WO2013180256A1 - 消毒された肝臓の製造方法 - Google Patents
消毒された肝臓の製造方法 Download PDFInfo
- Publication number
- WO2013180256A1 WO2013180256A1 PCT/JP2013/065144 JP2013065144W WO2013180256A1 WO 2013180256 A1 WO2013180256 A1 WO 2013180256A1 JP 2013065144 W JP2013065144 W JP 2013065144W WO 2013180256 A1 WO2013180256 A1 WO 2013180256A1
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- WIPO (PCT)
- Prior art keywords
- liver
- chlorine
- disinfectant
- bacteria
- freezing
- Prior art date
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/06—Freezing; Subsequent thawing; Cooling
- A23B4/08—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block
- A23B4/09—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block with direct contact between the food and the chemical, e.g. liquid N2, at cryogenic temperature
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/06—Freezing; Subsequent thawing; Cooling
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/06—Freezing; Subsequent thawing; Cooling
- A23B4/08—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/24—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/20—Meat products; Meat meal; Preparation or treatment thereof from offal, e.g. rinds, skins, marrow, tripes, feet, ears or snouts
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Definitions
- the present invention relates to a disinfection treatment method for providing safe raw food. Specifically, the present invention relates to a method for sterilizing food poisoning bacteria present in food by washing, disinfecting, and freezing the food in hot water. The present invention also relates to a method for producing a sterilized food.
- Liver contains a lot of vitamin A, vitamin B group, iron and folic acid. Above all, folic acid and iron have the function of helping hematopoiesis, and are said to be ideal foods for those who need large amounts of iron, such as anemia prevention and pregnant women.
- the liver's high-quality protein is a food that has been eaten since ancient times, such as a diet for liver disease, because it regenerates hepatocytes, and vitamins and minerals activate liver function. Above all, raw liver is a particularly nutritious food because it does not inactivate nutrients such as vitamins due to heat compared to heated liver.
- Campylobacter a food poisoning bacterium
- enterohemorrhagic Escherichia coli hereinafter referred to as EHEC
- EHEC enterohemorrhagic Escherichia coli
- Contamination due to bile moving from the intestinal tract to the liver by circulation is considered. Furthermore, the possibility of the transfer of bacteria into the liver by translocation of the bacteria from the intestinal tract via the portal vein is also considered. In fact, it is known that Campylobacter is present in bovine bile fluid.
- liver disinfection has been performed in order to improve safety as a food material.
- surface cleaning with tap water is mainly known, and in addition to this, there is a method of surface sterilization by immersion in functional water such as ozone water, ice water, and chlorine-based disinfectant.
- functional water such as ozone water, ice water, and chlorine-based disinfectant.
- chlorinated disinfectants sodium hypochlorite is the most widely used one.
- Sodium hypochlorite is industrially produced by absorbing chlorine gas in a sodium hydroxide (NaOH) solution, and a commercially available sodium hypochlorite concentrated solution has a pH of 12 to 12% containing free effective chlorine.
- a strong alkaline solution of .5 to 13.5.
- HOCl hypochlorite
- OCl - alkaline Dissociative
- OCl 2 dissolved chlorine
- HOCl, OCl - is either Cl 2 has a germicidal efficacy, Cl 2 is fast speed disappears scattered, unstable because usually HOCl, OCl - pH region ions are mainly Disinfection and sterilization is performed in Further, OCl - does not pass through the cell wall and cell membrane, have an oxidizing action from the outside of the cell membrane, by damaging, it exerts a bactericidal effect. On the other hand, HOCl is thought to exert a bactericidal effect by passing through the microbial cell wall and exerting an oxidizing action on essential tissues such as enzymes and nucleic acids present in the cytoplasm and plasma membrane.
- HOCl the general public to HOCl is, OCl - there is a strong sterilizing power than. Therefore, in order to increase the sterilization effect, the abundance ratio of HOCl by electrolyzing dilute saline and hydrochloric acid to produce HOCl, and diluting sodium hypochlorite and acidic solution in tap water A two-component method, etc., that increases the temperature is used.
- Chlorine disinfectant is recognized as a food additive and is used in various places such as fresh food.
- bactericidal effect is extremely reduced when organic substances are mixed.
- surface cleaning with a hypochlorous acid solution confirms bacteria even after disinfection, and the bactericidal effect is not always sufficient (Non-Patent Documents 1 and 2).
- these methods are sterilization of only the liver surface, it was impossible to kill bacteria in the bile duct and blood vessels and organic matter in the bile duct.
- the present invention has been made in view of the above situation. It is an object of the present invention to provide a method for sterilizing microorganisms present in a non-human animal raw edible liver (liver) and a method for producing a sterilized raw edible liver. Another object of the present invention is to provide a sterilized liver.
- the present inventors have examined whether or not the cross-contamination that occurs during the dismantling of cattle can be disinfected by an appropriate method to kill bacteria existing in the liver. In addition, it was examined whether or not bacteria existing in the liver could be killed by disinfecting the bile duct and / or portal vein by an appropriate disinfection method and further freezing the liver. Specifically, the present inventors artificially attached enterohemorrhagic Escherichia coli or Campylobacter to the liver surface, and examined the bactericidal effect of the chlorine-based disinfectant. As a result, the present inventors have found that the bacteria existing on the liver surface can be sterilized by the chlorine-based disinfectant.
- the present inventors artificially contaminated enterohemorrhagic Escherichia coli or Campylobacter into the bile duct and portal vein of the liver, and examined the washing effect with hot water and the bactericidal effect of the chlorine-based disinfectant.
- bacteria present in the liver are also present in the liver by washing the bile duct and / or portal vein with boiling water and then disinfecting the bile duct and / or portal vein and liver surface using a chlorine-based disinfectant. It was found that the bacteria can be sterilized.
- the present inventors have found that the disinfection effect can be further enhanced by freezing the liver after disinfection with a chlorine-based disinfectant.
- the present inventors examined the bactericidal effect by adding a disinfecting step in which a chlorine-based disinfectant penetrates into the liver after disinfection of the liver surface. As a result, it was found that the bactericidal effect can be remarkably enhanced. In addition, it has been found that if the frozen liver is thawed at a temperature in ice water, microorganisms inside and on the surface of the liver can be reduced more effectively.
- a method for producing a liver in which microorganisms are sterilized comprising the following steps; (A) high pressure injection of boiling water into the biliary and / or portal vein of the removed non-human animal liver, and (b) both the intrabiliary and portal vein of the liver in step (a) or A process of injecting a chlorine-based disinfectant into either one. [2] The method according to [1], further comprising a step of allowing a chlorine-based disinfectant to penetrate into the liver. [3] The method according to [1] or [2], further comprising a step of freezing the liver.
- microorganism according to [7], wherein the microorganism causing food poisoning is selected from the group consisting of Campylobacter, pathogenic E. coli, Salmonella, Shigella, Aeromonas, Staphylococcus, and hepatitis E virus Method.
- the present invention a method for sterilizing microorganisms existing in the liver, bile duct or blood vessel is provided.
- this method it became possible to reduce the risk of food poisoning due to liver raw diet caused by microorganisms such as Campylobacter and pathogenic E. coli.
- the method of the present invention is useful for preventing microbial contamination of the liver due to crossings from intestinal contents that occur during cattle dissection.
- the present invention has established a technique that enables removal of food poisoning bacterial microorganisms such as Campylobacter and pathogenic Escherichia coli present in the bile duct and portal vein in a state where the liver can eat raw food.
- the present invention can provide a safe liver from which microorganisms are removed not only on the liver surface but also on the inside.
- a chlorine-based disinfectant there is a problem that the sterilization effect is extremely reduced due to organic substances adhering to the liver.
- the microorganisms remaining in the liver can be further sterilized by freezing and thawing the liver after sterilization with a chlorine-based disinfectant.
- FIG. 7 It is a figure which shows the number of each block of the liver left lobe used in Example 7.
- FIG. It is a figure which shows the number of each block of the liver left lobe used in Examples 9-11. It is a figure which shows the relationship between the chemical equilibrium of free effective chlorine, and pH.
- the present invention relates to a method for producing a liver in which microorganisms are sterilized including the following steps.
- the present invention also relates to a method for sterilizing microorganisms present in the liver.
- A a step of high-pressure injection of boiling water into the biliary and / or portal vein of the removed non-human animal liver, and
- a liver whose surface and / or inside is sterilized can be produced.
- the method of the present invention can sterilize microorganisms existing on and / or inside the liver.
- “sterilization” can also be expressed as “sanitization”.
- microorganisms to be sterilized include bacteria, viruses, protozoa and the like. Therefore, “sterilization” in the present invention includes not only killing bacteria but also killing viruses and protozoa.
- bacteria, viruses, and protozoa to be sterilized are not particularly limited, but include bacteria, viruses, and protozoa that cause infectious diseases such as food poisoning.
- Disinfection is a treatment method used to reduce the number of living microorganisms, and sterilization in the present invention includes disinfection.
- infectious bacteria and toxin-type bacteria are known. Infectious bacteria • Campylobacter spp. • Escherichia spp. (Pathogenic E. coli) -Salmonella spp.-Shigella spp.-Vibrio spp.-Aeromonas spp.-Plesiomonas spp.-Yersinia spp.-Listeria spp.-Cronobacter spp.-Citrobacter spp.-Enterobacter spp. Examples include, but are not limited to, genus, Brucella, Helicobacter, Serratia, and the like. On the other hand, examples of the toxin-type bacteria include, but are not limited to, Staphylococcus spp., Clostridium spp., Bacillus spp.
- viruses that causes food poisoning examples include, but are not limited to, norovirus, rotavirus, adenovirus, astrovirus, hepatitis A virus, hepatitis E virus, and the like.
- Campylobacter bacteria are pathogens of humans and wild or domestic animals, and are causative agents of miscarriages, enteritis in animals, and enteritis in humans.
- Campylobacter jejuni and Campylobacter coli are known as causative bacteria of Campylobacter infection, and these bacteria have been designated as food poisoning bacteria (Japanese Patent Application Laid-Open (JP-A)). 62-228096, JP-A-2-84200).
- Campylobacter is classified into 15 bacterial species and 9 subspecies. Among them, C. jejuni accounts for 95-99% of the bacterial species isolated from human diarrhea, and other bacterial species such as C. coli are several percent (Japanese Patent Laid-Open No. 2-154700). C. coli has a very high colonization rate in pigs. In recent years, Campylobacter infectious diseases have been increasing with the increase in imported meat mainly in Southeast Asia. In particular, there has been a rapid increase in cases of infection caused by chicken-related foods whose consumption has increased as substitutes for beef due to problems such as BSE and O157. is doing.
- C. fetus (Campylobacter fetus) was known as a miscarriage of sheep and cattle, it has recently been reported to be involved in human miscarriage and premature labor.
- C. fetus infections caused by live consumption of liver and beef contaminated with C. fetus may present symptoms such as sepsis and meningitis.
- the most important source of infection for humans is chicken that has a high concentration in the intestinal tract. (Japanese Patent Laid-Open No. 3-112498)
- E. coli are harmless, but some are harmful to humans as pathogenic E. coli.
- pathogenic Escherichia coli subject to sterilization in the present invention ⁇ Enterohemorrhagic Escherichia coli (EHEC) (O26, O103, O111, O128, O145, O157, etc.), ⁇ Enteropathogenic E.
- EEC Intestinal invasive Escherichia coli
- EIEC Intestinal invasive Escherichia coli
- ETEC Toxigenic E. coli
- ETEC O6, O8, O11, O15, O25, O27, O29, O63, O73, O78, O85, O114, O115, O128, O139, O148, O149, O159, O166, O169, etc.
- EAggEC Intestinal aggregating E. coli
- Salmonella is a kind of intestinal bacteria that mainly inhabit the digestive tract of humans and animals, and some of them are pathogenic by infecting humans and animals.
- Salmonella spp. That are pathogenic to humans those that cause infectious food poisoning include S. typhimurium S. y Typhimurium and S. enteritidis S. Enteritidis. It is a typical infectious food poisoning, and its main symptoms are abdominal pain, vomiting, digestive symptoms such as diarrhea (sometimes mucous stool), fever (high fever), etc. May become severe. Rarely, endotoxin-related sepsis may result in death.
- Hepatitis E is a viral acute hepatitis conventionally referred to as orally transmitted non-A non-B hepatitis, and its pathogen is hepatitis E virus (HEV).
- HEV hepatitis E virus
- the fatality rate of hepatitis E is said to be 10 times that of hepatitis A, and it can reach 20% in pregnant women. Animals such as pigs, deer, and wild boars are possessed, and recently, the route of infection by eating such as raw stab and leve stab of these animals has been pointed out.
- High-pressure injection of hot water into the bile duct or portal vein of a non-human animal can be performed, for example, with a pressurized spray gun. That is, a nozzle is inserted into the common bile duct on the gallbladder side, and the end of the bile duct toward the intestinal tract is sealed with a ligature or a stopper, a finger or the like, and then injected. Since the bile duct branches into the left lobe side and the right lobe side when entering the liver from the common bile duct, after inserting the nozzle to the back of each branch, change the angle and inject evenly into the left lobe side and the right lobe side.
- a nozzle is inserted into the bile duct toward the intestinal tract to seal the common bile duct on the gallbladder side, and then injected from the opposite direction (see FIGS. 5 and 6).
- the injection into the portal vein is excised, the nozzle is injected from the released portal vein, and the nozzle and portal vein are clamped, fixed with fingers, etc. so as not to leak, and injected.
- the temperature and amount of hot water injected into the bile duct and portal vein of a non-human animal are preferably set within a range in which the liver does not undergo heat degeneration. Examples of such a temperature range include, but are not limited to, 55 ° C. to 90 ° C., preferably 70 ° C. to 85 ° C., and more preferably 80 ° C. to 85 ° C.
- the amount of hot water to be injected can be, for example, 100 to 300 ml, preferably 150 to 250 ml, more preferably 100 to 200 ml, but is not limited thereto.
- the hot water can be injected at once or divided into a plurality of times.
- the injection time per injection can be, for example, 1 to 20 seconds, preferably 3 to 15 seconds, and more preferably 5 to 10 seconds, but is not limited thereto.
- Hot water is preferably injected at high pressure.
- Preferable pressures include, but are not limited to, for example, 0.05 to 0.5 MPa or 0.1 to 0.4 MPa, particularly preferably 0.2 to 0.3 MPa.
- the chlorine-based disinfectant can also be injected, for example, with a pressure spray gun.
- the specific method is as described above.
- the injection amount per injection can be, for example, 100 to 2,000 ml, preferably 150 to 1,000 ml, more preferably 200 to 500 ml, but is not limited thereto.
- the injection of the chlorine-based disinfectant can be performed at once or divided into a plurality of times.
- the injection time per injection can be, for example, 1 to 20 seconds, preferably 3 to 15 seconds, and more preferably 5 to 10 seconds, but is not limited thereto.
- non-human animals in the present invention include, but are not limited to, cows, pigs, chickens, duck, horses, goats, sheep, rabbits, deer, rabbits, dogs, whales, dolphins and the like.
- the liver in the present invention is preferably a liver of a non-human animal in which bile ducts and / or portal veins are not cut off, but is not particularly limited.
- the chlorine-based disinfectant used in the present invention is not limited as long as it can reduce microorganisms present in the liver.
- the chlorine-based disinfectant refers to a drug containing effective chlorine effective for disinfection.
- Chlorine-based disinfectants include disinfectants containing compounds that generate hypochlorite and hypochlorite ions when dissolved in water, such as sodium hypochlorite and calcium hypochlorite, strong acid water, Disinfection agents containing hypochlorous acid and hypochlorite ions, chlorine dioxide water, stable complex chlorine preparations, etc. are known by utilizing electrolysis such as chlorous acid water and electrolytic hypochlorous water. If so, an appropriate disinfectant can be selected based on its composition and properties.
- a food additive type disinfectant it is preferable to use a food additive type disinfectant.
- medical agent examples include alcohol-based disinfectants (eg, ethanol), organic acids and salts thereof, chemicals containing inorganic acids and salts thereof, surfactants, electrolytic acid water, ozone water, and the like.
- alcohol-based disinfectants eg, ethanol
- organic acids and salts thereof e.g, ethanol
- chemicals containing inorganic acids and salts thereof chemicals containing inorganic acids and salts thereof, surfactants, electrolytic acid water, ozone water, and the like.
- the chlorine-based disinfectant injected into the bile duct and the chlorine-based disinfectant injected into the portal vein may be the same or different.
- the concentration of the chlorine-based disinfectant include 50 to 2,000 ppm, 100 to 2,000 ppm, preferably 100 to 1,000 ppm, more preferably 100 to 500 ppm, 200 to 500 ppm, and particularly preferably 200 to 400 ppm. However, it is not limited to these.
- the injection amount per one time can be, for example, 50 to 2,000 ml, preferably 150 to 1,000 ml, more preferably 200 to 500 ml, but is not limited thereto.
- the injection time can be, for example, 1 to 20 seconds, preferably 3 to 15 seconds, more preferably 5 to 10 seconds, but is not limited thereto.
- the liver after disinfection with a chlorine-based disinfectant. Washing can be performed using tap water.
- the surface can be cleaned with sufficient running water.
- the injection amount can be, for example, 100 to 300 ⁇ ml, preferably 150 to 250 ⁇ ml, more preferably 100 to 200 ⁇ ml, but is not limited thereto.
- the cleaning can be performed at once or divided into a plurality of times.
- the injection time per injection can be, for example, 1 to 20 seconds, preferably 3 to 15 seconds, and more preferably 5 to 10 seconds, but is not limited thereto.
- the number of microorganisms contained in the bile fluid in the liver or gallbladder is reduced as compared with that before the injection of hot water and the chlorine-based disinfectant, it can be determined that the bacteria have been sterilized.
- Methods for detecting Campylobacter bacteria and pathogenic E. coli are well known.
- a person skilled in the art can use a commercially available detection reagent such as immunochromatography or ELISA in addition to a method of culturing, a method of quantifying the number of bacteria in bile by PCR, real time PCR, etc.
- the number of microorganisms contained in the bile fluid can be measured, it is not limited to these methods.
- Campylobacter bacteria is mCCDA medium, blood agar medium, skilow medium, etc.
- pathogenic E. coli for example, SMAC medium, CT-SMAC medium, LB medium, etc.
- Salmonella for example, DHL agar medium, MLCB agar medium, SS agar medium and the like can be used, but are not limited thereto.
- a person skilled in the art can select and carry out an optimal detection method by referring to documents such as “Toda Shinbacteriology, Nanzan-do ISBN978-4-525-16013-5”.
- the presence or absence of sterilization can be determined using the shape of the colony as an index. That is, in the present invention, even when the number of bacteria does not change due to the injection of hot water and a chlorine-based disinfectant, it can be determined that the bactericidal effect by Escherichia coli was seen when the colony shape changed from the smooth type to the rough type. .
- Step of freezing the liver In addition to the chlorine disinfection treatment, the effect of sterilizing and extinguishing microorganisms damaged by hot water treatment and chlorine-based disinfectant treatment can be further enhanced by freezing the liver after vacuum packaging. .
- the method of the present invention may further include the following steps in addition to the steps (a) and (b) described above.
- the microorganisms damaged by the steps (a) and (b) can be completely sterilized.
- Disinfection with a chlorine-based disinfectant can be performed by, for example, sprinkling the above-mentioned disinfectant with a spray bottle or the like.
- the chlorine-based disinfectant those described above can be used at the above-mentioned concentrations.
- freezing of the liver is, for example, in the range of -196 ° C to -1 ° C, preferably -80 ° C to -5 ° C, particularly preferably -40 ° C to -10 ° C, more preferably -30 ° C to -20 ° C.
- freezing conditions include -196 ° C. and 28 hours, but are not limited thereto.
- the present invention can include a step of thawing frozen liver. Furthermore, the present invention can include a step of storing the thawed liver.
- the liver is preferably frozen and stored in a sealed container such as a nylon bag and then vacuum-packed using a general vacuum packaging machine or the like. It is a surprising effect that the bactericidal action against microorganisms such as Escherichia coli and Campylobacter which can generally be cryopreserved is enhanced by freezing after heat treatment and chlorination and storage in ice water.
- the excised non-human animal liver tissue including the non-human animal liver, bile duct, portal vein and the like can be obtained through a general distribution channel. Considering microbial contamination in the distribution process, liver tissue as clean as possible is preferable. Further, it is preferable to treat the liver tissue by the method described in the present invention at the time of disassembly.
- the liver surface may additionally include a disinfecting step for allowing the chlorine-based disinfectant to penetrate into the liver.
- a chlorine-based disinfectant-containing liquid-absorbing material a sheet having properties of moisture absorption / liquid-absorbing
- a chlorine-based disinfectant-containing liquid-absorbing material a sheet having properties of moisture absorption / liquid-absorbing
- a chlorine-based disinfectant can be added directly to the liver surface.
- moisture-absorbing and liquid-absorbing materials examples include food dehydration sheets (Drip Keeper (registered trademark, distributor: Nippon Vilene Co., Ltd.), Pichit (registered trademark, distributor: Okamoto Corporation), Cerakeeper (distributor: Show Co., Ltd.)
- the penetration can be performed in a vacuum packaging container.
- the liver can be frozen after an additional disinfection step.
- Examples of the disinfectant used in the additional disinfecting step may include those described above.
- the disinfectant used for disinfection of the bile duct and portal vein may be the same or different.
- the concentration of the chlorine-based disinfectant may be 100 to 2,000 ppm, 200 to 2,000 ppm, 500 to 2,000 ppm, 200 to 500 ppm, but is not limited to these.
- the amount of the penetrant can be 1 to 50 ml, preferably 5 to 30 ml, more preferably 8 to 20 ml per 50 g of the sample, but is not limited thereto.
- the immersion time can be, for example, 10 to 60 seconds, preferably 20 to 50 seconds, and more preferably 30 to 40 seconds, but is not limited thereto.
- the liver obtained by such a method is a liver in which microorganisms (particularly microorganisms causing food poisoning) are sterilized on the surface and / or inside thereof. That is, the present invention also provides a liver tissue whose surface and / or inside is obtained by the method of the present invention.
- the liver tissue of the present invention is a liver tissue that is different from a liver tissue containing microorganisms such as Campylobacter and pathogenic Escherichia coli, and has no risk of food poisoning due to a raw diet. Such liver tissue is preferably distributed in a sealed state.
- a vacuum pack containing liver tissue can be processed under ultrahigh pressure conditions of 2000 to 4500 atm.
- the liver tissue of the present invention is, for example, a washed liver itself, or a liver divided into a left lobe, a right lobe, or the like, or a block-shaped liver cut out of an edible portion, sealed with a nylon bag or vacuum-packed Can be distributed. At that time, it can be distributed as a set with meat other than the liver.
- this invention provides the preservation
- the preservation of the liver can be performed, for example, in the range of 0 to 4 ° C., preferably in the temperature range of ice water (eg, ⁇ 1 ° C. to 1 ° C., particularly preferably 0 ° C.), but is not limited thereto.
- the preservation method of the present invention microorganisms causing food poisoning can be further reduced, and the risk of infection of microorganisms can be reduced.
- the liver can be safely stored and distributed without impairing the texture and taste. It should be noted that all prior art documents cited in the present specification are incorporated herein by reference.
- Example 1 Liver surface decontamination experiment
- Surface contamination countermeasure Assuming cross contamination by intestinal contents at the time of dismantling, certain intestinal hemorrhagic Escherichia coli or Campylobacter is artificially contaminated on the liver surface, and chlorinated disinfectant is sterilized. The effect was examined.
- Method 1. Intestinal hemorrhagic Escherichia coli O157 Sakai (hereinafter referred to as EHEC O157) and Campylobacter jejuni 81-176 (hereinafter referred to as C.
- jejuni 81-176) in the logarithmic growth phase were adjusted to approximately 10 5 and 10 6 cfu / ml in PBS, respectively. Adjusted. In order to count colonies after adjustment, the cells were cultured in a medium suitable for each. EHEC O157 was obtained from the Institute for Microbial Diseases, Osaka University (purchase number: RIMD0509952), and C. jejuni 81-176 was obtained from the American Type Culture Collection (ATCC) (purchase number: BAA-2151). 2. Each surface of the beef liver was cut into blocks with a knife or boiled and sterilized with alcohol, or a scalpel so that the surface was approximately 10 cm square. 3. 1 ml of the bacterial mixture was applied evenly. 4.
- the water wash group is sprinkled with about 10 ml of water, and the disinfection group is sprayed with about 10 ml of the chlorine-based disinfectant Telluron Bleach (ADEKA Clean Aid Co., Ltd.) (400 ppm, 800 ppm). Let stand for a minute. 5.
- the surface of each block was reciprocated twice at a constant pressure using sterilized puss, etc., and 100 cm 2 was wiped off.
- the stock solution was diluted with stock solution, diluted 10-fold and diluted 100-fold, and 100 ⁇ L each was spread on 3 sheets of SMAC medium and mCCDA medium, and cultured.
- SMAC medium was cultured at 37 ° C for 1 day under aerobic conditions, and mCCDA medium was cultured at 37 ° C for 2 days under slightly aerobic conditions (5% O 2 , 10% CO 2 , 85% N 2 ). -Like colonies and C. jejuni-like colonies were counted.
- Method 1 After aseptically collecting bile, the inside of the bile duct of the beef liver from which the gallbladder and pancreas were excised was gently washed and kept at about 40 ° C. The collected bile was inoculated into CT-SMAC medium and mCCDA medium, and the number of viable bacteria in the background was counted. In addition, the number of bacteria was measured by real time PCR as necessary. 2.
- EHEC O157 10 5 cfu / ml and 10 6 cfu / ml of C. jejuni 81-176 were injected into the liver from the bile duct into the liver. 3. After standing for 30 minutes, about 150 ml of hot water (about 85 ° C.) was injected into the bile duct with high pressure (0.2 Mpa) and washed. This operation was repeated again. Further, about 50 ml of hot water (about 85 ° C.) was injected into the portal vein under high pressure (0.2 Mpa) and washed.
- the front, back and cut portions of the cut block were aseptically trimmed, and about 50 g (4 cm ⁇ 4 cm ⁇ 3 cm) of about 2 from the center of the block were cut out aseptically. 6).
- One of about 50 g of the cut sample was not frozen and an equal volume of PBS was added as it was, and a stomacher treatment was performed for 30 seconds to prepare an equal volume of brewed liquid. 7).
- the other sample was frozen at -35 ° C and stored for 28 hours. Thereafter, it was thawed gently at 4 ° C., stored at 4 ° C. for 16 hours, and then a squeezed solution was prepared in the same manner as described above. 8).
- CT-SMAC medium is aerobic condition at 37 ° C for 1 day
- mCCDA medium is microaerobic condition (5% O 2 , 10%
- the cells were cultured at 42 ° C. for 2 days with CO 2 , 85% N 2 ), and the obtained EHEC-like colonies and C. jejuni-like colonies were counted.
- Example 3 Flavor, etc. Fresh beef liver was treated with untreated product (surface washed with tap water, refrigerated at 4 ° C) and according to the example (85 ° C hot water wash 150 ml 2 times, 200 ppm Shiten clean 200 ml injection, surface disinfection with 400 ppm telluron bleach, cut into blocks, vacuum-packed to about -0.07 to -0.09 MPa with vacuum packaging machine, frozen at -35 ° C and then 4 ° C 11 people (10 men, 1 woman) taste the processed product, and the color of the processed liver, texture (whether it has a crisp feeling), taste, smell (chlorine odor, etc.) ) Items were evaluated in the following five stages and scored.
- Example 4 Effect of chlorinated disinfectant in the presence of organic matter
- Shiten clean and portal vein washing solution (mainly containing blood) were mixed 1: 1 and allowed to react at room temperature for 10 minutes. Then, after neutralizing chlorine with 10 times the amount of sodium thiosulfate, 100 ⁇ l of EHEC O157 was inoculated into SMAC medium and C. jejuni 81-176 was inoculated into mCCDA medium, and the number of colonies obtained was calculated.
- Example 5 Rate of bacteria reduction when stored in ice water After aseptically collecting bile, the bile duct of the bovine liver from which the gallbladder and pancreas were excised was gently washed and kept at about 40 ° C. The collected bile was inoculated into CT-SMAC medium and mCCDA medium, and the number of viable bacteria in the background was counted. In addition, the number of bacteria was measured by real time PCR as necessary. Then, a 10 5 cfu / ml of EHEC O157 in the liver from bile duct to 10 6 cfu / ml of C. jejuni 81-176 was poured into 20 ml bile duct.
- the liver was allowed to stand for 30 minutes and sprayed with 400 ppm telluron bleach and 80% ethanol to disinfect the liver surface. Thereafter, two blocks of about 10 cm 3 were cut out from the left lobe using a knife sterilized with hot water and alcohol or a scalpel. The front, back, and cut portions of the cut block were aseptically trimmed, and about 50 g (4 cm x 4 cm x 3 cm) of about 3 from the center of the block were cut aseptically. One of the three cut out samples was added with an equal amount of PBS without being subjected to a freezing treatment, and subjected to a stomacher treatment for 30 seconds to prepare an equal amount of squeezed liquid. One of the remaining two samples was frozen at ⁇ 35 ° C.
- Results Table 4 shows the average number of colonies of EHEC O157 in the liver and the degree of colony reduction (no freezing / 4 ° C., no freezing / ice water) after each treatment. Although there were individual differences in the group without sterilization, an average of about 80 to about 150 cfu of EHEC O157 was present in the liver. In addition, the number of bacteria was not significantly different from the group that was not disinfected even when disinfected with a chlorine-based disinfectant, and about 44 to 805 cfu of EHEC O157 was detected. After sterilization, the damage to EHEC O157 due to freezing was examined. In the non-disinfected group, the number of bacteria decreased from about 1/12 to 1/16 in the 4 ° C storage group.
- the number of bacteria in the group preserved in ice water decreased from 1/30 to 1/180 compared to the untreated group.
- the chlorination 100 ppm, 200 ml treatment group there was no significant difference in the bacterial count reduction rate by freezing (storage at 4 ° C and storage in ice water) compared to the untreated group.
- the 200 ppm and 400 ppm treated groups showed a decrease in the number of bacteria due to freezing compared to the untreated group, and the decrease rate in the 4 ° C. storage group was 57 to 1000 or more.
- the rate of decrease increased in almost all groups, with some exceptions, and in some groups the number of bacteria was zero even at low concentrations such as 100 ppm and 200 ppm.
- Table 5 shows the average number of colonies in C. jejuni in the liver and the degree of colony reduction (no freezing / 4 ° C., no freezing / ice water) after each treatment.
- C. jejuni 81-176 there was no significant difference in the degree of colony reduction between the non-disinfected group and the chlorine-based disinfected group, but in almost all groups, storage in ice water was more effective than 4 ° C. Showed a high reduction rate.
- Example 6 Confirmation of Variation and Uniformity of Internal Contamination by Artificially Ingested Bacteria
- the bacteria were artificially ingested into the liver and the distribution of the bacteria at each site was confirmed.
- Methods EHEC O157 3 ⁇ 10 6 cfu / ml was injected into the liver from the bile duct into the 20 ml bile duct. The liver surface was then disinfected by spraying 400 ppm telluron bleach and 80% ethanol. Next, as shown in FIG. 7, a block of about 100 cm 3 was cut out from the left lobe of the liver and the bottom of the rectangular lobe using a hot water disinfected and alcohol disinfected knife or a scalpel.
- the front, back, and cut portions of the cut block were aseptically trimmed, and about 50 g (4 cm ⁇ 4 cm ⁇ 3 cm) was cut from the center of the block.
- Add an equal volume of PBS to about 50 g of the cut sample perform a stomacher treatment for 30 seconds, prepare an equal volume of squeezed solution, and inoculate 100 ⁇ l of the obtained squeeze liquid on each CT-SMAC medium,
- the cells were cultured at 37 ° C. for 1 day under aerobic conditions, and the resulting EHEC-like colonies were counted to confirm the dispersion and uniformity of internal contamination by artificially ingested bacteria (Table 6).
- Example 7 Bactericidal effect of each chlorine-based disinfectant The bactericidal effect was examined using chlorine-based disinfectants having different properties that are generally commercially available.
- Method EHEC O157 3x10 6 cfu / ml is injected into the bile duct into the 20 ml bile duct, allowed to stand for 10 minutes, then about 150 ml of hot water (about 85 ° C) is injected into the bile duct with high pressure (0.2 Mpa) and washed. This operation was repeated again. Further, about 50 ml of hot water (about 85 ° C.) was injected into the portal vein under high pressure (0.2 Mpa) and washed.
- Example 8 Disinfection effect due to difference in freezing temperature and thawing temperature
- Method EHEC O157 1x10 6 cfu / ml is injected into the bile duct into the 20 ml bile duct, then about 150 ml of hot water (about 85 ° C) is injected into the bile duct with high pressure (0.2 Mpa) and washed again. Repeated. Furthermore, about 50 ml of hot water (about 85 ° C.) was injected into the portal vein at high pressure (0.2 Mpa) and washed.
- Vinac Vinac was injected into the bile duct with high pressure (0.2 Mpa) and disinfected. After disinfection, the liver surface was disinfected by spraying 400 ppm telluron bleach and 80% ethanol. Then, using a knife or scalpel sterilized with hot water and alcohol, cut out about 80 cm 3 blocks from the left lobe (A to E) at 5 locations (A to E), and about 50 g (4 cm x 4 cm) from the center of each block. 4 cm x 3 cm).
- Vacuum-packed samples are divided into three groups, which are frozen at -30 ° C and -196 ° C without freezing, frozen and stored for 28 hours, and then thawed gently in ice water (0 ° C) and 4 ° C, Stored at 0 ° C. and 4 ° C. for 16 hours, respectively. The group without freezing immediately moved to the next operation.
- Example 9 Additional disinfection effect by freezing process and chlorine-based disinfectant-containing hygroscopic / liquid-absorbing material
- the sterilizing effect of freezing process and additional disinfection by using a chlorine-based disinfectant-containing hygroscopic / liquid-absorbing material was examined.
- Method After injecting 20 ml of EHEC O157 1x10 7 cfu / ml into the bile duct, inject about 150 ml of hot water (about 85 ° C) into the bile duct with high pressure (0.2 Mpa), wash, and repeat this operation. It was. Furthermore, about 50 ml of hot water (about 85 ° C.) was injected into the portal vein at high pressure (0.2 Mpa) and washed.
- Example 10 Additional disinfection effect by adding chlorine-based disinfectant
- the bactericidal effect of additional disinfection by adding a chlorine-based disinfectant was examined.
- Method After injecting 20 ml of EHEC 1x10 7 cfu / ml into the bile duct about 150 ml of hot water (about 85 ° C) was injected into the bile duct with high pressure (0.2 Mpa), and this operation was repeated. . Further, about 50 ml of hot water (about 85 ° C.) was injected into the portal vein under high pressure (0.2 Mpa) and washed.
- Example 11 Additional disinfection effect by stable complex chlorine preparation
- the disinfection effect of the additional disinfection by K guard which is a stable complex chlorine preparation was examined.
- Method After injecting 20 ml of EHEC O157 1x10 7 cfu / ml into the bile duct, inject about 150 ml of hot water (about 85 ° C) into the bile duct with high pressure (0.2 Mpa), wash, and repeat this operation. It was. Further, about 50 ml of hot water (about 85 ° C.) was injected into the portal vein under high pressure (0.2 Mpa) and washed.
- K-guard trade name, Eight Knot Co., Ltd.
- the liver surface was disinfected by spraying 400 ppm of the chlorine disinfectant Telluron Bleach and 80% ethanol.
- a knife or surgical scalpel sterilized with boiling water, cut out about 100 cm 3 blocks from the left lobe (A to C) at three locations (A to C), and about 50 g (4 cm x 4 cm) from the center of each block. Two samples (4 cm ⁇ 3 cm) were cut out (FIG. 9).
- the present invention is considered to be an invention that greatly contributes to conventional processing, distribution and sales methods of food, particularly raw meat such as raw liver.
- a method for sterilizing microorganisms existing in the liver, bile duct or blood vessel is provided.
- the method of the present invention is useful in the field of providing foods with reduced risk of food poisoning or distributing such foods.
- the liver obtained according to the method of the present invention did not show any influence on taste and the like. Therefore, the present invention is useful in the fields of processing, distribution and sales of raw meat. It is also useful for protecting Japanese food culture.
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Abstract
Description
具体的には本発明者らは、腸管出血性大腸菌あるいはカンピロバクターを肝臓表面に人為的に付着させ、塩素系消毒剤の殺菌効果を検討した。その結果本発明者らは、塩素系消毒剤により、肝臓表面に存在する細菌を殺菌できることを見出した。
また本発明者らは、腸管出血性大腸菌あるいはカンピロバクターを肝臓の胆管内および門脈に注入して人為的に汚染させ、熱湯による洗浄効果および塩素系消毒剤の殺菌効果を検討した。その結果、肝臓内部に存在する細菌についても、胆管および/または門脈を熱湯洗浄し、その後胆管および/または門脈と肝臓表面を塩素系消毒剤を使用して消毒することにより、肝臓に存在する細菌の殺菌が可能なことを見出した。さらに本発明者らは、塩素系消毒剤による消毒後、肝臓を冷凍処理することにより、さらに殺菌効果を高めることが可能なことを見出した。
加えて、本発明者らは肝臓表面消毒後に肝臓に塩素系消毒剤を浸透させる消毒工程を追加することによる殺菌効果を検討した。その結果、殺菌効果を顕著に高めることが可能なことを見出した。
また冷凍された肝臓の解凍を氷水中の温度で行えば、より効果的に肝臓内部や表面の微生物を減少させることが可能なことを見出した。
〔1〕以下の工程を含む、微生物が殺菌された肝臓の製造方法;
(a)摘出された非ヒト動物の肝臓の胆管内および門脈の両方またはいずれか一方に熱湯を高圧注入する工程、および
(b)工程(a)の肝臓の胆管内および門脈の両方またはいずれか一方に塩素系消毒剤を注入する工程。
〔2〕肝臓に塩素系消毒剤を浸透させる工程をさらに含む、〔1〕に記載の方法。
〔3〕肝臓を冷凍する工程をさらに含む、〔1〕または〔2〕に記載の方法。
〔4〕以下の工程をさらに含む、〔1〕に記載の方法;
(a)肝臓表面を塩素系消毒剤により消毒する工程、および
(b)工程(a)の肝臓を冷凍する工程。
〔5〕肝臓を冷凍する工程の前に、肝臓に塩素系消毒剤を浸透させる工程をさらに含む、〔4〕に記載の方法。
〔6〕肝臓を解凍する工程をさらに含む、〔3〕~〔5〕のいずれかに記載の方法。
〔7〕微生物が食中毒を引き起こす微生物である、〔1〕~〔6〕のいずれかに記載の方法。
〔8〕食中毒を引き起こす微生物がカンピロバクター、病原性大腸菌、サルモネラ属菌、シゲラ属菌、エロモナス属菌、スタフィロコッカス属菌、E型肝炎ウィルスからなる群より選択される、〔7〕に記載の方法。
〔9〕〔1〕~〔8〕のいずれかに記載の方法によって得られる、殺菌された肝臓組織。
あるいは本発明は、肝臓の生食が可能な状態で、胆管内や門脈内に存在するカンピロバクターや病原性大腸菌などの食中毒細菌微生物の除去を可能とする技術を確立した。本発明により、肝臓表面のみならず、内部についても微生物が除かれた安全な肝臓を提供することができる。
塩素系消毒剤による殺菌については、肝臓に付着した有機物により殺菌効果が極端に減少するという問題があった。しかし本発明の方法では、塩素系消毒剤による殺菌の後、肝臓を冷凍し、解凍することにより、肝臓に残存する微生物をさらに殺菌することができる。
(a)摘出された非ヒト動物の肝臓の胆管内および/または門脈に熱湯を高圧注入する工程、および
(b)工程(a)の肝臓の胆管内および/または門脈に塩素系消毒剤を注入する工程。
本発明の方法を使用することにより、その表面および/または内部が殺菌された肝臓を製造することができる。また本発明の方法によって、肝臓表面および/または内部に存在する微生物を殺菌することができる。本発明において「殺菌」は「除菌」と表現することもできる。
感染型細菌としては
・カンピロバクター属菌
・エシェリヒア属菌(病原性大腸菌)
・サルモネラ属菌
・シゲラ属菌
・ビブリオ属菌
・エロモナス属菌
・プレシオモナス属菌
・エルシニア属菌
・リステリア属菌
・クロノバクター属菌
・シトロバクター属菌
・エンテロバクター属菌
・プロテウス属菌
・プロビデンシア属菌
・ブルセラ属菌
・ヘリコバクター属菌
・セラチア属菌
などを例示することができるがこれらに限定されない。
一方、毒素型細菌としては
・スタフィロコッカス属菌
・クロストリジウム属菌
・バシラス属菌
などを例示することができるがこれらに限定されない。
・ノロウィルス
・ロタウィルス
・アデノウィルス
・アストロウィルス
・A型肝炎ウィルス
・E型肝炎ウィルス
などを例示することができるがこれらに限定されない。
・クリプトスポリジウム
・サイクロスポラ
・クドア(住肉胞子虫)
・アメーバ赤痢
・ザルコシスティス
などを例示することができるがこれらに限定されない
・腸管出血性大腸菌(EHEC)(O26、O103、O111、O128、O145、O157など)、
・腸管病原性大腸菌(EPEC)(O6、O44、O55、O86、O111、O114、O119、O125、O127、O128、O142、O158など)、
・腸管侵入性大腸菌(EIEC)(O28ac、O112、O121、O124、O136、O143、O144、O152、O164など)、
・毒素原性大腸菌(ETEC)(O6、O8、O11、O15、O25、O27、O29、O63、O73、O78、O85、O114、O115、O128、O139、O148、O149、O159、O166、O169など)、
・腸管凝集性大腸菌(EAggEC)(O44、0104、O127、O128など)
などが挙げられるがこれらに限定されない。
・shitenクリーン(販売元:株式会社Shiten)
・Vアイナック(販売元:株式会社ルピナス、登録商標)
・テルロンブリーチ(販売元:ADEKAクリーンエイド株式会社、登録商標)
・ミルトン(販売元:キョーリン製薬、登録商標)
・ピュリファンP(販売元:健栄製薬株式会社、登録商標)
・テキサント(販売元:日本ケミファ株式会社、登録商標)
・ハイポライト(販売元:サンケミファ株式会社、登録商標)
・ピューラックス(販売元:株式会社オーヤラックス、登録商標)
・ヤクラックスD(販売元:ヤクハン製薬株式会社)
・ハイクロソフト水(製造器販売元:株式会社オーク)
・クレベリン(販売元:大幸薬品株式会社、登録商標)
・エヴァ水(販売元:エヴァテック株式会社、登録商標)
・Kガード(販売元:エイトノット株式会社)
などが市販されており、容易に入手することが可能である。あるいは、塩素系消毒剤を調製し、それを使用することもできる。塩素系消毒剤は、例えば次亜塩素酸ナトリウムや次亜塩素酸カルシウムなどを使用して、当業者に周知の方法によって調製することができる。
胆管に注入する塩素系消毒剤と門脈に注入する塩素系消毒剤は同じものであっても異なるものであってもよい。
カンピロバクター属細菌および病原性大腸菌の検出方法は周知である。例えば当業者であれば、培養による方法、PCR、real time PCR等により胆汁液中の菌数を定量する方法の他、イムノクロマト法、ELISA等の市販の検出試薬を使用することにより、肝臓や胆嚢内の胆汁液に含まれる微生物数を計測することができるが、これらの方法に限定されない。培養による方法の場合、カンピロバクター属細菌の検出は例えばmCCDA培地、血液寒天培地、スキロー培地などを、病原性大腸菌の場合は例えばSMAC培地、CT-SMAC培地、LB培地などを、サルモネラ属菌の場合は例えば、DHL寒天培地、MLCB寒天培地、SS寒天培地などを使用することができるがこれらに限定されない。
当業者であれば、例えば「戸田新細菌学、南山堂ISBN978-4-525-16013-5」などの文献を参照することにより、最適な検出方法を選択し、実施することができる。
(c)肝臓を冷凍する工程
塩素消毒処理に加え、肝臓を真空包装後に冷凍処理することにより、熱湯処理および塩素系消毒剤処理によりダメージを受けた微生物の殺菌、消滅効果をより高めることができる。
(c)肝臓表面を塩素系消毒剤により消毒する工程、および
(d)工程(c)の肝臓を冷凍する工程。
一般に凍結保存が可能とされている大腸菌やカンピロバクター等の微生物に対する殺菌作用が、熱処理および塩素処理後の冷凍および氷水中での保存により増強されることは驚くべき効果である。
吸湿・吸液材としては例えば食品用脱水シート(ドリップキーパー(登録商標、販売元:日本バイリーン株式会社)、ピチット(登録商標、販売元:オカモト株式会社)、セラキーパー(販売元:株式会社ショーテック)等が用いられるが、これらと同様の性質を持つものであれば特に限定されない。あるいは、浸透(浸漬)は真空の包装容器の中で行うこともできる。
本発明においては、追加の消毒工程の後、肝臓を冷凍することができる。
塩素系消毒剤の濃度は、100~2,000 ppm、200~2,000 ppm、500~2,000 ppm、200~500 ppmなどを採用することができるがこれらに限定されない。また浸透液の量は検体50 gあたり1~50 ml、好ましくは5~30 ml、さらに好ましくは8~20 mlとすることができるがこれらに限定されない。
また浸漬時間は、例えば10~60秒、好ましくは20~50秒、さらに好ましくは30~40秒とすることができるがこれらに限定されない。
本発明の肝臓組織は、例えば、洗浄された肝臓そのもの、もしくは左葉、右葉などに小分けした肝臓、もしくは可食部を切り出したブロック状の肝臓としてナイロンバック等で密封、もしくは真空包装した形で流通させることができる。その際、肝臓以外の食肉とセットで流通させることができる。
(a)本明細書に記載の方法により殺菌された肝臓を取得する工程、
(b)工程(a)の肝臓を氷水中の温度下において保存する工程。
肝臓の保存は、例えば0~4 ℃の範囲、好ましくは氷水中の温度の範囲(例えば-1 ℃~1 ℃、特に好ましくは0 ℃)で行うことができるがこれに限定されない。本発明の保存方法を使用することにより、食中毒を引き起こす微生物がさらに減少し、微生物の感染リスクを低減させることができる。加えて、食感、味覚を損なうことなく安全に肝臓を保存、流通させることができる。
なお本明細書において引用された全ての先行技術文献は、参照として本明細書に組み入れられる。
実施例1 肝臓表面汚染除去実験
表面汚染対策:解体時の腸管内容物による交差汚染を想定して、一定の腸管出血性大腸菌あるいはカンピロバクターをレバー表面に人為的に汚染させ、塩素系消毒剤の殺菌効果を検討した。
方法
1.対数増殖期の腸管出血性大腸菌O157 Sakai株(以下EHEC O157)とカンピロバクター・ジェジュニ 81-176株(以下C. jejuni 81-176)をそれぞれPBSにて約105、106 cfu/mlとなるように調整した。調整後コロニーを計測するためそれぞれに適した培地にて培養した。なお、EHEC O157は大阪大学微生物病研究所(購入番号:RIMD0509952)、C. jejuni 81-176はAmerican Type Culture Collection(ATCC)(購入番号:BAA-2151)より入手した。
2.牛レバーの表面それぞれ、約10 cm 四方になるように煮沸およびアルコールにて滅菌した包丁、もしくは外科用メスにてブロック状に切り取った。
3.菌の混液1 mlずつを均等に塗り付けた。
4.30分間静置後、水洗浄群は約10 mlの水、消毒群は塩素系消毒剤テルロンブリーチ(ADEKAクリーンエイド株式会社)(400 ppm, 800 ppm)約10 mlを霧吹きで振りかけ、30分間静置した。
5.それぞれのブロックの表面を滅菌プース等を用いて一定圧力で2往復、100 cm2を拭き取り、拭き取ったプース等をPBSに懸濁し、揉み出した。揉みだし液を原液、10倍希釈、100倍希釈し、それぞれ100 μLずつを各3枚のSMAC培地とmCCDA培地上に広げ、それぞれ培養した。SMAC培地は好気条件下で37 ℃、1日、mCCDA培地は微好気条件(5 % O2, 10 % CO2, 85 % N2)で37 ℃、2日培養し、得られたEHEC様コロニーとC. jejuni様のコロニーを計測した。
EHEC O157、C. jejuni 81-176の105 cfu/ml塗布群では400 ppmの塩素系消毒剤により表面汚染菌が完全に除去できた。また、EHEC O157の106 cfu/ml塗布群でも400 ppmの塩素系消毒剤によりEHEC O157は約1/2000、800 ppmでは約1/10000まで除菌できた。C. jejuni 81-176の106 cfu/ml塗布群においても400 ppmでは約1/1000に、800 ppmでは完全に除菌する事ができた(図1)。
内部汚染対策:一定の腸管出血性大腸菌あるいはカンピロバクターを肝臓の胆管内に注入して人為的に汚染させ、温湯による洗浄効果および塩素系消毒剤の殺菌効果を検討した。
方法
1.無菌的に胆汁を採取した後、胆嚢、膵臓を切除した牛肝臓の胆管内を緩やかに洗浄し、約40 ℃に保った。採取した胆汁はCT-SMAC培地、mCCDA培地に接種し、バックグラウンドの生菌数を計測した。また、必要に応じてreal time PCRにより、菌数を測定した。
2.胆管から肝臓内にEHEC O157 105 cfu/mlとC. jejuni 81-176 の106 cfu/mlを20 ml胆管内に注入した。
3.30分間静置後、約150 mlの温湯(約85 ℃)を胆管内に高圧注入(0.2 Mpa)し洗浄した。この操作を再度繰り返した。さらに、門脈内に約50 mlの温湯(約85 ℃)を高圧注入(0.2 Mpa)し洗浄した。その後、約200 mlもしくは500 mlの塩素系消毒剤Shitenクリーン(株式会社Shiten)(400 ppmおよび2,000 ppm)を胆管内に高圧注入(0.2 Mpa)し消毒した。さらに約200 mlのShitenクリーンを門脈内に高圧注入(0.2 Mpa)し消毒した。
4.消毒後30分間静置後、400 ppmのテルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。
5.熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて左葉から約10 cm3程度のブロックを2カ所切り出した。切り出したブロックの表面、裏面、切断部分を無菌的にトリミングし、ブロックの中心部から約50 g (4 cm x 4 cm x 3 cm)を2つ、滅菌的に切り出した。
6.切り出した約50gのサンプルのうち一方は凍結させずそのまま等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製した。
7.もう一方のサンプルについては-35 ℃で凍結させ、28時間保存した。その後、4 ℃で緩やかに解凍し、16時間4℃で保存した後、上記と同様の方法で揉み出し液を作製した。
8.揉みだし液100 μlをそれぞれCT-SMAC培地とmCCDA培地上に接種し、CT-SMAC培地は好気条件下で37 ℃、1日、mCCDA培地は微好気条件(5 % O2, 10 % CO2, 85 % N2)で42 ℃、2日培養し、得られたEHEC様コロニーとC. jejuni様のコロニーを計測した。
消毒を行わない群では個体差はあるが、平均で46.7~111.8 cfuのEHEC O157が肝臓内に存在した。また、塩素系消毒剤により消毒を行っても菌数は消毒しなかった群と大きな差は認められず、12.3~570.0 cfu のEHEC O157が検出された。しかし、消毒しない群のコロニー形状と消毒群のコロニー形状を比較したところ、無消毒群ではEHEC O157に典型的なスムースなコロニー形状を示していたが、塩素系消毒処理群では表面が粗くなったラフ型のコロニー形状を示していた(図2)。
C. jejuni 81-176を用いた投与実験では、塩素系消毒剤による消毒と冷凍により、完全に0 cfuにはならなかったが、コロニー数の減少度合(冷凍無/冷凍後)は無消毒群の約9から18と比べ、塩素系消毒処理群では約7から66と無消毒群と同等か、それ以上の菌の減少効果が認められた。
新鮮な牛生レバーについて、未処理品(水道水による表面洗浄の後、4 ℃で冷蔵)と実施例に従って処理した(85 ℃温湯洗浄150 ml 2回、200 ppm のShitenクリーン200 mlの注入、400 ppm のテルロンブリーチによる表面消毒、ブロックに切り分け後、真空包装機にて真空度約-0.07~-0.09 MPa程度に真空包装した後に-35 ℃凍結の後、4 ℃で緩やかに解凍)処理品を11名(男性10名、女性1名)に試食してもらい、処理した生レバーの色、食感(ぷりぷり感があるか)、美味しさ、臭い(塩素臭等)の項目について次の5段階で評価し、採点を受けた。
良い。ほとんど変わらない。違いがわからない。 ・・・・・ 5 点
やや良い。違いがわずかにあるが、気にはならない。・・・・・ 4 点
普通。違いが少しあるが、喫食には問題ない。 ・・・・・ 3 点
やや劣る。明らかに違いがあり、気になる ・・・・・ 2 点
劣る。明らかに違いがある。喫食をためらう ・・・・・ 1 点
11名とも形状、臭い、味覚、食感等の品質に対する悪影響を認めず、商品価値の高い物であると判定した(表3)。また、処理済み生レバーの試食後の感想として・処理済みのほうが、レバー独特のにおいがなく、食べやすい・熟成している・おいしい・あまみがある・逆にいい・くせがない・もちもちしている、といった味や風味、食感の向上を訴えた感想が見られた。
方法
200 ppmのShitenクリーンと門脈洗浄液(主に血液を含む)を1:1混合し、10分間室温で反応させた。その後、10倍量のチオ硫酸ナトリウムで塩素を中和した後、100 μlをEHEC O157はSMAC培地、C. jejuni 81-176はmCCDA培地に植菌し、得られたコロニー数を算出した。
EHEC O157、C.jejuni 81-176とも、濃い濃度(0.8 mg/ml)の門脈洗浄液(血液タンパク)が存在している場合、塩素系消毒剤の効果が減弱した。
一方、1/4希釈した門脈洗浄液 (血液 タンパク量0.2 mg/ml)の場合、消毒効果が見られ、約5 x 106 cfuのEHEC O157、C.jejuni 81-176 が検出限界である0 cfu(100 cfu/ml以下)まで減少した(図3、4)。
方法
無菌的に胆汁を採取した後、胆嚢、膵臓を切除した牛肝臓の胆管内を緩やかに洗浄し、約40 ℃に保った。採取した胆汁はCT-SMAC培地、mCCDA培地に接種し、バックグラウンドの生菌数を計測した。また、必要に応じてreal time PCRにより、菌数を測定した。次に、胆管から肝臓内にEHEC O157の 105 cfu/mlとC. jejuni 81-176の 106 cfu/mlを20 ml胆管内に注入した。30分間静置後、約150 mlの温湯(約85 ℃)を胆管内に高圧注入(0.2 Mpa)し洗浄し、この操作を再度繰り返した。さらに、門脈内に約50 mlの温湯(約85 ℃)を高圧注入(0.2 Mpa)し洗浄した。
その後、約200 mlのShitenクリーン(400 ppm、200 ppm、100 ppm)を胆管内に高圧注入(0.2 Mpa)し消毒した。さらに約200 mlのShitenクリーン(400 ppm、200 ppm、100 ppm)を門脈内に高圧注入(0.2 Mpa)し消毒した。消毒後30分間静置し、400 ppmのテルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。その後、熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて左葉から約10 cm3程度のブロックを2カ所切り出した。切り出したブロックの表面、裏面、切断部分を無菌的にトリミングし、ブロックの中心部から約50 g (4 cm x 4 cm x 3 cm)を3つ、滅菌的に切り出した。切り出した3つのサンプルのうち1つは凍結処理を行わずに等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製した。また、残り2つのサンプルのうち一方は、-35 ℃で凍結させ28時間保存後、4 ℃で緩やかに解凍し、4 ℃で16時間保存した。さらにもう一方は、凍結・解凍後、氷水中で40時間保存した。これらは保存後、上記と同様の方法で揉み出し液を作製した。得られた揉みだし液100 μlをそれぞれCT-SMAC培地とmCCDA培地上に接種し、CT-SMAC培地は好気条件下で37 ℃、1日、mCCDA培地は微好気条件(5 % O2, 10 % CO2, 85 % N2)で42 ℃、2日培養し、得られたEHEC様コロニーとC. jejuni様のコロニーを計測し、温湯による洗浄効果および塩素系消毒剤の内部汚染に対する殺菌効果を検討した。
実験に影響を与える可能性のある肝臓については実施例2と同様の方法で棄却を行った。
表4に各処理を行った肝臓内EHEC O157の平均コロニー数およびコロニー数減少度合(冷凍無/4 ℃、冷凍無/氷水中)を示す。消毒を行わない群では個体差はあるが、平均で約80~約150 cfuのEHEC O157が肝臓内に存在した。また、塩素系消毒剤により消毒を行っても菌数は消毒しなかった群と大きな差は認められず、約44~805 cfu のEHEC O157が検出された。
消毒後、冷凍によるEHEC O157へのダメージを検討したところ、無消毒群では4 ℃保存群で菌数が約1/12から1/16へと減少した。さらに氷水中で保存した群では菌数が無処理群に比べ1/30から1/180に減少した。一方、塩素消毒100 ppm、200 ml処理群では無処理群と比べ冷凍(4 ℃保存および氷水中での保存)による菌数減少率に、大きな差は見られなかった。しかしながら、200 ppm、400 ppm処理群では無処理群と比べ冷凍による菌数の減少が見られ、4 ℃保存群の減少率は57から1000以上を示した。さらに氷水中での保存により、一部を除いて、ほぼ全ての群で減少率は増加し、塩素濃度100 ppm や200 ppmといった低い濃度でも菌数が0となる群も認められた。
消毒効果を正確に測定するため、人工的に肝臓に菌を摂取させ各部位における菌の分布を確認した。
方法
胆管から肝臓内にEHEC O157 3x106 cfu/mlを20 ml胆管内に注入した。その後、400 ppmのテルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。次に図7に示す様に肝臓の左葉および方形葉下部9カ所から熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて約100 cm3程度のブロックを切り出した。切り出したブロックの表面、裏面、切断部分を無菌的にトリミングし、ブロックの中心部から約50 g (4 cm x 4 cm x 3 cm)を切り出した。切り出した約50 gのサンプルに等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製し、得られた揉みだし液100 μlをそれぞれCT-SMAC培地上に接種し、好気条件下で37 ℃、1日培養し、得られたEHEC様コロニーを計測し、人工的に摂取した菌による内部汚染のばらつきおよび均一性を確認した(表6)。
EHEC O157を胆管内に注入し、その分布を観察したところ、部位によって若干の差は見られるが、ほぼ全域に肝臓1 g あたり約105 cfuの菌が均一に分布していることが分かった。
一般的に市販されている性質の異なる塩素系消毒剤を使用して殺菌効果の検討を行った。
方法
胆管内にEHEC O157 3x106 cfu/mlを20 ml胆管内に注入し、10分間静置後、約150 mlの温湯(約85 ℃)を胆管内に高圧注入(0.2 Mpa)し洗浄し、この操作を再度繰り返した。さらに、門脈内に約50 mlの温湯(約85 ℃)を高圧注入(0.2 Mpa)し洗浄した。
その後、約500 mlの塩素系消毒剤Vアイナック(登録商標、株式会社ルピナス)(500 ppm)(表8)、エヴァ水(登録商標、エヴァテック株式会社)(500 ppm)(表9)、テルロンブリーチ(500 ppm)(表10)を胆管内に高圧注入(0.2 Mpa)し消毒した。さらに約300 mlのVアイナックを注入後、約300 mlのエヴァ水を注入した群(表11)も検討した。消毒剤注入後、400 ppmの塩素系消毒剤テルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。その後、熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて左葉から約100 cm3程度のブロックを4カ所(A~D)切り出し、各ブロックの中心部から約50 g (4 cm x 4 cm x 3 cm)を2サンプル(例えばA1, A2)、切り出した(図8)。そのうち一方は-35 ℃で急速凍結し28時間保存し、その後、4 ℃で緩やかに解凍し、16時間4 ℃に保存した。切り出した約50 gのサンプルに等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製した。得られた揉みだし液100 μlをそれぞれCT-SMAC培地上に接種し、好気条件下で37 ℃、1日培養し、得られたEHEC様コロニーを計測し、塩素系消毒剤の内部汚染に対する殺菌効果を検討した(表8~11)。
注入した菌の殺菌効果をそれぞれ性質の異なる塩素系消毒剤、Vアイナック(CCクラスター製剤)、エヴァ水(緩衝法次亜塩素酸水)、テルロンブリーチ(塩素系除菌・漂白剤)を用いて検討した。結果、肝臓の個体差は見られるが、塩素系消毒剤による単独消毒では、どの塩素系消毒剤を用いても肝臓1 gあたりの菌数を約1/2~1/10程度低下させる程度であった。しかしながら、塩素処理に凍結処理を加えると、すべての群において約1/1000~1/5000程度にまで低下させることができ、もっとも消毒効果が強かった例では肝臓1 gあたり10個程度の菌数にまで低下させることが出来た。また、異なる塩素系消毒剤の併用による効果を検討したが、CCクラスター製剤であるVアイナックと緩衝法次亜塩素酸水であるエヴァ水との併用では特に相乗効果等は認められなかった。
各塩素系消毒剤の特徴を表7に示した。
冷凍温度および解凍温度の違いによる消毒効果の影響を検討した。
方法
胆管内にEHEC O157 1x106 cfu/mlを20 ml胆管内に注入し、その後、約150 mlの温湯(約85 ℃)を胆管内に高圧注入(0.2 Mpa)し洗浄し、この操作を再度繰り返した。さらに、門脈内に約50 mlの温湯(約85℃)を高圧注入(0.2 Mpa)し洗浄した。
次に、約500 mlの Vアイナックを胆管内に高圧注入(0.2 Mpa)し消毒した。消毒後、400 ppmのテルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。その後、熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて左葉から約80 cm3程度のブロックを5カ所(A~E)切り出し、各ブロックの中心部から約50 g(4 cm x 4 cm x 3 cm)を切り出した。切り出したサンプルを無菌袋に入れ、卓上真空包装機(DZ-300、株式会社アスクワークス)を用いて約30秒~1分かけて真空度約-0.1 MPa付近(約-0.07~-0.09 MPa程度)になるまで真空にし、真空包装した。真空包装した各サンプルを冷凍なし、-30 ℃、-196 ℃で急速凍結する3群に分け、冷凍した後28時間保存し、その後、氷水中(0 ℃)と4 ℃で緩やかに解凍し、16時間それぞれ0 ℃、4 ℃に保存した。冷凍処理しない群はすぐに次の操作に移った。解凍した約50 gの各サンプルに等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製した。得られた揉みだし液100 μlをそれぞれCT-SMAC培地上に接種し、好気条件下で37 ℃、1日培養し、得られたEHEC様コロニーを計測し、冷凍温度および解凍温度による影響を検討した(表12)。
冷凍温度と解凍温度の違いによる消毒効果の影響を調べたところ、凍結温度の違いによる差はあまり認められなかったが、-196 ℃で凍結した群の菌数が低い傾向が見られた。解凍温度の違いでは、一部効果が見られない群があるが、4 ℃での解凍より氷水中(0 ℃)での解凍が効果的である傾向が認められた。一方、消毒処理を行っていない群では、冷凍、解凍によって1/10程度の菌数減少効果しか見られなかった。
凍結工程および塩素系消毒剤含有吸湿・吸液材の使用による追加消毒の殺菌効果を検討した。
方法
胆管内にEHEC O157 1x107 cfu/mlを20 ml胆管内に注入した後、約150 mlの温湯(約85℃)を胆管内に高圧注入(0.2 Mpa)し洗浄し、この操作を再度繰り返した。さらに、門脈内に約50 mlの温湯(約85℃)を高圧注入(0.2 Mpa)し洗浄した。
その後、約500 mlのエヴァ水を胆管内に高圧注入(0.2 Mpa)し消毒した。消毒後、400 ppmのテルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。その後、熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて左葉から約80 cm3程度のブロックを3カ所(A~C)切り出し、各ブロックの中心部から約50 g(4 cm x 4 cm x 3 cm)のサンプルを2か所切り出した(図9)。切り出したサンプルを無菌袋に入れ、さらに約10 mlの500 ppmのエヴァ水を含ませた吸湿・吸液材((ドリップキーパー、約9 cm x 15 cm)(登録商標、日本バイリーン株式会社))を入れ、卓上真空包装機を用いて約30秒~1分かけて真空度約-0.1 MPa付近(約-0.07~-0.09 MPa程度)になるまで真空にし、真空包装した。真空包装した各サンプルを-30 ℃で急速凍結し、28時間保存後、4 ℃で緩やかに16時間かけて解凍した。冷凍処理しない群はすぐに次の操作に移った。解凍した約50 ggの各サンプルに等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製した。得られた揉みだし液100 μlをそれぞれCT-SMAC培地上に接種し、好気条件下で37 ℃、1日培養し、得られたEHEC O157様コロニーを計測し、冷凍温度およびドリップキーパーによる影響を検討した(表13)。
消毒剤注入、凍結・解凍による消毒工程に加え、肉汁などの吸収材としてよく用いられるドリップキーパーに塩素系消毒剤を含ませたものを用いて肝臓肉片に対する追加消毒を試みた。結果、塩素系消毒剤含有ドリップキーパーによる追加消毒により、通常の消毒より1/2~1/5まで低下させることが出来た。また、消毒剤注入後、凍結処理していないサンプルに対しても塩素系消毒剤含有ドリップキーパーによる追加消毒によって菌数が減少する傾向が見られた。
塩素系消毒剤添加による追加消毒の殺菌効果を検討した。
方法
胆管内にEHEC 1x107 cfu/mlを20 ml胆管内に注入した後、約150 mlの温湯(約85 ℃)を胆管内に高圧注入(0.2 Mpa)し洗浄し、この操作を再度繰り返した。さらに、門脈内に約50 mlの温湯(約85 ℃)を高圧注入(0.2 Mpa)し洗浄した。
その後、約500 mlのエヴァ水(2,000 ppm)を胆管内に高圧注入(0.2 Mpa)し消毒した。消毒後、400 ppmの塩素系消毒剤テルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。その後、熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて左葉から約100 cm3程度のブロックを3カ所(A~C)切り出し、各ブロックの中心部から約50 g(4 cm x 4 cm x 3 cm)のサンプルを2か所切り出した(図9)。切り出したサンプルを無菌袋に入れ、さらに約10 mlの2,000 ppmのエヴァ水を入れ、卓上真空包装機を用いて約30秒~1分かけて真空度約-0.1 MPa付近(約-0.07~-0.09 MPa程度)になるまで真空にし、真空包装した。真空包装した各サンプル-30 ℃で急速凍結し、28時間保存後、4 ℃で緩やかに16時間かけて解凍した。冷凍処理しない群はすぐに次の操作に移った。解凍した約50 gの各サンプルに等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製した。得られた揉みだし液100 μlをそれぞれCT-SMAC培地上に接種し、好気条件下で37 ℃、1日培養し、得られたEHEC様コロニーを計測し、エヴァ水による追加消毒効果を検討した(表14)。
塩素系消毒剤含有吸液・吸湿材による追加消毒の代わりに塩素系消毒剤を添加し、真空包装を行ったところ、同様の効果が認められ、1/2から1/10程度まで菌数が低下した。
安定型複合塩素製剤であるKガードによる追加消毒の殺菌効果を検討した。
方法
胆管内にEHEC O157 1x107 cfu/mlを20 ml胆管内に注入した後、約150 mlの温湯(約85 ℃)を胆管内に高圧注入(0.2 Mpa)し洗浄し、この操作を再度繰り返した。さらに、門脈内に約50 mlの温湯(約85 ℃)を高圧注入(0.2 Mpa)し洗浄した。
次に、約500 mlのKガード(商品名、エイトノット株式会社)(200 ppm)を胆管内に高圧注入(0.2 Mpa)し消毒した。消毒後、400 ppmの塩素系消毒剤テルロンブリーチと80 %エタノールをスプレーし、肝臓表面を消毒した。その後、熱湯消毒、およびアルコール消毒した包丁もしくは外科用メスを用いて左葉から約100 cm3程度のブロックを3カ所(A~C)切り出し、各ブロックの中心部から約50 g(4 cm x 4 cm x 3 cm)のサンプルを2か所切り出した(図9)。切り出したサンプルを無菌袋に入れ、さらに約10 mlのKガード(200 ppm)を入れ、卓上真空包装機を用いて約30秒~1分かけて真空度約-0.1 MPa付近(約-0.07~-0.09 MPa程度)になるまで真空にし、真空包装した。真空包装した各サンプル-30 ℃で急速凍結し、28時間保存後、4 ℃で緩やかに16時間かけて解凍した。冷凍処理しない群はすぐに次の操作に移った。解凍した約50 gの各サンプルに等量のPBSを加え、30秒間ストマッカー処理を行ない、等量揉み出し液を作製した。得られた揉みだし液100 μlをそれぞれCT-SMAC培地上に接種し、好気条件下で37 ℃、1日培養し、得られたEHEC様コロニーを計測し、Kガードによる消毒および追加消毒効果を検討した(表15)。
安定型複合塩素製剤と呼ばれる塩素系消毒剤Kガードについても他の消毒剤と同様の効果が認められるかを検討した。結果、Kガードにも強い殺菌効果が認められた。
さらに、食品、特に牛肝臓(胆汁内)には比較的高濃度(106 cfu/ml以上)の大腸菌様細菌やカンピロバクターが存在する個体がまれに認められた。高濃度の菌が存在する場合、消毒効果が減弱する可能性もある。生食やレアステーキなどのそれほど強い加熱を行わない調理法に利用する食材においては、何らかの方法で食材に存在する菌数をモニタリングする必要があると思われる。肝臓の場合、胆汁内の菌数を直接培養、PCR等で菌数をモニタリングするのが最適であると考えられた。
また、本発明の方法に沿って消毒、冷凍、解凍した食材は味覚等に何の影響も見られなかったうえに、食材の欠点である臭み、くせ等の減弱が見られ、むしろ味や風味が向上することから、本発明は食品、特に生レバー等の生食用食肉の従来の加工、流通、販売法に大きく資する発明であると考えられる。
本発明の方法に従って得られる肝臓は、味覚等に何らの影響も現れなかった。従って本発明は、生食用食肉の加工、流通、販売の分野において有用である。
また日本の食文化を守るうえでも有用である。
Claims (9)
- 以下の工程を含む、微生物が殺菌された肝臓の製造方法;
(a)摘出された非ヒト動物の肝臓の胆管内および門脈の両方またはいずれか一方に熱湯を高圧注入する工程、および
(b)工程(a)の肝臓の胆管内および門脈の両方またはいずれか一方に塩素系消毒剤を注入する工程。 - 肝臓に塩素系消毒剤を浸透させる工程をさらに含む、請求項1に記載の方法。
- 肝臓を冷凍する工程をさらに含む、請求項1または2に記載の方法。
- 以下の工程をさらに含む、請求項1に記載の方法;
(a)肝臓表面を塩素系消毒剤により消毒する工程、および
(b)工程(a)の肝臓を冷凍する工程。 - 肝臓を冷凍する工程の前に、肝臓に塩素系消毒剤を浸透させる工程をさらに含む、請求項4に記載の方法。
- 肝臓を解凍する工程をさらに含む、請求項3から5のいずれかに記載の方法。
- 微生物が食中毒を引き起こす微生物である、請求項1~6のいずれかに記載の方法。
- 食中毒を引き起こす微生物がカンピロバクター、病原性大腸菌、サルモネラ属菌、シゲラ属菌、エロモナス属菌、スタフィロコッカス属菌、E型肝炎ウィルスからなる群より選択される、請求項7に記載の方法。
- 請求項1~8のいずれかに記載の方法によって得られる、殺菌された肝臓組織。
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AU2013268382A AU2013268382B2 (en) | 2012-06-01 | 2013-05-31 | Method for producing disinfected liver |
US14/404,627 US20150147447A1 (en) | 2012-06-01 | 2013-05-31 | Method for producing detoxified liver |
MX2014014677A MX362860B (es) | 2012-06-01 | 2013-05-31 | Metodo para producir higado desinfectado. |
JP2014518747A JP6087915B2 (ja) | 2012-06-01 | 2013-05-31 | 消毒された肝臓の製造方法 |
US16/257,766 US11160286B2 (en) | 2012-06-01 | 2019-01-25 | Method for producing disinfected liver |
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Citations (2)
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JPH1189509A (ja) * | 1997-09-19 | 1999-04-06 | Prima Meat Packers Ltd | 食肉の処理方法 |
JP2004016189A (ja) * | 2002-06-20 | 2004-01-22 | S Foods Inc | 家畜内臓肉の処理方法 |
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US3819329A (en) * | 1971-05-11 | 1974-06-25 | Morton Norwich Products Inc | Spray sanitizing system with electrolytic generator |
US4663173A (en) * | 1985-04-09 | 1987-05-05 | Campbell Soup Company | Hot solution injection |
US6010729A (en) * | 1998-08-20 | 2000-01-04 | Ecolab Inc. | Treatment of animal carcasses |
US6569482B2 (en) * | 1998-10-30 | 2003-05-27 | Excel Corporation | Method for surface treating animal tissue |
US20020122854A1 (en) * | 1998-10-30 | 2002-09-05 | Bio-Cide International, Inc. | Acidified metal chlorite solution for disinfection of beef, pork and poultry |
US7077967B2 (en) * | 2000-02-18 | 2006-07-18 | Zentox Corporation | Poultry processing water recovery and re-use process |
US6964788B2 (en) * | 2001-05-07 | 2005-11-15 | Steris Inc. | System for handling processed meat and poultry products |
EP2314165B1 (en) * | 2004-12-23 | 2014-10-01 | Albemarle Corporation | Microbiocidal control in the processing of meat-producing four-legged animals |
US20110200688A1 (en) * | 2010-02-16 | 2011-08-18 | Harvey Michael S | Methods and compositions for the reduction of pathogenic microorganisms from meat and poultry carcasses, trim and offal |
WO2011118821A1 (ja) * | 2010-03-23 | 2011-09-29 | 国立大学法人宮崎大学 | 真空及び共振型超音波処理による食品材料における微生物の制御方法及び制御装置 |
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JPH1189509A (ja) * | 1997-09-19 | 1999-04-06 | Prima Meat Packers Ltd | 食肉の処理方法 |
JP2004016189A (ja) * | 2002-06-20 | 2004-01-22 | S Foods Inc | 家畜内臓肉の処理方法 |
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US20190159468A1 (en) | 2019-05-30 |
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US20150147447A1 (en) | 2015-05-28 |
MX362860B (es) | 2019-02-20 |
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AU2013268382A1 (en) | 2015-01-15 |
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