, 1983; Taylor et al., 1985) and also on
Leuconostoc mesenteroides and L. monocytogenes (Gill and
Holley, 2003). Addition of nitrite also increased the anti-listeria
activity of bacteriocinogenic lactobacilli in meat (Hugas et al.,
1996) and the activities of enterocin EJ97 against L. monocytogenes, Bacillus coagulans and Bacillus macroides (García
et al., 2003, 2004a,b) and enterocin AS-48 against B. cereus
(Abriouel et al., 2002).
Organic acids and their salts can potentiate the activity of
bacteriocins greatly, while acidification enhances the antibacterial activity of both organic acids and bacteriocins (Jack et al.,
1995; Stiles, 1996). The increase in net charge of bacteriocins at
low pH might facilitate translocation of bacteriocin molecules
through the cell wall. The solubility of bacteriocins may also
increase at lower pH, facilitating diffusion of bacteriocin molecules. Buncic et al. (1995) found that the sensitivity of
L. monocytogenes to nisin (400 IU/ml) increased in combination
with lactate. Further reports have confirmed the increased
antibacterial activity of nisin in combination with sodium lactate
in several food systems (Scannell et al., 1997; Nykänen et al.,
2000; Long and Phillips, 2003; Ukuku and Fett, 2004). A nisinsorbate combination showed increased activity against Listeria
(Avery and Buncic, 1997), and B. licheniformis (Mansour et al.,
1998). In the production of Ricotta-type cheeses, the combination of nisin with acetic acid and sorbate controlled L. monocytogenes contamination over a long period storage (70 days) at
6–8 °C (Davies et al., 1997). Lacticin 3147 activity also increased in combination with sodium lactate or sodium citrate
(Scannell et al., 2000b), as well as did pediocin AcH activity in
combination with sodium diacetate and sodium lactate (Uhart
et al., 2004). Activity of enterocin AS-48 against B. cereus in
Fig. 2. Application of bacteriocins as part of hurdle technology.
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
rice gruel was also potentiated by sodium lactate (Grande et al.,
2006). Lactic acid, sodium lactate and peracetic acid (as well as
several other chemical compounds) increased AS-48 activity for
decontamination of L. monocytogenes in sprouts (Cobo Molinos
et al., 2005). When nisin and pediocin were used in combination
with chemical compounds to combat L. monocytogenes in fresh
produce, best results were obtained for nisin with phytic acid
(Bari et al., 2005).
Chelating agents permeate the outer membrane (OM) of
Gram-negative bacteria by extracting Ca2+ and Mg2+ cations
that stabilize lipopolysaccharide of this structure, allowing bacteriocins to reach the cytoplasmic membrane (Stevens et al.,
1991; Vaara, 1992; Schved et al., 1994; Helander et al., 1997).
The enhanced effect of chelators such as EDTA, disodium
pyrophosphate, trisodium phosphate, hexametaphosphate or
citrate and bacteriocins against Gram-negative bacteria has been
demonstrated for nisin both under laboratory conditions and in
foods (Stevens et al., 1991; Cutter and Siragusa, 1995a,b;
Carneiro De Melo et al., 1998; Boziaris and Adams, 1999; Fang
and Tsai, 2003). Brochrocin C and enterocin AS-48 also showed
increased antimicrobial activity on EDTA-treated Gram-negative bacteria (Abriouel et al., 1998; Gao et al., 1999; Ananou et
al., 2005). Sensitization of Gram-negative bacteria to bacteriocins by other chelators such as maltol or ethyl maltol has been
reported (Schved et al., 1996). Sodium lactate or sodium citrate
in combination with nisin showed increased antimicrobial
activity against Arcobacter butzleri in chicken due to their
chelating effect (Long and Phillips, 2003). Chelating agents can
also enhance the activity of bacteriocins on Gram-positive
bacteria. A combination of nisin and sodium polyphosphate
showed an increased activity against L. monocytogenes (Buncic
et al., 1995). More recently, an increased activity of chrisin (a
commercial nisin preparation) in combination with EDTA has
been reported against several Gram-positive bacteria (Gill and
Holley, 2003). The combination of sodium tripolyphosphate and
enterocin EJ97 showed increased activity against B. coagulans
and B. macroides (García et al., 2003, 2004a).
Other antimicrobial compounds such as ethanol can act synergistically with nisin to reduce the survival of L. monocytogenes
(Brewer et al., 2002). Sub-lethal concentrations of nisin
(30 IU/ml) and monolaurin (100 μg/ml) in combination acted
synergistically on B. licheniformis vegetative cells and spore
outgrowth in milk (Mansour et al., 1999). Synergism was also
observed for the sucrose fatty acid esters sucrose palmitate
and sucrose stearate and nisin against several strains of
L. monocytogenes, B. cereus (cells and spores), L. plantarum
and Staphylococcus aureus, but not against Gram-negative
bacteria (Thomas et al., 1998). Reuterin also showed a significant synergistic effect on L. monocytogenes and a slight
additive effect on S. aureus after in combination with nisin
(100 IU/ml), although the antimicrobial effect of reuterin
against Gram-negative pathogens was not enhanced, though
(Arqués et al., 2004).
Essential oils and their active components, the phenolic
compounds are also attractive natural preservatives (Burt, 2004).
When used in combination with bacteriocins, the dose of added
phenolic compounds could be lowered thereby decreasing their
57
impact on the food flavour and taste. Nisin acted synergistically
with carvacrol, eugenol or thymol against B. cereus and/or
L. monocytogenes (Pol and Smid, 1999; Periago et al., 2001;
Yamazaki et al., 2004). Combinations of nisin with carvacrol,
eugenol, or thymol resulted in synergistic action against Bacillus
subtilis and Listeria innocua, while nisin and cinnamic acid had
synergistic activity against L. innocua, but only additive against
B. subtilis (Olasupo et al., 2004). Carvacrol (0.5 mM) was used
to enhance the synergy found between nisin and a pulsed electric
field treatment (PEF) against vegetative cells of B. cereus in milk
(Pol et al., 2001a,b). The combination of nisin and cinnamon
accelerates death of Salmonella Typhimurium and Escherichia
coli O157:H7 in apple juice (Yuste and Fung, 2004). The natural
variant nisin Z also acted synergistically with thymol against
L. monocytogenes and B. subtilis (Ettayebi et al., 2000). The
antimicrobial activity of enterocin AS-48 against S. aureus cell
in vegetable sauces was potentiated significantly in combination
with the phenolic compounds carvacrol, geraniol, eugenol,
terpineol, caffeic acid, p-coumaric acid, citral and hydrocinnamic acid (Grande et al., 2007).
Combinations of bacteriocins have also been tested in order
to increase their antimicrobial activities. The simultaneous use of
nisin with pediocin AcH (Hanlin et al., 1993) or with leucocin
F10 (Parente et al., 1998) as well as lactacin B or lactacin F with
nisin or pediocin AcH, and lactacin 481/pediocin AcH (MuletPowell et al., 1998) provides a greater antibacterial activity than
each bacteriocin separately. Simultaneous or sequential additions of nisin (50 IU/ml) and curvaticin 13 (160 AU/ml) also
induced a greater inhibitory effect against L. monocytogenes
than each bacteriocin individually (Bouttefroy and Millière,
2000). The simultaneous use of two or more bacteriocins could
be useful not only to lower the added bacteriocin doses, but also
to avoid regrowth of bacteriocin-resistant/adapted cells. However, development of cross-resistance should be carefully
considered, especially for the combinations of bacteriocins
belonging to the same class, as it has been shown that a general
mechanism may account for high-level resistance to class IIa
bacteriocins in L. monocytogenes (Gravesen et al., 2002a,b).
An increased antibacterial activity can also be achieved
by combination of bacteriocins with other (non-bacteriocin)
antimicrobial proteins or peptides. Nisin and lysozyme acted
synergistically against Gram-positive bacteria, including spoilage lactobacilli and S. aureus (Chung and Hancock, 2000;
Nattress and Baker, 2003). The spectrum of the nisin-lysozyme
combination could be extended to Gram-negative bacteria by
addition of chelating agents (Gill and Holley, 2000) The combined addition of nisin and the lactoperoxidase system (LPS) had
a strong antimicrobial effect against L. monocytogenes Ohio and
(to a lesser extent) L. monocytogenes Scott A in skim milk after
24 h at 30 °C (Zapico et al., 1998), against L. monocytogenes
ATCC 15313 in skim milk at 25 °C for at least 15 days
(Boussouel et al., 2000), and also against fish spoilage flora
(Elotmani and Assobhei, 2004). Lactoferrin (and its partialhydrolysis derivative lactoferricin) is another natural protein
(which is found in milk and other secretions) with antimicrobial
activity due to its iron-binding capacity and polycationic nature
(Ellison, 1994). The combination of nisin and lactoferrin showed
58
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
increased antilisterial activity when tested in culture media
(Branen and Davidson, 2004). Synergy between eukaryotic
antimicrobial peptides and bacteriocins has also been investigated. The antimicrobial activity of pleurocidin (an antimicrobial peptide from fish) against E. coli was greatly enhanced by
pediocin PA-1, sakacin P, and curvacin A (Lüders et al., 2003).
Although no potential application has been proposed for the
observed synergy, the available synthetic pleurocidin could
probably be used in combination with bacteriocins for selective
inhibition of Gram-negative bacteria in fish.
3.2. Bacteriocins and heat treatments
Bacteriocins can be used to reduce the intensity of heat
treatments in foods without compromising microbial inactivation. Nisin and heat act synergistically against L. plantarum and
L. monocytogenes (Mahadeo and Tatini, 1994; Ueckert et al.,
1998), reducing the heat resistance of L. monocytogenes in milk
(Maisnier-Patin et al., 1995) and in cold-pack lobster meat
(Budu-Amoako et al., 1999). Nisin-resistant L. monocytogenes
cells grown in the presence of nisin were more sensitive to heat at
55 °C than wild-type cells (Modi et al., 2000). The efficacy of
enterocin AS-48 was higher on S. aureus cells sub-lethally
injured by heat due to the lower concentration of remaining
viable cells and to the cell damage induced by the heat treatment
(Ananou et al., 2004). Bacteriocins can also provide an
additional protection during food storage against proliferation
of endospores surviving heat treatments. Moreover, it has been
demonstrated that the intensity of heat treatments against
bacterial endospores can be lowered in combination with nisin
as well as with enterocin AS-48 (Beard et al., 1999; Wandling et
al., 1999; Grande et al., 2006), saving costs in the heat treatment
and decreasing the impact of heat on the food. Sub-lethal heat
has been shown to sensitize Gram-negative bacteria to several
bacteriocins such as nisin or pediocin AcH (Kalchayanand et al.,
1992; Boziaris et al., 1998), enterocin AS-48 (Abriouel et al.,
1998; Ananou et al., 2005), or jenseniin G (Bakes et al., 2004),
extending their spectrum of action. Highest sensitization was
reported for combined treatments of bacteriocins, heat and a
chelating agent (Abriouel et al., 1998; Ananou et al., 2005).
3.3. Bacteriocins and modified atmosphere packaging
Modified atmosphere packaging (MAP) is frequently used in
the food industry to prolong the shelf life of perishable food
products. MAP may be defined as “the enclosure of food products in gas-barrier materials, in which the gaseous environment
has been changed” (Young et al., 1988). Prolongation of the shelf
life of food by MAP is based on retardation of intrinsic food
changes and inhibition of spoilage microbiota. In a modified
atmosphere, the dissolved CO2 will determine growth inhibition
of microorganisms (Devlieghere et al., 1998). Gram-negative
bacteria are generally more sensitive to CO2, while lactic acid
bacteria are much more resistant (Farber, 1991; Church, 1994).
Since Gram-negative bacteria are usually not sensitive to
bacteriocins, MAP and bacteriocins are therefore two complementary hurdles of advantage to food spoilage.
Fang and Lin (1994a,b) found that growth of L. monocytogenes was completely inhibited on pork immersed in 10 IU/ml
nisin and packed in 80% CO2/20% air during 30 days of storage
at 4 °C. Activity of nisin as well as ALTA™ 2341 against
L. monocytogenes also increased in cold smoked salmon packaged under vacuum as well as under a 100% CO2 atmosphere
(Nilsson et al., 1997; Szabo and Cahill, 1999). A cocktail of
seven L. monocytogenes isolates was inhibited by nisin as well
as by ALTA™ 2341 under a 100% CO2 atmosphere, but not
under 100% N2, or 40% CO2/60% N2 (Szabo and Cahill, 1998).
It has been reported that nisin and CO2 atmosphere acted
synergistically on the cytoplasmic membrane of L. monocytogenes by enhancing membrane permeabilization (Nilsson et al.,
2000).
3.4. Bacteriocins and pulsed electric fields
Pulsed electric field (PEF) technology is a non-thermal process where microbial inactivation is achieved by application of
high-voltage pulses between a set of electrodes (Vega-Mercado
et al., 1997). The effects of PEF resemble bacterial electroporation, but the higher intensity of this treatment causes severe
damage to the bacterial cell membrane. Although this
technology can only be applied to pumpable food products, it
has gained attraction in recent years as an individual treatment
or in combination with other hurdles such as bacteriocins. Since
most bacteriocins act on the bacterial cytoplasmic membrane,
the combined application of bacteriocins and PEF is expected to
elicit increased bactericidal effects. Moreover, bacteriocins
could also provide an additional hurdle against survivors from
PEF treatments, such as sub-lethally injured cells or bacterial
endospores (Fig. 3). PEF could also be applied to extend the
antimicrobial spectrum of bacteriocins, since PEF disrupts the
bacterial outer membrane allowing bacteriocin molecules to
reach the bacterial cytoplasmic membrane target (Fig. 3). The
efficacy of the combined treatments of PEF and bactericiocins
in food preservation depends on several factors related to the
PEF treatment (such as field strength, number of pulses, wave
form or pulse duration), the food microbial load, composition
and physiological stage, the added bacteriocin, and other
environmental factors (Wouters et al., 2001; Bendicho et al.,
2002; Heinz et al., 2002). All of these may have an influence on
the numbers and types of bacteria surviving the combined
treatment and, most important, their proliferation during the
shelf life period of the processed food. For this reason, particular applications of bacteriocins and PEF must be studied in
detail for each type of food and target bacteria.
Several investigations have thrown light on the effectiveness
of combined treatments of bacteriocins and PEF treatments in
food systems (Table 2). Exposure of L. innocua to nisin in
liquid whole egg following PEF treatment exhibited an additive
effect on inactivation of the microorganism (Calderón-Miranda
et al., 1999a). A synergistic effect was observed as the electric
field intensity, number of pulses and nisin concentration
increased both in liquid whole egg and in skim milk
(Calderón-Miranda et al., 1999a,b). L. innocua treated by
PEF-nisin in skimmed milk exhibited an increase in the cell wall
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
59
Fig. 3. Effects of pulsed electric fields in combination with bacteriocins on microbial populations.
roughness, cytoplasmic clumping, leakage of cellular material,
and rupture of the cell walls and cell membranes (CalderónMiranda et al., 1999c). It has been reported recently that the
efficiency of the combined treatment of nisin and PEF in liquid
whey protein concentrate was strongly dependent on the
sequence of application, since exposure to nisin after PEF
produced a lower effect on L. innocua inactivation. This
behaviour was mainly attributed to changes in the cell envelope
and to modifications of the medium caused by PEF application
(Gallo et al., 2007).
The application of nisin clearly enhanced the lethal effect of
PEF treatment on other Gram-positive bacteria such as Micrococcus luteus cells in phosphate buffer (Dutreux et al., 2000),
L. plantarum in model beer (Ulmer et al., 2002), S. aureus in
skim milk (Sobrino-Lopez and Martin Belloso, 2006), and
vegetative cells of B. cereus (Pol et al., 2000). The synergy
between nisin and PEF treatment against resting vegetative cells
of B. cereus was enhanced by carvacrol (0.5 mM). Nisin
showed less activity against B. cereus in milk compared to
buffer (Pol et al., 2001a).
Bacterial endospores are refractile to PEF treatments, and
incorporation of bacteriocins into the food may provide an
additional hurdle against surviving endospores. Treatment of
B. cereus spores with nisin and/or PEF treatment did not lead to
direct inactivation of the spores or increased heat sensitivity as a
result of sub-lethal damage. In contrast, germinating spores
were found to be sensitive to PEF treatment. Nisin treatment
was more efficient than PEF treatment for inactivating
germinating spores (Pol et al., 2001b). Accordingly, bacteriocins could be applied as an additional hurdle against
endospores surviving PEF treatment, provided that bacteriocin
molecules remain active in the treated food.
Several studies have shown that the efficacy of PEF against
Gram-negative bacteria can be enhanced by nisin. When PEF
treatment was applied to Salmonella cells in orange juice in the
presence of nisin (100 U/ml), lysozyme (2,400 U/ml), or a
mixture of nisin (27.5 U/ml) and lysozyme (690 U/ml), cell
viability loss was increased by an additional 0.04 to 2.75 log
cycles. The combination of nisin and lysozyme had a more
pronounced bactericidal effect (by at least 1.37 log cycles) than
either nisin or lysozyme alone (Liang et al., 2002). Similarly,
PEF treatment combined with cinnamon or nisin triggered cell
death of E. coli O157:H7 in fresh apple cider, resulting in a
reduction in viable counts of 6 to 8 log cycles (Iu et al., 2001).
Although nisin was totally inactivated by PEF treatment in
simulated milk ultrafiltrate media, a 4-log cycle reduction of
inoculated E. coli cells was accomplished with nisin (ca. 1,000
IU/ml) and three pulses of 11.25 kV/cm or 500 IU/ml for five
pulses of the same intensity (Terebiznik et al., 2000). Nisin-PEF
inactivation of E. coli in simulated milk ultrafiltrate media was
enhanced by water activity reduction. Decreasing water activity
to 0.95 with NaCl and applying PEF at 5 kV/cm (a non-lethal
intensity when no other hurdle is used) with the further addition
of nisin (1200 IU/ml) resulted in a 5-log cycle reduction of the
bacterial population (Terebiznik et al., 2002). Nisin also increased PEF effectiveness against Pseudomonas aeruginosa.
At high PEF intensities (i.e., 11 kV/cm), the inhibitory effect of
nisin increased with the number of pulses applied (Santi et al.,
2003). In conclusion, addition of nisin can improve the efficacy of PEF treatments against vegetative cells of foodborne
Table 2
Reported effects on the application of nisin and pulsed-electic fields for bacterial
inactivation
Observed effects
References
–Increased inactivation of M. luteus in phosphate Dutreux et al. (2000)
buffer
–Increased inactivation of P. aeruginosa
Santi et al. (2003)
–Increased inactivation of S. aureus in skim milk
Sobrino-Lopez and
Martin Belloso (2006)
–Increased inactivation of L. innocua in liquid
Calderón-Miranda
whole egg, skim milk, and liquid whey
et al., 1999a,b,c; Gallo
protein concentrate
et al., 2007
–Increased inactivation of E. coli in simulated
Terebiznik et al., 2000,
milk ultrafiltrate media. Observed synergism
2002
with reduced water activity
–Increased inactivation of B. cereus vegetative
Pol et al., 2000, 2001a,b
cells (more efficient in buffer than in skim milk).
Observed synergism with carvacrol
–Inactivation of Salmonella in orange juice.
Liang et al. (2002)
Observed synergism with lysozyme
–Inactivation of E. coli O157:H7 in fresh apple
Iu et al. (2001)
cider. Observed synergism with cinnamon
–Inactivation of L. plantarum in model beer
Ulmer et al. (2002)
60
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
pathogenic and spoilage Gram-positive and Gram-negative
bacteria. Presumably, similar effects are to be expected for other
bacteriocins, although experimental data are needed to support
this conclusion. The stability of bacteriocins to PEF treatments
is another issue to be addressed, since remaining bacteriocin
activity may provide an important hurdle against survivors
(such as bacterial endospores) after PEF treatment.
3.5. Bacteriocins and high hydrostatic pressure (HHP)
High hydrostatic pressure (HHP) is an innovative food
processing and preservation method that causes injury and
killing of microbial cells (Kalchayanand et al., 1994; Farkas and
Hoover, 2000; Patterson, 2000; Ray, 2002). A variety of
pressure-treated products, such as ready-to-to eat chicken meat,
sliced ham, fresh whole oysters, jams, fruit juices, and
guacamole, are now commercially available. During pressurization, the disruption of H-bonds, ionic bonds and hydrophobic
interactions of the macromolecules adversely affects their
structures and functions (Hoover, 1993). The sub-lethal damage
is initiated by membrane phase transitions (Kato and Hayashi,
1999), affecting mainly ATP-generating and transport proteins.
Cell death caused by HHP increases with pressure and so does
the synergism with bacteriocins. Since most bacteriocins act on
the bacterial cytoplasmic membrane it can be hypothesized that
the observed synergy between bacteriocins and HHP results
from cumulative damage to this structure. However, bacteriocin–membrane interaction during phase transition at high pressure has never been studied. A tailing effect is often observed
after application of HHP treatments (Kalchayanand et al.,
1998a), indicating that cell death occurs as a consequence of
multiple events or cumulative cell damage. Sub-lethally injured
vegetative cells surviving HHP treatment may develop pressure
resistance, as has been previously reported for E. coli (GarcíaGraells et al., 1998) and L. monocytogenes (Karatzas and
Bennik, 2002). The increased cell damage caused by combined
treatments of HHP and bacteriocins could prevent the tailing
effect, providing an additional hurdle against selection of pressure-resistant vegetative cells.
The bactericidal effect of HHP (as well as its negative effects
on food constituents) increases along with the temperature,
which determines the different modalities of treatment, eg. cold
HHP pasteurisation (ca. 5 °C), HPP-assisted pasteurisation (ca.
40 °C), or HHP-assisted sterilisation (ca. 90 °C). The food pH is
also an influencing factor, and bacteria are usually more resistant
to HHP in low acid foods. Addition of bacteriocins could improve the efficacy of HHP treatments in foods, compensating for
the required increase in pressure or temperature.
The bacterial type and physiological stage (e.g. vegetive cells
or endospores) may have great influence on HHP efficacy (Chen
et al., 2006). Although bacteriocins are generally inactive on
Gram-negative bacteria, HHP transiently sensitizes Gram-negative bacteria through outer membrane damage, increasing the
possibilities for application of bacteriocins in food preservation
(Kalchayanand et al., 1994; Hauben et al., 1996; Masschalck
et al., 2001; Black et al., 2005). HHP also induces a more
persistent sensitisation of Gram-negative bacteria to small dif-
fusible antimicrobial molecules (García-Graells et al., 1998,
2000), which may act synergistically with other hurdles as well.
Bacterial endospores are resistant to HHP treatments currently
applied to foods (Smelt, 1998), although HHP treatments can
induce endospore germination. Addition of bacteriocins as a
second hurdle against surviving endospores could also improve
the safety and shelf life of HHP-processed foods (Shearer et al.,
2000).
Several studies have described the combined effects of bacteriocins and HHP on bacteria (Table 3). Nisin in combination with HHP showed strong synergistic effects against
L. plantarum, E. coli and L. monocytogenes (ter Steeg et al.,
1999; Farkas et al., 2003). HHP transiently sensitized Gramnegative bacteria to nisin, and a mechanism of pressure-promoted uptake of antimicrobial proteins and peptides was
proposed to explain this sensitisation (Masschalck et al., 2001).
Pediocin AcH also increased decimal reductions caused by
HHP on food spoilage and pathogenic bacteria in peptone
solution (Kalchayanand et al., 1998b), and increased cell lysis in
L. mesenteroides through cell wall degradation (Kalchayanand
et al., 2002). The synergistic activity of pediocin AcH in combination with nisin was greatly enhanced during pressurization
(Kalchayanand et al., 1998a,b, 2004a). A combination of nisin/
pediocin AcH added to the plating medium was also effective on
killing Clostridium spores induced to germinate by HHP
treatment (Kalchayanand et al., 2004b).
A key issue in the efficacy of HHP treatments is the protective
effect afforded by the food. For example, milk and meat (having
a complex composition as well as a pH closer to neutrality) exert
a grater protection against HHP. Inactivation of E. coli MG1655
was reduced from 7 logs at 400 MPa in phosphate buffer to only
3 logs at 700 MPa in milk, but addition of lysozyme (400 μg/ml)
and nisin (400 IU/ml) to the milk increased the lethality of
treatment (Garcia-Graells et al., 1999). Combining HHP and
nisin (500 IU/ml) also increased inactivation of bacteria associated with milk (Black et al., 2005). Similarly, the combination
of lacticin 3147 and HHP increased the viability loss of S. aureus
and L. monocytogenes in milk and in whey (Morgan et al.,
2000), and residual lacticin still showed inhibitory effect in the
food, inactivating and preventing growth of sub-lethally injured
cells. Since HHP may have unwanted effects on milk
components (Trujillo et al., 2002), bacteriocin-HHP treatments
could serve to decrease the intensity of HHP treatment without
compromising microbial inactivation.
Combined bacteriocin-HHP treatments have successfully
applied for inhibition of foodborne pathogens in cheese. Survival of L. monocytogenes Scott A in cheeses made from raw
milk that were previously inoculated with nisin and other
bacteriocin-producer strains decreased as the intensity of HHP
treatment increased (Arqués et al., 2005). Nisin-HHP has also
shown to increase inactivation of Bacillus and Clostridium endospores (Roberts and Hoover, 1996; Stewart et al., 2000). In
Mato' cheese, combining nisin with high pressure improved
the biocidal effect on spores and aerobic mesophilic bacteria
(Capellas et al., 2000). In model cheeses submitted to a
germination cycle of 60 MPa at 30 °C for 210 min, followed
by a vegetative cells destruction cycle of 300 or 400 MPa at
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
61
Table 3
Reported effects on the application of bacteriocins and high tydrostatic pressure (HHP) for bacterial inactivation
Bacteriocin
Observed effects
Nisin
Increased inactivation of B. coagulans, B. subtilis and Clostridium spores
References
Roberts and Hoover, 1996;
Stewart et al., 2000
Increased bactericidal activity and spectrum (E. coli, S. enteritidis, S. typhimurium, S. sonnei, S. flexneri, Masschalck et al. (2001)
P. fluorescens and S. aureus)
Increased inactivation of bacteria associated with milk (E. coli, P. fluorescens, L. innocua, and Garcia-Graells et al., 1999;
L. viridescens)
Black et al., 2005
Increased sensitivity of pressure-resistant E. coli
Garcia-Graells et al. (1999)
Strong synergistic effects against L. plantarum and E. coli at reduced temperature
ter Steeg et al. (1999)
Improved bactericidal effect on spores and aerobic mesophilic bacteria in cheese
Capellas et al. (2000)
Increased inactivation of B. cereus spores and inhibition of the surviving fraction in cheese
López-Pedemonte et al. (2003)
Increased inactivation of L. monocytogenes Scott A in cheese inoculated with a nisin-producing strain Arqués et al. (2005)
Increased inactivation of E. coli and L. innocua in liquid whole egg
Ponce et al. (1998)
In a meat model system, nisin reduced viable counts of E. coli, reduced growth of S. aureus, and Garriga et al. (2002)
suppressed slime-producing bacteria
Pediocin AcH
Increased inactivation of food spoilage and pathogenic bacteria (S. aureus, L. monocytogenes, Kalchayanand et al. (1998b)
S. typhimurium, E. coli O157:H7, L. sakei, L. mesenteroides, S. liquefaciens, and P. fluorescens)
suspended in peptone water
Increased cell lysis through cell wall degradation in L. mesenteroides
Kalchayanand et al. (2002)
Reduction of L. monocytogenes viable counts and inhibition of proliferation during storage in meat model Garriga et al. (2002)
system
Pediocin AcH + nisin
Increased inactivation of S. aureus, L. monocytogenes Scott A, Salmonella Thyphimurium and E. coli Kalchayanand et al., 1998a,b,
O157:H7
2004a
Killing of Clostridium spores induced to germinate
Kalchayanand et al. (2004b)
Sakacin K, enterocins A Reduction of L. monocytogenes viable counts and inhibition of proliferation during storage in meat model Garriga et al. (2002)
and B
system
Lacticin 3147
Increased inactivation of L. monocytogenes and S. aureus in milk and in whey
Morgan et al. (2000)
30 °C for 15 min the presence of nisin significantly increased
inactivation of B. cereus spores and reduced proliferation of the
surviving fraction measured 24 h and 15 d after HHP treatment
(López-Pedemonte et al., 2003).
In liquid whole egg, HHP inactivation of E. coli and
L. innocua improved significantly with nisin addition. A reduction of almost 5 log units in E. coli counts and more than 6 log
units for L. innocua was reported at 450 MPa and 5 mg/l of
nisin, and the two microorganisms were not detectable after
one month of storage at 4 °C (Ponce et al., 1998).
The application of HHP to fresh meat products may result in a
cooked-like aspect, and sometimes the products may develop a
rubbery consistency (Murano et al., 2002). Bacteriocin addition
could improve the efficacy of HHP treatments at lower pressure
without adverse effects on meat (Hugas et al., 2002). In a meat
model system, while added bacteriocins (enterocins A and B,
sakacin K, pediocin AcH or nisin) had no influence on killing
and survival of Salmonella enterica, a greater inactivation of
E. coli (N6 logs) was reported in the presence of nisin (Garriga
et al., 2002). Nisin also kept staphylococci at significant low
levels during storage and reduced slime-producing LAB below
the detection limit. Although the bacteriocin-HHP treatment
failed to inhibit proliferation of L. monocytogenes during storage in the case of nisin, counts of listeria remained below
detection levels in samples treated with sakacin K, enterocins A
and B as well as pediocin AcH (Garriga et al., 2002). These
results clearly indicate that each particular bacteriocin may show
different effects in HHP-combined treatments depending on the
target bacteria. Presumably, the combination of more than one
bacteriocin and HHP could provide better results when the target
consists of a mixture of different bacteria with varying sensitivities to bacteriocins.
High pressure homogeneisation (HPH) is another processing
treatment applied to certain types of food. It was observed
that E. coli became sensitive to lysozyme and nisin during
homogeneisation at pressures above 150 MPa when these
compounds were added before the HPH treatment, but cells
treated by HPH remained insensitive for lysozyme and nisin
added after that treatment (Diels et al., 2005). Similarly to HHP,
HPH may sensitise E. coli to lysozyme and nisin by inducing a
transient permeabilisation of the outer membrane that does not
involve a physical disruption and that is immediately repaired
after the process. Accordingly, the risks of foodborne pathogens
in HPH-treated foods could be lowered by an optimised incorporation of bacteriocins in foods before HPH treatment.
3.6. Bacteriocins and other non-thermal treatments
In spite of the many other non-thermal treatments currently
under study for food processing application, only a few reports
have been published on their combination with bacteriocins.
Irradiation offers a great potential for application in food preservation (Farkas, 1998, 2006). The spectrum of applications of
food irradiation could be expanded in combination with bacteriocins, especially if the radiation dose can be lowered, since
low-dose gamma irradiation has less unwanted effects on food
and may also have better acceptance among consumers. It was
reported that the combined application of pediocin (as ALTA™
2341) and low-dose irradiation (2.3 kGy) had an increased antimicrobial effect on L. monocytogenes on frankfurters (Chen et al.,
62
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
2004). One of the main limitations of gamma irradiation is
the increased resistance of bacterial endospores (Farkas, 1998),
requiring the application of combined treatments for a higher
effectiveness. In sous vide meals inoculated with spores of
psychrotrophic B. cereus, a combined treatment of nisin, heat
(90 °C, 10 min) and gamma irradiation (5 kGy) markedly reduced
the number of survivors for at least 42 days at an abuse temperature of 10 °C (Farkas et al., 2002).
Other non-thermal treatments such as pulsed magnetic fields
(PMF) have also been tested in combination with bacteriocins.
However, a combined treatment of PMF with EDTA and nisin
had no effect on E. coli (San Martín et al., 2001).
4. Bacteriocins and recent advances in molecular biology
and genome studies
Advances in molecular biology and molecular microbial
ecology have provided new valuable tools to study microorganisms in food ecosystems, such as the determination of their
bacteriocinogenic potential, the capacity for proliferation and
inhibition of unwanted bacteria, or the response to stress factors.
The distribution of bacteriocin-encoding genes among food
isolates is a common issue that can be easily be resolved by
molecular techniques. As an example, Maldonado et al. (2002)
established that the plantaricin S operon is widely distributed
among wild-type L. plantarum strains from olive fermentations
by PCR amplification and hybridization with specific probes.
Similarly, Ben Omar et al. (2004) analyzed the incidence of
structural genes for several known bacteriocins among food
isolates of Enterococcus faecalis and E. faecium by PCR amplification, and Faye et al. (2004) studied the distribution of
propionicin T1 genes among propionibacteria in a similar way.
PCR amplification with specific primers for bacteriocin genes
may be used to follow the predominance of an inoculated strain
in food fermentation, as shown for sakacin-P producing L. sakei
during production of fermented sausages (Urso et al., 2006a) and
L. gasseri K7 in semi-hard cheese (Matijašiæ et al., 2007).
Similarly, multiplex PCR targeting both bacteriocin genes and
species-specific genes can serve for precise identification of
bacteriocinogenic strains in a single analysis. Nisin-producing
lactococci and divercin 41 producing C. divergens V41 were
identified rapidly by this procedure (Moschetti et al., 2001;
Connil et al., 2002). Multiplex PCR was also used successfully
to follow implantation of V41 strain in cold smoked salmon
(Connil et al., 2002). Other procedures such as analysis of
RAPD-PCR profiles (Ryan et al., 2001; Matijašiæ et al., 2007),
REP-PCR (Foulquié-Moreno M.R., Verluyten J., Vancanneyt
M., Adriany T., Leroy F., Swings J., and De Vuyst L., unpublished) or PFGE analysis (Moschetti et al., 2001) have also been
used to evaluate strain implantation in food systems and the
impact of bacteriocinogenic starters on the food quality from the
microbiological point of view. A recent study addressed the use
of DGGE to evaluate simultaneously the impact of a bacteriocinogenic LAB on foodborne pathogens inoculated in a
fermented food as well as on the overall microbiological profile
of the fermentation (Díaz G., Ben Omar N., Abriouel H., Lucas
R., Martínez Cañamero M., and Galvez A, unpublished).
Real-time PCR could be used to follow survival of target
bacteria in the presence of added or in situ produced bacteriocin
while simultaneously determining the growth of bacteriocinogenic strains. This approach could provide additional data independent of the biases introduced by sample preparation
procedures and the pressure of selective media currently used for
enumeration of microorganisms. In other cases, it could solve
the problems of differentiation of closely related bacteria in
mixed populations. Grattepanche et al. (2005) used this method
to evaluate the impact of the nisin Z-producing L. lactis subsp.
lactis biovar. diacetylactis strain UL719 on Lactococcus
cremoris in milk fermented with mixed cultures, avoiding the
problem to distinguish the two bacterial populations in differential media when they were in the same order of magnitude.
Real-time PCR could also provide more precise information on
cells sub-lethally injured by bacteriocin molecules. Similarly,
DNA-based technology could be used to follow the expression
of bacteriocin genes in food systems, as well as the influence of
environmental conditions on gene expression and the stress
response of target bacteria to the produced or added bacteriocin
in food. In a recent work, expression of sakacin P structural gene
sspA by the L. sakei strain I151 in sausages was studied in order
to determine the influence of the production procedure for fermented sausages on bacteriocin production (Urso et al., 2006b).
This alternative method could overcome the biases introduced
by the currently used extraction procedures on the determination
of the amounts of bacteriocin produced in the food.
The use of fluorescence-based technology (such as fluorescence in-situ hybridisation—FISH—, confocal laser microscopy, or fluorescence ratio imaging microscopy—FRIM—) could
also provide valuable information, for example, on the distribution of bacteriocinogenic strains within the food matrix
(Fernández de Palencia et al., 2004) or the heterogeneous response of bacterial populations to bacteriocins (Hornbæk et al.,
2006). Other suggested applications could be to study the
distribution of target bacteria in the food both in the absence and
in the presence of bacteriocin pressure and the generation of
gradients and protected niches as a function of bacteriocin concentration. These techniques may facilitate the study of the
effects of bacteriocins in food systems at the level of single
cells, providing a completely new scenario picture of bacteriocin
effects.
Another line of research on bacteriocin application may rise
from genomic studies. Classical methods for detection of produced bacteriocins may underestimate the bacteriocinogenic
potential of LAB due to several factors such as the influence of
environmental conditions on bacteriocin production, the inducible character of many bacteriocins, and the loss of the
production capacity (which may be caused by gene mutation,
gene loss or genetic rearrangements). However, the analysis of
complete genomes may reveal the presence of potential bacteriocin genes and new bacteriocins independently of the producer capacity of strains (Nes and Johnsborg, 2004). As an
example, in silico analyses of the probiotic strain Lactobacillus
acidophilus NCFM predicted a chromosomal locus for lactacin
B, a class II bacteriocin (Altermann et al., 2005). Similarly,
while a limited number of bacteriocins have been described in
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
Streptococcus thermophilus (Gilbreth and Somkuti, 2005 and
references cited therein), analysis of the genome sequences
available revealed two loci predicted to be related to bacteriocin production (Hols et al., 2005). The first locus (lab, for
lantibiotic) contains genes that are similar to genes generally
found in lantibiotic production loci. However, it is not clear
whether this locus is really involved in lantibiotic production.
The second locus is similar to the bacteriocin-like peptide (blp)
locus described in S. pneumoniae and displays the typical
characteristics of a Class II bacteriocin locus. The blp loci of the
S. thermophilus strains also contain genes encoding bacteriocinlike peptides (i.e., lpD, blpU, blpE, blpF in strain LMD9, blpU,
blpK in strain LMG18311, and blpK in strain CNRZ1066 (Hols
et al., 2005). However, nonsense mutations detected in some of
the regulatory genes indicate that this locus is not functional at
least in some strains. The data available suggest that bacteriocin
loci in S. thermophilus could be exploited by genetic engineering
in order to develop new bacteriocinogenic strains of technological interest. Similarly, genome sequencing of L. sakei 23 K has
revealed that this strain carries a 6.5 kb region containing part
of the spp gene cluster responsible for sakacin P production
(Møretrø et al., 2005) described previously in L. sakei Lb674
(Hühne et al., 1996). Since spp homologue genes (mainly sppK
and sppR homologues) seem to be widely distributed among
L. sakei, it has been suggested that these genes could be used as a
fingerprinting region in typing methods and/or as a marker for
L. sakei strains (Møretrø et al., 2005).
Many bacteriocins are encoded by small genes that are often
omitted in the annotation process of bacterial genomes (De Jong
et al., 2006). In addition, bacteriocins and their accessory proteins are often encoded by poorly conserved ORFs. The identification of genes that are functionally similar but have limited
or no sequence homology is often a problem in genome data
mining. A web server (BAGEL) that identifies putative bacteriocin ORFs in a DNA sequence using novel, knowledgebased bacteriocin databases and motif databases has been recently created (De Jong et al., 2006). BAGEL is freely accessible
at: http://bioinformatics.biol.rug.nl/websoftware/bagel. Hopefully, this software will help researchers to identify novel bacteriocin-related genes in the upcoming LAB genome sequences.
A recent data mining study demonstrated that the genome of
Pediococcus pentosaceus ATCC 25745 contains a gene cluster
that resembles a regulated bacteriocin system (Diep et al.,
2006). The gene cluster has an operon-like structure consisting
of a putative pediocin-like bacteriocin gene (termed penA),
a potential immunity gene (termed peiA) as well as genetic
determinants involved in bacteriocin transport and regulation.
Nevertheless, the accessory gene involved in transport and the
inducer gene involved in regulation are missing, which makes
this bacterium a poor bacteriocin-producer. Cloning of penA
and peiA in a L. sakei host that contains the complete apparatus
for gene activation, maturation and externalization of bacteriocins confirmed the production of this new and potent bacteriocin, termed penocin A (Diep et al., 2006).
Based on the genome sequences and gene identification, it
will be possible to develop adequate methodologies (such as
microarray technology) to study the global response of bacteria
63
to bacteriocins. In one example, by using microarrays for global
analysis of gene expression, it has been shown that lactococcin
972 induces a cell-envelope stress response in L. lactis mediated
by the two-component system KinD/LlrD (Martínez et al.,
2006). Transcriptome analysis can also reveal new and relevant
data on the biology of bacteriocins such as the mechanisms
of resistance/adaptation. Strains of L. lactis that are naturally
adapted or transiently resistant to nisin can be readily obtained
by subcultivation with sub-lethal nisin amounts. The adaptation
is lost upon subcultivation without nisin. Transcriptome analysis
of adapted strains revealed a significant up/down regulation of
95 genes belonging to several main functional categories: (i) cell
wall synthesis, (ii) central and energy metabolism, (iii)
phospholipid- and fatty acid metabolism, (iv) gene regulation,
(v) transport, (vi) stress, and (vii) miscellaneous or unknown
functions (Kok et al., 2005). The comparative transcriptome
analysis of nisin-sensitive and nisin-resistant L. lactis concluded
that nisin resistance is a complex phenotype, involving various
different mechanisms, mainly (i) preventing nisin from reaching
the cytoplasmic membrane, (ii) reducing the acidity of the
extracellular medium, thereby stimulating the binding of nisin to
the cell wall, (iii) preventing the insertion of nisin into the
membrane, and (iv) possibly transporting nisin across the
membrane or extruding nisin out of the membrane (Kramer
et al., 2006). Studies similar to these are clearly needed in order
to elucidate the mechanisms that may be involved in adaptation
of foodborne pathogens upon exposure to bacteriocin pressure in
food systems.
Overall, the set of methodologies that have emerged in recent
years provide an arsenal of barely unexplored tools that could
expand the potential of bacteriocinogenic strains for food application and improve our understanding on the global effects of
bacteriocins in food ecosystems, allowing a more rational application of these natural antimicrobial hurdles in foods.
5. Conclusions
A large number of bacteriocins from LAB have been characterized to date, and many different studies have indicated the
potential usefulness of bacteriocins in food preservation. Bacteriocins are a diverse group of antimicrobial proteins/peptides,
and therefore are expected to behave differently on different
target bacteria and under different environmental conditions.
Since the efficacy of bacteriocins in foods is dictated by environmental factors, there is a need to determine more precisely the
most effective conditions for application of each particular
bacteriocin. The use of novel preservation technologies offers
new opportunities for application of bacteriocins as part of
hurdle technology, as has been demonstrated for PEF and HHP.
However, the combined application of many other technologies (such as ultrasonication, irradiation, microwave and ohmic
heating, or pulsed light) still remains unexplored. Bacteriocinogenic cells may also act as living factories in foods. The antimicrobial effects of bacteriocins and bacteriocinogenic cultures
in food ecosystems must be understood in terms of microbial
interactions. The application of a microbial ecology approach
may provide a more realistic portrait of the complex interactions
64
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
occurring in food systems that ultimately lead to microbial
inactivation, survival or adaptation to environmental stress. The
use of molecular microbial ecology methodologies may help to
understand better the biology of bacteriocins and food microbial
ecosystems at cellular and molecular levels. The study of
bacterial genomes and other related aspects such as global gene
expression analysis can provide valuable information on the
bacteriocinogenic potential of LAB. Genomic studies can also
throw light on other issues of great interest in food application
such as the global response of ecosystems to bacteriocins or the
development of adaptation or resistance.
References
Aasen, I.M., Markussen, S., Møretrø, T., Katla, T., Axelsson, L., Naterstad, K.,
2003. Interactions of the bacteriocins sakacin P and nisin with food
constituents. International Journal of Food Microbiology 87, 35–43.
Abriouel, H., Valdivia, E., Gálvez, A., Maqueda, M., 1998. Response of Salmonella choleraesuis LT2 spheroplasts and permeabilized cells to the bacteriocin
AS-48. Applied and Environmental Microbiology 64, 4623–4626.
Abriouel, H., Maqueda, M., Gálvez, A., Martínez-Bueno, M., Valdivia, E.,
2002. Inhibition of bacterial growth, enterotoxin production, and spore
outgrowth in strains of Bacillus cereus by bacteriocin AS-48. Applied and
Environmental Microbiology 68, 1473–1477.
Altermann, E., Russell, W.M., Azcarate-Peril, M.A., Barrangou, R., Buck, B.L.,
McAuliffe, O., Souther, N., Dobson, A., Duong, T., Callanan, M., Lick, S.,
Hamrick, A., Cano, R., Klaenhammer, T.R., 2005. Complete genome
sequence of the probiotic lactic acid bacterium Lactobacillus acidophilus
NCFM. Proceedings of the National Academy of Sciences of the USA 102,
3906–3912.
Ananou, S., Valdivia, E., Martínez Bueno, M., Gálvez, A., Maqueda, M., 2004.
Effect of combined physico-chemical preservatives on enterocin AS-48
activity against the enterotoxigenic Staphylococcus aureus CECT 976
strain. Journal of Applied Microbiology 97, 48–56.
Ananou, S., Gálvez, A., Martínez-Bueno, M., Maqueda, M., Valdivia, E., 2005.
Synergistic effect of enterocin AS-48 in combination with outer membrane
permeabilizing treatments against Escherichia coli O157:H7. Journal of
Applied Microbiology 99, 1364–1372.
Arqués, J.L., Fernández, J., Gaya, P., Nuñez, M., Rodríguez, E., Medina, M., 2004.
Antimicrobial activity of reuterin in combination with nisin against food-borne
pathogens. International Journal of Food Microbiology 95, 225–229.
Arqués, J.L., Rodríguez, E., Gaya, P., Medina, M., Nuñez, M., 2005. Effect of
combinations of high-pressure treatment and bacteriocin-producing lactic
acid bacteria on the survival of Listeria monocytogenes in raw milk cheese.
International Dairy Journal 15, 893–900.
Avery, S.M., Buncic, S., 1997. Antilisterial effects of a sorbate-nisin combination
in vitro and on packaged beef at refrigeration temperature. Journal of Food
Protection 60, 1075–1080.
Aymerich, M.T., Artigas, M.G., Garriga, M., Monfort, J.M., Hugas, M., 2000.
Effect of sausage ingredients and additives on the production of enterocins A
and B by Enterococcus faecium CTC492. Optimization of in vitro production
and anti-listerial effect in dry fermented sausages. Journal of Applied
Microbiology 88, 686–694.
Bakes, S.H., Kitis, F.Y.E., Quattlebaum, R.G., Barefoot, S.F., 2004. Sensitization
of Gram-negative and Gram-positive bacteria to jenseniin G by sublethal
injury. Journal of Food Protection 67, 1009–1013.
Barefoot, S.F., Chen, Y.R., Hughes, T.A., Bodine, A.B., Shearer, M.Y., Hughes,
M.D., 1994. Identification and purification of a protein that induces
production of the Lactobacillus acidophilus bacteriocin lactacin B. Applied
and Environmental Microbiology 60, 3522–3528.
Bari, M.L., Ukuku, D.O., Kawasaki, T., Inatsu, Y., Isshiki, K., Kawamoto, S.,
2005. Combined efficacy of nisin and pediocin with sodium lactate, citric
acid, phytic acid, and potassium sorbate and EDTA in reducing the Listeria
monocytogenes population of inoculated fresh-cut produce. Journal of Food
Protection 68, 1381–1387.
Ben Omar, N., Castro, A., Lucas, R., Abriouel, H., Yousif, N.M., Franz, C.M.,
Holzapfel, W.H., Perez-Pulido, R., Martinez-Cañamero, M., Galvez, A.,
2004. Functional and safety aspects of Enterococci isolated from different
Spanish foods. Systematic and Applied Microbiology 27, 118–130.
Beard, B.M., Sheldon, B.W., Foegeding, P.M., 1999. Thermal resistance of
bacterial spores in milk-based beverages supplemented with nisin. Journal of
Food Protection 62, 484–491.
Bendicho, S., Barbosa-Cánovas, G.V., Martín, O., 2002. Milk processing by
high intensity pulsed electric fields. Trends in Food Science & Technology
13, 195–204.
Bhunia, A.K., Johnson, M.C., Ray, B., Kalchayanand, N., 1991. Mode of action
of pediocin AcH from Pediococcus acidilactici H on sensitive bacterial
strains. Journal of Applied Bacteriology 70, 25–33.
Black, E.P., Kelly, A.L., Fitzgerald, G.F., 2005. The combined effect of high
pressure and nisin on inactivation of microorganisms in milk. Innovative
Food Science and Emerging Technologies 6, 286–292.
Boussouel, N., Mathieu, F., Revol-Junelles, A.-M., Millière, J.-B., 2000. Effects
of combinations of lactoperoxidase system and nisin on the behaviour of
Listeria monocytogenes ATCC 15313 in skim milk. International Journal
of Food Microbiology 61, 169–175.
Bouttefroy, A., Millière, J.-B., 2000. Nisin-curvaticin 13 combinations for
avoiding the regrowth of bacteriocin resistant cells of Listeria monocytogenes ATCC 15313. International Journal of Food Microbiology 62,
65–75.
Bouttefroy, A., Mansour, M., Linder, M., Milliere, J.-B., 2000. Inhibitory combinations of nisin, sodium chloride, and pH on Listeria monocytogenes
ATCC 15313 in broth by an experimental design approach. International
Journal of Food Microbiology 54, 109–115.
Boziaris, I.S., Adams, M.R., 1999. Effect of chelators and nisin produced in situ
on inhibition and inactivation of Gram negatives. International Journal of
Food Microbiology 53, 105–113.
Boziaris, I.S., Humpheson, I., Adams, M.R., 1998. Effect of nisin on heat injury
and inactivation of Salmonella enteritidis PT4. International Journal of Food
Microbiology 43, 7–13.
Branen, J.K., Davidson, P.M., 2004. Enhancement of nisin, lysozyme, and
monolaurin antimicrobial activities by ethylenediaminetetraacetic acid and
lactoferrin. International Journal of Food Microbiology 90, 63–74.
Brewer, R., Adams, M.R., Park, S.F., 2002. Enhanced inactivation of Listeria
monocytogenes by nisin in the presence of ethanol. Letters in Applied
Microbiology 34, 18–21.
Budu-Amoako, E., Ablett, R.F., Harris, J., Delves-Broughton, J., 1999. Combined
effect of nisin and moderate heat on destruction of Listeria monocytogenes in
cold-pack lobster meat. Journal of Food Protection 62, 46–50.
Buncic, S., Fitzgerald, S., Bell, C.M., Hudson, R.G., 1995. Individual and
combined listericidal effects of sodium lactate, potassium sorbate, nisin and
curing salts at refrigeration temperatures. Journal of Food Safety 15,
247–264.
Burt, S., 2004. Essential oils: their antibacterial properties and potential
applications in foods — a review. International Journal of Food Microbiology
94, 223–253.
Calderón-Miranda, M.L., Barbosa-Canovas, G.V., Swanson, B.G., 1999a.
Inactivation of Listeria innocua in liquid whole egg by pulsed electric fields
and nisin. International Journal of Food Microbiology 51, 7–17.
Calderón-Miranda, M.L., Barbosa-Canovas, G.V., Swanson, B.G., 1999b.
Inactivation of Listeria innocua in skim milk by pulsed electric fields and
nisin. International Journal of Food Microbiology 51, 19–30.
Calderón-Miranda, M.L., Barbosa-Canovas, G.V., Swanson, B.G., 1999c.
Transmission electron microscopy of Listeria innocua treated by pulsed
electric fields and nisin in skimmed milk. International Journal of Food
Microbiology 51, 31–38.
Capellas, M., Mor-Mur, M., Gervilla, R., Yuste, J., Guamis, B., 2000. Effect of
high pressure combined with mild heat or nisin on inoculated bacteria and
mesophiles of goats' milk fresh cheese. Food Microbiology 17, 633–641.
Carneiro De Melo, A.M.S., Cassar, C.L., Miles, R.J., 1998. Trisodium
phosphate increases sensitivity of Gram-negative bacteria to lysozyme and
nisin. Journal of Food Protection 61, 839–844.
Chen, H., Hoover, D.G., 2003. Bacteriocins and their food applications.
Comprehensive Reviews in Food Science and Food Safety 2, 82–100.
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
Chen, C.-M., Sebranek, J.G., Dickson, J.S., Mendonca, A.F., 2004. Combining
pediocin with post-packaging irradiation for control of Listeria monocytogenes on frankfurters. Journal of Food Protection 67, 1866–1875.
Chen, H., Guan, D., Hoover, D.G., 2006. Sensitivities of foodborne pathogens to
pressure changes. Journal of Food Protection 69, 130–136.
Chumchalová, J., Josephsen, J., Plocková, J.M., 1998. The antimicrobial
activity of acidocin CH5 in MRS broth and milk with added NaCl, NaNO3
and lysozyme. International Journal of Food Microbiology 43, 33–38.
Chung, W., Hancock, R.E.W., 2000. Action of lysozyme and nisin mixtures
against lactic acid bacteria. International Journal of Food Microbiology 60,
25–32.
Church, N., 1994. Developments in modified-atmosphere packaging and related
technologies. Trends in Food Science & Technology 5, 345–352.
Cleveland, J., Montville, T.J., Nes, I.F., Chikindas, M.L., 2001. Bacteriocins:
safe, natural antimicrobials for food preservation. International Journal of
Food Microbiology 71, 1–20.
Cobo Molinos, A., Abriouel, H., Ben Omar, N., Valdivia, E., Lucas, R., Maqueda,
M., Martínez Cañamero, M., Gálvez, A., 2005. Effect of immersion solutions
containing enterocin AS-48 on Listeria monocytogenes in vegetable foods.
Applied and Environmental Microbiology 71, 7781–7787.
Connil, N., Prévost, H., Dousset, X., 2002. Production of biogenic amines and
divercin V41 in cold smoked salmon inoculated with Carnobacterium
divergens V41, and specific detection of this strain by multiplex-PCR.
Journal of Applied Microbiology 92, 611–617.
Cotter, P.D., Hill, C., Ross, R.P., 2005. Bacteriocins: developing innate
immunity for food. Nature Reviews Microbiology 3, 777–788.
Coventry, M.J., Gordon, J.B., Alexander, M., Hickey, M.W., Wan, J., 1996. A
food-grade process for isolation and partial purification of bacteriocins of
lactic acid bacteria that uses diatomite calcium silicate. Applied and
Environmental Microbiology 62, 1764–1769.
Cutter, C.N., Siragusa, G.R., 1995a. Population reductions of Gram-negative
pathogens following treatments with nisin and chelators under various
conditions. Journal of Food Protection 58, 977–983.
Cutter, C.N., Siragusa, G.R., 1995b. Treatments with nisin and chelators to
reduce Salmonella and Escherichia coli on beef. Journal of Food Protection
58, 1028–1030.
Daeschel, M.A., Mcguire, J., Almakhla, H., 1992. Antimicrobial activity of
nisin adsorbed to hydrophilic and hydrophobic silicon surfaces. Journal of
Food Protection 55, 731–735.
Davies, E.A., Adams, M.R., 1994. Resistance of Listeria monocytogenes to the
bacteriocin nisin. International Journal of Food Microbiology 21, 341–347.
Davies, E.A., Bevis, H.E., Delves-Broughton, J., 1997. The use of the
bacteriocin, nisin, as a preservative in ricotta-type cheeses to control the
food-borne pathogen Listeria monocytogenes. Letters in Applied Microbiology 24, 343–346.
Dawson, P.L., Carl, G.D., Acton, J.C., Han, I.Y., 2002. Effect of lauric acid and
nisin-impregnated soy-based films on the growth of Listeria monocytogenes
on turkey bologna. Poultry Science 81, 721–726.
Dawson, P.L., Harmon, L., Sotthibandhu, A., Han, I.Y., 2005. Antimicrobial
activity of nisin-adsorbed silica and corn starch powders. Food Microbiology 22, 93–99.
Deegan, L.H., Cotter, P.D., Hill, C., Ross, P., 2006. Bacteriocins: biological
tools for bio-preservation and shelf-life extension. International Dairy
Journal 16, 1058–1071.
Degnan, A.J., Luchansky, J.B., 1992. Influence of beef tallow and muscle on the
antilisterial activity of pediocin AcH and liposome-encapsulated pediocin
AcH. Journal of Food Protection 55, 552–554.
De Jong, A., van Hijum, S.A., Bijlsma, J.J., Kok, J., Kuipers, O.P., 2006.
BAGEL, a web-based bacteriocin genome mining tool. Nucleic Acids 34
(Web Server issue), W273–W279.
Delgado, A., Brito, D., Peres, C., Arroyo-López, F.N., Garrido-Fernández, A.,
2005. Bacteriocin production by Lactobacillus pentosus B96 can be
expressed as a function of temperature and NaCl concentration. Food
Microbiology 22, 721–726.
Devlieghere, F., Debevere, J., van Impe, J., 1998. Concentration of carbon
dioxide in the water-phase as a parameter to model the effect of a modified
atmosphere on microorganisms. International Journal of Food Microbiology
43, 105–113.
65
Devlieghere, F., Vermeiren, L., Debevere, J., 2004. New preservation
technologies: possibilities and limitations. International Dairy Journal 14,
273–285.
Diels, A.M.J., De Taeye, J., Michiels, C.W., 2005. Sensitisation of Escherichia
coli to antibacterial peptides and enzymes by high-pressure homogenisation.
International Journal of Food Microbiology 105, 165–175.
Diep, D.B., Godager, L., Brede, D., Nes, I.F., 2006. Data mining and
characterization of a novel pediocin-like bacteriocin system from the
genome of Pediococcus pentosaceus ATCC 25745. Microbiology 152,
1649–1659.
Drider, D., Fimland, G., Héchard, Y., McMullen, L.M., Prévost, H., 2006. The
continuing story of class IIa bacteriocins. Microbiology and Molecular
Biology Reviews 70, 564–582.
Dutreux, N., Notermans, S., Góngora-Nieto, M.M., Barbosa-Cánovas, G.V.,
Swanson, B.G., 2000. Effects of combined exposure of Micrococcus luteus to
nisin and pulsed electric fields. International Journal of Food Microbiology 60,
147–152.
Dykes, G.A., Hastings, J.W., 1998. Fitness costs associated with class IIa
bacteriocin resistance in Listeria monocytogenes B73. Letters in Applied
Microbiology 26, 5–8.
Ellison, R.T., 1994. The effects of lactoferrin on gram-negative bacteria. In:
Hutchens, T.W., Lönnerdal, B., Rumball, S. (Eds.), Lactoferrin-Structure
and Function. Plenum Press, New York, pp. 71–87.
Elotmani, F., Assobhei, O., 2004. In vitro inhibition of microbial flora of fish by
nisin and lactoperoxidase system. Letters in Applied Microbiology 38,
60–65.
El-Ziney, M.G., Debevere, J., Jakobsen, M., 2000. Reuterin. In: Naidu, A.S. (Ed.),
Natural Food Antimicrobial Systems. CRC Press, London, pp. 567–587.
Ercolini, D., Storia, A., Villani, F., Mauriello, G., 2006. Effect of a bacteriocinactivated polythene film on Listeria monocytogenes as evaluated by viable
staining and epifluorescence microscopy. Journal of Applied Microbiology
100, 765–772.
Ettayebi, K., El Yamani, J., Rossi-Hassani, B., 2000. Synergistic effects of nisin
and thymol on antimicrobial activities in Listeria monocytogenes and Bacillus subtilis. FEMS Microbiology Letters 183, 191–195.
Fang, T.J., Lin, L.-W., 1994a. Growth of Listeria monocytogenes and Pseudomonas fragi on cooked pork in a modified atmosphere packaging/nisin
combination system. Journal of Food Protection 57, 479–485.
Fang, T.J., Lin, L.-W., 1994b. Inactivation of Listeria monocytogenes on raw
pork treated with modified atmosphere packaging and nisin. Journal of Food
and Drug Analysis 2, 189–200.
Fang, T.J., Tsai, H.-C., 2003. Growth patterns of Escherichia coli O157:H7
in ground beef treated with nisin, chelators, organic acids and their
combinations immobilized in calcium alginate gels. Food Microbiology 20,
243–253.
Farber, J.M., 1991. Microbiological aspects of modified-atmosphere packaging
technology—a review. Journal of Food Protection 54, 58–70.
Farkas, J., 1998. Irradiation as a method for decontaminating food: a review.
International Journal of Food Microbiology 44, 189–204.
Farkas, J., 2006. Irradiation for better foods. Trends in Food Science &
Technology 17, 148–152.
Farkas, D.F., Hoover, D.G., 2000. High pressure processing. Journal of Food
Science 65, 47–64 (Supplement).
Farkas, J., Polyák-Fehér, K., Andrássy, É., Mészáros, L., 2002. Improvement of
microbiological safety of sous-vide meals by gamma radiation. Radiation
Physics and Chemistry 63, 345–348.
Farkas, J., Andrassy, E., Meszaros, L., Simon, A., 2003. Increased salt- and
nisin-sensitivity of pressure-injured bioluminescent Listeria monocytogenes.
Acta Microbiologica et Immunologica Hungarica 50, 331–337.
Faye, T., Brede, D.A., Langsrud, T., Nes, I.F., Holo, H., 2004. Prevalence of the
genes encoding propionicin T1 and protease-activated antimicrobial peptide
and their expression in classical propionibacteria. Applied and Environmental Microbiology 70, 2240–2244.
Fernández de Palencia, P., de la Plaza, M., Mohedano, M.L., Martinez-Cuesta, M.C.,
Requena, T., López, P., Peláez, C., 2004. Enhancement of 2-methylbutanal
formation in cheese by using a fluorescently tagged Lacticin 3147 producing
Lactococcus lactis strain. International Journal of Food Microbiology 93,
335–347.
66
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
Fimland, G., Johnsen, L., Dalhus, B., Nissen-Meyer, J., 2005. Pediocin-like
antimicrobial peptides (class IIa bacteriocins) and their immunity proteins:
biosynthesis, structure, and mode of action. Journal of Peptide Science 11,
688–696.
Foulquié Moreno, M.R., Sarantinopoulos, P., Tsakalidou, E., De Vuyst, L.,
2006. The role and application of enterococci in food and health.
International Journal of Food Microbiology 106, 1–24.
Franklin, N.B., Cooksey, K.D., Getty, K.J., 2004. Inhibition of Listeria
monocytogenes on the surface of individually packaged hot dogs with a
packaging film coating containing nisin. Journal of Food Protection 67,
480–485.
Gallo, L.I., Pilosof, A.M.R., Jagus, R.J., 2007. Effect of the sequence of nisin
and pulsed electric fields treatments and mechanisms involved in the
inactivation of Listeria innocua in whey. Journal of Food Engineering 79,
188–193.
Gao, Y., van Belkum, M.J., Stiles, M.E., 1999. The outer membrane of gramnegative bacteria inhibits antibacterial activity of brochocin-C. Applied and
Environmental Microbiology 65, 4329–4333.
García, M.T., Ben Omar, N., Lucas, R., Pérez-Pulido, R., Castro, A., Grande, M.J.,
Martínez-Cañamero, M., Gálvez, A., 2003. Antimicrobial activity of enterocin
EJ97 on Bacillus coagulans CECT 12. Food Microbiology 20, 533–536.
García, M.T., Lucas, R., Abriouel, H., Ben Omar, N., Pérez, R., Grande, M.J.,
Martínez-Cañamero, M., Gálvez, A., 2004a. Antimicrobial activity of
enterocin EJ97 against ‘Bacillus macroides/Bacillus maroccanus’ isolated
from zucchini purée. Journal of Applied Microbiology 97, 731–737.
García, M.T., Martínez Cañamero, M., Lucas, R., Ben Omar, N., Pérez Pulido,
R., Gálvez, A., 2004b. Inhibition of Listeria monocytogenes by enterocin
EJ97 produced by Enterococcus faecalis EJ97. International Journal of
Food Microbiology 90, 161–170.
García-Graells, C., Hauben, K., Michiels, C.W., 1998. High pressure inactivation
and sublethal injury of pressure-resistant Escherichia coli mutants in fruit
juices. Applied and Environmental Microbiology 64, 1566–1568.
Garcia-Graells, C., Masschalck, B., Michiels, C.W., 1999. Inactivation
of Escherichia coli in milk by high-hydrostatic-pressure treatment in
combination with antimicrobial peptides. Journal of Food Protection 62,
1248–1254.
García-Graells, C., Valckx, C., Michiels, C.W., 2000. Inactivation of Escherichia
coli and Listeria innocua in milk by combined treatment with high hydrostatic
pressure and the lactoperoxidase system. Applied and Environmental
Microbiology 66, 4173–4179.
Garriga, M., Aymerich, M.T., Costa, S., Monfort, J.M., Hugas, M., 2002.
Bactericidal synergism through bacteriocins and high pressure in a meat
model system during storage. Food Microbiology 19, 509–518.
Ghalfi, H., Allaoui, A., Destain, J., Benkerroum, N., Thonart, P., 2006.
Bacteriocin activity by Lactobacillus curvatus CWBI-B28 to inactivate
Listeria monocytogenes in cold-smoked salmon during 4 degrees C storage.
Journal of Food Protection 69, 1066–1071.
Gilbreth, S.E., Somkuti, G.A., 2005. Thermophilin 110: a bacteriocin of
Streptococcus thermophilus ST110. Current Microbiology 51, 175–182.
Gill, A.O., Holley, R.A., 2000. Inhibition of bacterial growth on ham and
bologna by lysozyme, nisin and EDTA. Food Research International 33,
83–90.
Gill, A.O., Holley, R.A., 2003. Interactive inhibition of meat spoilage and
pathogenic bacteria by lysozyme, nisin and EDTA in the presence of nitrite
and sodium chloride at 24 °C. International Journal of Food Microbiology
80, 251–259.
Grande, Ma.J., Lucas, R., Abriouel, H., Valdivia, E., Ben Omar, N., Maqueda,
M., Martínez-Bueno, M., Martínez-Cañamero, M., Gálvez, A., 2006.
Inhibition of toxicogenic Bacillus cereus in rice-based foods by enterocin
AS-48. International Journal of Food Microbiology 106, 185–194.
Grande, Ma.J., Lucas, R., Abriouel, H., Valdivia, E., Ben Omar, N., Maqueda,
M., Martínez-Cañamero, M., Gálvez, A., 2007. Treatment of vegetable
sauces with enterocin AS-48 alone or in combination with phenolic
compounds to inhibit proliferation of Staphylococcus aureus. Journal of
Food Protection 70, 405–411.
Grattepanche, F., Lacroix, C., Audet, P., Lapointe, G., 2005. Quantification by
real-time PCR of Lactococcus lactis subsp. cremoris in milk fermented by a
mixed culture. Applied Microbiology and Biotechnology 66, 414–421.
Gravesen, A., Jydegaard Axelsen, A.-M., Mendes da Silva, J., Hansen, T.B.,
Knøchel, S., 2002a. Frequency of bacteriocin resistance development and
associated fitness costs in Listeria monocytogenes. Applied and Environmental Microbiology 68, 756–764.
Gravesen, A., Ramnath, M., Rechinger, K.B., Andersen, N., Jansch, L.,
Hechard, Y., Hastings, J.W., Knochel, S., 2002b. High-level resistance to
class IIa bacteriocins is associated with one general mechanism in Listeria
monocytogenes. Microbiology 148, 2361–2369.
Guerra, N.P., Macias, C.L., Agrasar, A.T., Castro, L.P., 2005. Development of a
bioactive packaging cellophane using Nisaplin as biopreservative agent.
Letters in Applied Microbiology 40, 106–1610.
Guinane, C.M., Cotter, P.D., Hill, C., Ross, R.P., 2005. Microbial solutions to
microbial problems; lactococcal bacteriocins for the control of undesirable
biota in food. Journal of Applied Microbiology 98, 1316–1325.
Hanlin, M.B., Kalchayanand, N., Ray, P., Ray, B., 1993. Bacteriocins of lactic
acid bacteria in combination have greater antibacterial activity. Journal of
Food Protection 56, 252–255.
Harris, L.J., Fleming, H.P., Klaenhammer, T.R., 1991. Sensitivity and resistance
of Listeria monocytogenes ATCC 19115, Scott A, and UAL500 to nisin.
Journal of Food Protection 54, 836–840.
Hauben, K.J.A., Wuytack, E.Y., Soontjens, C.C.F., Michiels, C.W., 1996. High
pressure transient sensitization of Escherichia coli to lysozyme and nisin by
disruption of outer membrane permeability. Journal of Food Protection 59,
350–355.
Heinz, V., Alvarez, I., Angersbach, A., Knorr, D., 2002. Preservation of liquid
foods by high intensity pulsed electric fields-basic concepts for process
design. Trends in Food Science & Technology 12, 103–111.
Helander, I.M., von Wright, A., Mattila-Sandholm, T.-M., 1997. Potential of
lactic acid bacteria and novel antimicrobials against Gram-negative bacteria.
Trends in Food Science & Technology 8, 146–150.
Henning, S., Metz, R., Hammes, W.P., 1986. New aspects for the application of
nisin to food products based on its mode of action. International Journal of
Food Microbiology 3, 135–141.
Himmelbloom, B., Nilsson, L., Gram, L., 2001. Factors affecting production of
an antilisterial bacteriocin by Carnobacterium piscicola strain A9b in
laboratory media and model fish systems. Journal of Applied Microbiology
91, 506–513.
Hols, P., Hancy, F., Fontaine, L., Grossiord, B., Prozzi, D., Leblond-Bourget, N.,
Decaris, B., Bolotin, A., Delorme, C., Ehrlich, S.D., Guédon, E., Monnet, V.,
Renault, P., Kleerebezem, M., 2005. New insights in the molecular biology
and physiology of Streptococcus thermophilus revealed by comparative
genomics. FEMS Microbiology Reviews 29, 435–463.
Holtzel, A., Ganzle, M.G., Nicholson, G.J., Hammes, W.P., Jung, G., 2000. The
first low-molecular-weight antibiotic from lactic acid bacteria: reutericyclin,
a new tetramic acid. Angewandte Chemie. International Edition 39,
2766–2768.
Holzapfel, W.H., 2002. Appropriate starter culture technologies for small-scale
fermentation in developing countries. International Journal of Food
Microbiology 75, 197–212.
Holzapfel, W.H., Geisen, R., Schillinger, U., 1995. Biological preservation of
foods with reference to protective cultures, bacteriocins and food-grade
enzymes. International Journal of Food Microbiology 24, 343–362.
Hoover, D.G., 1993. Pressure effects on biological systems. Food Technology
47, 150–155.
Hornbæk, T., Brocklehurst, T.F., Budde, B.B., 2004. The antilisterial effect of
Leuconostoc carnosum 4010 and leucocins 4010 in the presence of sodium
chloride and sodium nitrite examined in a structured gelatin system.
International Journal of Food Microbiology 92, 129–140.
Hornbæk, T., Brockhoff, P.B., Siegumfeldt, H., Budde, B.B., 2006. Two
subpopulations of Listeria monocytogenes occur at subinhibitory concentrations of leucocin 4010 and nisin. Applied and Envirnomental Microbiology 72, 1631–1638.
Hugas, M., Neumeyer, B., PagCs, F., Garriga, F., Hammes, W.P., 1996. Die
antimikrobielle Wirkung von Bakteriozin-bildenden Kulturen in Fleischwaren: IT) Vergleich des Effektes unterschiedlicher bakteriozinbildender
Laktobazillen auf Listerien in Rohwurst. Fleischwirtschaft 76, 649–652.
Hugas, M., Garriga, M., Monfort, J.M., 2002. New mild technologies in meat
processing: high pressure as a model technology. Meat Science 62, 359–371.
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
Hühne, K., Axelsson, L., Holck, A., Kröckel, L., 1996. Analysis of the sakacin P
gene cluster from Lactobacillus sakei Lb674 and its expression in sakacinnegative L. sakei strains. Microbiology 142, 1437–1448.
Iu, J., Mittal, G.S., Griffiths, M.W., 2001. Reduction in levels of Escherichia
coli O157:H7 in apple cider by pulsed electric fields. Journal of Food
Protection 64, 964–969.
Jack, R.W., Tagg, J.R., Ray, B., 1995. Bacteriocins of Gram positive bacteria.
Microbiological Reviews 59, 171–200.
Jung, D.-S., Bodyfelt, F.W., Daeshel, M.A., 1992. Influence of fat and
emulsifiers on the efficacy of nisin in inhibiting Listeria monocytogenes in
fluid milk. Journal of Dairy Science 75, 387–393.
Jydegaard, A.-M., Gravesen, A., Knøchel, S., 2000. Growth condition-related
response of Listeria monocytogenes 412 to bacteriocin inactivation. Letters
in Applied Microbiology 31, 68–72.
Kalchayanand, N., Hanlin, M.B., Ray, B., 1992. Sublethal injury makes Gramnegative and resistant Gram-positive bacteria sensitive to the bacteriocins,
pediocin AcH and nisin. Letters in Applied Microbiology 16, 239–243.
Kalchayanand, N., Sikes, A., Dunne, C.P., Ray, B., 1994. Hydrostatic pressure
and electroporation have increased bactericidal efficiency in combination
with bacteriocins. Applied and Environmental Microbiology 60, 4174–4177.
Kalchayanand, N., Sikes, A., Dunne, C.P., Ray, B., 1998a. Factors influencing
death and injury of foodborne pathogens by hydrostatic pressure pasteurization. Food Microbiology 15, 207–214.
Kalchayanand, N., Sikes, A., Dunne, C.P., Ray, B., 1998b. Interaction of
hydrostatic pressure, time and temperature of pressurization and pediocin
AcH on inactivation of foodborne bacteria. Journal of Food Protection 61,
425–431.
Kalchayanand, N., Frethem, C., Dunne, P., Sikes, A., Ray, B., 2002. Hydrostatic
pressure and bacteriocin-triggered cell wall lysis of Leuconostoc mesenteroides. Innovative Food Science and Emerging Technologies 3, 33–40.
Kalchayanand, N., Dunne, C.P., Sikes, A., Ray, B., 2004a. Viability loss and
morphology change of foodborne pathogens following exposure to
hydrostatic pressures in the presence and absence of bacteriocins.
International Journal of Food Microbiology 91, 91–98.
Kalchayanand, N., Dunne, C.P., Sikes, A., Ray, B., 2004b. Germination induction
and inactivation of Clostridium spores at medium-range hydrostatic pressure
treatment. Innovative Food Science and Emerging Technologies 5, 277–283.
Karatzas, K.A.H., Bennik, M.H., 2002. Characterization of a Listeria
monocytogenes Scott A isolate with intolerance towards high hydrostatic
pressure. Applied and Environmental Microbiology 68, 3183–3189.
Kato, M., Hayashi, R., 1999. Effects of high pressure on lipids of biomembranes
for understanding high-pressure-induced biological phenomena. Bioscience,
Biotechnology and Biochemistry 68, 1321–1328.
Klaenhammer, T.R., 1993. Genetics of bacteriocins produced by lactic acid
bacteria. FEMS Microbiology Reviews 12, 39–86.
Ko, S., Janes, M.E., Hettiarachchy, N.S., Johnson, M.G., 2001. Physical and
chemical properties of edible films containing nisin and their action against
Listeria monocytogenes. Journal of Food Science 66, 1006–1011.
Kok, J., Buist, G., Zomer, A.L., van Hijum, S.A.F.T., Kuipers, O.P., 2005.
Comparative and functional genomics of lactococci. FEMS Microbiology
Reviews 29, 411–433.
Kramer, N.E., van Hijum, S.A., Knol, J., Kok, J., Kuipers, O.P., 2006. Transcriptome
analysis reveals mechanisms by which Lactococcus lactis acquires nisin
resistance. Antimicrobial Agents and Chemotherapy 50, 1753–1761.
Kumar, C.G., Anand, S.K., 1998. Significance of microbial biofilms in food
industry: a review. International Journal of Food Microbiology 42, 9–27.
Larsen, A.G., Vogensen, F.K., Josephsen, J., 1993. Antimicrobial activity of
lactic acid bacteria isolated from sourdoughs: purification and characterization of bavaricin A, a bacteriocin produced by Lactobacillus bavaricus
MI401. Journal of Applied Bacteriology 75, 113–122.
Leal-Sánchez, M.V., Jiménez-Díaz, R., Maldonado-Barragán, A., GarridoFernández, A., Ruiz-Barba, J., 2002. Optimization of bacteriocin production
by batch fermentation of Lactobacillus plantarum LPCO10. Applied and
Environmental Microbiology 68, 4465–4471.
Lee, S., Iwata, T., Oyagi, H., 1993. Effects of salts on conformational change of
basic amphipathic peptides from β-structure to α-helix in the presence of
phospholipid liposomes and their channel-forming ability. Biochimica et
Biophysica Acta 1151, 75–82.
67
Leistner, L., 1978. Hurdle effect and energy saving. In: Downey, W.K. (Ed.),
Food Quality and Nutrition. Applied Science Publishers, London, UK,
pp. 553–557.
Leistner, L., 1999. Combined methods for food preservation. In: Shafiur
Rahman, M. (Ed.), Handbook of Food Preservation. Marcel Dekker, New
York, pp. 457–485.
Leistner, L., 2000. Basic aspects of food preservation by hurdle technology.
International Journal of Food Microbiology 55, 181–186.
Leistner, L., Gorris, L.G.M., 1995. Food preservation by hurdle technology.
Trends in Food Science & Technology 6, 41–46.
Leroy, F., De Vuyst, L., 1999. The presence of salt and a curing agent reduces
bacteriocin production by Lactobacillus sakei CTC 494, a potential starter for
fermented sausage. Applied and Environmental Microbiology 65, 5350–5356.
Leroy, F., De Vuyst, L., 2003. A combined model to predict the functionality of
the bacteriocin-producing Lactobacillus sakei strain CTC 494. Applied and
Environmental Microbiology 69, 1093–1099.
Leroy, F., De Vuyst, L., 2005. Simulation of the effect of sausage ingredients and
technology on the functionality of the bacteriocin-producing Lactobacillus
sakei CTC 494 strain. International Journal of Food Microbiology 100,
141–152.
Leroy, F., Verluyten, J., De Vuyst, L., 2006. Functional meat starter cultures for
improved sausage fermentation. International Journal of Food Microbiology
106, 270–285.
Liang, Z., Mittal, G.S., Griffiths, M.W., 2002. Inactivation of SalmonellaTyphimurium in orange juice containing antimicrobial agents by pulsed electric
field. Journal of Food Protection 65, 1081–1087.
Long, C., Phillips, C.A., 2003. The effect of sodium citrate, sodium lactate and
nisin on the survival of Arcobacter butzleri NCTC 12481 on chicken. Food
Microbiology 20, 495–502.
López-Pedemonte, T., Roig-Sagués, A.X., Trujillo, A.J., Capellas, M., Guamis,
B., 2003. Inactivation of spores of Bacillus cereus in cheese by high
hydrostatic pressure with the addition of nisin or lysozyme. Journal of Dairy
Science 86, 3075–3081.
Luchansky, J.B., Call, J.E., 2004. Evaluation of nisin-coated cellulose casings
for the control of Listeria monocytogenes inoculated onto the surface of
commercially prepared frankfurters. Journal of Food Protection 67,
1017–1021.
Lüders, T., Birkemo, G.A., Fimland, G., Nissen-Meyer, J., Nes, I.F., 2003. Strong
synergy between a eukaryotic antimicrobial peptide and bacteriocins from
lactic acid bacteria. Applied and Environmental Microbiology 69, 1797–1799.
Lungu, B., Johnson, M.G., 2005. Fate of Listeria monocytogenes inoculated
onto the surface of model Turkey frankfurter pieces treated with zein
coatings containing nisin, sodium diacetate, and sodium lactate at 4 ºC.
Journal of Food Protection 68, 855–859.
Magnusson, J., Schnürer, J., 2001. Lactobacillus coryniformis subsp. coryniformis
strain Si3 produces a broad-spectrum proteinaceous antifungal compound.
Applied and Environmental Microbiology 67, 1–5.
Mahadeo, M., Tatini, S.R., 1994. The potential of nisin to control Listeria
monocytogenes in poultry. Letters in Applied Microbiology 18, 323–326.
Maisnier-Patin, S., Tatini, S.R., Richard, J., 1995. Combined effect of nisin and
moderate heat on destruction of Listeria monocytogenes in milk. Lait 75,
81–91.
Maldonado, A., Ruiz-Barba, J.L., Floriano, B., Jimenez-Diaz, R., 2002. The
locus responsible for production of plantaricin S, a class IIb bacteriocin
produced by Lactobacillus plantarum LPCO10, is widely distributed among
wild-type Lact. plantarum strains isolated from olive fermentations.
International Journal of Food Microbiology 77, 117–124.
Maldonado, A., Ruiz-Barba, J.L., Jimenez-Diaz, R., 2004. Production of
plantaricin NC8 by Lactobacillus plantarum NC8 is induced in the presence
of different types of gram-positive bacteria. Archives of Microbiology 181,
8–16.
Mansour, M., Linder, M., Millière, J.-B., Lefebvre, G., 1998. Combined effects
of nisin, lactic acid and potassium sorbate on Bacillus licheniformis spores
in milk. Le Lait 7, 117–128.
Mansour, M., Amri, D., Bouttefroy, A., Linder, M., Milliere, J.B., 1999.
Inhibition of Bacillus licheniformis spore growth in milk by nisin,
monolaurin, and pH combinations. Journal of Applied Microbiology 86,
311–324.
68
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
Martínez, B., Bravo, D., Rodriguez, A., 2005. Consequences of the development
of nisin-resistant Listeria monocytogenes in fermented dairy products.
Journal of Food Protection 68, 2383–2388.
Martínez, B., Zomer, A.L., Rodríguez, A., Kuipers, O.P., 2006. Cell wall
synthesis inhibition by the antimicrobial peptide Lcn972 is sensed by the
lactococcal two-component system KinD/LlrD. FOODMICRO 2006. Food
safety and biotechnology: diversity and global impact. The 20th International ICFMH Symposium, Bologna (Italy), August 29-September 2, p. 87.
Book of Abstracts.
Masschalck, B., Van Houdt, R., Michiels, C.W., 2001. High pressure increases
bactericidal activity and spectrum of lactoferrin, lactoferricin and nisin.
International Journal of Food Microbiology 64, 325–332.
Matijašić, B.B., Rajšp, M.K., Perko, B., Rogelj, I., 2007. Inhibition of Clostridium tyrobutyricum in cheese by Lactobacillus gasseri. International
Dairy Journal 17, 157–166.
Mattila, K., Saris, P., Tyopponen, S., 2003. Survival of Listeria monocytogenes
on sliced cooked sausage after treatment with pediocin AcH. International
Journal of Food Microbiology 89, 281–286.
Mauriello, G., Ercolini, D., La Storia, A., Casaburi, A., Villani, F., 2004.
Development of polythene films for food packaging activated with an
antilisterial bacteriocin from Lactobacillus curvatus 32Y. Journal of Applied
Microbiology 97, 314–322.
Mauriello, G., De Luca, E., La Storia, A., Villani, F., Ercolini, D., 2005.
Antimicrobial activity of a nisin-activated plastic film for food packaging.
Letters in Applied Microbiology 41, 464–469.
Mazzotta, A.S., Montville, T.J., 1997. Nisin induces changes in membrane fatty
acid composition of Listeria monocytogenes nisin-resistant strains at 108 °C
and 30 °C. Journal of Applied Microbiology 82, 32–38.
Mazzotta, A.S., Crandall, A.D., Montville, T.J., 1997. Nisin resistance in
Clostridium botulinum spores and vegetative cells. Applied and Environmental Microbiology 63, 2654–2659.
Ming, X., Daeschel, M.A., 1993. Nisin resistance of foodborne bacteria and the
specific resistance responses of Listeria monocytogenes Scott A. Journal of
Food Protection 56, 944–948.
Ming, X., Weber, G.H., Ayres, J.W., Sandine, W.E., 1997. Bacteriocins applied
to food packaging materials to inhibit Listeria monocytogenes on meats.
Journal of Food Science 62, 413–415.
Modi, K.D., Chikindas, M.L., Montville, T.J., 2000. Sensitivity of nisin-resistant
Listeria monocytogenes to heat and the synergistic action of heat and nisin.
Letters in Applied Microbiology 30, 249–253.
Møretrø, T., Naterstad, K., Wang, E., Aasen, I.M., Chaillou, S., Zagorec, M.,
Axelsson, L., 2005. Sakacin P non-producing Lactobacillus sakei strains
contain homologues of the sakacin P gene cluster. Research in Microbiology
156, 949–960.
Morgan, S.M., Galvin, M., Kelly, J., Ross, R.P., Hill, C., 1999. Development of
a lacticin 3147-enriched whey powder with inhibitory activity against
foodborne pathogens. Journal of Food Protection 62, 1011–1016.
Morgan, S.M., Ross, R.P., Beresford, T., Hill, C., 2000. Combination
of hydrostatic pressure and lacticin 3147 causes increased killing of
Staphylococcus and Listeria. Journal of Applied Microbiology 88,
414–420.
Moschetti, G., Blaiotta, G., Villani, F., Coppola, S., 2001. Nisin-producing
organisms during traditional ‘Fior di latte’ cheese-making monitored by
multiplex-PCR and PFGE analyses. International Journal of Food Microbiology 63, 109–116.
Mulet-Powell, N., Lacoste-Armynot, A.M., Vinas, M., Simeon De Buochberg,
M., 1998. Interactions between pairs of bacteriocins from lactic bacteria.
Journal of Food Protection 61, 1210–1212.
Muñoz, A., Maqueda, M., Gálvez, A., Martínez-Bueno, M., Rodriguez, A.,
Valdivia, E., 2004. Biocontrol of psychrotrophic enterotoxigenic Bacillus
cereus in a non fat hard type cheese by an enterococcal strain-producing
enterocin AS-48. Journal of Food Protection 67, 1517–1521.
Murano, E., Hugas, M., Garriga, M., Monfort, J.M., 2002. New mild
technologies in meat processing: high pressure as a model Technology.
Meat Science 62, 359–371.
Natrajan, N., Sheldon, B.W., 2000. Efficacy of nisin-coated polymer films to
inactivate Salmonella typhimurium on fresh broiler skin. Journal of Food
Protection 63, 1189–1196.
Nattress, F.M., Baker, L.P., 2003. Effects of treatment with lysozyme and nisin
on the microflora and sensory properties of commercial pork. International
Journal of Food Microbiology 85, 259–267.
Nes, I.F., Johnsborg, O., 2004. Exploration of antimicrobial potential in LAB by
genomics. Current Opinion in Biotechnology 15, 100–104.
Neysens, P., De Vuyst, L., 2005. Carbon dioxide stimulates the production of
amylovorin L by Lactobacillus amylovorus DCE 471, while enhanced
aeration causes biphasic kinetics of growth and bacteriocin production.
International Journal of Food Microbiology 105, 191–202.
Neysens, P., Messens, W., De Vuyst, L., 2003. Effect of sodium chloride on
growth and bacteriocin production by Lactobacillus amylovorus DCE 471.
International Journal of Food Microbiology 88, 29–39.
Nilsson, L., Huss, H.H., Gram, L., 1997. Inhibition of Listeria monocytogenes
on cold-smoked salmon by nisin and carbon dioxide atmosphere.
International Journal of Food Microbiology 38, 217–227.
Nilsson, L., Chen, Y., Chikindas, M.L., Huss, H.H., Gram, L., Montville, T.J.,
2000. Carbon dioxide and nisin act synergistically on Listeria monocytogenes. Applied and Environmental Microbiology 66, 769–774.
Nykänen, A., Weckman, K., Lapvetelainen, A., 2000. Synergistic inhibition of
Listeria monocytogenes on cold-smoked rainbow trout by nisin and sodium
lactate. International Journal of Food Microbiology 61, 63–72.
Olasupo, N.A., Fitzgerald, D.J., Narrad, A., Gasson, M.J., 2004. Inhibition of Bacillus subtilis and Listeria innocua by nisin in combination with some naturally
occurring organic compounds. Journal of Food Protection 67, 596–600.
O'Mahony, T., Rekhif, N., Cavadini, C., Fitzgerald, G.F., 2001. The application
of a fermented food ingredient containing ‘variacin’, a novel antimicrobial
produced by Kocuria varians, to control the growth of Bacillus cereus in
chilled dairy products. Journal of Applied Microbiology 90, 106–114.
O'Sullivan, L., Ross, R.P., Hill, C., 2002. Potential of bacteriocin-producing
lactic acid bacteria for improvements in food safety and quality. Biochimie
84, 593–604.
Parente, E., Giglio, M.A., Riccardi, A., Clementi, F., 1998. The combined effect
of nisin, leucocin F10, pH, NaCl and EDTA on the survival of Listeria
monocytogenes in broth. International Journal of Food Microbiology 40,
65–75.
Patterson, M., 2000. High pressure treatments of foods. In: Robinson, R.K.,
Batt, C.A., Patel, P.D. (Eds.), Encyclopedia of food microbiology. Academic
Press, San Diego, pp. 1059–1065.
Peláez, C., Requena, T., 2005. Exploiting the potential of bacteria in the cheese
ecosystem. International Dairy Journal 15, 831–844.
Periago, P.M., Palop, A., Fernandez, P.S., 2001. Combined effect of nisin,
carvacrol and thymol on the viability of Bacillus cereus heat-treated
vegetative cells. Food Science and Technology International 7, 487–492.
Pol, I.E., Smid, E.J., 1999. Combined action of nisin and carvacrol on Bacillus
cereus and Listeria monocytogenes. Letters in Applied Microbiology 29,
166–170.
Pol, I.E., Mastwijk, H.C., Bartels, P.V., Smid, E.J., 2000. Pulsed-electric field
treatment enhances the bactericidal action of nisin against Bacillus cereus.
Applied and Environmental Microbiology 66, 428–430.
Pol, I.E., Mastwijk, H.C., Slump, R.A., Popa, M.E., Smith, E.J., 2001a.
Influence of food matrix on inactivation of Bacillus cereus by combinations
of nisin, pulsed electric field treatment, and carvacrol. Journal of Food
Protection 64, 1012–1018.
Pol, I.E., van Arendonk, W.G., Mastwijk, H.C., Krommer, J., Smid, E.,
Moezelaar, R., 2001b. Sensitivities of germinating spores and carvacroladapted vegetative cells and spores of Bacillus cereus to nisin and pulsedelectric-field treatment. Applied and Environmental Microbiology 67,
1693–1699.
Ponce, E., Pla, R., Sendra, E., Guamis, B., Mor-Mur, M., 1998. Combined effect
of nisin and high hydrostatic pressure on destruction of Listeria innocua and
Escherichia coli in liquid whole egg. International Journal of Food
Microbiology 43, 15–19.
Ray, B., 2002. High hydrostatic pressure: microbial inactivation and food
preservation. In: Britton, G. (Ed.), The Encyclopedia of Environmental
Microbiology. Wiley, New York, pp. 1552–1563.
Rayman, K., Aris, B., Hurst, A., 1981. Nisin: possible alternative or adjuct to
nitrite in the preservation of meats. Applied and Environmental Microbiology 41, 375–380.
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
Rayman, K., Malik, N., Hurst, A., 1983. Failure of nisin to inhibit outgrowth
of lostridium botulinum in a model cured meat system. Applied and
Environmental Microbiology 46, 1450–1452.
Rekhif, N., Atrih, A., Lefebvre, G., 1994. Selection and properties composition
of spontaneous variants of Listeria monocytogenes ATCC 15313 resistant to
different bacteriocins produced by lactic acid bacteria strains. Current
Microbiology 28, 237–241.
Roberts, C.M., Hoover, D.G., 1996. Sensitivity of Bacillus coagulans spores to
combinations of high hydrostatic pressure, heat, acidity and nisin. Journal of
Applied Bacteriology 81, 363–368.
Robertson, A., Tirado, C., Lobstein, T., Jermini, M., Knai, C., Jensen, J.H., FerroLuzzi, A., James, W.P.T. (Eds.), 2004. Food and health in Europe: a new basis
for action. WHO Regional Publications, European Series, vol. 96. Geneva.
Rodríguez, J.M., Martinez, M.I., Horn, N., Dodd, H.M., 2003. Heterologous
production of bacteriocins by lactic acid bacteria. International Journal of
Food Microbiology 80, 101–116.
Ross, R.P., Sporns, P., Dodd, H.M., Gasson, M.J., Mellon, F.A., McMullen, L.M.,
2003. Involvement of dehydroalanine and dehydrobutyrine in the addition of
glutathione to nisin. Journal of Agriculture and Food Chemistry 51,
3174–3178.
Ross, R.P., Stanton, C., Hill, C., Fitzgerald, G.F., Coffey, A., 2000. Novel
cultures for cheese improvement. Trends in Food Science & Technology 11,
96–104.
Ross, R.P., Morgan, S., Hill, C., 2002. Preservation and fermentation: past,
present and future. International Journal of Food Microbiology 79, 3–16.
Ross, A.I.V., Griffiths, M.W., Mittal, G.S., Deeth, H.C., 2003. Combining
nonthermal technologies to control foodborne microorganisms. International
Journal of Food Microbiology 89, 125–138.
Ryan, M.P., Ross, R.P., Hill, C., 2001. Strategy for manipulation of cheese flora
using combinations of lacticin 3147-producing and-resistant cultures.
Applied and Environmental Microbiology 67, 2699–2704.
San Martín, F., Harte, F.M., Lelieveld, H., Barbosa-Canovas, G.V., Swanson, Barry
G., 2001. Inactivation effect of an 18-T pulsed magnetic field combined with
other technologies on Escherichia coli. Innovative Food Science and Emerging
Technologies 2, 273–277.
Santi, L., Cerrutti, P., Pilosof, A.M., de Huergo, M.S., 2003. Optimization of the
conditions for electroporation and the addition nisin for Pseudomonas
aeruginosa inhibition. Revista Argentina de Microbiología 35, 198–204.
Scannell, A.G.M., Hill, C., Buckley, D.J., Arendt, E.K., 1997. Determination of
the influence of organic acids and nisin on shelf-life and microbiological
safety aspects of fresh pork sausage. Journal of Applied Microbiology 83,
407–412.
Scannell, A.G.M., Hill, C., Ross, R.P., Marx, S., Hartmeier, W., Arendt, E.K.,
2000a. Development of bioactive food packaging materials using immobilised bacteriocins Lacticin 3147 and Nisaplins. International Journal of Food
Microbiology 60, 241–249.
Scannell, A.G., Ross, R.P., Hill, C., Arendt, E.K., 2000b. An effective lacticin
biopreservative in fresh pork sausage. Journal of Food Protection 63,
370–375.
Schillinger, U., Geisen, R., Holzapfel, W.H., 1996. Potential of antagonistic
microorganisms and bacteriocins for the biological preservation of foods.
Trends in Food Science & Technology 71, 58–64.
Schved, F., Henis, Y., Juven, B.J., 1994. Response of spheroplasts and chelatorpermeabilized cells of Gram-negative bacteria to the action of the bacteriocins
pediocin SJ-1 and nisin. International Journal of Food Microbiology 21,
305–314.
Schved, F., Pierson, M.D., Juven, B.J., 1996. Sensitization of E. coli to nisin by
maltol and ethyl maltol. Letters in Applied Microbiology 22, 189–191.
Shearer, A.E.H., Dunne, C.P., Sikes, A., Hoover, D.G., 2000. Bacterial spore
inhibition and inactivation in foods by pressure, chemical preservatives, and
mild heat. Journal of Food Protection 63, 1503–1510.
Sip, A., Grajek, W., Boyaval, P., 1998. Enhancement of bacteriocin production
by Carnobacterium divergens AS7 in the presence of a bacteriocin-sensitive
strain Carnobacterium piscicola. International Journal of Food Microbiology 42, 63–69.
Siragusa, G.R., Cutter, C.N., Willett, J.L., 1999. Incorporation of bacteriocin in
plastic retains activity and inhibits surface growth of bacteria on meat. Food
Microbiology 61, 229–235.
69
Smelt, J.P.P.M., 1998. Recent advances in the microbiology of high pressure
processing. Trends in Food Science & Technology 9, 152–158.
Sobrino-Lopez, A., Martin Belloso, O., 2006. Enhancing inactivation of Staphylococcus aureus in skim milk by combining high-intensity pulsed
electric fields and nisin. Journal of Food Protection 69, 345–353.
Stergiou, V.A., Thomas, L.V., Adams, M.R., 2006. Interactions of nisin with
glutathione in a model protein system and meat. Journal of Food Protection
69, 951–956.
Stevens, K.A., Sheldon, B.W., Klapes, N.A., Klaenhammer, T.R., 1991. Nisin
treatment for inactivation of Salmonella species and other gram-negative
bacteria. Applied and Environmental Microbiology 57, 3613–3615.
Stewart, C.M., Dunne, C.P., Sikes, A., Hoover, D.G., 2000. Sensitivity of spores
of Bacillus subtilis and Clostridium sporogenes PA3679 to combinations of
high hydrostatic pressure and other processing parameters. Innovative Food
Science and Emerging Technologies 1, 49–56.
Stiles, M.E., 1996. Biopreservation by lactic acid bacteria. Antonie van
Leeuwenhoek 70, 331–345.
Szabo, E.A., Cahill, M.E., 1998. The combined effects of modified atmosphere,
temperature, nisin and ALTA™ 2341 on the growth of Listeria
monocytogenes. International Journal of Food Microbiology 43, 21–31.
Szabo, E.A., Cahill, M.E., 1999. Nisin and ALTATM 2341 inhibit the growth of
Listeria monocytogenes on smoked salmon packaged under vacuum or
100% CO2. Letters in Applied Microbiology 28, 373–377.
Taylor, J.I., Somer, E.B., Kruger, L.A., 1985. Antibotulinal effectiveness of
nisin-nitrite combinations in culture medium and chicken frankfurter
emulsions. Journal of Food Protection 48, 234–249.
Terebiznik, M.R., Jagus, R.J., Cerrutti, P., De Huergo, M., Pilosof, A.M., 2000.
Combined effect of nisin and pulsed electric fields on the inactivation of
Escherichia coli. Journal of Food Protection 63, 741–746.
Terebiznik, M.R., Jagus, R.J., Cerrutti, P., De Huergo, M., Pilosof, A.M., 2002.
Inactivation of Escherichia coli by a combination of nisin, pulsed electric
fields, and water activity reduction by sodium chloride. Journal of Food
Protection 65, 1253–1258.
ter Steeg, P.F., Hellemons, J.C., Kok, A.E., 1999. Synergistic actions of nisin,
sublethal injury pressure, and reduced temperature on bacteria and yeast.
Applied and Environmental Microbiology 65, 4148–4154.
Thomas, L.V., Wimpenny, J.W.T., 1996. Investigation of the effect of combined
variations in temperature, pH, and NaCl concentration on nisin inhibition of
Listeria monocytogenes and Staphylococcus aureus. Applied and Environmental Microbiology 62, 2006–2012.
Thomas, L.V., Davies, E.A., Delves-Broughton, J., Wimpenny, J.W.T., 1998.
Synergistic effect of sucrose fatty acid ester on nisin inhibition of Grampositive bacteria. Journal of Applied Microbiology 85, 1013–1022.
Thomas, L.V., Clarkson, M.R, Delves-Broughton, J., 2000. Nisin. In: Naidu,
A.S. (Ed.), Natural food antimicrobial systems. CRC Press, Boca-Raton,
FL, pp. 463–524.
Todorov, S., Onno, B., Sorokine, O., Chobert, J.M., Ivanova, I., Dousset, X.,
1999. Detection and characterization of a novel antibacterial substance
produced by Lactobacillus plantarum ST 31 isolated from sourdough.
International Journal of Food Microbiology 48, 167–177.
Trujillo, A.J., Capella, M., Saldo, J., Gervilla, R., Guamis, B., 2002.
Applications of high-hydrostatic pressure on milk and dairy products: a
review. Innovative Food Science and Emerging Technologies 3, 295–307.
Työppönen, S., Petäjä, E., Mattila-Sandholm, T., 2003. Bioprotectives and
probiotics for dry sausages. International Journal of Food Microbiology 83,
233–244.
Ueckert, J.E., ter Steeg, P.F., Coote, P.J., 1998. Synergistic antibacterial action of
heat in combination with nisin and magainin II amide. Journal of Applied
Microbiology 85, 487–494.
Uhart, M., Ravishankar, S., Maks, N.D., 2004. Control of Listeria monocytogenes with combined antimicrobials on beef franks stored at 4 degrees
C. Journal of Food Protection 67, 2296–2301.
Ukuku, D.O., Fett, W.F., 2004. Effect of nisin in combination with EDTA,
sodium lactate, and potassium sorbate for reducing Salmonella on whole
and fresh-cut cantaloupe. Journal of Food Protection 67, 2143–2150.
Ulmer, H.M., Heinz, V., Gänzle, M.G., Knorr, D., Vogel, R.F., 2002. Effects of
pulsed electric fields on inactivation and metabolic activity of Lactobacillus
plantarum in model beer. Journal of Applied Microbiology 93, 326–335.
70
A. Gálvez et al. / International Journal of Food Microbiology 120 (2007) 51–70
Urso, R., Rantsiou, K., Cantoni, C., Comi, G., Cocolin, L., 2006a. Technological
characterization of a bacteriocin-producing Lactobacillus sakei and its use in
fermented sausages production. International Journal of Food Microbiology
110, 232–239.
Urso, R., Rantsiou, K., Cantoni, C., Comi, G., Cocolin, L., 2006b. Sequencing
and expression analysis of the sakacin P bacteriocin produced by a Lactobacillus sakei strain isolated from naturally fermented sausages. Applied
Microbiology and Biotechnology 71, 480–485.
Vaara, M., 1992. Agents that increase the permeability of the outer membrane.
Microbiological Reviews 56, 395–411.
Vega-Mercado, H., Martín-Belloso, O., Qin, B.-L., Chang, F.J., Góngora-Nieto,
M.M., Barbosa-Cánovas, G.V., Swanson, B.G., 1997. Non-thermal food
preservation: pulsed electric fields. Trends in Food Science & Technology 8,
151–157.
Verluyten, J., Schrijvers, V., Leroy, F., De Vuyst, L., 2002. Modelling the
behaviour of the potential meat starter culture Lactobacillus curvatus LTH
1174 as influenced by different environmental factors important for
European sausage fermentations. Proceedings of the 18th International
ICFMH Symposium FOOD MICRO 2002, pp. 167–172. Lillehammer,
Norway, August 18–23.
Verluyten, J., Messens, W., De Vuyst, L., 2003. The curing agent sodium nitrite,
used in the production of fermented sausages, is less inhibiting to the
bacteriocin-producing meat starter culture Lactobacillus curvatus LTH 1174
under anaerobic conditions. Applied and Environmental Microbiology 69,
3833–3839.
Verluyten, J., Messens, W., De Vuyst, L., 2004a. Sodium chloride reduces
production of curvacin A, a bacteriocin produced by Lactobacillus curvatus
strain LTH 1174, originating from fermented sausage. Applied and
Environmental Microbiology 70, 2271–2278.
Verluyten, J., Leroy, F., De Vuyst, L., 2004b. Effect of different spices used in
production of fermented sausages on growth of and curvacin A production
by Lactobacillus curvatus LTH 1174. Applied and Environmental
Microbiology 70, 4807–4813.
Vignolo, G., Fadda, S., de Kairuz, M.N., de R. Holgado, A.P., Oliver, G., 1998.
Effects of curing additives on the control of Listeria monocytogenes by
lactocin 705 in meat slurry. Food Microbiology 15, 259–264.
Wandling, L.R., Sheldon, B.W., Foegeding, P.M., 1999. Nisin in milk sensitizes
Bacillus spores to heat and prevents recovery of survivors. Journal of Food
Protection 62, 492–498.
WHO, 2002. Food safety strategic planning meeting: report of a WHO strategic
planning meeting, WHO headquarters, Geneva, Switzerland, 20–22 February
2001 (http://whqlibdoc.who.int/hq/2001/WHO_SDE_PHE_FOS_01.2.pdf).
World Health Organization, Geneva.
Wouters, P., Álvarez, I., Raso, J., 2001. Critical factors determining inactivation
kinetics by pulsed electric field food processing. Trends in Food Science &
Technology 12, 112–121.
Yamazaki, K., Yamamoto, T., Kawai, Y., Inoue, N., 2004. Enhancement of
antilisterial activity of essential oil constituents by nisin and diglycerol fatty
acid ester. Food Microbiology 21, 283–289.
Yang, R.G., Johnson, M.G., Ray, B., 1992. Novel method to extract large
amounts of bacteriocins from lactic acid bacteria. Applied and Environmental Microbiology 58, 3355–3359.
Young, L.L., Reverie, R.D., Cole, A.B., 1988. Fresh red meats: A place to apply
modified atmospheres. Food Technology 42, 64–66, 68–69.
Yuste, J., Fung, D.Y., 2004. Inactivation of Salmonella typhimurium and
Escherichia coli O157:H7 in apple juice by a combination of nisin and
cinnamon. Journal of Food Protection 67, 371–377.
Zapico, P., Medina, M., Gaya, P., Nuñez, M., 1998. Synergistic effect of nisin
and the lactoperoxidase system on Listeria monocytogenes in skim milk.
International Journal of Food Microbiology 40, 35–42.
Zhou, X.X.L., Wei Fen Li, W.F., Ma, G.X., Pan, Y.J., 2006. The nisin-controlled
gene expression system: construction, application and improvements.
Biotechnology Advances 24, 285–295.