Method for removing clostridium perfringens spores and use thereof

Abstract

The application discloses a method for removing Clostridium perfringens spores and application thereof. The method comprises the following steps: (1) mixing a nutrient germinant and a Clostridium perfringens spore solution to obtain a mixed solution; (2) heat stimulating the mixed solution obtained in step (1); (3) performing ultrasonic wave assisted nutrient induction treatment on the heat stimulated mixed solution; (4) performing ultrahigh pressure treatment on the ultrasonic wave assisted nutrient induction treated mixed solution; and (5) performing heat treatment on the ultrahigh pressure treated mixed solution. The method can not only achieve a removal effect of 6log CFU / g or more than 6log CFU / mL of Clostridium perfringens spores, but also shorten the whole spore removal process to 40-80 min.

Description

Technical Field

[0001] This invention belongs to the field of safety control technology in food processing, and relates to a method for removing Clostridium perfringens spores and its application. Background Technology

[0002] *C. perfringens* is one of the main spore-forming pathogens causing foodborne gastroenteritis outbreaks in chilled cooked meat products. *C. perfringens* spores have the highest contamination rate in meat and meat products, easily contaminating both raw meat and chilled cooked meat products. In recent years, the detection rate of *C. perfringens* spores in both raw and cooked meat has been high, with some chilled cooked meats showing a detection rate exceeding 50%. *C. perfringens* spores are extremely heat-resistant (D... 100℃ =124 min), and under pasteurization conditions of 68-72℃, it is actually heat-activated. If heat-activated C. perfringens spores are not cooled in time or properly refrigerated, they have the opportunity to germinate and grow into a large number of C. perfringens vegetative cells. C. perfringens vegetative cells can survive in the human stomach, and then form spores in the human small intestine. The process of spore formation is the process of synthesizing heat-sensitive C. perfringens enterotoxin, thus leading to the outbreak of foodborne diseases.

[0003] To control bacterial spores in low-acid foods, the commonly used treatment methods are as follows: First, ultra-high temperature sterilization (121.1℃ or 115℃) directly inactivates the spores. Second, pasteurization is first used to kill any vegetative cells and pathogens that may be present in the food, while simultaneously using single or combined methods such as low water activity, pH value, refrigeration, and bacteriostatic agents and bactericides to inhibit spore germination and growth, thus ensuring that the number of spore-forming bacteria in the food remains within safe limits throughout the shelf life. Clearly, the former treatment method requires a high heat load and can negatively impact food quality attributes (such as vitamin content, color, and flavor). While the latter method can better maintain food quality, it requires artificially altering the food's physical properties (low water activity and high pH) or adding preservatives (potassium sorbate, sodium lactate, etc.) to inhibit spore proliferation during storage. In recent years, a spore removal method known as the germination-inactivation strategy has gained significant attention. Compared to the commonly used methods of killing or inhibiting spores, the germination-inactivation strategy does not directly kill or inhibit spores, but first induces them to germinate and form vegetative cells, and then inactivates them at sub-thermal lethal temperatures. This method is beneficial for maintaining food quality and preserving nutrients, and is of great significance for the sterilization of food, especially low-acid foods.

[0004] Although the germination-inactivation strategy has obvious advantages and potential as a mild and effective spore removal technique, the most important factor in achieving the desired removal effect is the effectiveness of spore germination.

[0005] Therefore, there is an urgent need to provide a method that has a good spore removal effect, a high removal rate, and a short processing time, so as to maintain the quality of livestock and poultry meat products to the greatest extent and reduce problems such as increased hardness and juice loss in livestock and poultry meat products. Summary of the Invention

[0006] To address the shortcomings of existing technologies and practical needs, this invention provides a method for removing *C. perfringens* spores and its application. The method employs a synergistic induction approach to improve the germination rate of *C. perfringens* spores, specifically following this sequence: first, adding a certain concentration of nutrient inducer; second, applying heat stimulation; third, using ultrasound-assisted nutrient induction; and finally, applying ultra-high pressure synergistic induction. After induction using the above methods, heat treatment is used to achieve the purpose of removing *C. perfringens* spores. This method can not only achieve a removal effect of *C. perfringens* spores of 6 log CFU / g or 6 log CFU / mL or higher, but also shorten the entire spore removal process time to within 40–80 minutes.

[0007] To achieve this objective, the present invention adopts the following technical solution:

[0008] In a first aspect, the present invention provides a method for removing Clostridium perfringens spores, the method comprising the following steps:

[0009] (1) Mix the nutrient germination agent and the sample to obtain a mixed solution;

[0010] (2) The mixed solution obtained in the thermal stimulation step (1);

[0011] (3) The mixed solution after heat stimulation was subjected to ultrasound-assisted nutrient induction treatment;

[0012] (4) The mixed solution after ultrasonic-assisted nutrient induction treatment was subjected to ultra-high pressure treatment;

[0013] (5) Heat treatment is performed on the mixed solution after ultra-high pressure treatment.

[0014] This invention employs a synergistic induction method to improve the germination rate of *C. perfringens* spores. The specific steps are as follows: first, a certain concentration of nutrient inducer is added; second, heat stimulation is applied; third, ultrasound-assisted nutrient induction is used; and finally, ultra-high pressure synergistic induction is applied. After induction using the above methods, heat treatment is used to remove *C. perfringens* spores. This treatment method not only achieves a spore removal effect of 6 log CFU / g or 6 log CFU / mL or higher, but also shortens the entire spore removal process to within 40–80 minutes.

[0015] The "germination-inactivation" strategy is a novel method for addressing bacterial spores in low-acid foods, but achieving ideal removal still depends on the germination effect. It is generally believed that spore induction follows these facts: first, hyperdormant spores exist under any treatment method, and the number of hyperdormant spores is related to the treatment conditions; second, the hyperdormant spore population is not constant under different induction methods. Based on this, this invention proposes a method for removing *C. perfringens* spores, with the following main innovations: First, the addition of a nutrient inducer, combined with heat stimulation, ultrasound assistance, and ultra-high pressure treatment, synergistically induces the germination of *C. perfringens* spores; second, ultrasound-assisted nutrient induction is placed between heat stimulation and ultra-high pressure treatment to significantly improve the germination effect of *C. perfringens* spores; third, ultrasound assistance can significantly shorten the germination time of this stage (without ultrasound assistance) of the nutrient induction phase alone.

[0016] Preferably, the nutrient germination agent in step (1) includes a nutrient germination solution, which includes KCl and L-cysteine.

[0017] Preferably, the concentration ratio of KCl to L-cysteine ​​in the nutrient germination solution is 1:(0.5-2).

[0018] The specific point values ​​from 0.5 to 2 mentioned above can be 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.2, 1.4, 1.6, 1.8, 1.9, 2, etc.

[0019] Preferably, the concentration of the nutrient germination solution in the sample is 50–100 mmol / L or 50–100 mmol / g.

[0020] The specific point values ​​in the range of 50 to 100 mmol / L can be selected as 50 mmol / L, 51 mmol / L, 52 mmol / L, 54 mmol / L, 56 mmol / L, 60 mmol / L, 70 mmol / L, 80 mmol / L, 90 mmol / L, 98 mmol / L, 99 mmol / L, 100 mmol / L, etc.

[0021] The specific point values ​​in the above 50-100 mmol / g range can be 50 mmol / g, 51 mmol / g, 52 mmol / g, 54 mmol / g, 56 mmol / g, 60 mmol / g, 70 mmol / g, 80 mmol / g, 90 mmol / g, 98 mmol / g, 99 mmol / g, 100 mmol / g, etc.

[0022] Preferably, the thermal stimulation in step (2) includes subjecting the mixed solution obtained in step (1) to a water bath.

[0023] Preferably, the water bath temperature is 70-80°C and the time is 10-20 minutes.

[0024] The specific point values ​​in the above 70-80℃ range can be selected as 70℃, 71℃, 72℃, 73℃, 74℃, 75℃, 76℃, 77℃, 78℃, 79℃, 80℃, etc.

[0025] The specific time values ​​in the above 10-20 min range can be selected as 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, 16 min, 17 min, 18 min, 19 min, 20 min, etc.

[0026] Preferably, the ultrasonic power of the ultrasonic-assisted nutrient induction treatment is 200-400W, and the ultrasonic duty cycle is 40-60%.

[0027] The specific point values ​​in the 200-400W range can be selected as 200W, 201W, 202W, 210W, 250W, 200W, 320W, 360W, 380W, 390W, 400W, etc.

[0028] Preferably, the temperature of the ultrasound-assisted nutrient induction treatment is 50–60°C.

[0029] The specific point values ​​in the above 50-60℃ range can be selected as 50℃, 51℃, 52℃, 53℃, 54℃, 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, etc.

[0030] Preferably, the pressure of the ultra-high pressure treatment is 500-600 MPa.

[0031] The specific point values ​​in the 500-600MPa range can be selected as 500MPa, 510MPa, 520MPa, 530MPa, 540MPa, 550MPa, 560MPa, 570MPa, 580MPa, 590MPa, 600MPa, etc.

[0032] Preferably, the ultra-high pressure treatment is performed at a temperature of 50–60°C for 10–20 minutes.

[0033] The specific point values ​​in the above 50-60℃ range can be selected as 50℃, 51℃, 52℃, 53℃, 54℃, 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, etc.

[0034] The specific time values ​​in the above 10-20 min range can be selected as 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, 16 min, 17 min, 18 min, 19 min, 20 min, etc.

[0035] Preferably, the heat treatment includes a water bath.

[0036] Preferably, the water bath temperature is 80-90°C and the time is 10-20 minutes.

[0037] The specific point values ​​in the above 80-90℃ range can be selected as 80℃, 81℃, 82℃, 83℃, 84℃, 85℃, 86℃, 87℃, 88℃, 89℃, 90℃, etc.

[0038] The specific time values ​​in the above 10-20 min range can be selected as 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, 16 min, 17 min, 18 min, 19 min, 20 min, etc.

[0039] Secondly, the present invention provides the application of the method described in the first aspect in food production, wherein the food includes meat products.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] (1) The present invention adopts a synergistic induction method, which can significantly improve the removal effect of C. perfringens spores and achieve a removal rate of more than 6 log CFU / g spores;

[0042] (2) The synergistic induction method of this invention can significantly shorten the germination time, and the time range of the entire spore removal process is 40 min to 80 min;

[0043] (3) The maximum processing temperature of the heat stimulation and heat treatment of the present invention is 90°C, which does not exceed 100°C. It is very suitable for the processing of meat products (especially livestock and poultry meat products). It can maintain the quality of livestock and poultry meat products to the greatest extent and reduce problems such as increased hardness and increased juice loss in livestock and poultry meat products. In addition, the heat stimulation, ultrasonic assistance, ultra-high pressure treatment, nutrient agents and heat treatment methods used all have advantages such as high safety and wide application. Attached Figure Description

[0044] Figure 1 This is a schematic diagram of the germination process of C. perfringens spores according to the present invention. Detailed Implementation

[0045] To further illustrate the technical means and effects of this invention, the following description, in conjunction with embodiments and accompanying drawings, provides a further explanation of the invention. It is understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it.

[0046] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0047] Example 1

[0048] C. perfringens was cultured and expanded in liquid thioglycolate (FTG) medium until the number of vegetative cells reached 9 log CFU / mL. Subsequently, it was inoculated into spore-forming broth (DS) medium for sporulation. High-purity C. perfringens spores (spore content exceeding 95%) were obtained by centrifugation and washing, and their quantity was collected to approximately 8.5 log CFU / mL.

[0049] Preparation of PBS buffer samples: Each sample bag contains 10 mL of nutrient germination medium (KCl solution and L-cysteine ​​solution, 1:1 volume), 1 mL of spore solution, and 4 mL of PBS buffer. The concentration of the nutrient germination medium (KCl solution and L-cysteine ​​solution, 1:1 volume) is 50 mmol / L, and the final spore count is approximately 7.5 log CFU / mL. Samples are vacuum-packed at a vacuum level of 2000 Pa.

[0050] PBS buffer samples obtained by heat stimulation (80℃ / 10min) were then subjected to ultrasound-assisted nutrient induction (300W power, 55℃, 50% duty cycle) followed by high-pressure treatment (550MPa / 55℃ / 20min), and then heat treatment (80℃ / 20min). Finally, the decrease in spore count was measured.

[0051] Example 2

[0052] C. perfringens was cultured and expanded in liquid thioglycolate (FTG) medium until the number of vegetative cells reached 9 log CFU / mL. Subsequently, it was inoculated into spore-forming broth (DS) medium for sporulation. High-purity C. perfringens spores (spore content exceeding 95%) were obtained by centrifugation and washing, and their quantity was collected to approximately 8.5 log CFU / mL.

[0053] Preparation of PBS buffer samples: Each sample bag contains 10 mL of nutrient germination medium (KCl solution and L-cysteine ​​solution, volume 1:0.5), 1 mL of spore solution, and 4 mL of PBS buffer. The concentration of the nutrient germination medium (KCl solution and L-cysteine ​​solution, volume 1:0.5) is 50 mmol / L, and the final spore count is approximately 7.5 log CFU / mL. Samples are vacuum-packed at a vacuum level of 2000 Pa.

[0054] PBS buffer samples obtained by heat stimulation (70℃ / 20min) were then subjected to ultrasound-assisted nutrient induction (200W power, 50℃, 40% duty cycle) followed by high-pressure treatment (500MPa / 50℃ / 20min), and then heat treatment (80℃ / 20min). Finally, the decrease in spore count was measured.

[0055] Example 3

[0056] C. perfringens was cultured and expanded in liquid thioglycolate (FTG) medium until the number of vegetative cells reached 9 log CFU / mL. Subsequently, it was inoculated into spore-forming broth (DS) medium for sporulation. High-purity C. perfringens spores (spore content exceeding 95%) were obtained by centrifugation and washing, and their quantity was collected to approximately 8.5 log CFU / mL.

[0057] Preparation of PBS buffer samples: Each sample bag contains 10 mL of nutrient germination medium (KCl solution and L-cysteine ​​solution, volume 1:2), 1 mL of spore solution, and 4 mL of PBS buffer. The concentration of the nutrient germination medium (KCl solution and L-cysteine ​​solution, volume 1:2) is 100 mmol / L, and the final spore count is approximately 7.5 log CFU / mL. Samples are vacuum-packed at a vacuum level of 2000 Pa.

[0058] PBS buffer samples obtained by heat stimulation (80℃ / 10min) were then subjected to ultrasound-assisted nutrient induction (400W power, 60℃, 60% duty cycle) followed by high-pressure treatment (600MPa / 60℃ / 10min), and then heat treatment (90℃ / 10min). Finally, the decrease in spore count was measured.

[0059] Example 4

[0060] The difference from Example 1 is that the sample is a chicken product sample, with each bag containing 10g of chicken product sample, specifically including 5mL of nutrient germination solution (KCl solution and L-cysteine ​​solution, volume 1:1), 1mL of spore solution, and 4g of chicken. The concentration of the nutrient germination solution is 50mmol / L, and the final spore count is approximately 7.5log CFU / g. The sample is vacuum-packed at a vacuum level of 2000Pa.

[0061] Example 5

[0062] The only difference from Example 1 is that the volume ratio of KCl solution to L-cysteine ​​solution in the nutrient germination solution is 1:0.2.

[0063] Example 6

[0064] The only difference from Example 1 is that the volume ratio of KCl solution to L-cysteine ​​solution in the nutrient germination solution is 1:5.

[0065] Comparative Example 1

[0066] The only difference from Example 1 is that the nutrient germination solution does not contain L-cysteine.

[0067] Comparative Example 2

[0068] The only difference from Example 1 is that the nutrient germination solution does not contain KCl.

[0069] Comparative Example 3

[0070] The only difference from Example 1 is that no nutrient germination solution was added.

[0071] Comparative Example 4

[0072] The only difference from Example 1 is that ultrasound-assisted nutrient induction treatment and ultra-high pressure treatment were not performed.

[0073] Comparative Example 5

[0074] The only difference from Example 1 is that ultrasonic treatment and ultra-high pressure treatment were not performed, and nutritional induction (55℃ / 60min) was performed after thermal stimulation, followed by heat treatment.

[0075] Comparative Example 6

[0076] The only difference from Example 1 is that the ultra-high pressure treatment is changed to low ultra-high pressure treatment, with a pressure of 150 MPa, 55°C, and 20 min.

[0077] Comparative Example 7

[0078] The only difference from Example 1 is that ultrasonic nutrient induction treatment was not performed.

[0079] Comparative Example 8

[0080] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, nutrient induction at 55℃ / 60min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, and heat treatment at 80℃ / 20min.

[0081] Comparative Example 9

[0082] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, nutrient induction at 55℃ / 60min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, and heat treatment at 80℃ / 20min.

[0083] Comparative Example 10

[0084] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, nutrient induction at 55℃ / 60min, and heat treatment at 80℃ / 20min.

[0085] Comparative Example 11

[0086] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, nutrient induction at 55℃ / 60min, and heat treatment at 80℃ / 20min.

[0087] Comparative Example 12

[0088] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, nutrient induction at 55℃ / 60min, and heat treatment at 80℃ / 20min.

[0089] Comparative Example 13

[0090] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, nutrient induction at 55℃ / 60min, and heat treatment at 80℃ / 20min.

[0091] Comparative Example 14

[0092] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, nutrient induction at 55℃ / 60min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, and heat treatment at 80℃ / 20min.

[0093] Comparative Example 15

[0094] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, nutrient induction at 55℃ / 60min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, and heat treatment at 80℃ / 20min.

[0095] Comparative Example 16

[0096] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, nutrient induction at 55℃ / 60min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, and heat treatment at 80℃ / 20min.

[0097] Comparative Example 17

[0098] The difference from Example 1 is that the treatment methods are as follows: heat stimulation at 80℃ / 10min, nutrient induction at 55℃ / 60min, low ultra-high pressure treatment at 150MPa / 55℃ / 20min, ultra-high pressure treatment at 550MPa / 55℃ / 20min, and heat treatment at 80℃ / 20min.

[0099] Test Example 1

[0100] Statistics on the decrease in spore count.

[0101] The results are shown in Table 1.

[0102] Table 1

[0103]

[0104]

[0105] The results in the table show that: Examples 1-3, using the method of the present invention to remove *C. perfringens* spores from PBS buffer samples or chicken samples, effectively removed *C. perfringens* spores. Example 4, using the method of the present invention to remove *C. perfringens* spores from chicken samples, yielded similar results to Example 1, effectively removing *C. perfringens* spores. Examples 5-6 demonstrate that precise volume ratios of KCl solution and L-cysteine ​​solution effectively ensure spore removal. Comparative Examples 1-3, compared to Example 1, show that the selection of specific nutrient germination solutions such as KCl solution and L-cysteine ​​solution affects the results and effectively ensures spore removal. Comparative Examples 4-7, compared to Example 1, show that a single step can also remove *C. perfringens* spores, but the removal effect is not good; the synergistic method of the present invention ensures the best *C. perfringens* spore removal effect. Comparative Examples 8-17, compared to Example 1, show that the synergistic method with a specific sequence of the present invention yields the best treatment effect.

[0106] Test Example 2

[0107] Statistics on the time required to remove C. perfringens spores.

[0108] The results are shown in Table 2.

[0109] Table 2

[0110]

[0111]

[0112] The results in the table show that: Examples 1-3, using the method of the present invention to remove Clostridium perfringens spores from PBS buffer samples or chicken samples, can be completed within 40-80 minutes. Example 4, using the method of the present invention to remove Clostridium perfringens spores from chicken samples, yielded similar results to Example 1, with the time also controlled within 40-80 minutes. Comparative Examples 8-11, compared to Example 1, demonstrate that ultrasound-assisted nutrient induction significantly shortens the induction time compared to nutrient induction alone, reducing it from 60 minutes for nutrient induction alone to 20 minutes with ultrasound assistance, while maintaining essentially the same effect. Comparative Examples 12-17, compared to Example 1, show that even if changing the specific order of experimental steps and adding experimental steps can achieve a spore count reduction similar to that of Examples 1-4, the required time would reach 130 minutes. This further demonstrates that the present invention, by using a specific order and specific steps, can significantly improve the spore count reduction, shorten the time, and reduce costs.

[0113] In summary, the method of the present invention can not only achieve the removal effect of Clostridium perfringens spores of 6 log CFU / g or 6 log CFU / mL or higher, but also shorten the time of the entire spore removal process to within 40 to 80 minutes.

[0114] The applicant declares that the detailed method of the present invention is illustrated by the above embodiments, but the present invention is not limited to the above detailed method, that is, it does not mean that the present invention must rely on the above detailed method to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A method for removing Clostridium perfringens spores, characterized in that, The method includes the following steps: (1) Mix the nutrient germination agent and the sample to obtain a mixed solution; (2) The mixed solution obtained in step (1) of thermal stimulation; (3) The mixed solution after heat stimulation was subjected to ultrasound-assisted nutrient induction treatment; (4) The mixed solution after ultrasonic-assisted nutrient induction treatment was subjected to ultra-high pressure treatment; (5) Heat treatment of the mixed solution after ultra-high pressure treatment; The nutrient germination agent mentioned in step (1) includes a nutrient germination solution, which is KCl and L-cysteine; The concentration ratio of KCl to L-cysteine ​​in the nutrient germination solution is 1:(0.5~2); The concentration of the nutrient germination solution in the sample is 50-100 mmol / L or 50-100 mmol / g; The ultrasonic power of the ultrasonic-assisted nutrient induction treatment is 200~400 W, and the ultrasonic duty cycle is 40~60%. The temperature for the ultrasound-assisted nutrient induction treatment is 50~60℃. The pressure of the ultra-high pressure treatment is 500~600 MPa; The ultra-high pressure treatment is performed at a temperature of 50~60℃ for 10~20 minutes.

2. The method according to claim 1, characterized in that, The thermal stimulation in step (2) includes subjecting the mixed solution obtained in step (1) to a water bath.

3. The method according to claim 2, characterized in that, The water bath temperature is 70~80℃, and the time is 10~20min.

4. The method according to claim 1, characterized in that, The heat treatment includes a water bath.

5. The method according to claim 4, characterized in that, The water bath temperature is 80~90℃, and the time is 10~20min.

6. The application of the method of any one of claims 1-5 in food production, wherein the food includes meat products.

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