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A Review on Utilization of Ozone in Food preservation

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 A Review on Utilization of Ozone in Food
preservation
H.Jegadeeshwar1, S.Vijay Shankar2, J.Premkumar3, T.V.Ranganathan4
Department of Food Processing and Engineering, Karunya University, Coimbatore 641114, India
Corresponding Author: premkumarj@karunya.edu.in

Abstract: Current sanitization technologies are crucial to maintaining

Ozone is a strong oxidant and potent disinfecting agent. Ozone
use may have many advantages in the food industry. There are
numerous application areas of ozone in food industry such as
food surface hygiene, sanitation of food plant equipment, reuse of
waste water, pest control in stored product and disinfection of
packaging materials. Ozone has a strong microbicidal action
against bacteria, fungi, parasites and viruses. Ozone application
is gaining attention due to the fact of zero residues on the
product and no need of aeration to remove the gas. This interest
was accompanied by a US governmental approval of ozone for
the safe use, in gaseous and aqueous phases as antimicrobial
agent on food. This paper presents available literature on ozone
in relation to its physico-chemical properties, microbial
destruction, food properties and ozone applicability, applications
in food industries.
Key words: ozone, antimicrobial action, food properties, ozone
applications, regulations, limitations and toxicity
I.INTRODUCTION
Consumers are increasingly aware of food borne illness and
risks associated with consumption of food. The food industry
is devoting considerable resources and expertise to the
production of wholesome and safe products to fulfil the
consumer expectations. Production of safe food includes
scrutinizing materials entering the food chain, suppressing
microbial growth and reducing the food borne pathogens by
processing which prevents post-processing contamination.
The presence of a processing unit operation aiming at
microbial destruction is of primary importance to achieve
safety and stability of food. Thermal treatments are
traditionally applied to pasteurize or sterilize food, generally
at the expense of its sensory and nutritional qualities. As
consumers increasingly believe fresh food is always healthier
than heat treated food, the industry is in need to seek the
alternative technologies to maintain most of the fresh
attributes, safety and storage stability of food [1]
the quality and enhancing the safety of fresh agricultural
commodities. These technologies were failed to meet the
consumer’s expectation due to the recent disease outbreaks in
US and many European countries. Safety of produce, which is
commonly treated with chlorine, is currently questionable
because of frequent disease outbreaks associated with these
products. Chemical disinfectants such as chlorine and
hypochlorite’s are commonly used as sanitizing agents in the
food industry. Treatments with 50-100 ppm free chlorine
solutions reduce initial contamination of vegetables, [2], but
these levels of the sanitizer may lead to discoloration and
production of off-flavours in fresh produce [3]. Food and food
related substances treated with chlorine could increase the
chance of cancer development and many other health related
issues. [4] [5] [6].
High pressure processing, pulsed electric field and high
intensity pulsed light are some of the emerging technologies.
Attention is now focused on ozone as powerful sanitizer that
may meet expectations of the industry. Ozone finds wide
application as a powerful disinfectant in many direct and
indirect food oriented operations and various other
environmental applications. Ozone as an oxidant has
numerous potential applications in the food industry because
of its advantages over traditional food preservation techniques
and superior antimicrobial properties.[7]
Ozone is an effective, chlorine alternative, sanitizer with
superior antimicrobial properties [8]. It is capable of
inactivating bacterial spores, molds, yeasts, protozoan cysts
and viruses at relatively low concentration and in short
exposure time when applied to pure cell suspensions [9].
Ozone has been tested on nearly every type of food during
storage and processing to improve the safety and to extend the
shelf-life of these products. The ability of ozone to inactivate
contaminant micro flora on food is variable; in some
instances, however, ozone decreased food micro flora more
than 5 log units [10].
 Ozone not only inactivates microbial contaminants, but also
reduces the level of pesticides, such as azinphosmethyl,
captan, formethanate-HCl and ethylenethiourea, on fresh
produce [11][12]. Ozonation reduces the amount of chemical
oxygen demand (COD) and biological oxygen demand (BOD)
of water used in washing and processing [13]. Thus, use of
ozone minimizes the accumulation of inorganic waste in the
environment [14]. Moreover, rapid decomposition to oxygen
and lack of toxic residues make ozone a favourable
environment friendly sanitizer. Consequently, this article
reviews the physico-chemical properties, microbial
destruction, food properties and ozone applicability, food
applications, regulatory issues, limitations and toxicity of
ozone.
II. PHYSICO-CHEMICAL PROPERTIES OF OZONE
Generally, ozone is formed in the stratosphere region
between the range of 15-35 km altitude by the action of short
ultraviolet (UV) solar radiation (<240 nm) on molecular
oxygen and a small portion of ozone (nearly, 10%) is
transported to the troposphere (<15 km altitude), but only a
small concentration of ozone occurs naturally at the Earth’s
surface [15].
Large amounts of the gas can be synthesized by generators
for industrial use. Ozone is a triatomic molecule (O 3) that is
considered to be an allotropic modification of oxygen. Ozone
has its own unique molecular structure containing resonance
hybrid of the four canonical forms having delocalized bonding
with mass of 48.
Pure ozone is a pale blue gas and bluish liquid with pungent
and characteristic odor. Ozone prevails in a gaseous state at
room and refrigeration temperature and it is partially soluble
in water. Ozone has an oxidation-reduction potential of 2.07 V
[16], which makes it the strongest oxidant currently available
for food applications. The density of ozone in the gaseous
state is 2.14 gL-1 at 0˚C and 101.3kPa [15], which is greater
than that of air (1.28 gL-1 ) under similar conditions.
Ozone is more stable in the gaseous than in the aqueous
phase [17]. Stability of dissolved ozone is affected by its
concentration, pH of the aqueous medium, exposure to UV
radiation, presence of radical scavengers, application of
turbulence, temperature and presence of organic matter and
heavy metal ions.
Except gold, platinum and iridium, ozone oxidizes metals to
oxides of the metals in their highest oxidation states. Ozone
converts oxides to peroxides, sulphides to sulphates, carbon to
carbon dioxide, and NH3 to NH4NO3, ozone at high
concentrations corrodes equipment, but such high
concentrations are found only inside the generator or in the
ozone-to-water contacting system. Materials used in food
processing are usually compatible with ozone at low
concentrations. Stainless steel is corroded less by ozone than
by chlorine [18]. Glass-lined steel [19] and steel with a
phosphate inside surface [20] are resistant to ozone and
suitable for maximum half-life storage of ozone.
Common plastics used in food processing are generally
resistant to ozone: these include polychlorotrifluoroethylene
(EFTFE), polydichlorofluoroethylene (PDFE),
polytetrafluoroethylene (PTFE), polyvinylidenefluoride
(PVDF), polyvinylchloride (PVC) and silicon tubing and
gaskets [19] [21].
Rubber in seals, pipes and other components reacts actively
with ozone, leading to a total disintegration into powder.
Synthetic rubbers, however, are resistant to ozone [14].
Lubricants like, fluorinated hydrocarbon have a good
resistance to ozone, but polymers of
monochlorotrifluoroethylene are the most better one than
other lubricants. The sealing materials on doors and windows
of fruit storage rooms should be made of ozone-resistant
synthetic materials [22].
III.MICROBIAL DESTRUCTION
Although ozone is highly effective against microorganisms
in pure cell suspensions, it is unlikely to be used directly in
food containing high ozone-demand materials. Organic
constituents of such food compete with microorganisms for
ozone, and thus high doses of this agent may be needed for
effective elimination of microorganisms.
Ozone, whether it’s in gaseous or in aqueous form which is
effective against numerous bacteria, molds, yeasts, parasites
and viruses at a low dose with short contact time [23] [24].
Sensitivity of ozone may vary according to the type of
microorganisms. Ozone was tested recently against Gram-
positive vegetative cells, bacterial spores, mold conidiospores
and yeast ascospores that are commonly isolated from fruits
which are known to spoil fruit juices. Several researchers
found that bacterial spores are the most resistant and bacterial
vegetative cells are the most sensitive to ozone [25]. Among
eight Bacillus spp. Tested, spores of B.stearothermophilus
were most resistant, while spores of B.cereus were most
sensitive to ozone.
The physiological status of the treated microorganism may
affect its susceptibility to ozone. Exponential-phase cells are
less resistant to ozone than are cells from the stationary phase.
The resistance of microorganism to ozone is greater with
natural micro flora on food than with microorganisms
frequently cultured in the laboratory [26].
ET.Ryser compared the efficacy of sanitizers against natural
and artificial contaminants on produce. In produce inoculation
studies, ozone (3ppm) treatments for 5 min decreased the
micro ial population 5 log units. n contrast, naturally
occurring ackground populations generally decreased 3 log
units after similar treatments. Dry microbial cells are
extremely resistant to gaseous ozone.
Ozone causes damage to the following cellular constituents:
1. Unsaturated lipids in the microbial cell envelope; 2. The
lipo-polysaccharides layer of Gram-negative bacteria; 3.
Intracellular enzymes and 4. Microbial genetic materials.
Earlier studies suggest that ozone reacts with microbial cell
membranes [27]. Ozone is believed to cause the oxidation of
lipids on the cell envelope of bacteria [28] [29]. Further,
oxidation leads to leakage of intracellular cell contents,
damage of genetic material and death [30] [31]. Ozone reacts
with cell dehydrogenase [32], DNA [33] and RNA [34].
Electron microscopic analysis revealed damage to cellular
structures after ozone treatment [9] [35][36]. Damage was
more pronounced in Gram-negatives, P.flurescens and E.coli
O157:H7, than in Gram-positives, Leuconostoc mesnteroides
and Listeria monocytogenes [9]. When treated with ozone
under similar conditions, Gram-positive bacteria seemed to
lose only some mucoid material outside the cell wall, whereas
Gram-negative cells tended to collapse outer cell membrane
and lose cellular fluids and components. Therefore, ozone at
low concentrations damages the outer membrane of Gram-
negative bacteria and thus causes dramatic changes in cell
structure. Similar concentrations of ozone cause less visible
damage to the cell wall of Gram-positive bacteria is evident,
but the agent causes intracellular damage and effectively
inactivates these cells [9].
Tests on injury of microorganisms by ozone varied
depending on the type of microorganisms, ozone
concentration and exposure time [9]. Maximum injury
occurred at ozone concentrations that caused mild lethality.
Ozonation is combined with pulsed electric field (PEF)
resulted in a synergistic lethal effect against E.coli O157:H7
and L.monocytogenes [37]. The combination of ozone and
PEF treatment cause cell injury followed rapid inactivation of
injured cells.
IV.FOOD PROPERTIES AND OZONE APPLICABILITY
Commodities with different chemical composition required
different ozone dose for effective sanitization. Fresh meat,
which contains high fat contents, for example, requires more
ozone than do fruits and vegetables, which contain low fat and
high carbohydrates. The nature of the surface of food
contributes substantially to the efficacy of ozone treatment.
Studies on microbial destruction of poultry carcasses revealed
that only small reduction in carcass microbial counts during
washing and ozonation. Strongly attached surface
microorganisms and those attached to areas that are not freely
exposed to ozone cannot be eliminated by mere dipping in
ozonated water. Therefore, when ozone is applied in food
processing, good contact between the sanitizer and the target
microorganisms on the treated food should be ensured.
Gaseous ozone treatment was not effective in decreasing
infection in inoculated wounds in apples. Only 3.8 mg L-1
aqueous ozone for 10 min is required to inactivate spores of
B.cinerea on the surface of non-injured tomato fruit but in
case of injured tomato fruit spores were not inactivated [38].
Plant tissues and extracellular biochemicals at wound sites
react with ozone and render it ineffective. Therefore, the
composition, surface texture, release of exudates and injuries
and wounds of food commodities are taken into account for
ozone treatment.
V.APPLICATION OF OZONE IN FOOD INDUSTRY
A. Meat and poultry
Shelf-life of meat and poultry depends on count and diversity
of the microbial population.. Meat and poultry products carry
the pathogenic microorganisms such as Campylobacter spp.,
Salmonella spp., L.monocytogenes and pathogenic E.coli.
Food contamination with these microorganisms can occur at
multiple steps along the food chain including slaughtering,
handling, storage and distribution [39].
Studies indicated that, variable results were obtained for
decontamination of beef and beef brisket fat by ozone and
also, the application of gaseous ozone minimized or prevented
growth of microorganisms on the meat surface [40].
Numerous investigators have demonstrated the microbicidal
efficacy and safety of ozone for use in washing poultry
carcasses, reconditioning poultry chiller water and sanitizing
hatchery equipments [41].
Salmonella enteric ser. Caused Enteritidis in shell eggs
which has serious public health implications. Several thermal
and chemical treatments have been developed to control or
eliminate this pathogen in eggs. Shell eggs were externally
contaminated with S.Enteritidis contained 106 CFU g-1 . Eggs
were treated with gaseous or liquid ozone for 1-20 min, at 4-
25˚C and 0-15 psi (0-100kPa). Gaseous ozone treatment
without pressure decreased the count of S.Enteritidis on shells
by 2.2-2.7 log units. Contaminated eggs were treated with
gaseous ozone for 10 min at 22-25˚C and 15 psi decreased
Salmonella population by more than 5 log units [42].
B. Fruits and Vegetables
Food production chain involved in number of steps with
multiple potential sources of contamination at each step. Main
causes for pre-harvest contamination of fresh fruits and
vegetables are dirty irrigation water, manure fertilizer and
improper working hygiene. Following harvest, improper
handling and storage use of contaminated wash water,
processing equipment and transportation facilities and cross
contamination from other produce contribute to the microbial
hazards associated with fresh fruits and vegetables. Following
cutting, shredding and slicing of fresh-cut fruits and
vegetables, the loss of surface integrity can lead to penetration
and rapid growth of microorganisms [43]. The following are
examples of studies and successful attempts to sanitize fresh
produce with ozone.
1. Apples
Apples are subject to contamination with pathogenic
microorganism on the farm. Use of contaminated apples to
produce unpasteurized apple juice and cider resulted in E.coli
O157:H7 food borne infections [44] [45] [46]. Inoculated
apples with E.coli O157:H7 prior to treatment with an
aqueous solution containing 21-28 mg L-1 ozone. Apples
washed in bubbled ozone were more effective than by dipping
in pre-ozonated water during decontamination treatment of
apples. Maximum decreases in surface counts of E.coli
O157:H7 were 2.6-3.7 log units, compared E.coli to unwashed
inoculated controls. However, counts of O157:H7 decreased
by less than 1 log unit in the stem calyx region by the ozone
treatment.
2. Date fruits and dried figs
Ozone was applied in gaseous form at three concentrations
(1, 3 and 5 ppm) for four different periods (15, 30, 45 and 60
min) on Iranian date fruits and the reduction in the total
bacterial count, Coliform, Staphylococcus aureus as well as
yeast/mold counts were examined. Treated samples shown the
lower microbial count than the untreated ones. Minimum of
one hour ozone treatment at 5 ppm indicated the considerable
reduction of coliform and S.aureus of date fruits but longer
exposure times are required for elimination of the total
mesophilic bacteria as well as yeast/mold [47].
Studies on measuring the effectiveness of gaseous ozone and
ozonated water on microbial flora and aflatoxin B 1 content of
dried figs. Dried figs were exposed to 13.8 mg L-1 ozone gas
and 1.7 mg L-1 ozonated water for 7.5, 15 and 30 min. Before
and after ozone treatments molds on dried figs were isolated
and identified. In both ozone treatments, AMB (Aerobic
Mesophilic Bacteria) was not exactly inactivated whereas
E.coli was completely destroyed at 7.5 min. Coli form and
yeast were also destroyed at 7.5 and 15 min in ozonated water.
Dried figs were artificially contaminated with aflatoxin B 1
samples also treated with gaseous ozone and ozonated water
for 30, 60 and 180 min. Results indicated that gaseous ozone
was more effective than ozonated water for reduction of
aflatoxin B1[48].
3.Lettuce, Spinach and Parsley
Tests on ozone against natural contaminants in fresh lettuce
and results were compared with those obtained from chlorine
treatment [26]. Ozone (1.3 mM) or chlorine (1mM)
inactivated mesophilic and psychrotrophic bacteria by 1.4 and
1.8 log units in 3 min, respectively. In another experiment,
shredded lettuce was treated with gaseous ozone, or mixed
with aqueous solution of ozone (1:20 w/w) with or without
bubbles. Results show that bubbling gaseous ozone in water,
combined with stirring, is the most effective ozonation
method for shredded lettuce. In a recent study, the application
of 2 ppm ozonated water treatment for 2 min was found to be
the optimum processing conditions for ozone disinfection of
green leaf lettuce than the chlorine and organic acid
treatments in maintaining the sensory quality during the 9
days of storage period [49].
Research on quality of fresh pre-packaged spinach
inoculated with Escherichia coli O157:H7 6460 treated in
packaging with ozone generated in air and oxygen. Samples
were treated for 5 min and stored at room temperature (22˚C)
or refrigeration (5˚C) for 0.5, 2 and 24 h. All treated samples
showed reductions in E.coli O157:H7 populations with the
largest reductions (3-5 log10 CFU/leaf) after 24 h of storage.
[50].
Vegetables like lettuce, spinach and parsley were treated
with aqueous ozone (12 mg L-1) , gaseous ozone (950 µL L-1,
20 min) and chlorinated water (100 mg L-1) to inactivate the
inoclulated Escherichia coli and Listeria innocua. Chlorine
and ozone washes resulted in average log reductions of
2.9±0.1 and 2.0±0.3 for E.coli, while the efficiency of ozone
(2.2±0.1 log) was very close to that of chlorine (2.3±0.1 log)
on L.innocua. Gaseous ozone treatment indicated the 1.0-1.5
log reductions in the numbers of both microorganisms but
caused significant losses in important bioactive compounds of
parsley [51].
4.Alfalfa sprouts
Alfalfa sprouts received great attention in recent years owing
to the incidence of pathogens in this product and associated
disease outbreaks. Treatment of alfalfa sprouts with ozone for
5 min decreased their microbial load by 1.2 log units. An
additional 0.8 log unit decrease in population was observed
when ozonation was accompanied with agitation. As an
alternative ozonation process, alfalfa seeds were pre-treated
with ozone and the microbial count during seed germination
and growth was monitored. Additionally, the sprouts were
watered twice daily using ozonated water during the growth
period. he initial micro ial count on the seeds was 105 CFU
g-1 and ozone treatment decreased the count y 2 log units.
he difference in count etween ozone-treated and non-treated
seeds diminished during the growth period and the population
in oth treatments reached 109 CFU g-1 after 4 days of seed
incubation.
An end treatment consisting of bubbling ozone into the
sprouts for 2.5 min decreased the microbial count from 5.0 x
109 to2.0 x 107 CFU g-1. Alternatively, the sprouts were
placed in ozonated water (30-32 ppm) and stirred for 20 min;
the average count decreased to 4.8 x 107 CFU g-1.[52].
C. Fruit Juice Ingredients
Survival of heat-resistant bacteria and fungi during
pasteurization of juice can be a serious concern to citrus
processors, ozone is potentially useful in inactivating heat-
resistant spores in juice components with low ozone demand.
Therefore, the efficacy of ozone against spores of
Alicyclobacillus acidocaldarius, Neosartorya fischeri and
Zygosaccharomyces bailii, which are known to be problematic
in juices, was tested in selected juice ingredients [53]. Fruit
juice components, high fructose corn syrup (HFCS), orange
juice concentrate (OJC), and pine apple juice concentrate
(PJC), spiked with cells or spores (c, 10 7 mL-1), were treated
with gaseous ozone. The sensitivity of spores to ozone varied
depending on the type of spores and the juice component.
Inactivation of 5 log units of A.acidocaldarius spores require
0.31, 0.28 and 0.41 mg ozone per mL of spore suspension in
HFCS, OJC and PJC respectively. The ozone dose for
inactivating 5 log units of N.fischeri spores was 0.12-0.51 mg
mL-1. In order to achieve a decrease of 5 log units of Z.bailii
spores, 0.04-0.24 mg mL-1 was needed, depending on the juice
component. Inactivation of Z.bailii spores by ozone was faster
in HFCS than in other juice components even though HFCS
has higher solids than the other components. In conclusion,
the ozone dosage from 0.04 to 0.5 mg mL-1 decreases up to 5-
log unit of A.acidocaldarius, and N.fischeri spores were three
to four times more resistant to ozone than were Z.bailii spores.
Apple juice samples were ozonated with concentration (1-
4.8% w/w) and processing time (0-10 min). Treated apple
juice colour, rheological properties and phenol content were
observed to be significantly influenced by ozonation [54].
D. Fish processing and storage
Ozone was tested for decontaminating shrimp, mussels and
various fish such as jack mackerel, sockeye salmon, Japanese
flounder. The antimicrobial efficacy of ozone was equal to or
better than that of chlorine in some applications studied [55]
[56]. Ozone reduced disease incidence in hatcheries with less
mortality and shorter growth cycles. The desirability of ozone
use in processing fish products for human consumption limits
until the development of adverse sensory effects and harmful
oxidation by-products. Ozone treatment of shrimp meat
extract, however, was ineffective against microorganisms in
the product [57]. Ozone may have reacted with ozone-demand
substances in the extract, instead of microbial cells.
E. Dry food and food ingredients
Gaseous ozone can eliminate Bacillus spp. and Micrococcus
spp., which are dominant in cereal grains, peas, beans and
spices, by up to 3 log units, depending on concentration,
temperature and relative humidity [58]. The surface area of
the dry food is an important factor for the effectiveness of
ozone treatment. Cereal flour and ground pepper, for example,
require higher concentration of ozone and longer contact time
than whole cereal and pepper to achieve the same degree of
microbial inactivation [59] [60]. In addition to antimicrobial
effects, ozone destroys or greatly reduces aflatoxins from
peanut [61]. and cottonseed meal [62] and oxidizes odors
produced during dehydration of onions and garlic [63]. Ozone
treatment may cause lipid oxidation, decrease amino acid (e.g.
thiamine) content and essential oil content and contribute
negative effects on the sensory quality of some dry food.
F. Food Proteins
Ozonation of proteins cause oxidation of the polypeptide
backbone of the protein, peptide bond cleavage, protein-
protein cross-linking and a range of amino acid side chain
modifications. Amino acids were oxidized by reactive oxygen,
the major aromatic amino acids tyrosine, tryptophan,
phenylalanine, the sulphur containing amino acids cysteine,
methionine and aliphatic amino acids arginine, lysine, proline
and histidine appear especially sensitive to oxidation. The
sequence of amino acids that are condensed in a chain by
amide bonds, representing main chain backbone of proteins.
Helices, sheets, coils and folded branches which define the
secondary and tertiary structure of proteins seem to be
modified by the reaction of ozone with proteins. The action of
ozone converts the thiol group of cysteine into disulphides
which denature the protein and change its solubility [64].
Proteins, as macromolecules, perform important roles in
functionality in food systems. The growing demand for
proteins as important ingredients in many food formulations
or in pharmaceutical and industrial mixtures has created a
necessity for proteins with specific and consistent functional
properties [65].
Protein sample like whey protein isolates and egg white
proteins were treated with ozone gas (4.5 ppm in aqueous
medium, 15 min at 8˚C) to study the functional properties like
foaming and emulsion. Foaming ability and foam stability of
WPI and egg white proteins increased. Increased ozone
concentration increases the foam ability. However, ozone
treatment reduced the emulsion stability of WPI and solubility
of both WPI and egg white proteins; whereas, the percentage
of reactive amino acids to ozone in egg white proteins seemed
to be higher than that in WPI. Therefore, ozonation represents
an interesting alternative technique for enhancement of
functional properties and controlled dosage prevents the
undesirable oxidation of proteins [66].
G. Packaging material and food contact surfaces
Food packaging materials contain less number of microbial
contaminants, but they may survive conventional
decontamination processes and cause food spoilage.
Currently, sterilization of packaging materials is achieved by
several methods including heat, hydrogen peroxide and UV
radiation [67] [68] [69]. Sterilization by hydrogen peroxide is
a tedious and variable method [68], and unacceptable levels of
hydrogen peroxide residues may remain and interact with
some of the polymer in the packaging material. Sanitization of
equipment and food-contact surfaces is essential for safety and
quality of processed food. When bio films develop on wet
food contact surfaces such as those made from stainless steel,
the microorganisms inside the bio films are usually protected
from sanitizers. Hence alternative methods for
decontamination of packaging materials and stainless steel are
being sought.
A multilaminated aseptic food packaging material and
stainless steel were treated with ozone to inactivate natural
contaminants; bacterial bio films (P.fluorescens) and dried
films of B.subtilis spores [70]. Sterilized multilaminated
packaging material, which contained a low natural, mostly
mesophilic, contaminants was achieved when it was treated
with 5.9 mg L-1 ozone in water for 1 min. Counts of bacteria
in dried films decreased by 4.6-5.1 log units for the
multilaminated packaging material and 5.5 log units for
stainless steel when treated with 5 mg L-1 aqueous ozone for 1
min. Dried films of spores (108 per 6.3 cm2 surface) decreased
below detection (> 10 spores per 6.3 cm2 surface) by
application of 13 mg L-1 aqueous ozone for the multilaminated
packaging material and 8 mg L-1 in the case of the stainless
steel. Repeated exposure to ozone and agitation during the
treatment were needed to decrease effectively the population
in bio films. Ozone inactivated P.fluorescens in bio films
more effectively on stainless steel than on the multilaminated
packaging material. It is concluded that ozone is an effective
sanitizer with potential applications in the decontamination of
packaging materials and equipment food-contact surfaces.
H. Pesticides on agricultural commodities
Use of pesticides on fruits and vegetables is crucial for
insuring high quality products. Residues of these pesticides
could be removed to assure the health of consumers. The use
of chlorine, ozone, and chlorine dioxide and peracetic acids as
postharvest treatments has been effective in remediating
several different pesticides on apples [11] [71] [12]. These
same treatments also have the added benefit of reducing
microbial populations on the surface of fruits and vegetables
by up to 5 log units [24]. Organic food processors and
consumers of organic food also are concerned about the
presence of chemical residues, including chlorine and
chlorinated by-products. However, these residues levels are
likely reduced by ozonation in solution.
Degradation of the four pesticides (methyl-parathion,
parathion, diazinon and cypermethrin) by dissolved ozone
[72]. Solid-phase micro-extraction (SPME) and GC-MS
results indicated that dissolved ozone (1.4 mg/l) was effective
to oxidize 60-99% of methyl-parathion, cypermethrin,
parathion and diazinon in aqueous solution in 30 min and the
degradation was mostly completed in the first 5 min. Ozone
was mostly effective in cypermethrin removal (>60%).
Ozone is an effective alternative to traditional insecticides, as
a viable bio-fumigant against a range of insect pests,
microorganisms and mycotoxins [73].
 VI. REGULATORY STATUS
Many European and Asian countries were permitted the use
of ozone. Ozone has been safely and effectively used in water
treatment for several decades in European countries [74].
Industrial use of ozone in the United States was limited to the
removal of metal ions, colour, taste and odors in water [75]. In
1975, the US-FDA permitted the use of gaseous ozone up to
0.1ppm in meat-aging coolers and in 1982, 0zone was
approved as a Generally Recognized As Safe (GRAS)
substance for treatment of bottled water [76]. According to the
1982, other uses of ozone (e.g.in food) require a food additive
petition. Subsequently, the United States Department of
Agriculture (USDA) permitted the use of ozone in poultry
chiller water [77]. The Electric Power Research Institute and
Agriculture and Food Technology Alliance submitted a direct
food additive petition to FDA so that ozone could be used in
food processing without limitations [78]. In response to the
petition, the FDA amended the food additive regulations to
provide for the safe use of ozone in gaseous and aqueous
phases as an antimicrobial agent on food, including meat and
poultry [79]. This approval should boost a broad use of ozone
in food processing.
VII. LIMITATIONS, TOXICITY AND SAFETY
The reactivity of ozone and the level of product quality may
limit uses of this sanitizer in food processing. Sensory
attributes may be altered, depending on the chemical
composition of food, ozone dose and treatment conditions.
Surface oxidation of food by ozone results in discoloration,
undesirable odors and oxidative spoilage. Ozone decreased
vitamins and amino acids contents and increased lipid
oxidation and activity of some enzymes such as superoxide
dismutase, ascorbate peroxidise and glutathione reductase in
lettuce leaves [80].
The acute and chronic effects of excessive exposure to ozone
were investigated [81]. Small concentrations of ozone gas in
air (0.3 ppm) may cause discomfort to susceptible people. [82]
the reported data that shows as little as 0.02-0.04 mg L-1 can
be detected by humans and 0.1 mg L-1 is objectionable to all
normal humans because of irritation to the nose, throat and
eyes. The respiratory tract is the primary site of ozone
toxicity, which may cause pulmonary congestion. Symptoms
resulting from exposure to ozone include headaches and
dryness of the throat, nose and eyes [81] [83]. Some
researchers demonstrated that repeated exposures have
progressively lesser effects, suggesting that tolerance may
develop [84]. Results indicated that with an hour exposure
symptomatic, irritant, toxic and irreversible lethal effects can
be induced by ozone concentrations of 2, 4, 15 and 95 ppm,
respectively [85]. Ozone may generate toxic substance when
tissue proteins, unsaturated fatty acids or other components of
food are oxidized [86]. However, immersion of shrimp meat
in saline containing 5 mg L-1 ozone, for 120 min, did not
generate mutagens in the product [57].
In a recent direct food additive petition, [78] ozone can be
used comparatively safely in industrial applications.
According to the author, ozone is detected by human olfactory
senses at concentrations as low as 0.01 ppm, and higher
concentrations of the agent exert only temporary acute
symptoms in humans. Since ozone has a high oxidation
potential, effective disinfection can be accomplished with less
concentration and shorter exposure time, compared to other
oxidizing agents. Ozone is manufactured on-site at relatively
low concentrations and pressures; therefore, an uncontrolled,
widespread and immediate release of large quantities of ozone
is not possible. The relatively short half-life of ozone
minimizes the persistence of the gas in the environment, and
ozone breakdown product (diatomic oxygen) causes no harm
[78].
In the United States, current permissible exposure level-time
weighted average (PEL-TWA) for ozone exposure in the work
place environment is 0.1 ppm, as recommended by the
American Conference of Governmental Industrial Hygienists
[87]. And adopted by the United States Occupational Safety
and Health Administration (OSHA). Susceptible individuals
can be exposed continually to this ozone concentration during
a normal 8-h day/ 40-h working-week without adverse effects
[88]. The short term exposure limit is 0.3 ppm for an exposure
less than 15 min and four times per day.
VIII. CONCLUSION:
Increased global production, distribution and consumer
awareness on health benefits and risks associated with
consumption of food. Novel industrial applications and
increased demand of new alternative technologies made ozone
as a prominent technology and with new regulatory actions,
making its use in the food industry easier and applicable in a
wide range of cases. This review paper demonstrated that
ozone is an effective greener alternative to number of
conventional disinfection methods in wide range of food
processing operations. Ozone offers unique advantages for
food materials with minimal or desired effects on the
physicochemical properties. Further research is still needed to
explore new applications for ozone and to best utilize the
unique features of this sanitizer.
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