MXPA98008364A - Wide spectrum of prevention and removal of microbial food contamination through cuaterna ammonium compounds - Google Patents

Abstract

The present invention relates to a method for using quaternary ammonium compounds to inhibit the binding of and remove a broad spectrum of microbial contamination supported on the food of food products. The method uses quaternary ammonium compounds to inhibit the binding of and remove microorganisms such as Staphylococcus, Campylobacter, Arcobacter, Listeria, Aeromonas, Bacillus, Salmonella, non-toxin-producing Escherichia, and toxin-producing pathogenic Escherichia, such as O157: H7 fungi, such as Aspergillus flavus and Penicillium chrysogenum, and parasites, such as Entamoeba histolytica from a wide range of foods. The foods that can be treated by this method are meat, marine products, vegetables and fruit. One of the treatment methods is to atomize quaternary ammonium compounds in food products to prevent microbial contamination supported by broad-spectrum foods. New formulations of quaternary ammonium compounds, combined with glycerin and / or ethyl alcohol, provide a new concentrated formulation for industrial use and a diluted formulation for use in atomization methods.

Description

WIDE SPECTRUM OF PREVENTION AND REMOVAL OF MICROBIAL FOOD POLLUTION THROUGH QUATERNARY AMMONIUM COMPOUNDS BACKGROUND OF THE INVENTION This application is a continuation in part of the Serial US pending No. 08/631, 578 filed April 12, 1996, which is incorporated herein by reference in its entirety. 1. FIELD OF THE INVENTION The present invention relates generally to a method for preventing the growth of a wide range of microorganisms in food products. More specifically, the present invention relates to a method for using quaternary ammonium compounds (QACs) to prevent the growth of a broad spectrum of microorganisms in food products; such as, meat products, for example, poultry, beef, pork, lamb, venison and other edible meat products; marine products, for example, fish and seafood; fruits, vegetables and any other food product that can be treated using the aqueous treatment methods of the present invention without detrimentally affecting the appearance, texture and quality of the food. More specifically, the present invention relates to a method for using QACs to inhibit the binding of, remove and prevent the growth of microorganisms in food products. Particularly, the use refers to the effect of QACs on microorganisms that can cause contamination supported in foods. More particularly, these microorganisms include microorganisms of the genus Staphylococcus, Campylobacter, Arcobacter, Listeria, Aeromonas, Bacillus, Salmonella, Escherichia that does not produce toxins, and Escherichia that produces pathogenic toxins, such as O157: H7. More particularly, the present invention relates to an improved treatment method of atomizing QACs in food products to prevent the broad spectrum of microbial growth in these products. The present invention also relates to a formulation of QACs that makes the treatment method more accessible for commercial use in a food processing plant. 2. Description of the Prior Art The prevention of food borne diseases by microbial contamination is of vital importance to the food processing industry, regulatory agencies and consumers. A recent report from Food Safety & Inspection Service (FSIS) of the United States Department of Agriculture (Federal Register, February 3, 1995) estimates that about 2 million cases of foodborne illnesses are produced annually from microbial contamination in the United States, with an associated cost of about 1 trillion dollars. Microbial contamination supported by food occurs both before entering the processing facilities, and through cross-contamination in the processing environment. FSIS has instituted new Hazard Analysis and Critical Control Point (HACCP) requirements to reduce the occurrence and number of pathogens supported by food. These regulations must be met by the food processors. Although the means to achieve this microbial reduction is left to the discretion of the processor, FSIS expects that antimicrobial treatments will be an important component of the HACCP plans. The treatment methods of the present invention, which employ an aqueous formulation of QACs, are useful to meet the requirements of HACCP. In their efforts to provide a product completely free from microbial contamination, poultry and meat processors have encountered significant difficulties in removing microorganisms that adhere or bind vigorously to poultry and meat tissues, which are intended to be used as products. food. If contaminating microorganisms do not bind to the surface of the food, can be easily eliminated by rinsing. However, microorganisms that are tightly bound can not be removed by rinsing and are quite resistant to removal by chemical or physical means. Various chemical and physical methods have been proposed to reduce microorganisms in meat products, such as the use of chlorine or chlorine dioxide, ozone, hydrogen peroxide, lactic acid, sodium carbonate, trisodium phosphate, and electrical stimulation. Generally, these methods have shown limited effectiveness in reducing microbial contamination and can affect the physical appearance of meat products.

Contamination with Salmonella typhimurium has been of special interest in the poultry processing industry because the organism is frequently present in live birds. Poultry processors have had great difficulty in removing microorganisms, such as S. typhimurium, which bind or adhere to the tissues of poultry. It has been suggested that a variety of chemical and physical approaches be used during the processing of poultry to eliminate S. typhimurium contamination from carcasses and minimize cross-contamination between carcasses. Trisodium phosphate (TSP) has been used in the processing of poultry to suppress S. typhimurium; However, studies report conflicting results on the efficacy of TSP against Salmonella. As a result of its solubility in water, the TSP can be washed from the poultry, and thus, can not inhibit the binding of microorganisms. U.S. Patent No. 5,366,983, incorporated herein by reference, discloses a method for removing or preventing contamination of Salmonella from meat products by treatment with an effective amount of an aqueous QAC solution. Specifically, cationic quaternary ammonium surfactants, such as alkyl pyridinium, particularly cetylpyridinium chloride (CPC) and cetylpyridinium bromide (CPB), were effective in removing S. typhimurium from poultry. However, this patent does not disclose that QACs have a broader antimicrobial spectrum against other genera of food contaminating microorganisms than Salmonella. Furthermore, it does not suggest that this method of treatment would be effective in food products other than meat. The food substances differ chemically and physically by virtue of their protein content, porosity, lipophilicity, surface pH, water permeability, surface area, and net electrical charge of the surface. The porosity of the food could be important in the separation of bacteria while a hard, impermeable integument in a food substance could reduce the bacterial contamination of the food. All these chemical and physical differences between the food products make it difficult to predict whether a successful antimicrobial agent in meat products would suggest success in other food products, such as fruits, vegetables and marine products. For example, it is known that CPC binds to proteins; however, if the antimicrobial efficacy of CPC in food products was due in large part to protein ligation, then the present method for treating non-protein fruits and vegetables would not be expected to be successful. On the rise, foodborne illnesses caused by other pathogenic and pathogenic bacteria other than Salmonella have become a problem for food processors. A list of these bacteria with the products in which they have been identified is presented in Table 1: TABLE 1 INCIDENCE OF DECOMPOSITION AND PATHOGENIC BACTERIA Among these contaminating microorganisms listed in the table, Escherichia coli O157: H7 is of special interest due to its virulence, severity of the disease produced, and associated with mortality. E. coli O157: H7 produces strong "shiga-like" toxins that lead to blood coagulation abnormalities, kidney failure (hemolytic uraemic syndrome), and death. Even if the recovery of the watery disease is completed, 15-305 of people infected with haemolytic uremic syndrome will have evidence of chronic kidney disease. The risks associated with contamination with E. coli O157: H7 are compounded by their reported resistance to antibiotics. In 1993, between 8,000-16,000 cases of foodborne illnesses were produced by E. coii O157: H7 with an estimated cost between 0.2 and 0.5 billion dollars.

Another virulent food contaminant has been found, Listeria monocytogenes in meat, vegetables, and various dairy products.; and can cause sepsis, meningitis and disseminated abscesses. L. monocytogenes is a cold-tolerant microorganism capable of growing under refrigeration. In 1993, approximately 1,700 cases of food borne diseases were produced by L. monocytogenes at an estimated cost of between $ 01 and $ 0.2 trillion. Another microorganism of interest in the food industry is Aeromonas hydrophila which causes decomposition in the meat and food processing industry, and reduces the shelf life of these products. Currently, there are no known microbicide compounds which are effective in preventing and removing contamination in a wide range of food products against a broad spectrum of gram positive, gram negative, aerobic, facultative anaerobic and microaerophilic microorganisms. The present inventors have determined that QACs are effective against a broad spectrum of different microorganisms, which produce food borne diseases when they are linked to a wide range of food products. This sensitivity of a broad spectrum of pathogenic microorganisms could not have been predicted. The sensitivity of a microorganism to a particular microbial agent is not predictable from the sensitivity of other microorganisms to the same agent. It is believed that antiseptics or germicides have a continuous spectrum of activity, but the relative susceptibilities of different microorganisms should be considered. For example, germicide, hexachlorophene, is mainly effective against Gram-positive microorganisms, and cationic antiseptics are not effective against sporulating organisms. Some Gram-negative microorganisms, such as Pseudomonas cepacia, have been known to grow in solutions of the drug, benzalkonium chloride. Other bacteria that are capable of growing in 70% ethanol have been known (Harvey, S.C., Antimicrobial Drugs in Remington's Pharmaceutical Sciences, 18th Ed., Mack Publishing Co., pp. 1 163-1241 1990). With respect to the treatment of food products, it has been reported that Listeria is more resistant to the action of TSP than Salmonella or E. coli (Somers, EB et al., Int. J. Food Microbiol .. 22: 269-276 , 1994). Additionally, (Breen et al., J. Food Sciences, 60: 1991 -1996, 1995) it was demonstrated that TSP is much less effective in inhibiting the growth of Salmonella than it is in separating this organism. Similarly, the TSP has reduced the numbers of E. coli O157: H7 in chicken carcasses, but is not effective in inhibiting cross-contamination of this organism to other chickens. The present invention shows that QACs are effective against E. coli O157: H7 in suspension in liquids, to reduce the numbers of this bacterium when it is attached to food products, as well as to inhibit the binding of this bacterium to food products. It has been reported that E. coli O157: H7 shows resistance to broad spectrum antimicrobial agents, such as, tetracycline, streptomycin, sulfisoxazole (Kim et al., J. Infect. Dis. 170: 1606-1609, 1994) and oxytetracycline (Ciosek et al., Med. Weter. 40: 335.338: 1984). while the same agents are very active against species that do not produce regular toxins of £. coli Clearly, the effectiveness of an antimicrobial agent or biocide against a particular microorganism can not be predicted based on its effectiveness against a different microorganism. There are many factors to consider, such as microbial characteristics, which may play a role in the effectiveness of an antimicrobial agent against a particular microorganism. These characteristics include but are not limited to: (1) the degree of glycocalyx formation by a given species of bound microorganism, (2) the presence of a cellular envelope containing lipopolysaccharide and phospholipid in gram negative bacteria, (3) the presence of lipoprotein as in most enteric bacteria and Pseudomonas, and (4) the presence of porin protein channels, for example in E. coli and Salmonella (Fulton et al., Structure in Medical Microbioloqy, 3rd Ed., pp. 37-54, 1991). The food processing industry is in need of a more effective process for preventing the growth of a wide range of contaminating microorganisms in many different food products. This is especially true for microorganisms that are attached to the surfaces of food. As a result of increasing numbers of diseases caused by pathogenic microorganisms supported by food, the food processing industry requires more effective processes for the removal and prevention of a wider spectrum of microorganisms, and particularly for pathogenic microorganisms, such as Escherichia. that produces toxin, that is, E. coli O157: H7, which are known to cause serious human diseases as a result of food contamination. The present inventors have provided a method to prevent the growth of microorganisms in liquids associated with food products, an important objective to prevent cross-contamination within the processing plant; to remove attached microorganisms from food products, to inhibit the binding of microorganisms to food products; and to prevent the growth of microorganisms that remain attached to food products. In addition, the method of the present invention can be easily adapted for use in a food processing plant. Additionally, the present invention provides a concentrated QAC formulation for use in dilution for a working solution for use in the present method. A formulation of the present invention contains components that enhance solubility, which can also result in longer contact times of the formulation with the food product.

BRIEF DESCRIPTION OF THE INVENTION The present invention provides a method for preventing the growth of a broad spectrum of microorganisms in food products. The prevention of growth of microorganisms in food products is intended to provide a food product that is devoid of or contains minimum numbers of viable microorganisms that could cause disease in humans or animals, or decomposition of the food product before ingestion. The prevention of microorganism growth in food products is intended to include, but is not limited to the following mechanism: (1) removal of bound microorganisms from food products; (2) inhibition of binding of microorganisms to food products; (3) death or inactivation of bound microorganisms in food products; and (4) death or inactivation of microorganisms, which are not bound to the food product, but which are present in liquids associated with the food products during processing; such as, in cooling tanks. The microorganisms that are susceptible to QACs include microorganisms of the genus Staphylococcus, Campylobacter, Arcobacter, Listeria, Aeromonas, Bacillus, Salmonella, Escherichia that does not produce toxins, Escherichia that produces pathogenic toxins, and other microorganisms supported in food, which are capable of cause microbial contamination supported by food foods for human or animal consumption. Additional microorganisms, which are also susceptible to QACs, are fungi, such as Aspergillus flavum and Penicillium chrysogenum, and parasites, such as Entamoeba histolytica. The composition of the present invention comprises an effective amount that inhibits the microbial growth of QAC in an aqueous solution. The QACs of the present invention are effective in preventing the growth of a broad spectrum of pathogenic and decomposing microorganisms. QACs, particularly cetylpyridinium chloride (CPC), are especially effective in preventing the growth of a broad spectrum of microorganisms in a wide range of food products. The present invention has an important application in the food processing industry, as well as for home use. QACs are readily available and the cost of performing the method of the present invention is not expensive compared to existing antimicrobial processes. Unlike existing treatments that use, for example, TSP, the use of QACs does not alter the appearance, color, taste or texture of the food product. A range of concentrations of QACs is effective in preventing broad-spectrum microbial growth in food products. The QACs are safe, as shown by the lack of mutagenicity of CPC using the Ames assay. In addition, CPC is already approved for human use in products for oral ingestion in preparations, such as Cepacol® tablets, which are orally ingested in amounts of up to 20 mg per day. The present invention is also directed to a QAC formulation for use with the present method for the treatment of food products, in which, for example, CPC is formulated with agents that enhance solubility, such as ethyl alcohol and / or glycerin . The present invention is also directed to an improved method of contacting food products with QAC for a period of time of less than five minutes, even as short as 20 to 90 seconds, which results in important prevention of growth of microorganisms in the food products. The invention also includes an improved method of contacting QACs with food products by atomizing the compound in the food product. The atomization method can be performed using QAC in solution in water, or using the new formulation with QAC formulated with solubility enhancing agents. Additionally, the method of the present invention may optionally include a determination step before contacting the food product with the QACs, to determine the presence of microorganisms in the food prior to treatment. Any conventional method for rapidly determining the presence of microorganisms can be used as the determination step, which, for example, includes PRC and immunoassays. Additionally, the method of the present invention may optionally include a step to determine the presence of QACs on the surface of the food product after contacting the QACs. This determination can be made immediately after the contact step or after several washing steps. For example, the QAC can be extracted from the tissues of the food in a form suitable for the analysis of high performance liquid chromatography (HPLC). The method comprises ethanol extraction from the food tissue followed by solid phase extraction using a weak cation exchange column, which selectively separates the QACs from other compounds in the matrix that would otherwise interfere with the HPLC analysis. The HPLC assay for quantification of QAC residues employs a reverse phase cyano column and uses a QAC analog as an internal standard.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a bar graph showing the inhibition of the binding of E. coli O157: H7 to beef shoulder tissue after treatment with CPC. Fig. 2 is a bar graph showing the reduction of viable microorganisms in catfish skin after treatment with CPC in 5% aqueous glycerin in nonselective medium. Fig. 3 is a bar graph showing the reduction of viable S. typhimurium in catfish skin after treatment with CPC in 5% aqueous glycerin in selective medium. Fig. 4 is a bar graph showing the reduction of viable S. typhimurium in black grapes after treatment with CPC in 5% aqueous glycerin. Fig. 5 is a bar graph showing the reduction of viable S. typhimurium in broccoli after treatment with CPC in 5% aqueous glycerin.

DETAILED DESCRIPTION OF THE INVENTION The present invention is based on the determination that QACs can be used to treat a wide range of food products to reduce a broad spectrum of microbial contamination supported by foods in these products. The present invention is also based on the finding that QACs are effective in removing, killing, inactivating and inhibiting the binding of a wide range of pathogenic microorganisms supported in foods to food products. These microorganisms include, but are not limited to, bacteria belonging to the genera Salmonella, Staphylococcus, Campylobacter, Arcobacter, Listeria, Aeromonas, Bacillus, Escherichia that does not produce toxins, and the Escherichia species that produce virulent toxins, such as, . coli O157: H7; fungi, such as, Aspergillus Flavus and Penicillium chrysogenum; and parasites, such as Entamoeba histolytica. The compositions of the present invention comprise an effective amount of QAC in an aqueous solution. The QAC is selected from the group consisting of alkyl pyridinium, tetraalkylammonium and acylaliccyclic ammonium salts. The alkylpyridinium is represented by the structural formula (I): CH3-CCH2) ft-? - \ - f where n is 9-21; and X is a halide. The tetra-alkylammonium is represented by the structural formula (I I): CH3 - < CH2 > "- H- to 8 where n is 9-21; R is selected from the group consisting of CH3 and C2H5; and X is a halide.

The alkylacyclic ammonium salts are represented by the structural formula (III): CH, - < CH2) - H CCH ^ where n is 9-21; Z is 4-5; R is selected from the group consisting of CH3 and C2HS; and X is a halide. A variety of QACs, which are all active surface cationic agents; that is, surfactants, were evaluated for their effectiveness in removing microorganisms bound from various foods, as well as to inhibit the union of microorganisms: Of the QACs studied, cetylpyridinium chloride (CPC) was the most effective and will be used in the examples discussed below, but are not intended to limit the use of QACs to CPCs within the meaning of the present invention, because other members of QACs also have similar properties against pathogenic microorganisms supported in foods. The present invention is further based on the determination that the contact time of QACs with food products in the dipping process can be reduced to 1 minute still results in significant inhibition of microorganism binding, for microorganisms supported in food including Salmonella, which is a significant improvement and a commercial advantage in the industrial use of this process.

The present invention is also based on the determination that a new method for atomizing QACs under various pressures in food products for 20 to 90 seconds, significantly reduces the viable microorganisms supported in the foods in these products. The present invention is also based on the determination that a new formulation of QACs in solutions containing varying concentrations of at least one solubility enhancing agent, such as ethyl alcohol or glycerin, is useful for the treatment of food products. This is particularly true when QAC solutions are to be used in combination with salt, are highly concentrated, or will be subjected to low temperatures, such as inside food processing plants. This new formulation allows concentrated QACs to be stored and can be easily diluted then for use in the antimicrobial treatment process in the processing plant, instead of requiring powdered QACs to be mixed before use. This new formulation of QACs provides a facility to use concentrate, which is advantageous for industrial use. This new formulation of QAC can be used both in the standard immersion method and in the new atomization method. The above-described aspects of the present invention are described in detail below with reference to Figs. fifteen. The examples set forth below serve to further illustrate the present invention in its preferred embodiments, and are not intended to limit the present invention. The examples use poultry, beef, catfish, broccoli and grapes as the food products treated in the method, but it is intended that the treatment of other food products, which are not adversely affected by the treatment process, also be covered by the present invention.

EXAMPLES The microorganisms used in the following examples are as follows: Staphylococcus aureus ATCC 29213, Campylobacter jejuni ATCC 29428, Escherichia coli (species that does not produce toxins) ATCC 25922; Escherichia coli O157: H7 (species that produces toxins) ATCC 43895, Arcobacter butzleri ATCC 49616, Listeria monocytogenes ATCC 49594, Aero monas hydrophila ATCC 49140, Bacillus cereus ATCC 49063, Salmonella typhimurium ATCC 14028 and NCTC 12023, and commercially available cultures of Aspergillus flavus and Penicillium chrysogenum.

Example 1 Bactericidal activity of quaternary ammonium compounds in suspension cultures (not bound to meat products) Minimum inhibitory concentration (MIC) of quaternary ammonium compounds The minimum inhibitory concentrations (MIC) for QAC were determined in Mueller Hinton broth (BBL Microbiology System) using the macrodilution method established by the 1987 National Committee for Clinical Laboratory Standards. Experiments were conducted by incubation for 16 hours at 37 ° C for Staphylococcus aureus, Escherichia coli O157: H7, Listeria monocytogenes, and Salmonella typhimurium. For incubations of Aeromonas hydrofhila, and Bacillus cereus were performed at 30 ° C. The MICs were determined by the lowest dilution with which there was no visible turbidity. Table 2 shows the data from the previous experiment: TABLE 2 MINIMUM INH CONDITIONING MINIMUM (MIC) Minimum bactericidal concentration (MIC) of quaternary ammonium compounds The minimum bactericidal concentrations (MBC) for QAC towards Campylobacter jejuni and Arcobacter butzleri were determined in broth Mueller Hinton (BBL Microbiology System) using the macrodilution method established by the 1987 National Committee for Clinical Laboratory Standards. The experiments were conducted by microaerophilic incubation at 37 ° C for 48 hours. An aliquot of each dilution was plated on agar and incubated under microaerophilic conditions at 37 ° C for 48 hours. The MBCs were determined as the lowest dilution without growth. Table 3 shows the data of the previous experiment: TABLE 3 BACTERICIDE MINIA CONCENTRATION (MBC) MIC and MBC data show that CPC is effective against a wide range of microorganisms.

Activity of quaternary ammonium compounds in plankton cells A 16-hour culture of each E.coli O157: H7 was centrifuged in trypticase soy broth (1 5,000 rpm, 10 min, 4 ° C). After removal of the supernatant, the pellet was washed with 10 ml of 0.04 M potassium phosphate buffer (PPB, pH 7.0), and suspended in PPB to a final suspension of 1-2 X 10 9 cells / ml. Aliquots (1.0 ml) were centrifuged (14,000 rpm, 3 min), and the supernatants were removed. Each pellet was suspended either in 1 ml of an aqueous solution of various concentrations (100-1,000 μg / ml) of test composition (CPC) or 1 .0 ml of PPB, vortexed (30 sec), incubated for 1 min at 25 ° C, and centrifuged (14,000 rpm, 3 min). After removal of the supernatant, each pellet was suspended in 0.5 ml of PPB. Cells from each sample were counted using aliquots of 0.05 ml per duplicate and standard serial dilution techniques in trypticase soy agar, and the data were recorded as average colony forming units (CFU) / ml. The results of the previous experiment show that Complete reduction of viable E. coli O157: H7 in suspension was achieved at all tested CPC concentrations (100, 250, 500 and 1000 μg / ml). The results of this experiment are particularly significant for the prevention of cross contamination with E. coli O157: H7 in industrial meat processing. As discussed above, this species of E. coli that produces toxins shows resistance to many broad spectrum antimicrobial agents. These results provide evidence that the treatment of meat products with QAC will prevent a contaminated piece of meat from contaminating other non-contaminated parts, because QAC will kill the organism in the liquid, which is the transfer agent responsible for cross-contamination .

^ Example 2 Effects of quaternary ammonium compounds on the reduction of viable bacteria bound to chicken skin Chicken skin (2.5 x 2.5 cm) cut from a thigh, sterilized by a dose of 45KGy of irradiation from an electron source, they were placed with the epidermis up in each cavity of six-pocket tissue culture plate. Each piece of skin was inoculated with 5 ml of saline buffered with 0.008 M phosphate (PBS, pH 7.2) containing 6-8 x 103 CFU / ml bacteria, with the exception of the control group of support that was treated with only 5 ml. of PBS. The plates were incubated (30 min, 35 ° C), and each piece of skin was rinsed (2X, 5 ml PBS) to freely remove the bound (unattached) microorganisms. Each inoculated skin was treated with 5 ml of CPC containing PBS. Three pieces of skin were used for each concentration of CPC, including one in which the skins were treated with only 5 ml of PBS (concentration 0). Plates were incubated with shaking (100 rpm) for 30 min at 25 ° C. After incubation, each piece of skin was rinsed (5 ml of PBS), placed in a sterile plastic bag containing 80 ml of saline or 1% peptone, and homogenized for 2 minutes using a laboratory mixer (Stomacher ® 400, Seward Medical, London, England). Three aliquots of the homogenate (1 ml) were emptied into plates and incubated (37 ° C, 18-24 h). Bacterial colonies were counted, corrected by dilution, and reported as CFU / skin.

These studies show the reduction in viable bacteria (Salmonella typhimurium, Staphylococcus aureus, Campylobacter jejuni, Escherichia coli (species that does not produce toxins) and Escherichia coli O157: H7) after treatment with concentrations of 50 to 1000 ppm of CPC. Higher concentrations of CPC up to 8,000 ppm were tested against Escherichia coli O157: H7, and it was found to reduce the number of bacteria bound to below 0.1%. These studies show a significant inhibition of the growth of these five bacteria in chicken skin.

EXAMPLE 3 Effects of quaternary ammonium compounds on the inhibition of bacterial binding to chicken skin Chicken skin (2.5 x 2.5 cm) cut from a thigh, sterilized by a 45KGy dose of irradiation from an electron source, were placed with the epidermis up in each cavity of six-pocket tissue culture plate. Each piece of skin was inoculated with 5 ml of saline buffered with 0.008 M phosphate (PBS, pH 7.2) containing CPC. Three pieces of skin were used for each concentration of the test compound, including one in which the skins were only treated with 5 ml of PBS (concentration 0). The plates were incubated with shaking (100 rpm) for several times (1 min or 10 min) at 25 ° C. The incubation solution was removed by aspiration, and the skins were rinsed (5 ml PBS) containing 6-8 x 103 CFU / ml of bacteria. After incubation, each piece of skin was rinsed (2X, 5 ml of PBS), to freely remove the bound (unattached) microorganisms, placed in a sterile plastic bag containing 80 ml of saline or 1% peptone, and homogenized for 2 minutes using a laboratory mixer (Stomacher® 400, Seward Medical, London, England). Three aliquots of the homogenate (1 ml) were drained and incubated (37 ° C, 18-24 h). Bacterial colonies were counted, corrected by dilution, and reported as CFU / skin. These studies show the inhibition of the binding of bacteria (Salmonella typhimurium, Staphylococcus aureus, Campylobacter jejuni, Escherichia coli (non-toxin producing species) and Escherichia coli O157: H7) to chicken skin after treatment with concentrations of 50 to 1000 ppm of CPC. The data in these studies show that pre-treating chicken skin with CPC significantly inhibits the binding of these microorganisms to chicken skin. Treatment of chicken skin with CPC for only 1 minute results in significant inhibition of S. typhimurium binding at 500 ppm and 1000 ppm. This shorter contact time of QAC with meat products supports using shorter contact times than those previously reported as effective. Usually, cooling tank dives can be up to 60 minutes, but the data presented here supports that a shorter immersion or contact time can be used, which still results in a significant reduction in the number of viable microorganisms. The contact step of the present invention can be performed for about 20 seconds to about 60 minutes; however, the present invention also discloses a method of a contact time of less than less than 10 minutes, preferably about 20 seconds to about 9 minutes, more preferably about 20 seconds to about 5 minutes, and most preferably about 20 seconds. up to about 90 seconds.

EXAMPLE 4 Effects of Quaternary Ammonium Compounds in the Reduction of Viable Bacteria Attached to a Beef Steak Fillet 82.5 x 2.5 cm resilient tissue squares approximately 0.5 cm thick, sterilized by a 45 KGy dose of irradiation from an electron source, they were placed in each cavity of the six-pocket tissue culture plate. Each piece of tissue was inoculated with 5 ml of saline solution buffered with 0.008 M phosphate (PBS, pH 7.2) containing 6-8 x 103 CFU / ml of bacteria with the exception of the support control group that was treated with only 5 ml. of PBS. The plates were incubated (30 min, 35 ° C), and each square was rinsed (2X, 5 ml of PBS9 to freely remove the bound (non-bound) migroorganisms.) The inoculated squares were treated with 5 ml of PBS containing the CPC. The three pieces of tissue were used for each concentration of the test compound, including one in which the squares were treated with only 5 ml of PBS (concentration 0.) The plates were incubated with shaking (100 rpm) for 30 min at 25 min. C. After incubation, each square was rinsed (5 ml of PBS), placed in a sterile plastic bag containing 50 ml of 1% peptone and homogenized for 2 minutes using a laboratory mixer (Stomacher® 400, Seward Medical, London, England) Three aliquots of the homogenate (1 ml) were incubated and incubated (37 ° C, 18-24 h) Bacterial colonies were counted, corrected for dilution, and reported as CFU / square. The results of this est udio show a reduction in viable Escherichia coli O157: H7 after treatment with concentrations of 50 to 1000 ppm CPC.

EXAMPLE 5 Effects of Quaternary Ammonium Compounds on the Inhibition of Bacterial Binding to Beef Stem Tissue Resilient tissue (2.5 x 2.5 cm) approximately 0.5 cm thick, sterilized by a dose of 45 KGy of irradiation at From an electron source, they were placed in each cavity of the six-pocket tissue culture plate. Each piece of tissue was treated with 5 ml of saline buffered with 0.008 M phosphate (PBS, pH 7.2) containing CPC. Three pieces of beef tissue were used for each concentration of the test compound, including one in which the squares were treated with only 5 ml of PBS (concentration 0). The culture plates were incubated with shaking (100 rpm) for 10 minutes at 25 ° C. The incubation solution was removed by aspiration, and the squares were rinsed (5 ml of PBS), and then incubated (30 min, 35CC) with 5 ml of PBS containing 6-8 x 103 CFU / ml of bacteria. After incubation, each piece of tissue was rinsed (2X, 5 ml of PBS), to freely remove the bound microorganisms (not bound), placed in a sterile plastic bag containing 50 ml of 1% peptone, and homogenized during 2 minutes using a laboratory mixer (Stomacher® 400, Sewared Medical, London, England). Three aliquots of the homopolyzed (1 ml) were plated and incubated (37 ° C, 18-24 h). Bacterial colonies were counted, corrected by dilution and reported as CFU / square. The results of this study show the inhibition of binding of Escherichia coli O157: H7 after treatment with 50 to 1000 ppm of CPC with a reduction of 76% in the number of bacteria bound to the beef at concentrations of 1000 ppm CPC. Fig. 1 shows the results of a separate test using higher CPC concentrations and the same experimental procedure. At 20,000 ppm CPC, the bacteria were completely inhibited from binding to the res.

EXAMPLE 6 Atomization of chicken precooled with 0.1% cetylpyridinium chloride A spray test chamber was designed and constructed for use in a pilot poultry processing plant. The atomization test system consisted of a test chamber, an atomizing water storage tank, a pressure pump, a filter, pressure regulators, a plastic spray chamber with eight nozzles located on four sides, and a used water collector. There were three nozzles in each of the pipes for front and rear atomization. A nozzle for upper atomization and a nozzle for lower atomization was used. The dimensions of the camera are preferably 91.44 x 91.44 x 91.44 centimeters. With a high pressure booster pump, the pressure could be adjusted between 0-9,842 kg / cm2. The distance between the atomization nozzles and the chicken carcass was 30.48-38.1 centimeters. The upper nozzle was used to atomize the interior of the chicken carcass. Flat cone atomization nozzles (1 / 8TK-SS1, Spraying Systems Co.) were used. The atomization solution in the storage tank was pumped to the pressure regulator, and then atomized through the nozzles in the chamber. In the atomization chamber, several atomization layers consisting of nozzles and stainless steel pipes were installed, and the chamber was covered with plastic sheets to prevent chemical entrainment. A ring was used to hang the chicken carcass in the chamber. The pre-chilled chicken carcasses were obtained from a local poultry processing plant. They were taken from the end of a disemboweling processing line, transported to the research laboratory and used immediately for testing. The time that elapsed between the processing plant and the research laboratory was less than half an hour. The temperature of the chicken carcasses was in the range of 32 to 37 ° C.

Chicken carcasses were inoculated by spraying 1 ml of S. typhimurium at 1 x 106 CFU / ml and then incubated at room temperature for 30 min. The inoculated chicken carcasses were rinsed by spraying running water at 2,109 kg / cm2 and 22 ° C for 5 seconds to freely wash the bound Salmonella cells. Then each cadaver was hung in the atomization chamber and atomized with one of the test compounds. After atomization, each chicken carcass was rinsed with running water for 20 seconds. The chicken carcasses were then washed with buffered peptone water in a plastic bag on an automatic shaker to obtain samples for microbial analysis. The skin color of the chicken was examined visually by comparing the treated birds with the test compounds, such as QACs, with untreated birds. CPC was used at a concentration of 1000 ppm at different atomization pressures and durations. The atomization water temperature was set at room temperature of 22 ° C. The pressures were set at 2,109, 3,515 and 8,436 kg / cm2, and duration at 30 and 90 seconds. Three replications were made for each trial. The reduction of S. typhimurium in chicken carcasses was compared between the groups atomized with the test compound, atomized with water and not atomized. After the spray treatments, each carcass was mechanically agitated with 100 ml of buffered peptone water (BPW) for 1 min, and then the wash water was collected. The samples were diluted, enriched, placed on XLT agar or Petrifilm (3M, Inc., St. Paul, MN for total aerobic count plates) and incubated for 18-24 hours at 37 ° C. Then the colony forming units were counted. The number of bound bacteria was calculated using a most probable number technique. The most likely numbers of total aerobic plate counts and Salmonella were made for each corpse using the wash water samples. An analysis of variance was used to analyze the experimental data to determine any important difference between the treatment groups and controls (SAS / STAT User's Guide, SAS Institute, Inc., Cary, NC 1993). The results of this experiment show that 30 and 90 seconds of atomization of 1000 ppm CPC solution at pressures of 2,109, 3,515 and 8,436 kg / cm2 cause a significant reduction in the number of Salmonella in chicken carcasses. These data show that the atomization method is a viable alternate method to the standard method of immersion or submersion of chickens when sprayed for 30 seconds to 90 seconds with a pressure in the range of 2,109 to 8,436 kg / cm2 at CPC concentration at 1 %. It may be possible to use lower concentrations of CPC with varying spray pressures within the described range of 2,109 to 8,436 gk / cm2 or greater, and varying spray times to obtain the most efficient process, which results in significant reduction in the microorganisms supported in the food. The atomization method would be advantageous for use in industrial processes because many chicken carcasses could be automatically atomized for short periods of time and still result in significant reduction of pathogenic bacteria.

Example 7 Study of effective concentration and time of the effects of quaternary ammonium compounds on S. tvphimurium in chicken skin The effects of CPC on the inhibition and reduction of viable S. typhimurium in chicken skin were studied. The test solutions comprised various concentrations of CPC (Sigma Chemical Co., St.

Louis, MO) in 5% (v / v) glycerin in saline solution buffered with 0.008 M phosphate, pH 7.2 (PBS). The solutions were prepared by dissolving the appropriate amounts of CPC in the glycerin-PBS mixture. Squares of skin (2.5 x 2.5 cm) of thighs of unprocessed chickens, freshly frozen, were sterilized by a dose of 45 kGy irradiation (electron beam of a linear accelerator, lowa State University). The source of S. typhimurium was the ATCC species # 14028 or the NCTC species # 12023). All colony counts were performed on tryptic soy agar plates (TSA, DIFCO, Detroit, MI). The storage of Salmonella was in TSA. The inoculum preparation was carried out as follows.

A flask containing 50 ml of tryptic soy broth was inoculated with S. typhimurium from a single colony and then incubated (37 ° C) with shaking (150 rpm) overnight. An aliquot of the culture was washed with 9 ml of PBS (4800 rpm, 10 min) twice. The pellet was resuspended in PBS to obtain a final cell concentration (spectrophotometrically, 420 nm) from 1 to 2 x 106 colony forming units (CFU) per ml. Chicken skin was removed from the thighs and the epidermis was placed upward in each cavity of six-well tissue culture plates. Skin pieces were inoculated with 5 ml of PBS containing 1 to 2 x 106 CFU of S. typhimurium by me, with the exception of the control control group that was treated with only 5 ml of PBS. The culture plates with the skin pieces were incubated (30 min, 35 ° C), and then the incubator solution was removed by aspiration. The inoculated skins were treated with 5 ml of the test solution. Three-piece skin sets were used for each concentration of test solution, including a set in which the skins were treated only with 5 ml of 5% (v / v) glycerin in PBS (concentration 0). Plates were incubated at 25 ° C with shaking (100 rpm) for 1, 3 or 10 min. After incubation, each piece of chicken was rinsed with suction (5 ml of PBS), placed in a sterile plastic bag containing 50 ml of 0.1% (w / v) peptone, and homogenized for 2 minutes using a laboratory mixer Stomacher® 400 (Seward Medical Co., London, England). One corner of the bag was aseptically cut and the entire contents were transferred to a sterile centrifuge tube, which was then centrifuged for 10 min (12,000rpm, 20 ° C). The pellet was resuspended in 5 ml 0.1% (w / v) peptone / water. One ml of the appropriate dilution was tripped on TSA agar in triplicate and then incubated at 37 ° C for 24 hours, after which the colonies were counted, corrected for dilution, and reported as CFU / skin. The results show that the reduction of Salmonella was dependent on both the CPC concentration and the exposure time. A 5 logio decontamination was almost achieved when dealing with CPC solutions of 4000 and 8000 ppm for contact times as low as 3 min. Skin squares were placed with the epidermis facing up in each cavity of six-pocket tissue culture plates. The skin pieces were treated with 5 ml of test solution. Three-piece sets of skin were used for each concentration of test solution, including a set in which the skins were treated with only 5 ml of 5% (v / v) glycerin in PBS (concentration 0). The culture plates with the skin pieces were incubated at 25 ° C with shaking (100 rpm) for 1, 3 or 10 min. The incubation solution was removed by aspiration, and the skins were rinsed (5 ml of PBS) and then incubated (30 min., 35 ° C) with 5 ml of PBS containing 1 to 2 x 106 CFU of S. typhimurium. my. After incubation, each piece of skin was rinsed with aspiration (5 ml of PBS), placed in a sterile plastic bag containing 50 ml of 0.1% (w / v) peptone, and homogenized for 2 minutes using a mixer of laboratory Stomacher® 400. Three aliquots of the homogenates (1 ml) were plated on TSA agar and incubated at 37 ° C for 24 h, and then the colonies were counted, corrected for dilution, and reported as log10. CFU / skin. The results indicate that the prevention of contamination of Salmonella by pretreatment with CPC also showed concentration and time dependence. The most marked effects were observed for pre-treatment times of 10 minutes, where an inhibition of 4.9 log10 of Salmonella binding was shown at a concentration of 8,000 ppm.

This result is important because the prevention of cross contamination is of vital importance in food processing. The log10 CFU / skin values for the controls were within the range of 4.61 to 5.03. The differences between treated samples and controls were analyzed using ANOVA followed by multiple range analysis of Newman-Keuls, and were statistically significant (p < 0. 01). In another atomization experiment, a 3.3 log10 reduction of Salmonella was obtained, after a 90 second spraying in chicken carcasses with a 5,000 ppm CPC solution.

EXAMPLE 8 Effects of quaternary ammonium compounds in the reduction of viable Listeria monocvtogenes bound to chicken skin The steps of Example 2 were followed, except that L. monocytogenes was used to inoculate the chicken skin and the medium in the plastic bag used in the Stomacher 400 contained 0.1% peptone. At CPC concentrations of 2000 ppm or greater, there was a reduction of more than 4 log10 in L. monocytogenes.

EXAMPLE 9 Effects of Quaternary Ammonium Compounds on the Inhibition of Viable Listeria Monocytokines Binding to Chicken Skin The steps of Example 3 were followed, except that L. monocytogenes was used to inoculate the chicken skin and the medium in the bag. The plastic used in the Stomacher 400 contained 0.1% peptone. The results of this study show a reduction of 82% of bacteria bound to 50 ppm, reduction of 92% to 100 ppm, and reduction of 100% to 500 and 1000 ppm.

Example 10 Effects of quaternary ammonium compounds on the reduction of viable Salmonella Typhimurium bound to catfish, black grapes, and broccoli The effects of CPC on the reduction of viable S. typhimurium in catfish, black grapes and broccoli were studied. The test solutions comprised various concentrations of CPC (Sigma Chemical Co., St. Louis, MO) in 5% (v / v) glycerin in 0.008 M phosphate buffered saline, pH 7.2 (PBS). The solutions were prepared by dissolving the appropriate amounts of CPC in the glycerin-PBS mixture. The food samples were small intact mushrooms, small intact black grapes, florets of broccoli, and squares of catfish skin (2.5 x 2.5 cm) extracted from freshly thawed, unprocessed catfish. The fruits and vegetables were purchased from a local grocery store, while the fish was shipped frozen from a local cat vendor. The source of S. typhimurium was the ATCC species # 14028 or the NCTC species # 12023). All colony counts were performed using XLD selective Salmonella agar plates (DIFCO, Detroit, MI). Additionally, in the catfish experiments, the total aerobic colon counts were performed using a non-selective medium, tryptic soy agar (TSA: DIFCO, Detroit, MI). The storage of Salmonella was in TSA.

The inoculum preparation for S. typhimurium was performed as described in Example 7 above. The food samples were placed in each cavity of the six-pocket tissue culture plates. The samples were then inoculated with 5 ml of PBS containing 1 to 2 x 106 CFU of S. typhimurium per my, with the exception of the control control group that was treated with only 5 ml of PBS. The culture plates with the food samples were incubated (30 min, 35 ° C), and then the incubator solution was removed by aspiration. The inoculated samples were treated with 5 ml of the test solution. Sets of three food samples were used for each concentration of test solution, including a set in which the food samples were treated with only 5 ml of 5% (v / v) glycerin in PBS (concentration 0). The plates were incubated at 25 ° C with shaking (100 rpm) for 3 min. After incubation, each food sample was prepared and placed in a plastic bag for use with the Stomacher ® laboratory mixer as described in Example 7 above. A corner of the bag was aseptically cut and the entire contents transferred to a sterile centrifuge tube, which was then centrifuged for 10 min (12,000 rpm, 20 ° C). The pellet was resuspended in 5 ml of 0.1% (w / v) peptone / water. One ml of the appropriate dilution was plated on XLD agar for grape and broccoli experiments and on XLD agar as TSA for catfish in triplicate. After incubation at 37 ° C for 24 hours, the colonies were counted, corrected for dilution, and reported as CFU / skin for catfish and as CFU / gram for the other food samples. The results of these experiments are shown in Figs. 2-5. As the catfish was not irradiated, Fig. 2 shows the total aerobic bacterial count in non-selective medium, while Fig. 3 shows only Salmonella counts.

Example 11: Atomization effect of quaternary ammonium compounds in the reduction of viable bacteria in whole chicken. These experiments tested whether the effect of atomizing QACs on whole chicken carcasses using a commercial spray would have on the reduction of viable bacteria. Bacterial inoculant solutions were made as follows: E. coli (ATTC # 25922) was grown in brain heart infusion broth (BHI) for 20-24 h and then diluted to a 1: 1000 concentration by adding 0.5 ml of culture. from E. coli to 500 ml of physiological saline solution (PSS). S. typhimurium was grown in BHI for 20-24 h and then diluted to a 1: 5000 concentration by adding 0.1 ml of S. typhimurium culture to 500 ml of physiological saline (PSS). The CPC treatment solution was prepared at a concentration of 5,000 ppm. Pre-cooled chicken carcasses were obtained from a local poultry processor plant for each trial. The corpses were placed in a row of rings, 1 ml of the bacterial inoculant solution was atomized in the corpse's chest, and 1 ml was atomized in the back. The bacteria were allowed to bind for 30 min at room temperature. After the union, the corpses were rinsed in the ring line with running water for 20 seconds. The corpses were divided into groups of ten. For each run, there was a group of ten chickens that were sprayed only with tap water but for S. typhimurium, there was also a group that was not sprayed after inoculation to evaluate the effect of spraying. For all bacteria, a group of carcasses was treated with the JohnsonMR scrubber for 20 seconds at 4,218 kg / cm2 with 35 cups of tap water. After rinsing, the bodies were allowed to settle for 90 seconds, and then rinsed with 20 cups of tap water for 20 seconds at 5,624 kg / cm2. This rinse cycle was repeated either two or three times. The interval of each rinse was also 90 seconds. Another group of corpses was treated with 5,000 ppm CPC for 20 seconds at 4,218 kg / cm2 in the JohnsonMR washer, then allowed to settle for 90 seconds, and then rinsed with 20 cups of tap water for 20 seconds at 5.624 kg / cm2. This rinse cycle was repeated either two or three times. After treatment, the corpses were placed in plastic bags and 100 ml of 0.1% buffered peptone water (BPW) was added to each bag. The bags were mechanically shaken and the rinse was collected for the most probable number (MPN) technique. Petrifilm R was also used for the evaluation of total aerobic plate counts (TPC). C. pre-existing C. jejuni (not inoculated) was enumerated by the MPN technique and E. coli by Petrifilm ™.

The results presented below show that CPC treatment is effective in reducing the number of C. jejuni, E. coli and S. typhimurium. The wash water for S. typhimurium runs was tested and it was found that CPC in the wash water reduced Salmonella by 1 log. Thus, the death data presented below for Salmonella can be reduced by 1 log.

Example 12 Effect of quaternary ammonium compounds on food-supported fungi This study tested the effect of CPC on food-supported fungi. Diagonal cultures of Aspergillus flavus were marked and Penicillium chrysogenum on potato dextrose agar plates (PDA). Thirty minutes after inoculation or 24 h after inoculation (and incubation at room temperature, two round filters (7 mm in diameter) were placed on the surface of each plate, CPC solutions of 200 ppm, 1000 ppm were added. 5000 ppm, and ,000 ppm or distilled and deionized water (DD) to the filters, 10 μl per filter. All plates were incubated with the lid up at room temperature for 48 hours. The diameters of the inhibition rings were measured. The results presented below show that CPC is effective against fungi supported in food.

CPC is effective against fungi supported by the tested foods.

Formulations of quaternary ammonium compounds When a composition is used in an industrial process, it is preferable to work only with small volumes of liquid concentrates instead of large volumes of liquid solupiones. A QAC formulation has been developed that allows QAC concentrations up to 1000-fold higher than those currently available in a formulation made in water alone. This formulation, which contains at least one solubility enhancing agent, provides a soluble concentrate for easy dilution to the final concentration for use in large-scale industrial processing. The enhanced solubility agent works to maintain the solubility of the QAC, so that it does not precipitate in the solution. Any compatible solubility agent can be used, but ethyl alcohol or glycerin are preferred. The formulation may contain ethyl alcohol, glycerin or both. This formulation contains about 100,000 ppm to about 300,000 ppm of QAC, about 0% to about 49% ethyl alcohol and about 0 to 20% glycerin in water. A preferred formulation contains about 150,000 ppm to about 250,000 ppm of QAC, about 10% to about 40% ethyl alcohol and about 0.5 to about 10% glycerin in water. More preferably, the concentration of ethyl alcohol can vary from about 15% to about 30% and the glycerin concentration can vary from about 0.5 to about 5%. Preferably, this formulation contains about 200,000 ppm of QAC, about 20% ethyl alcohol and about 1% glycerin. This formulation is particularly useful as a concentrate to be added to storage tanks, for use in food immersion treatment with QAC, but is also useful in a spray method at a final concentration of approximately 5,000 ppm QAC. A second formulation was developed to increase the contact time of a QAC solution in food products during processing, particularly when delivered by a spray method. This formulation potentially allows a longer contact time of the QAC with the product without any additional step, which would increase the processing time. The formulation, by virtue of its properties, potentially increases the antimicrobial effectiveness of the process. This formulation preferably contains about 50 ppm to about 20,000 ppm of QAC, and at least one solubility enhancing agent selected from about 0 to about 10% ethyl alcohol and about 0 to about 20% glycerin or both. More preferably, this formulation contains about 50 ppm to about 5,000 ppm, about 0 to about 10% ethyl alcohol, and about 1.0% to about 10% glycerin in water. Most preferably, this formulation contains about 500 ppm to about 5,000 ppm of QAC, 0 to 10% of ethyl alcohol and about 1.0 to about 5% of glycerin in water, and more preferably about 1.0 to 3% of glycerin. The above description of the preferred embodiments of the present invention was presented for illustrative purposes and is not intended to limit the invention to the specific compositions used in the examples, because various modifications to the disclosed invention are possible in light of the above teachings. The present invention is based on the discovery that QACs prevent and significantly reduce bacterial contamination by a broad spectrum of microbial contamination supported in foods than previously known. The invention is intended to cover alternatives, modifications and equivalents, which may be included within the spirit and scope of the invention, as defined by the appended claims.

Claims (31)

1. A method for preventing or reducing microbial contamination supported on food in poultry or meat products, comprising: contacting said poultry or meat product with an effective amount of a quaternary compound for less than about 10 minutes to avoid the growth of microorganisms supported in food, or reduce microorganisms supported in viable foods, responsible for said microbial contamination of said poultry or meat product, wherein said quaternary ammonium compound is an alipyridinium salt or a salt of Alkylammonium and said microorganisms supported in food are bacteria, fungi, or parasites and said bacteria are selected from the group consisting of Salmonella, Staphylococcus, Campylobacter, Arcobacter, Listeria, Aeromonas, Bacillus, Escherichia non-toxin producing, Escherichia that produces pathogenic toxins and combinations thereof.

2. The method of claim 1, wherein said pathogenic toxin producing Escherichia is Escherichia coli O157: H7.

3. The method of claim 1, wherein said quaternary ammonium compound is an alkyl pyridinium halide or an alkylammonium halide.

4. The method of claim 3, wherein said alkylpyridinium halide is cetylpyridinium chloride.

5. The method of claim 1, wherein said effective amount of quaternary ammonium compound ranges from about 50 to 20,000 parts per million.

The method of claim 1, wherein said effective amount of quaternary ammonium compound ranges from about 500 to 5,000 parts per million.

The method of claim 1, wherein said contact comprises immersing said poultry or meat product in said quaternary ammonium compound.

The method of claim 1, wherein said contacting comprises atomizing said poultry or meat product with said quaternary ammonium compound.

The method of claim 7 or 8, wherein said sufficient contact time period ranges from about 20 seconds to less than about 10 minutes.

The method of claim 9, wherein said sufficient contact time period ranges from about 20 seconds to about 5 minutes.

The method of claim 10, wherein said sufficient contact time period ranges from about 20 seconds to about 90 seconds.

The method of claim 1, further comprising determining the presence of said quaternary ammonium compound in said meat product after said contact step.

13. A method for preventing or reducing microbial contamination borne on foods in marine, vegetable or fruit products, comprising: contacting said marine, vegetable or fruit products with an effective amount of a quaternary ammonium compound for a sufficient period of time for avoid the growth of microorganisms supported in food, or reduce the microorganisms supported by viable foods, responsible for said microbial contamination in said marine products, vegetables or fruits.

The method of claim 13, wherein said microorganisms are bacteria, fungi or parasites.

15. The method of claim 14, wherein said bacteria are selected from the group consisting of Salmonella, Staphylococcus, Camylobacter, Arcobacter, Listeria, Aeromonas, Bacillus, non-toxin producing Escherichia, and pathogenic toxin-producing Escherichia.

16. The method of claim 15, wherein said pathogenic toxin producing Escherichia is Escherichia coli O157: H7.

The method of claim 13, wherein said quaternary ammonium compound is an alkyl pyridinium salt or an alkylammonium salt.

18. The method of claim 17, wherein said quaternary ammonium compound is alkyl pyridinium halides or alkylammonium halides.

The method of claim 18, wherein said alkylpyridinium halide is cetylpyridinium chloride.

20. The method of claim 13, wherein said effective amount of microbial growth inhibitor of quaternary ammonium compound ranges from about 50 to 20,000 parts per million.

The method of claim 20, wherein said effective amount of inhibiting the microbial growth of quaternary ammonium compound ranges from about 500 to 5,000 parts per million.

22. The method of claim 13, wherein said contact comprises immersing said marine, vegetable or fruit product in said quaternary ammonium compound.

23. The method of claim 13, wherein said contacting comprises atomizing said marine, vegetable or fruit product with said quaternary ammonium compound.

The method of claim 22 or 23, wherein said sufficient contact time period ranges from about 20 seconds to less than about 10 minutes.

25. The method of claim 24, wherein said sufficient contact time period ranges from about 20 seconds to about 5 minutes.

26. The method of claim 25, wherein said sufficient contact time period ranges from about 20 seconds to about 90 seconds.

27. The method of claim 13, further comprising determining the presence of said quaternary ammonium compound in said food product after the contacting step.

28. A concentrated composition for use after dilution in the removal and inhibition of the binding of food-borne microorganisms in food products, comprising: a concentrated amount of a quaternary ammonium compound ranging from about 100,000 parts per million to about 300,000 parts per million; and at least one solubility enhancing agent.

29. The composition of claim 28, wherein said solubilizing enhancing agent is ethyl alcohol or glycerin.

The composition of claim 29, wherein said ethyl alcohol is present from about 0 to about 49% and said glycerin is present from about 0 to about 20% glycerin.

31. A composition for the removal and inhibition of the binding of food-borne microorganisms in food products, comprising: an effective amount of a quaternary ammonium compound ranging from about 50 parts per million to about 20,000 parts per million, and at least one solubility enhancing agent selected from the group consisting of glycerin, ranging from about 0 to about 20%, and ethyl alcohol, ranging from about 0 to about 10%.