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US Patent 6485769 – Food disinfection using ozone
US Patent Issued on November 26, 2002

Abstract
Methods and apparatus are provided for decreasing the bacteria count of a
food commodity without affecting its overall organoleptic quality (taste,
odor, and color). This is accomplished using a treatment fluid comprising
ozone, which is injected into a treatment chamber containing the food
commodity. Some water is preferably added to obtain better contact of the
ozone with the food by forming a thin film of ozonated water on the food
surface. Spices and/or other ingredients may preferably be added with the
water. The food is placed in a tumbler and the tumbler is set in motion.
During treatment good contact between the treatment fluid and the food
commodity is obtained by reversibly oscillating the tumbler. A log
reduction of 40% or more in bacteria count may be obtained as compared
without the ozone.
Claims
What is claimed is:
1. A method of decreasing bacterial count in raw meat, the method
comprising the steps of:
(a) placing the meat in a container, and sealing the container;
(b) evacuating the container to a vacuum of at least about 2 inches Hg to
about 25 inches Hg;
(c) generating a fluid stream containing ozone;
(d) contacting the meat with the fluid stream containing ozone while
agitating the container, said contacting being for a time sufficient to
reduce the bacterial count by at least 40% compared to without ozone.
2. The method of claim 1, wherein the fluid stream containing ozone is
injected into the container until a pressure ranging from just below to
just above 1 atmosphere pressure is reached in the container.
3. The method of claim 1, wherein a small amount of water is injected into
the container just prior to or during step (d) so that a thin film of
ozonated water is produced on the meat surface.
4. The method of claim 1 further comprising a step (e) of evacuating the
container to remove residual ozone from the container.
5. The method of claim 1, wherein steps (b) and (c) are performed
simultaneously.
6. An apparatus for use in decreasing the bacterial count in raw meat, the
apparatus comprising a container which may be sealed after the meat is
placed therein, the container capable of being agitated with agitation
means; a conduit leading to an evacuating means, the evacuating means
being able to provide a vacuum to the container of at least about 10
inches of mercury, preferably at least 25 inches of mercury; an ozone
generating device connected to the container via injector means able to
provide an ozone containing fluid stream to the container; and vent means
for venting the ozone containing fluid until it is needed
7. A method for reducing microorganisms on a food commodity comprising:
providing a tumbler;
introducing an aqueous solution into the tumbler;
placing the food commodity in the tumbler;
introducing a fluid comprising ozone into the tumbler; vibrating the
tumbler in oone of a vertical direction, a horizontal direction and both a
vertical direction and horizontal direction; and maintaining said tumbler
vibration for a time sufficient to reduce the bacterial count by at least
40% compared to without ozone.
8. The method of claim 7, wherein introducing a fluid comprising ozone
comprises introducing at least about 0.001 gram of ozone per gram of food
commodity.
9. The method of claim 7, wherein introducing a fluid comprising ozone
comprises introducing at least about 0.5 mg of ozone per gram of food
commodity.
10. The method of claim 7, wherein introducing a fluid comprising ozone
comprises introducing at least about 1.0 mg of ozone per gram of food
commodity.
11. The method of claim 7, wherein introducing a fluid comprising ozone
comprises introducing at least about 2.0 mg of ozone per gram of food
commodity.
12. The method of claim 7, wherein introducing a fluid comprising ozone
comprises introducing at about 0.4 to about 0.6 mg of ozone per gram of
food commodity.
13. The method of claim 7, wherein the predetermined period of time
comprises about 2 minutes to about 360 minutes.
14. The method of claim 7, wherein introducing an aqueous solution
comprises introducing water.
15. The method of claim 14, wherein introducing an aqueous solution
comprises introducing an aqueous ozone solution.
16. The method of claim 14, wherein introducing an aqueous solution
comprises introducing about 0.001 to about 0.5 grams of water per gram of
food commodity.
17. The method of claim 7, wherein providing a tumbler comprises providing
a tumbler having a rotational axis, and wherein setting the tumbler in
motion comprises rotating about the rotational axis.
18. The method of claim 17, wherein rotating the tumbler comprises rotating
the tumbler at about 0.5 to about 30 revolutions per minute.
19. The method of claim 17, wherein rotating the tumbler comprises
alternatingly rotating the tumbler in a clockwise direction and in a
counterclockwise direction at about 1 to about 30 revolutions per minute.
20. The method of claim 19, wherein each revolution comprises one clockwise
rotation and one counterclockwise rotation, and wherein a clockwise
rotation and a counter clockwise rotation comprise about a 180°
rotation of the tumbler with respect to a reference point at a perimeter
of the tumbler.
21. The method of claim 7, wherein providing a tumbler comprises providing
a tumbler having a shaft, and wherein setting the tumbler in motion
comprises axially oscillating the tumbler along the shaft.
22. The method of claim 7, wherein the tumbler is maintained at a
temperature of about -200° C. to about 50° C.
23. The method of claim 22, wherein the tumbler is maintained at a
temperature of about 0.1° C. to about 25° C.
24. The method of claim 7, wherein introducing an aqueous solution and
introducing a fluid comprising ozone are performed simultaneously after
placing the food commodity in the tumbler.
25. The method of claim 7, wherein placing the food commodity in the
tumbler comprises placing a food commodity selected from the group
consisting of meat, poultry, fish, seafood, fruits and vegetables.
26. A method for reducing microorganisms on a food commodity comprising:
providing a tumbler having a rotational axis;
purging the tumbler with a purge gas;
placing the food commodity in the tumbler;
introducing about 0.125 to about 2.0 grams of a gas comprising ozone per
gram of meat and introducing an aqueous solution into the tumbler;
rotating the tumbler about the rotational axis; and
purging the tumbler with the purge gas.
27. The method of claim 26, wherein placing the food commodity in the
chamber comprises dividing the food commodity into portions and placing
one or more portions into chambers of a fixture and inserting the fixture
into the tumbler.
28. The method of claim 26, wherein the purge gas is selected from the
group consisting of nitrogen, carbon dioxide, air, argon and mixtures
thereof.
29. The method of claim 26, wherein the pressure within the tumbler is
controlled to a vacuum pressure of about 2 to about 25 inches Hg while
rotating the tumbler.
30. The method of claim 26, wherein placing the food commodity in the
tumbler comprises placing a food commodity selected from the group
consisting of meat, poultry, seafood, fruits and vegetables.
31. The method of claim 26, wherein rotating the tumbler comprises
alternatingly rotating the tumbler in a clockwise direction and a
counterclockwise direction for about 2 minutes to about 360 minutes.
Description
FIELD OF THE INVENTION
The present invention relates, in general, to methods for treating food
products and, more particularly, to methods for reducing pathogenic
microorganism populations on food commodities during food processing.
BACKGROUND
Microbial outgrowth is a primary cause of food spoilage. The presence of
pathogenic microorganisms on food products can potentially led to
food-borne outbreaks of disease and can cause significant economic loss to
food processors. The need to delay the onset of spoilage has lead the food
processing industry to seek effective means for disinfecting food products
in order to ensure food safety. Currently, food manufacturers use several
different technologies to eliminate, retard, or prevent microbial
outgrowth. For example, techniques such as heating, irradiation, and
application of chemical agents are currently in use. However, the
effectiveness of a particular technology can depend on the particular food
product and type of microorganism present on the food product.
Additionally, certain chemical agents can have a deleterious effect on
human health. For example, chlorine has been widely used as a sanitizer
for many years, however, chlorine can produce toxic by-products, such as
chloramines and trihalomethanes.
Another widely used chemical agent is ozone (O3). Ozone is a very
strong oxidizing agent, having an oxidation potential more than 1.5 times
that of chlorine and approximately 1.3 times that of hydrogen peroxide.
Ozone is normally produced by irradiating an oxygen-containing gas with
ultraviolet light or corona discharge. Ozone has been widely used as a
disinfectant in the food industry for many years. Processes have been
developed that use gaseous ozone to sterilize and disinfect food products.
Although applying gaseous ozone to food products can be an effective means
of controlling microbial outgrowth, an effective method of applying the
ozone to the food product must be available.
To address the public health concern associated with food contamination,
development of more effective processes to ensure safe and wholesome food
production has become a main strategy for the food industry. Processes for
the separate application of ozone and steam in a vacuum and pressure
regulated environment have been developed to improve the effectiveness of
ozone at killing the bacteria present on food. In addition, continuous
processes have been developed that include spraying ozone gas and a
mixture of ozone and water directly onto animal carcasses immediately
after slaughter.
Despite recent development of ozone application technology, food
contamination by pathogenic microorganisms continues to be a significant
health problem. According to recent statistics from Centers for Disease
Control (CDC), there are approximately 76 million cases of food borne
illness in the United States annually. The most common food-associated
pathogens are: Norwalk-like viruses, Campylobacter jejuni, and Salmonella.
Escherichia coli 0157:H7 and Listeria monocytogenes can also cause severe
illness. As the world population increases, the demand for processed food
will also increase and food borne illness is more likely to become an even
greater problem. To address this public health concern, development of
more effective processes to ensure safe and wholesome food production
continues to be an important objective of the food industry.
BRIEF SUMMARY
One primary objective of this invention is to treat a food commodity, such
as meat and, preferably, chicken breasts, with a fluid comprising ozone,
which can be a gas, a liquid, or liquid and gas mixture, in a sealed
container to obtain a lower bacterial count without affecting its
organoleptic properties (overall quality of taste, odor, and color). The
amount of ozone injected is preferably as small as possible, while at the
same time, showing a significant log reduction on bacteria count. Some
water is preferably added to obtain better contact of ozone with the food
commodity by forming a thin film of ozonated water on the surface of the
food commodity. Spices and/or other ingredients may be added with the
water.
In one preferred method, the invention generally includes, placing a food
commodity, such as meat, poultry, fish, seafood, fruits and vegetables in
a sealed container, such as a treatment chamber, which can be tumbler, or
an apparatus configured to receive a tumbler. A vacuum is generated in the
treatment chamber and either before or during vacuum generation, a
treatment fluid comprising ozone is produced to obtain a steady stream of
a treatment fluid flowing through a conduit, which is vented through an
exhaust system.
Once a vacuum of at least 2 inches Hg, preferably a vacuum of around 25
inches of Hg is reached, the treatment fluid is injected into the
treatment chamber. The treatment fluid is injected until the pressure
inside the chamber rises to about atmospheric pressure. In certain
preferred embodiments of the method, an excess of ozone is introduced in
the treatment chamber at a pressure slightly above atmospheric pressure,
or alternatively, a residual vacuum may be beneficial for certain
treatment processes.
In one embodiment of the invention, the treatment fluid contacts the food
commodity through action of a tumbler. The tumbler can function as the
treatment chamber, or the tumbler can be positioned within a treatment
chamber. Preferably, a small amount of water is injected to create a thin
water film rich in ozone. This treatment is carried out for a time
sufficient to obtain good disinfection, preferably about 50% reduction in
bacterial count, without affecting food quality (color, odor, taste).
The present invention contemplates a number of different motion patterns of
the tumbler. During the contact period, the tumbler is preferably rotated
about a rotation axis. Alternatively, the tumbler can be agitated by
alternatively rotating the tumbler about a rotational axis in both a
clockwise direction and a counterclockwise direction for a predetermined
period of time. In another embodiment of the invention, the tumbler is
axially oscillated along a shaft. In yet another embodiment, the tumbler
is vibrated in a vertical or horizontal reciprocating motion or both.
After the required time, the treatment fluid is purged from the treatment
chamber so that it can be opened safely. In one embodiment, the treatment
chamber and associated gas lines are purged by flushing with an inert gas,
such as carbon dioxide and the like.
BRIEF DESCRIPTION OF THE DRAWING
FIG. 1 illustrates a schematic diagram illustrating an apparatus arranged
in accordance with the invention;
FIG. 2 illustrates a schematic diagram of an apparatus arranged in
accordance with a more detailed embodiment the invention;
FIG. 3 illustrates a tumbler arranged in accordance with one embodiment of
the invention;
FIG. 4 is a plot illustrating microorganism population versus exposure time
for chicken coupons processed in accordance with the invention;
FIG. 5 is a plot illustrating microorganism population versus ozone
concentration for chicken coupons processed in;
FIG. 6 is a histogram illustrating the survival of microorganism
populations for various microorganisms on chicken coupons after treatment
in accordance with the invention;
FIGS. 7 and 8 are comparative histograms illustrating the survival of the
total microorganism population on chicken breasts processed in accordance
with the invention and in accordance with an ozone-free process.
DESCRIPTION OF PREFERRED EMBODIMENTS
Referring now to FIG. 1, an apparatus in accordance with one embodiment of
the invention is comprised of a sealed container 2, which takes feed from
an ozone generator 4, via conduit 6, ozone valve 8, fittings 9 and 10,
conduit 18, and valve 20. A fluid stream generated by ozone generator 4
includes from about 1% to about 20% ozone, when oxygen or air is fed to
the ozone generator 4. The ozonated fluid proceeds through valve 20,
through a fitting 24 having a negative pressure gauge 22, conduit 28, and
conduit 26 connected to the sealed container 2. The apparatus also
includes a vacuum supply valve 30, which is connected to a vacuum pump or
other means for creating a vacuum 32. Vacuum 32 exhausts into an ozone
destruction unit 34. A conduit 12 and valve 14 connects to an exhaust vent
16 through ozone destruction unit 34. A vent line 36 branches from conduit
6 at fitting 9 and is gated by valve 37 and is connected to an atmosphere
intake 38.
In order to practice the methods of the invention, one preferred method
comprises:
1. Creating a vacuum on a sealed container 2. Either before or during
vacuum generation, ozone is produced, either as a gas, a liquid or a
mixture of a gas and a liquid, to obtain a steady stream flowing through
conduit 6, which is vented through the exhaust through fittings 10, 12,
14, and 16.
2. Once a vacuum of at least 10 inches Hg, preferably a vacuum of around 25
inches of Hg is reached, vacuum valve 30 is closed, as well as ozone
exhaust valve 14, and the ozone containing fluid is injected into sealed
container 2 by opening rapidly valve 20 on the ozone line 18. Ozone
containing fluid is injected until the pressure inside sealed container 2
rises to about atmospheric pressure (about 1 atmosphere at sea level). In
certain preferred embodiments of the method, an excess of ozone with
pressure in the sealed container slightly above atmosphere or a residual
vacuum, may be beneficial for certain disinfection processes.
3. The ozone containing fluid is left in contact with the meat and
preferably a small amount of water is injected to create a thin water film
rich in ozone containing fluid. This condition is carried out for a time
sufficient to obtain good disinfection, preferably about 50% reduction in
bacterial count, without affecting meat quality (color, odor, taste).
During the contact period, the container is preferably agitated.
4. After the required time, vacuum pump 32 is turned on, valve 30 opened to
purge the ozone containing fluid from the container 2, and valve 37 is
opened to vent conduit 18 to atmospheric pressure so that sealed container
2 can be opened safely. The food commodities, such as a piece or pieces of
meat, are then shipped for bacterial analysis.
At the customer site, for example, sealed container 2 is preferably the
tumbler they are using to inject water and spices in batch processes on
large portions of chicken breasts to obtain the desired taste (these
tumblers can vary greatly in size).
In cases where an existing tumbler is to be employed, as described in the
previous paragraph, the door seal is preferably modified to include an
ozone compatible material, such as polyvinylfluoride (PVF),
polyvinylchloride (PVC), or Viton fluoroelastomer (Viton is a trademark of
DuPont de Nemours). The ozone would be injected through an ozone injector
as is commonly known in various arts, for example, the water purification
art.
As is commonly known, ozone generators, (such as those available from
Ozonia, North America, Elmwood Park, New Jersey), utilize an air or oxygen
feed gas, which converts from about 1% to about 20% of the oxygen to
ozone. For an oxygen feed gas to ozone generator 4, the stream 6 will
preferably have from about 1% to about 20% ozone, with the balance being
oxygen and traces of nitrogen and other air gases. The fluid produced by
the ozone generator can also contain one or more cryogens such as
nitrogen, argon, xenon, neon, helium, and the like.
An apparatus 40 in accordance with a more detailed embodiment of the
invention is illustrated in FIG. 2. Apparatus 40 includes, as major
components, those illustrated in the apparatus of FIG. 1, together with
additional components for pressurization and evacuation of the treatment
chamber. Apparatus 40 is merely an example of a laboratory-scale or pilot
plant system. Accordingly, a number of different equipment arrangements
for treating a food commodity in accordance with the invention are
possible. Those skilled in the art will appreciate that numerous
modifications can be made, and that an industrial-scale processing
facility may include other components, and that those components may be
arranged differently from that illustrated in FIG. 2.
In apparatus 40, a barrel-type treatment chamber 42 receives treatment gas
through a gas feed line 43 that is connected to a pressurized gas delivery
system 44. Gas feed line 43 is equipped with a ball valve 45. In one
embodiment of the invention treatment chamber 42 has a volume of about 5.6
liters.
Gas delivery system 44 includes an ozone generator 46 and pressure vessels
48 and 50. An exhaust system 52 is coupled to gas delivery system 44.
Exhaust system 52 includes an ozone analyzer 54 and gas scrubbers 56 and
58. A vacuum line 60 is connected to gas feed line 43 and operates to
regulate the pressure within treatment chamber 42.
Ozone generator 46 receives oxygen or air and outputs a gas comprising
ozone to pressure vessel 48 through an output line 62. Output line 62 is
equipped with a ball valve 63. Preferably, ozone generator 46 is a
water-cooled generator similar to that described in reference to FIG. 1.
In one embodiment of the invention, ozone generator 46 is an OZAT CFS-2
model generator (Ozonia Ltd., Switzerland). A desired gas pressure is
maintained in pressure vessel 48 by injecting carbon dioxide, or another
inert gas, into output line 62 immediately upstream from pressure vessel
48. Pressure vessel 48 is coupled to pressure vessel 50 through a gas line
64. Gas line 64 is equipped with a ball valve 66 that provides the
capability to isolate pressure vessel 48 from pressure vessel 50.
Ozone analyzer 54 monitors the ozone concentration in the treatment gas
within pressure vessel 50. In one embodiment, ozone analyzer 54 is a model
H I ozone analyzer (IN-USA, Needham, Mass.). Ozone analyzer 54 receives
sample portions of the treatment gas in pressure vessel 50 through a gas
sample line 68. Gas sample line 68 is equipped with both a ball valve 70
and a needle valve 72 to enable the periodic pressure controlled delivery
of gas samples to ozone analyzer 54. Ozone analyzer 54 outputs exhaust gas
to gas scrubber 58 though an exhaust line 74.
Gas scrubber 56 is connected to gas sample line 68 by a vent line 76. Vent
line 76 is equipped with a ball valve 78. Vent line 76 provides for the
release of treatment gas from pressure vessel 50. The gas pressure within
pressure vessel 50 can be controlled by properly adjusting valves 66 and
78 and by adjusting the volume of carbon dioxide or other inert gas
injected into pressure vessel 48.
The flow of treatment gas into and out of treatment chamber 42 is
controlled by the operation of valve 45 and valve 88. Vacuum line 60 is
connected to a vacuum pump 80 and enables the evacuation of gas feed line
43 and treatment chamber 42. A gas purge line 82 is connected to gas feed
line 43 at a point upstream from treatment chamber 42. The flow of an
inert purge gas, such as carbon dioxide and/or nitrogen, into gas feed
line 43 is controlled by operation of a ball valve 84. The treatment gas
in treatment chamber 42 can be evacuated to gas scrubber 56 through an
evacuation line 86. The flow of spent treatment gas through evacuation
line 86 is controlled by a ball valve 88. Treatment gas is distributed
within treatment chamber 42 through a manifold 90 positioned within
treatment chamber 42.
Additionally, water or an aqueous solution can be introduced into treatment
chamber 42 be means of a water supply 91 connected to treatment chamber
42. The water or aqueous solution can be introduced in treatment chamber
42 as a mist or liquid spray, or simply dispensed into a lower portion of
the treatment chamber. Further, the water or aqueous solution can be feed
into manifold 90 and mixed with the treatment gas prior to application
onto the food commodity.
Food commodities selected for treatment in treatment chamber 42 are
deposited within a tumbler 92. An exemplary embodiment of a pilot-plant or
laboratory scale tumbler 92 is illustrated in FIG. 3. In the illustrated
embodiment, tumbler 92 is a wire cage having a generally tubular shape
that contains a number of compartments 94. A shaft 96 connects tumbler 92
to an actuator (not shown). During food treatment, tumbler 92 is rotated
or oscillated about shaft 96, while the food product is subjected to
treatment gas dispensed through manifold 90. The tumbler 92 can be
operated in a variety of rotational modes, including 360° rotation,
agitational motion, partial rotation and the like. For example, tumbler 92
can turn in a full 360° rotation, or oscillate back and forth in a
series of partial rotations that can vary from a few degrees to
360°. In accordance with a preferred process embodiment of the
invention, tumbler 92 is alternatively rotated about shaft 96 in a
clockwise direction and then in a counterclockwise direction for a
predetermined period of time. Each rotation is preferably about
180° in each direction with respect to a reference point at the
perimeter of the tumbler.
Those skilled in the art will appreciate that the motion of tumbler 92 is
intended to assist in bringing treatment gas and liquid into contact with
the food product. Accordingly, the present invention contemplates a number
of different motion patterns of tumbler 92. For example, in another
embodiment of the invention, tumbler 92 is axially oscillated along shaft
96. In yet another embodiment, tumbler 92 is vibrated in a vertical or
horizontal reciprocating motion or both. All such motion patterns are
within the scope of the present invention.
In yet another embodiment of the invention, treatment chamber 42 itself can
receive the food commodity directly without the use of an internally
mounted device, such as tumbler 92. In this case, treatment chamber 42 can
be configured to have separate internal compartments or not depending upon
the particular food commodity. Those skilled in the art will appreciate
that a number of different commercially available massagers and tumblers
can be used with the present invention. For example, massager machines
having “T0, T1″ etc. model designations available from Lutetia
(Arnouville, France), “Reiser Ultra Vac” tumblers having capacities
ranging from 500 lbs. to 10,000 lbs. available from AMFEC (Hayward,
Calif.), tumblers having “MM-03, MM-10,” etc. model designations having
capacities ranging from 750 lbs. to 22,000 lbs. available from
Challenge-RMF Inc. (Grandview, Mo.), and “Tumbler BAMIX” available from
Armor Inox (Mauron, France), and the like.
In operation, desired ozone concentration and treatment gas pressures are
established in gas delivery system 44 by activating ozone generator 46 and
opening valves 63, 66 and 78. The desired ozone concentration in the
treatment gas is achieved by monitoring gas samples with ozone analyzer
54. Once the desired ozone concentration level in pressure vessels 48 and
50 is attained, ozone generator 46 is switched off.
The desired operating pressure is established in gas delivery system 44 by
closing valves 63 and 78 and injecting carbon dioxide into pressure vessel
48. Both pressure vessels 48 and 50 are pressurized to operating pressure.
The ozone concentration is periodically monitored and the concentration is
controlled by opening valve 78 and injecting carbon dioxide to reduce the
concentration, or supplying more ozone from ozone generator 46 to increase
the ozone concentration as needed.
A predetermined weight of a food commodity is placed into tumbler 92 and
the tumbler is sealed within treatment chamber 42. As described above,
water or aqueous solution is added to a desired level before, during or
after introduction of the treatment gas. In one embodiment, about 0.001 to
about 0.5 grams, and more preferably about 0.05 to about 0.15 grams of
water per gram of food commodity is introduced into treatment chamber 42.
Next, valves 45, 84 and 88 are closed and valve 81 is opened to draw a
vacuum in gas feed line 43 and in treatment chamber 42. Then, valve 81 is
closed and valve 45 is opened until treatment chamber 42 reaches
atmospheric pressure. Then, valve 45 is closed and the actuator is turned
on. In a preferred embodiment, the actuator is set to reciprocally rotate
tumbler 92 at a predetermined revolutions per minute (RPM). As described
above, the rotational motion can vary depending upon the particular
process. In one embodiment, tumbler 92 is rotated at about 1 to about 30
RPM. Alternatively, tumbler 92 can oscillate at about 1 to 30 RPM, where
one revolution is defined as one clockwise rotation of about 1800 and one
counterclockwise rotation of about 1800. The RPM of the tumbler can vary
from about 0.5 to about 30 RPM and, more preferably, from about 10 RPM to
about 20 RPM. The processing time can vary depending upon the amount of
the food commodity placed in the tumbler and the system operating
parameters. In a preferred method, processing is carried out for a
predetermined period of time that can range from about 2 to about 360
minutes. More preferably, the treatment time varies from about 2 minutes
to about 90 minutes. Once the processing is complete, treatment chamber 42
and gas feed line 45 are purged with inert gas by closing valve 45 and
opening valves 84 and 88.
Those skilled in the art will recognize that a wide variety of processing
conditions with treatment chamber 42 are possible through the operation of
apparatus 40. For example, the treatment of a food commodity within
treatment chamber 42 can be carried out under either pressurized or vacuum
atmospheric conditions and over a wide range of ozone concentration
levels. The operating pressure can be varied from below about 2 inches of
Hg to well over atmospheric pressure. In a preferred embodiment, the
treatment process is carried out at a vacuum pressure of about 2 to about
25 inches of Hg and, more preferably, about 10 to about 25 inches of Hg.
Additionally, the temperature of treatment chamber 42 can be controlled
through the use of heating and cooling systems (not shown) and can vary
over a wide range. For example, treatment chamber 42 can be regulated at a
temperature ranging from 30 about -200° C. to about 50° C.
More preferably, the temperature of treatment chamber 40 can be controlled
at a specific temperature within a range of from about 0.1° C. to
about 25° C. Further, water or aqueous solution can be applied to
the food commodity before, during or after application of treatment gas.
In a preferred embodiment, an ozone concentration and treatment gas
pressure are established in gas delivery system 44 that will permit
delivery of treatment gas containing a wide range of ozone concentration
levels. In one preferred embodiment, at least about 0.001 mg of ozone per
gram of food commodity to be supplied by delivery system 44. In another
embodiment, gas delivery system 44 supplies at least about 0.05 mg of
ozone per gram of food commodity. In yet another embodiment delivery
system 44 supplies at least about 1.0 mg of ozone per gram of food
commodity. In a still further embodiment delivery system 44 supplies at
least about 2.0 mg of ozone per gram of food commodity. In another
preferred embodiment, gas delivery system 44 supplies about 0.01 to about
2.0 mg of ozone per gram of food commodity. In a more preferred
embodiment, gas delivery system 44 supplies about 0.125 to about 2.0 mg of
ozone per gram of food commodity. In a still more preferred embodiment,
gas delivery system 44 supplies about 0.4 to about 0.6 mg of ozone per
gram of food commodity.
In a most preferred processing embodiment for a food commodity such as
poultry, the ozone concentration in the treatment gas is maintained at
about 0.5 mg/g meat and a treatment time of about 15 minutes is used.
Water is added to create a moisture content of about 13% by weight and a
tumbling speed of about 18-20 RPM is used. The treatment temperature is
preferably maintained at about 20° C. at atmospheric pressure.
Using the foregoing description, it is believed that those skilled in the
art can practice the invention to its fullest extent. Accordingly, the
following examples and description are merely intended to be explanatory
of the invention and not intended to limit the invention in any way
whatsoever.
EXAMPLE I
Several experiments were carried out to evaluate the efficacy of the
gaseous ozone process with moisture on the selected food borne pathogen,
i.e., Salmonella enteritidis and to assess the optimal ozone processing
parameters on other food borne pathogens (i.e., L. monocytogenes, E. coli
O157:H7, and generic E. coli) and spoilage microorganisms (i.e., lactic
acid bacteria and natural meat isolate).
I. Inoculum Preparation
Since spot inoculation represents the most likely contamination pattern in
nature, spot inoculation was used to inoculate the chicken coupons used in
the experiments described herein.
A. Generic E. coli
Each strain of a three-strain mixture of generic Escherichia Coli (all beef
isolates, obtained from University of Georgia) was maintained in tryptic
soy broth (TSB) (Difco Laboratories, Detroit, Mich.) and stored at about
4° C. between subcultures. To activate cultures prior to use, loop
transfers (1% inocula) were made for two consecutive days in TSB and
incubated at about 35° C. for about 16 to 20 hours. Cell counts in
the suspension generally ranged around 109 colony forming units per
milliliter (CFU/ml). The three strains were then combined at about equal
concentrations.
B. E. coli 0157:H7
Each strain of a two-strain mixture of Escherichia coli 0157:H7 (both beef
isolates, obtained from University of Georgia) was maintained in tryptic
soy broth (TSB) (Difco Laboratories, Detroit, Mich.) and stored at about
4° C. between subcultures. To activate cultures prior to use, loop
transfers (1% inocula) were made for two consecutive days in TSB and
incubated at about 35° C. for about 16 to 20 hours. Cell counts in
the suspension generally ranged around 109 CFU/ml. The two strains
were then combined at about equal concentrations.
C. Listeria monocytogenes
Four strains of Listeria monocytogenes (N-7031-Cabbage isolate;
N-7298-Clinical isolate; N-7325-ATCC; and N-7327-Radish isolate all
obtained from National Food Processors Association) were maintained in
tryptic soy broth (TSB) (Difco Laboratories, Detroit, Mich.) and stored at
40° C. between subcultures. To activate cultures prior to use, loop
transfers (1% inocula) were made for two consecutive days in TSB and
incubated at about 35° C. for about 16 to 20 hours. Cell counts in
the suspension generally ranged around 108 to 109 CFU/ml. The
four L. monocytogenes cultures were then combined at equal ratios.
D. Salmonella enteritidis
Salmonella enteritidis E565-88 (food isolate, obtained from University of
Georgia) was maintained in tryptic soy broth (TSB), and stored at about
4° C. between subcultures. To activate culture prior to use, loop
transfers (1% inocula) were made for two consecutive days in TSB and
incubated at about 35° C. for about 16 to 20 hours. Cell counts in
the suspension generally ranged around 109 CFU/ml.
E. Lactic Acid Bacteria (LAB)
Four strains of lactic acid bacteria (LAB) strains (HPS-Pediococcus sp;
LP-Pediococcus pentosaceous; LL2-Lactobacillus plantarum obtained from
Chr. Hansen, McFarland, Wis., and 8014-Lactobacillus plantarum ATCC 8014
obtained from ATCC, Manassas, Va.) were maintained in Lactobacilli MRS
broth (Difco Laboratories, Detroit, Mich.). They were stored at about
4° C. between subcultures. To activate cultures prior to use, loop
transfers (1% inocula) were made for two consecutive days in MRS broth and
incubated at about 35° C. for about 16 to 20 hours. Cell counts in
the suspension generally ranged around 108 to 109 CFU/ml. The
four strains were then combined at equal ratios.
F. Meat Spoilage Microorganisms
The culture of natural meat spoilage microorganisms was enumerated by
swabbing the beef surface and cultured the swab in tryptic soy broth (TSB)
(Difco Laboratories, Detroit, Mich.) at about 35° C. overnight. The
culture was stored at about 4° C. between subcultures. To activate
cultures prior to use, loop transfers (1% inocula) were made every 24
hours for two consecutive days in TSB and incubated at about 35° C.
for about 16 to 20 hours. Cell counts in the suspension generally ranged
around 108 to 109 CFU/ml.
II. Meat Sample Preparation
Irradiated boneless chicken breast samples were purchased from Royalty
Food, Inc. (Orlando, Fla.). The chicken samples were irradiated by Food
Technology Service, Inc. (Mulberry, Fla.), and normally came as frozen,
single lobe (4-oz portion). The microbial quality of an irradiated chicken
breast sample was negative with respect to Salmonella and about 2 log with
respect to Aerobic Plate Counts. The frozen samples were transferred to a
2° C. refrigerator and thawed overnight. The following day, each
single lobe breast sample was aseptically cut into a 2″×2″ coupon.
The coupons were stored in a freezer prior to use.
The frozen chicken breast coupons were thawed overnight before each
experiment. One 0.5 inch diameter Millipore, model AP10, filter disk
(Millipore Corporation, Bedford, Mass.) was placed on each coupon. Then,
about 0.1 ml of inoculum was placed onto the filter disk and then allowed
to set at room temperature for about 30 min to allow cell attachment. A
20-200 μl micropipetter (VWR Scientific, Chicago, Ill.) was used to
dispense the inoculum. The inoculated filter discs were removed from the
chicken coupons after about 30 min. Inoculated chicken coupons were
randomly grouped into trays (3/tray), weighed, and refrigerated before the
treatment.
III. Experiment Conditions
In accordance with the operational description of apparatus 40 set forth
above, ozone generator 46 was turned on and valves 63, 66 and 78 were
opened. Ozone generator 46 was turned off after pressure vessels 48 and 50
reached a desired ozone concentration. Then, valves 63 and 78 were closed
and valve 66 was allowed to remain open. The ozone concentration in the
treatment gas was monitored by ozone analyzer 54. Carbon dioxide
(CO2) was injected into pressure vessel 48 to pressurize both
pressure vessels 48 and 50 to about 30 psig. Then, valve 66 was closed and
valve 70 was opened to determine the ozone concentration in pressure
vessel 50. For the series of experiments described herein, the ozone
concentration in the treatment gas was adjusted to about 0, 0.125, 0.25,
0.5, 1, and 2 mg per gram of chicken (based on all three pieces).
The above procedure was followed prior to the addition of water and sealing
the reaction chamber. A tray of inoculated chicken coupons was then
removed from the refrigerator and the amount of water and ozone required
was calculated. The amount of sterile deionized water used was 13% by
weight for each set of three coupons, and the water was dispensed into
treatment chamber 42 using a 10-ml pipette. To ensure the even
distribution of water in the chamber, the water was equally divided
between the back, middle, and front of treatment chamber 42.
Then, the chicken coupons were loaded into the stainless steel wire cage
and each coupon was placed into a separate compartment in the reaction
chamber. In this design, no two chicken coupons could stack on each other
and each coupon received the maximum exposure to the treatment gas. The
cage was inserted into treatment chamber 42, and the chamber was sealed.
To initiate the process, valves 45, 84 and 88 were closed and 81 was opened
to draw a vacuum on gas feed line 43. Then vacuum pump 80 was turned on
until the vacuum gauge reached 24 inches Hg. In this experiment, a model
DAA-V17A-EB vacuum pump was used (Gast Manufacturing, Benton Harbor,
Mich.). Next, valve 81 was closed and the vacuum pump was shut off. Then,
valve 45 was opened until the pressure in vacuum line 60 reached about 0
psig. Valve 45 was then closed and the pneumatic actuator was turned on
and the timer was started. For the series of experiments described here,
the treatment times were about 0, 5, 8, 15, 30, and 45 minutes. Following
each experiment, the actuator was turned off and valves 84 and 88 were
opened, and treatment chamber 42 was flushed with carbon dioxide for about
20 seconds.
For each experiment, the motion of the tumbler was bidirectional, not
unidirectional like a typical tumbler. The tumbler rotated approximately
180 degrees in one direction and then rotated back the other direction.
Both rotations counted as one revolution. The number of revolutions per
minute (RPM) was 16 to 20 for these experiments. In all experiments, the
temperature of treatment chamber 42 was kept at about ambient temperature
(20° C.).
IV. Microbiological Analysis
Following treatment, each chicken coupon was put into a 24-oz sterile
Whirl-pak plastic bag (Nasco, Fort Atkinson, Wis.) with 90 ml of sterile
0.1% peptone water, and pummeled for 2 min at normal speed with a
Stomacher 400 Lab-blender. Each sample was then serially diluted. One
milliliter of sample from each dilution was plated on the appropriate
growth medium. Duplicates were made from each dilution. Lactic acid
bacteria (LAB) were plated on 3M Redigel™ MRS Test Media (Minnesota
Mining and Manufacturing Company, St. Paul, Minn.) and incubated at
35° C. in a CO2 incubator (5% CO2) for 48 h. Both E. Coli
0157:H7 and generic E. coli were plated on E. coli Count Plates
Petrifilm™ (3M, St. Paul, Minn.) and incubated at 35° C. for 48
h. L. monocytogenes were enumerated on PALCAM Agar (Difco Laboratories,
Detroit, Mich.) by spread plates and incubated at 35° C. for up to
5 days. S. enteritidis and natural meat spoilage microorganisms were
enumerated on Aerobic Count Plates Petrifilm™ (3M, St. Paul, Minn.) and
incubated at 35° C. for 48 h. The colonies were counted after the
incubation time. The count was recorded and expressed as CFU/g.
V. Effect of Treatment Gas on Microbial Inactivation
FIG. 4 shows the ozone biocidal efficacy for times ranging from 5 to 45
minutes for a fixed ozone concentration of 2 mg/g and with 13% by weight
added sterile moisture. The data shown in FIG. 4 indicates that the
microbial population decreases rapidly up to about 7 to 8 minutes and
decreases slowly at increasing treatment times greater than about 8
minutes.
FIG. 5 shows the biocidal efficacy for different ozone concentrations
ranging from about 0.125 to about 2 mg/g chicken with 13% added sterile
moisture at a fixed treatment time of about 15 minutes. The biocidal
efficacy increases rapidly with increasing ozone concentration and levels
off at about 0.5 mg/g chicken. No notable changes in texture or color of
the chicken coupons were observed with the various levels of ozone.
The data shown in FIGS. 6 and 7 illustrate the effectiveness of the
treatment process on the different microorganisms used in this experiment.
FIG. 7 is the subset data of FIG. 9 and shows the relative reduction
(initial counts minus survival) of different microbial populations on
chicken coupons treated with ozone. The microbial reduction in log units
for S. enteritidis, E. coli 0157:H7, generic E. coli, L. monocytogenes,
lactic acid bacteria, and natural meat spoilage microorganisms were 1.96,
2.88, 3.52, 2.83, 2.11, and 1.94, respectively. The data indicates S.
enteritidis had greater resistance to ozone inactivation among all
challenge microorganisms. However, other than S. enteritidis, natural
spoilage microorganisms (e.g., lactic acid bacteria and natural meat
isolate) were more resistant to ozone than pathogenic microorganisms.
The reduction in pathogenic bacteria shows that, based on a natural
microbial population and distribution, this inventive process can reduce
pathogenic bacteria, yet maintain a certain level of spoilage
microorganisms. This is an important result because natural spoilage
microflora also serve as a defense against subsequent pathogens that come
in contact with the food commodity.
In summary, the experimental results demonstrate that the gas treatment of
the invention with added moisture can significantly reduce pathogens on
the surface of the chicken coupons. The inventive process is able to
reduce S. enteritidis, E. coli 0157:H7, generic E. coli, L. monocytogenes,
lactic acid bacteria, and natural meat spoilage microorganisms levels by
1.96, 2.88, 3.52, 2.83, 2.11, and 1.94 log units, respectively.
EXAMPLE II
FIG. 7 illustrates in bar chart form comparative testing with and without
ozone in removing bacteria from chicken breasts. In Tests 1 and 2, the
chicken breasts were contacted with ozone gas for a period of 3 minutes,
while in Tests 3 and 4, the chicken breasts were contacted with ozone for
a period of 6 minutes. In Test 1, a 73% reduction with ozone is seen,
while in Test 2, a 48% reduction is seen. Test 3 showed a 40% reduction,
while Test 4 showed a 70% reduction in bacterial count. It may thus be
seen that a bacterial count reduction of at least 40% is obtainable with
as little as 6 minutes time contacted with ozonated gas.
FIG. 8 illustrates in bar chart form 7 other tests performed in accordance
with the present invention. Tests 1, 2, 3, and 4, exemplified in FIG. 8
resemble the tests carried out as explained previously for FIG. 2. Test 1
of FIG. 8 showed a 93% reduction, Test 2 showed an 83% reduction, Test 3
showed an 88% reduction, and Test 4 showed an 83% reduction in bacterial
count. Tests 5 and 6 illustrate the effect of a thin film of water in
helping the contact time for the ozone. Test 5 showed a 73% reduction in
bacterial count, while Test 6, the best test of all runs showed a 98%
reduction in bacterial count. Finally, Test 7 showed the comparison with
and without ozone gas treatment using residual vacuum.
From the above testing, it can be seen that great reduction in bacterial
count may be obtained simply by the use of either ozonated gas, or
ozonated water which appears as a thin film on the surface of the meat.
Thus it is apparent that there has been disclosed in accordance with the
invention, a method for food disinfection using gaseous ozone that fully
provides the advantages set forth above. Although particular embodiments
of the invention have been described, it will be apparent to one skilled
in the art that numerous modifications and variations can be made to the
presented embodiments, which still fall within the spirit and scope of the
invention. Accordingly, it is intended that all such variations and
modifications fall within the scope of the appended claims and equivalents
thereof.
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