JP2010202868A - Method for controlling biofilm, and method and device for measuring biofilm - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a technique to control bacteria suspended in food in a range with no influence on food taste, and to hamper formation of a biofilm in relation to production, storage, and distribution of the food. <P>SOLUTION: A bacteria controlling agent, formed by compounding a thiamine lauryl sulfate which is a vitamin B1 derivative, a sugar alcohol or an emulsifying agent or a lactate with calcium ion dissolved in brewed vinegar, not only inhibiting bacteria suspended in the food but also controls adhesion of the biofilm to a surface of food or a machine, an appliance, or a vessel involved therein, or to a fixed substrate in this environment, formation, proliferation or germination of the biofilm, and a device and a method for measuring the rise and fall of the biofilm and the bacteria in the process have been found. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a biofilm control method, a biofilm measurement method, and a measurement apparatus.

Bacteria, fungi, and yeast (hereinafter referred to as bacteria) involved in food preservation not only contaminate foods but cause rot and food poisoning, but also cause biofilm production.
Bacteria attach to manufacturing equipment and containers of food production facilities, and after attachment, discharge biopolymers of mucous substances such as polysaccharides and proteins to form a gel-like thin film, generating bacterial flora inside it To become a biofilm. This is a biofilm.
Biofilms adhere firmly to the surface of fixed substances, and the bacterial flora within them promotes growth and germination under suitable nutrient sources and environments. Based on this, corrosion and production of equipment and instruments such as stainless steel Spread the contamination again to the food inside.

JP 2003-325152 A JP 2005-210997 A JP2008-1000096 Introduction to Biofilms, 2007.10.9 Journal of the Japan Society for Antibacterial and Fungicidal Diseases 2008.8 Journal of the Japan Society for Antibacterial and Fungicidal Diseases 2008.9

Even if washing and cleaning are carried out at each stage of food storage, manufacturing, processing, distribution, and consumption, the bacteria in the food will adhere to the surrounding machinery and containers at the start of the work, and there will be bacterial flora there. A biofilm including the above is generated, and this serves as a starting point for the circulation in which the food in the process, the production machine and the like are contaminated again, thereby impairing the good hygiene conditions of the food.
In addition, it is difficult to sterilize and clean production machines and the like in operation, and there is a need for a technique for controlling contamination and proliferation under these conditions. Bacterial ecology that has been classified as neto or slime as part of bacterial contamination has been elucidated as a biofilm, and the first challenge is to control this, and the heavy use of chlorinated chemicals used for sterilization washing In order to accelerate the corrosion of stainless steel and other metals in machinery and equipment, it is also a challenge to control the biofilm and reduce the burden on excessive equipment for sterilization and cleaning.

The present invention has been made in view of such circumstances, and an object of the present invention is to provide a biofilm control agent capable of controlling the number of bacteria in a biofilm within a range that does not substantially affect the taste of food or the like. .
Another object of the present invention is to provide an apparatus and a method capable of measuring the number of bacteria in an object in a container and the number of bacteria in a biofilm.

The biofilm control agent of the first aspect of the invention is a brewing vinegar in which 1 to 7 parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons are dissolved in 100 parts by weight. It is a brewed vinegar and has a function of controlling airborne bacteria in foods by blending 0.01 to 12 parts by weight of thiamine auryl sulfate of vitamin B1 derivative.
Here, if one or more animal calcium ions are less than 1 part by weight, there is an adverse effect that there is no calcium ion antibacterial function. On the other hand, if the amount exceeds 7 parts by weight, all the added calcium precipitates and calcium ions are not generated, and the antibacterial function is lost.
Further, when thiamine auryl sulfate is added in an amount of less than 0.01 parts by weight, there is an adverse effect (no thiamine antibacterial function). Further, when thiamine auryl sulfate is added in an amount exceeding 12 parts by weight (additional thiamine does not dissolve but precipitates and the antibacterial function is lost), an adverse effect occurs.

The biofilm control agent of the second aspect of the invention is a brewing vinegar in which 1 to 7 parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons are dissolved in 100 parts by weight. It is a brewed vinegar, and has a function of controlling floating bacteria in foods by blending 3 to 100 parts by weight of one or two of the sugar alcohols sorbitol, xylitol and erythritol.
Here, when one or more animal calcium ions are less than 1 part by weight, there is a problem that the antibacterial function of calcium ions is absent. On the other hand, when the amount exceeds 7 parts by weight, all the added calcium precipitates and calcium ions are not generated, resulting in a problem that the antibacterial function is lost.
Furthermore, when one or two of the sugar alcohols sorbitol, xylitol, and erythritol are blended in less than 3 parts by weight, there is a disadvantage that the sugar alcohol does not have a penetrating function. Furthermore, if this is added in an amount exceeding 100 parts by weight, the adverse effect that the antibacterial function of the opposite calcium ions decreases.

The biofilm control agent according to the third aspect of the invention comprises 100 parts by weight of 1 to 7 parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons in brewed vinegar. It is a brewed vinegar, and has a function of controlling floating bacteria in foods, which is mixed with 1 to 50 parts by weight of one of sucrose fatty acid ester and glycerin fatty acid ester.
Here, if one or more of the animal calcium ions is less than 1 part by weight, there is an adverse effect that the antibacterial function of calcium ions is absent. On the other hand, when the amount exceeds 7 parts by weight, the added calcium precipitates, calcium ions are not generated and the antibacterial function is lost.
Furthermore, when one of sucrose fatty acid ester and glycerin fatty acid ester is blended in an amount of less than 5 parts by weight, there is an adverse effect that it is not emulsified and only the antibacterial function of calcium ions is not suitable for blending purposes. Furthermore, when this is compounded exceeding 50 parts by weight, the insoluble component of the fatty acid ester becomes a semi-solid form, resulting in a disadvantage that it cannot be used without being dispersed even when added to food.

The biofilm control agent according to the fourth aspect of the invention comprises 100 parts by weight of 1 to 7 parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons in brewed vinegar. The brewing vinegar has a function of controlling floating bacteria in foods in which 10 to 90 parts by weight of one or two kinds of potassium lactate or sodium lactate is blended.
Here, when one or more animal calcium ions are less than 1 part by weight, there is a problem that the antibacterial function of calcium ions is absent. On the other hand, when the amount exceeds 7 parts by weight (calcium is precipitated, calcium ions are not generated and the antibacterial function is lost).
Furthermore, when one kind of potassium lactate or sodium lactate is blended in an amount of less than 10 parts by weight, there is an adverse effect that only the antibacterial effect of calcium ions is not suitable for blending purposes. Furthermore, when it mix | blends exceeding 90 weight part, the bad effect that the antibacterial function of calcium ion will lose | disappear relatively and it will not become the objective of a mixing | blending will arise.

  The biofilm control agent of the present invention preferably has a function of controlling bacteria inside and outside the cuticle layer of plants.

  Moreover, the foodstuff of this invention adds the biofilm control agent of the 1st-4th viewpoint mentioned above.

  In addition, the apparatus or method of the present invention can be applied to metal, plastics, rubber, ceramics, wood, paper, glass, or fibers installed in a container, and the number of bacteria in the container and bacteria in the biofilm. Measure the number.

  When vitamin B1 is used in combination with calcium ion solution, the surface tension of water in food ingredients that come into contact with calcium ions is reduced by the interfacial action of vitamin B1, and B1 and calcium ions are sufficient on the surface of the substrate or biofilm. Can touch. And both antibacterial components coat the substrate surface and prevent the adhesion of bacteria, or contact the biofilm surface sufficiently and penetrate inside to reach the bacterial flora. Vitamin B1 has a destructive action on cell membranes, and calcium ions cause intracellular protein denaturation, each of which exhibits a specific antibacterial action to prevent bacteriostatic and growth.

  In addition, sugar alcohols such as sorbitol, xylitol or erythritol have a function of improving osmotic pressure, hydration and improving food properties, and have already been used as a non-cariogenic sweetener for teeth in the field of oral hygiene. Widely used in chewing gum and has the function of inhibiting the production of biofilms, which are biofilm plaques, in the oral cavity, so it can be dissolved in foods by dissolving sugar alcohols in calcium ion solution. Control the production of airborne bacteria and biofilm by them and external factors. Sugar alcohol has an inhibitor of biofilm production, and calcium ions penetrate the bacterial flora and exert antibacterial function against bacteria attached to a fixed substrate or by the mechanism of biofilm production inhibition These are controlled.

In addition, when sucrose fatty acid ester or glycerin fatty acid ester as an emulsifier is added to calcium ions, calcium ions can be intimately contacted with the surface of the biofilm by the function of wetting by emulsifying action. Has antibacterial action and controls it.
In addition, since potassium lactate or sodium lactate has a water regulation function of foods, a new function that effectively acts on the control of the bacterial flora in the biofilm was found by combining with calcium ions.
In addition, the combination of calcium ions and each of the above functional materials has the effect of adhering bacteria to food, airborne bacteria, food surfaces, and other fixed substrates in the machine, instrument, container, or environment where food comes into contact. And found that it inhibits the production of biofilm and prevents these processes.
Moreover, in order to grasp | ascertain this about the proliferation of floating bacteria and the production | generation of a biofilm, the apparatus and method mentioned later are provided.

Hereinafter, embodiments of the present invention will be described.
In the specification, the pH was measured using an EH1000 type manufactured by Line Seiki, the measurement time was 6 minutes, and the measured values were rounded off to the first decimal place. Bacteria test is performed on general live bacteria and displayed as the number of live bacteria.

  In contrast to the antibacterial function of brewed vinegar, the antibacterial effect is further enhanced by melting animal calcium ions at a certain concentration, and this has the function of inhibiting the production of vitamin B1 derivatives or biofilms having antibacterial and interface functions. We have found a control agent that contains sugar alcohols, or emulsifiers or lactates with antibacterial functions to control airborne bacteria in foods and prevent biofilm formation due to contamination.

As animal calcium ions, scallops were selected and baked at about 900 ° C. to obtain a powder of 150 ms. Vinegar is prepared brewing alcohol solution 100 parts by weight of the alcohol 24 g / water 76g concentration was brewed liquid by mixing equal amounts relative fermentative bacteria Asenobakuta 10 ⁹ bacteria count species vinegar.
The pH of the seed vinegar was 2.4 and the mixed solution was 4.1. When the mixture showed fermentation at room temperature 24 hours after starting fermentation at room temperature, 1 part of shellfish powder was added, the pH became 4.5, and then the shellfish powder when pH became 3 or less after 6 hours. One part was added. This was repeated every 6 to 8 hours. When the total amount of shellfish powder was 7 parts, the pH was 4.5, but when it became 8 parts, the pH started at 6.2 and precipitation started. In the part, precipitation further increased to pH 7.1, and then the fermentation was stopped without any change in pH. Although the process was 4 days, the period varies depending on the fermentation conditions.
In the course of fermentation, liquids with an addition amount of 1 part to 9 parts were separately collected to secure a test liquid. The calcium ion concentration is measured by the atomic absorption method.

There was a difference in antibacterial effect according to the melting amount of calcium ions, and this was measured by a turbidity test of water by bacterial growth to confirm this.
In the test, 680 g of mud with spinach is used, 7 kg of this secondary wash water is used as test water, 300 g of each is poured into a styrene cup, and 3 g of calcium ion solution each containing 1 to 8 parts of shellfish powder is added. It was added, covered with a wrap film, stored in a thermostatic chamber at 15 ° C., and the range of effective concentration for antibacterial effect was determined by turbidity after 48 hours.
The turbidity was measured by measuring a fluorescent lamp with a glass sample placed in a 300 ml cylinder fluoroscope turbidimeter. The initial turbidity in the control group was 1.4 degrees, and after that, it was 19.2 degrees.
0.5 part diluted 1 part of shellfish throwing amount with washing water of the previous period is 7.7 below the lower limit of the effect, 1 part 6.5 degrees or more 2 parts 4.5 parts 3 parts 3 1, 4 parts 2.2, 5 parts 1.6, 6 parts 1.4, 7 parts 5.1, 8 parts 18.9. From 7 parts 5.1 to 1 part 6.5 where the amount 7 of shellfish added was the melting limit, the effective range was 0.5 parts or less and 8 parts or more were out of the range.
As a result of measuring calcium ion by atomic absorption, 6 parts was 4.3 g / 100 g. From 1 part to 7 parts is calculated from the dry weight and calculated value of the precipitation residue, 1 part 0.9,
2 parts 1.6, 3 parts 2.3, 4 parts 3.1, 5 parts 3.7, 7 parts 5.9 g / 100 g, and the pH value shows the same change. The effective range was 9 g to 5.9 g. In the examples, 4.3 g / 100 g was used.

Table 1. Test of turbidity change due to melting amount of calcium ion in washing water of mud spinach, amount of shellfish used, pH, transparency

The culture medium for the bacterial culture test of the example was Petrifilm AC plate (manufactured by 3M) and a wiping set (manufactured by Eiken Kikai Co., Ltd.), and the culture conditions and test inspection were carried out in accordance with the Food Sanitation Law or Official Law. . The bacterial test detects the number of general viable bacteria and describes it as viable or viable count.
In the examples, in order to avoid the additive effect, the amounts of the test solution and the test agent to be blended are adjusted.
The present invention is not limited only to the following examples.

[Example 1]
The vitamin B1 derivative thiamine auryl sulfate (manufactured by Tanabe Seiyaku Co., Ltd.) is an active form of vitamin B1 and is used as a nutrient, but has a specific antibacterial function.
Antibacterial function is effective against gram-positive bacteria, heat-resistant spores, molds, and yeasts, as well as lactic acid bacteria that cause acidic spoilage. Technology development was carried out. It is also insoluble in the calcium ion solution described above.
Since vitamin B1 is readily soluble in aqueous alcohol, 7 parts of an aqueous solution of 75 g of alcohol / 100 g of water and 3 parts of vitamin vitamin B1 crystals dissolved therein are mixed with 90 parts of calcium ion solution and this is made in a hot water bath. The mixture was heated to 48 ° C. and stirred for 6 minutes at 40 rpm with a dosing mixer for dissolution.
This blended liquid has a blending ratio of calcium ions of 3.8 g / 100 g and vitamin B1 of 3 g / 100 g. When this mixed solution is stored in a refrigerator and there is no precipitation by visual observation after the passage of 3 days, 2 g of the liquid is dropped into 100 g of coffee extract to observe water solubility, and when observed, the solution disperses quickly and there is no precipitation. No precipitation occurred even after 1 day, and this was used as a test solution in the examples disclosed in the following patent publications.
JP 2005-211057, JP 2000-139409

Machinery and equipment for food production use stainless steel for most of its constituent materials, and it has been found that biofilms are easily generated on the surface of these machinery and equipment regardless of the degree of contact with food. Always exposed to pollution and corrosion hazards.
The formation and growth of biofilms on the surface of stainless steel plates that come into contact with foods are related to the growth of bacteria in foods, and a method to control each other's bacteria by simultaneously measuring this was investigated.
Four containers made of PVDC and having an outer shape of 14 cm × 23 cm and a depth of 12 cm were prepared. In order to measure the adhesion of bacteria or the formation of biofilm, the surface of 0.8mm thickness is processed into a circular plate with a diameter of 80mm on a DB specification, and a number corresponding to the number of days of measurement is prepared. A stand was prepared that stood up at 14 mm and did not contact each other but was electrically insulated. This is done by winding a vinyl bind wire 0.9mm around an iron pipe with an outer diameter of 5cm to make a coil with a winding number of 25, and using this as a gantry, it is installed on the bottom of the case and a stainless steel plate is placed between the coils. It was.

Area of the stainless steel plate is 100.48Cm ² becomes a double-sided in 50.24Cm ^2, inspection guidelines which wiping of the Food Sanitation Law bacteria, airborne fungi of the contents of the container by the apparatus and according to the area of 100 cm ² at 10cmX10cm Can measure bacteria and biofilm attached to the plate surface.
The shape may be a square or other shapes other than the circle, and the same number of samples as the number of samples is prepared.
The stainless steel plate and container are sterilized in a high-pressure sterilizer at 120 ° C for 20 minutes. The container shape may be rectangular parallelepiped or cylindrical, and the material is heat-resistant resin. The upper part is open or used for anaerobic bacteria. It is possible to measure the production of biofilm related to the fluid by providing the inlet and outlet holes at the diagonal of the container, and not limited to the above-mentioned dimensional materials. It can respond to demand, and this is a measuring container.

[Example 2]
A test was conducted to obtain an effective concentration range to be added to foods when using vitamin B1. The effective range was measured by the tendency of bacterial growth and inhibition from light pickled vegetables.
Five 830 g of unwashed cucumbers were cut into small pieces, immersed in 3.3 kg of water, and the soaked water was filtered with filter paper, and 300 g of each was dispensed into eight styrene cups as samples.
A solution obtained by dissolving 1 g of vitamin B1 crystals in an alcohol solution of 75 g of alcohol / 25 g of water is used as a test solution, and 3 g to 30 g of this solution is added to each cup. After 24 hours at a storage temperature of 15 ° C., a 300 ml cylinder fluoroscope is used. Turbidity was measured. It was set as 0 zone control, 1 zone 3g, 2 zone 6g, 3 zone 9g, 5 zone 15g, 7 zone 21g, 9 zone 27g, 10 zone 30g.

Table 2. Test of turbidity change by adding amount of vitamin B1 solution per 100g of pickled water (g):
The numerical value of 1st ward was significantly better than the control, and 10th ward had precipitation of vitamin B1, so that the effective range was from 1st ward to 9th ward.

Example 3
Field crop buckwheat has soil-derived bacteria. Using this as a sample, the behavior of bacteria and the production of biofilm caused by this were tested. 15 kg of tap water was poured into a 20-liter plastic bucket, 800 grams of domestic buckwheat flour was added, and the mixture was stirred for 12 minutes at 45 rpm for 12 minutes with an input mixer, and filtered through a filter cloth to obtain a sample solution. 3.2 kg of this was put into a bowl, the following samples were added, stirred and poured into each container.
For each container, 5 stainless plates were placed upright on the frame.
Container No. 11 in the test group is a control group, 12 is 32 g of calcium ion solution 4.3 g / 100 g solution, 13 is 1 g of vitamin B1 in 100 g of alcoholic solution of 75 g of alcohol / 25 g of water with 300 ppm of vitamin B1 as practical concentration. 96 g of the dissolved solution, and 14 g of the mixed solution of 16 g of calcium ion solution and 48 g of vitamin B1 solution were added.
This vitamin B1 solution is also used in the tests of the following examples.

The stainless steel plate was placed upright on a frame for each of the five plates, and the test solution was injected.
In each test section, 3.2 kg of test solution was stored and opened and stored at 15 ° C. in a heat insulation chamber, and a test was started to measure the growth of bacteria in the test solution and the formation of bacteria or biofilm adhering to the stainless steel plate. .
Collect 10 g of sample solution from each container every predetermined number of days, remove one stainless steel plate, wipe the test solution with an AC plate by a dilution method, and wipe the stainless steel plate with a cotton swab into a test tube containing 9 g of sterile water. The medium plate was similarly inoculated by the dilution method, cultured at 36 ° C. for 48 hours, and the number of each viable cell was counted. The initial number of viable bacteria in the sample solution was 4 × 10.
Test group 11 is control, 12. Calcium ion solution 13. Vitamin B1, 14 is a compounding agent

Table 3. Bacterial behavior and biofilm production test of buckwheat flour:
Upper A is the number of bacteria in the sample solution, Lower B is the number of bacteria on the stainless steel plate:
After 9th of 11A, it became impossible to measure due to corruption.
In Test Groups 12 and 13, the sample solution and the stainless steel plate are effective, but the calcium ion and vitamin B1 combination solution in Test Group 14 suppresses the growth of bacteria in the sample solution. Since it prevents the attachment of bacteria, the number of bacteria is low, controlling the attachment of bacteria and the production of biofilms.

Example 4
Prepare 1.6kg of 3 kinds of carrots, cabbage and cucumbers, which have been washed in advance, in a total of approximately the same amount, chopped within 2cm in width and 6cm in length, divided into 400g pieces, and stored in the test container. 3.2 kg of water was poured and stored at a storage temperature of 15 ° C. for 4 days to test the fate of bacteria attached to vegetables and the formation of a biofilm on a stainless steel plate.
10 g of vegetables were collected and the viable cell count was measured by a stomacher treatment and dilution method, and the stainless steel plate was wiped and cultured. The initial bacterial count of the control vegetables was 7 × 10 ² .
Test area 21. Control, 22. Calcium ion solution 32 g (1%) 23. 196 g (0.03%) vitamin B1 16 g (0.5%) calcium ion solution and 98 g (0.015%) vitamin B1 were added.

Table 4. Changes in vegetable bacteria and biofilm formation on stainless steel plates:
The top row is the number of bacteria in the vegetable, the bottom row is the number of bacteria on the stainless steel plate:
The control shows the number of spoilage bacteria in 3 days. 22 and 23 show the effect respectively, but the test group 24 shows the effect on both the vegetable and the stainless steel plate, and calcium ions permeate the cuticle layer covering the vegetable surface by the interface action of vitamin B1. In this way, the growth of bacteria in the vegetable tissue is suppressed, which confirms the effectiveness of both the combination agent penetrating into the cuticle layer of the plant epidermis and the control of biofilm formation on the stainless steel plate. .

Example 5
Use 180g starch and 20g salt for 1.8kg fish paste, knead with food mixer for 7 minutes, divide into 4 samples, add test solution to each test group except for control group, and knead again for 4 minutes. Molded into four kamaboko shapes. For molding, a wooden frame of 5 × 10 cm and a depth of 2 cm was used to form a plate and then wrapped with a wrap film. The weight of one piece was 120 g, which was steamed for 25 minutes, heat-dissipated, and stored in a heat-reservoir set at 15 ° C., and the number of bacteria in the sample, the biofilm on the surface, and the generation of bacterial flora were measured.

Test group 31 is a control, 32. In each test section, 1 g of calcium ion solution per 100 g of material, 33. 3 g of vitamin B1 solution, 34. A mixture of 0.5 g of calcium ion solution and 1.5 g of vitamin B1 solution was added. The sample was placed in a plastic bag, sealed, stored at 15 ° C. for 4 days, and measured. The initial bacterial count was a heated product and was 300 or less. A total of 100 cm ² of two 5 × 10 cm surfaces of the plate-like sample was wiped off as a sample for biofilm generation measurement. Next, 6 surfaces were thinly cut and only the contents were collected, and the contents were examined for bacteria.

Table 5. Kamaboko test:
The upper row is the number of bacteria inside, the lower row is the number of bacteria on the surface. Storage temperature: 15 ° C:
A viscous liquid leaching is observed on the lap film surface of the sample, and a biofilm is generated. Compared with the number of floating bacteria in the contents, the growth rate of biofilm production is high and the number of bacteria in the bacterial flora is large. It is assumed that the participation of moisture has an effect on the growth of bacteria, and it was confirmed that the calcium ion solution and vitamin B1 mixture were effective in controlling both floating bacteria and surface biofilm. .

Example 6
Bacon tended to grow a lot of bacteria on the slice surface, and the growth of both the slice surface and the contents of the bacteria was measured to test the production of the biofilm on the slice surface.
Prepare 2 kg of bacon blocks manufactured in the normal process and stored for 2 days at a storage temperature of 5 ° C., and divide them into 200 g samples to make 600 g of immersion water with the following test solutions added. Soaked.
It was taken out at an immersion time of 25 minutes and sliced to a thickness of about 6 mm, and each of the 5 sheets was stored as a unit in a plastic bag. The initial number of bacteria on the contents and surface of the bacon was 3 × 10. This was stored at a storage temperature of 15 ° C., and the contents and surface bacteria were measured over time. Collecting bacteria from the sample, making the wiping frame 2X5cm = 10cm ² to the slice plane of the surface, the dilution wiping both sides, contents will sample cut into strips wipe on both sides with gauze after wiping, stomacher process Incubation tests were carried out by the dilution method.

The test agent concentration is the control in the 41 section with respect to 100 g of the immersion water, and 42. Calcium ion solution 2 g, 43. Vitamin B1 3g, 44. Calcium ion 1g vitamin B1 1.5g.

Table 6. The upper part of the bacon preservation test is the number of bacteria inside, the lower part is the number of bacteria on the surface. Storage temperature: 15 ° C:
From the results of Table 6, it was confirmed that the test section 44 had an excellent effect.
Looking at the difference in the number of bacteria on the surface of the bacon and the contents, the growth of the biofilm on the surface is fast, and it is clear from the bacterial flora on the surface that the bacteria tend to enter the bacon and contaminate it, and the surface is slimy Technology can be provided for measures against biofilm produced in processed meat products such as bacon, ham and sausage.

Example 7
We tested the quality preservation of soy milk to measure the range of effective usage of sugar alcohol for quality preservation of food. 2.5 kg of soy milk immediately after production was prepared, and 300 g of each was dispensed into 8 cups. In the control group, no addition was made, and a total of 7 pieces were prepared including 6 pieces of sorbitol added from 3 g to 18 g every 3 g.
These were measured for changes in pH value due to bacterial growth after 24 hours at a storage temperature of 15 ° C. The initial pH value was 6.2.

Table 7. Soymilk preservation test:
In the control group, a decrease in pH due to the growth of lactic acid bacteria was observed, and in the added group, an effect was observed from the added amount of 3 g, and the same value was observed for 12 g or more.
As a result, the addition of 3 g or more is effective, and even if it is 18 g or more, there is no particular upper limit as a sweetener or an improving agent, and xylitol or erythritol can be similarly used as a sugar alcohol.

Example 8
The following tests were conducted on the effect of controlling floating bacteria and biofilms for three types of calcium ion solution and sugar alcohols. Prepare 26kg of the same amount of persimmon and orange juice at a concentration of 100g / kg of water, add 3.2kg each to a bowl container, add the following calcium ion solution and 3 sugar alcohols, and stir for 2 minutes. A total of 8 samples were prepared and dispensed into measurement containers to start the test.

The amount of the test agent used for 100 g of the material is 51 sections control, 52 calcium ion solution 2 g, sugar alcohol is 4 g of sorbitol, 4 g of xylitol, 4 g of erythritol (each manufactured by Nikken Kasei Co., Ltd) The amount was ½. Sugar alcohols were water-soluble, and there was no problem in blending with calcium ion solution. The initial bacterial count of fruit juice was 2 × 10.
51 in the test group is a control, 52 calcium ion solution, 53 sorbitol, 54 xylitol,
55 erythritol, 56 calcium ion solution / sorbitol combination, 57 calcium ion solution xylitol combination, 58 calcium ion solution / erythritol combination.

Table 8. Fruit juice test:
Storage temperature 15 ° C. A is fruit juice, B is a stainless steel plate. As a result, it was effective in controlling airborne bacteria and biofilms in a test area containing calcium ion solution, xylitol and erythritol.

Example 9
About three kinds of calcium ion solution and sugar alcohols, the effect of controlling floating bacteria and biofilm was tested with a measuring container for the production of floating bacteria and biofilm in custard sauce used for cakes and desserts.
A custard sauce of 25 kg was made using eggs, milk and sugar as ingredients, divided into 3 kg, placed in a stainless steel bowl, heated to about 50 ° C. with a hot water bath, added with the following test solution and stirred for 3 minutes to prepare a sample. The test agent was used in an amount of 2 g of calcium ion solution, sugar alcohol was 4 g of sorbitol, 4 g of xylitol, 4 g of erythritol, and the compounded solution was ½ each based on 100 g of material.
Test group 61 is a control, 62 calcium ion solution, 63 sorbitol, 64 xylitol,
65 erythritol, 66 calcium ionic liquid sorbitol blend, 67 calcium ionic liquid xylitol blend, 65 calcium ionic liquid erythritol blend This product was measured for 36 hours at a storage temperature of 15 ° C. due to the rapid growth of bacteria.

Table 9. Custard sauce test:
The content of A is effective, and as for B, the stainless steel plate calcium ion solution and erythritol are effective. However, depending on the application, xylitol or sorbitol other than erythritol can also be used.

Example 10
Three kinds of calcium ion solution and sugar alcohol were used in water sheep and tested for the production of internal floating bacteria and surface biofilm. Prepare 2.7 kg of raw agar and agar, heat and stir, then divide into 8 parts into 330 g, add the following test solution, stir again, and lay a wrap film in a wooden frame 5 × 10 cm deep 2 cm It was poured into a container and packaged after heat dissipation. Four pieces each having an average of about 80 g in the shape of a water sheep cage were made.
The amount of test agent used per 100 g of each category is 72 g calcium ion solution 2 g, 73 sorbitol 4 g, 74 xylitol 4 g, 75 erythritol 4 g, 76 calcium ion solution 1 g / sorbitol 2 g, 77 calcium ion solution 1 g xylitol 2 g, 78 calcium It was set as the mixing | blending of 1/2 each of ionic liquid 1g and erythritol 2g.
The water sheep was stored in a heat storage box at a storage temperature of 15 ° C., and suspended bacteria and biofilm on the wrap film surface were measured over the elapsed days.

Table 10. Water sheep test:
A contents, B surface:
“――――” was canceled due to corruption:
In the combination, calcium ion solution, erythritol, xylitol, and sorbitol were effective in that order, and the bacterial count increased with the leaching of water on the surface. Mucus was derived and biofilm was generated.

Example 11
The test which measures the range of the usage-amount was carried out about the stability function of the foodstuffs of sucrose fatty acid ester and glycerol fatty acid ester (made by Showa Chemical Co., Ltd.). To be used for the test, 80 g of sucrose fatty acid ester was dissolved in 720 g of warm water to prepare a test solution, 1.6 kg of cabbage was sliced and immersed in 4 kg of water, and 40 g of cabbage and 300 g of water for immersion were dispensed. Nine things were prepared. The amount of the above test solution was increased from 3 g to 3 g, and 15 g, 18 g, 21 g, and 24 g were added, and a control group was used.
After the sample was stored at a storage temperature of 15 ° C. for 24 hours, turbidity was measured with a 300 ml cylinder fluorometer.

Table 11. Cabbage test:
Emulsifier (g):
If it is 24 g or more, the taste of food deteriorates and there is an unpleasant odor, so it is not used and the use range is 6 to 21 g or less, and the same applies to glycerin fatty acid esters.

Example 12
A calcium ion solution was mixed with sucrose fatty acid ester and tested for control of airborne bacteria and biofilm formation in coffee beverages. Dissolve 300g of instant coffee powder and 380g of skimmed milk powder in 20kg of warm water to make a café au lait, put 3.2kg each into 6 balls, add the following test solution, stir in a whisk for 5 minutes and pour into a measuring container. And stored in a heat storage at a storage temperature of 15 ° C. Thereby, suspended bacteria in the liquid and bacteria on the stainless steel plate were measured, and the results are shown in Table 12. A test solution was prepared by placing 80 g of sucrose fatty acid ester in 720 g of warm water. Similarly, 800 g of a glycerin fatty acid ester was also used for the test.
The amount of the test agent used is 82. Calcium ion solution 2 g, 83. A sucrose fatty acid ester solution 6 g, 84 glycerin fatty acid ester solution 6 g, 85 calcium ion solution 1 g, sucrose fatty acid ester solution 3 g, 86 calcium ion solution 1 g, and glycerin fatty acid ester solution 3 g were used. The initial bacterial count was 4 × 10.

Table 12. Café Ore exam
Number of bacteria in A liquid, number of bacteria on B stainless steel plate Storage temperature 15 ℃
It is effective in controlling the production of airborne bacteria and biofilm in the test area containing glycerin fatty acid ester such as calcium ion solution and sucrose fatty acid ester, and other emulsifiers can also be used.

Example 13
Chickens are contaminated with Salmonella and Listeria, and are usually shipped to the market after being treated with a chlorine-based disinfectant for conservation. However, due to the influence of the chlorine agent, the flavor is impaired, or the processing machinery and equipment are corroded and the health of workers is damaged, so countermeasures are required.
The chicken half before disinfectant treatment was soaked in the liquid of each test section and tested for the production of fleshy airborne bacteria and epidermal biofilm. Bacteria over time were measured by comparison with calcium ion solution, sucrose fatty acid ester of emulsifier, glycerin fatty acid ester, and test solutions containing them, and the bacteria were measured over time. A is the meat quality and B is the epidermis. The weight of the chicken was 1.1 kg to 1.6 kg.

The following test agents were used for 2.4 kg of immersion water. For sucrose fatty acid ester and glycerin fatty acid ester, 80 g of solution was used for 720 g of warm water.
91 wards are control, 3 wards of 92 calcium ion solution, 0.6 gram of 93 sucrose fatty acid ester, 0.6 gram of 94 glycerin fatty acid ester, 1.5 gram of 95 calcium ion solution, 1.5 sucrose fatty acid ester. 3 g, 96 ward calcium ion solution 1.5 g and glycerin fatty acid ester 0.3 g.
The chicken was immersed for about 20 minutes, removed from the immersion water, placed in a plastic bag, and stored at a storage temperature of 15 ° C.
Samples were collected and tested by measuring cultures using 10 g of meat and a dilution method after the stomacher treatment, and the epidermis by a wiping dilution method using a 10 cm × 10 cm frame.
These results are shown in Table 10. By adding sucrose fatty acid or glycerin fatty acid to calcium ion solution, it was confirmed that both flesh and epidermis were controlled and bacteria were effective. The emulsifying action of the fatty acid ester promotes the effect of contact permeation of lipids and calcium ions in the chicken skin.

Table 13. Chicken test:
A meat quality B epidermis:
The initial bacterial count has a solid difference and the average value is 8 × 10. However, when immersed in the test agent, the bacterial count tends to decrease on the first day, and weakly resistant bacteria are sterilized.

Example 14
In order to make hamburger steak, 1.2kg of ground beef and pork are mixed with 200g of bread crumbs and mixed with 200g of bread, divided into 200g of each, mixed with test agent, and each 4 pieces wrapped in a wrap film. And stored at a storage temperature of 15 ° C. and tested for the number of bacteria in the contents after the elapsed days and the production of biofilm on the wrapping film in contact. As the sucrose fatty acid ester and glycerin fatty acid ester, a solution obtained by dissolving 80 g of each in 720 g of warm water was used.

The amount of the test agent used for 100 g of the material is as follows: test group 101 is the control, 102 is the calcium ion solution 2 g, 103 is the sucrose fatty acid ester solution 3 g, 104 is the glycerin fatty acid ester solution 3 g, and 105 is the calcium ion solution 1 g. Formulation of 1.5g of sucrose fatty acid ester solution, 106 groups, 1g of calcium ions and 1.5g of glycerin fatty acid ester solution, A in each group describes the test results of the dilution method, B is on the surface The contacted wrap film surface was collected by the wiping method, and the number of viable bacteria was counted by dilution culture. The results are shown in Table 11. Thus, it was possible to confirm the effect that the mixture of the calcium ion solution, the sucrose fatty acid ester and the glycerin fatty acid ester controls the floating bacteria in the contents and the biofilm on the wrap film surface. The initial bacterial count in the control group was 5 × 10.

Table 14. Minced meat putty test:
A is the contents, B is the surface Storage temperature 15 ℃

Example 15
The following tests were conducted on the control effect on the formation of floating bacteria and biofilm of koji by mixing potassium lactate or sodium lactate with calcium ion solution.
Potassium lactate 60g / water 40g solution (prepared by Kanto Chemical Co., Ltd.) Sodium lactate 50g / water 50g solution (Showa Chemical Co., Ltd.) is used. Emulsified liquid with a mixture ratio of lecithin 20g / water 80g to stabilize the test agent when blended. Was added to 80 g of the test agent.
The compounding ratio of the test agent was calcium ion 40, potassium lactate 40, or sodium lactate 40, and lecithin emulsion 20.

After 6 hours of soaking in 2 kg of brown rice with 22 kg of water, make rice cake in 50 minutes of rice cooking time. Divide this into 8 parts into 3.2 liters, separate into balls, add the following test agent and stir with a hand mixer for about 4 minutes. It was poured into a test container and stored at 15 ° C. in a thermostatic chamber.
The amount of the test agent used for 100 g of the material is 121 control group, 122. Calcium ion solution 1 g, 123. 1 g of potassium lactate solution, 124. 1 g of sodium lactate, 125. 13g calcium ion solution potassium lactate, 13g 126 calcium ion solution sodium lactate

Table 15. Test of brown rice bran:
Number of bacteria in A liquid, number of bacteria in B stainless steel plate Storage temperature 15 ℃:
In the test sections 105 and 106, the effect of suppressing bacteria was confirmed. Brown rice bran is a food with a lot of water, and it has traditionally been only retort processing. However, in order to stabilize the quality of this, the use of calcium lactate combined with potassium lactate or sodium lactate improves the taste and saves energy. Effective technology can be provided.

Example 16
Fresh fish salmon was processed into a fillet, and the effects of calcium ions and potassium lactate or sodium lactate and the above-mentioned test agent containing the same were tested on the control of fleshy airborne bacteria and epidermal biofilm formation after storage. An average of 4 pieces of fillet was set to 6 test sections and immersed in 1.7 kg of immersion water for 15 minutes and stored in a thermostatic chamber set at a storage temperature of 15 ° C. to measure the number of elapsed days of bacteria. 5 × 4 cm was used as the wiping frame for the epidermis, and the numerical value was converted to 5 times. As for the meat quality, 10 g from one piece was sampled, diluted with each other through a stomacher treatment, and the number of viable bacteria was measured.
Initial number of bacteria before immersion, meat quality 9 × 10 ^2, epidermis was 1X10 ^2.
The concentration of the test agent with respect to 100 g of immersion water is 132. Calcium ion solution 2 g, 133. Potassium lactate 2g, 134. Sodium lactate 2g, 135 calcium ion potassium lactate combination test agent 2.4g, 136 calcium ion sodium lactate combination test agent 2.4g, 131 control.

Table 16.鯖 Fillet test:
A Number of floating bacteria in flesh B Number of bacteria in epidermis
The growth rate of epidermis flora is large in comparison with meat quality, and the effect of controlling the biofilm of the epidermis and the growth of meat quality are controlled in parallel.

Example 17
The effect of a calcium ion solution, potassium lactate or sodium lactate and a test agent containing the same was tested on biofilm production and control in an instrument container in contact with airborne bacteria in meat extract soup.
400 g of beef streaked meat and 1.4 kg of chicken meat with a small amount of pepper were added, put in 25 kg of water and a pan, heated for about 2 hours and 30 minutes, made soup, filtered through a cloth, and used as a sample. Each test section was divided into 6 sections of 3.2 kg, placed in a ball, added with a test agent, stirred for 3 minutes, placed in a test container in the previous period, and stored in a thermostatic chamber set at a storage temperature of 15 ° C., and measured over time.
In each section A, the number of viable bacteria was measured by a soup dilution method, and B was wiped by a stainless steel plate. The results are shown in Table 17.

  The amount of the test agent used for the material is control in 141 ward, 142 is calcium ion solution 2 g, 143 potassium lactate solution 2 g, 144 sodium lactate solution 2 g, 145 calcium ion and potassium lactate combination test agent 2.4 g, 146 calcium ion Liquid and sodium lactate combination test agent 72.4g

Table 17. Meat soup test:
Number of bacteria in A soup B Number of bacteria in stainless steel plate Storage temperature 15 ° C

  From the test results, it was confirmed that the combination test agent functions effectively for the suppression of the floating bacteria in the soup and the formation of the biofilm on the stainless steel plate.

Claims (7)

One to seven parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons are dissolved in brewed vinegar to make 100 parts by weight of brewed vinegar, and this is a vitamin B1 derivative thiamine auril. A biofilm control agent having a function of controlling airborne bacteria in foods containing 0.01 to 12 parts by weight of sulfate. One to seven parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons are dissolved in brewed vinegar to make 100 parts by weight of brewed vinegar, and sugar alcohols sorbitol, xylitol, A biofilm control agent having a function of controlling airborne bacteria in foods by blending 3 to 100 parts by weight of one or two of erythritol. One to seven parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons are dissolved in brewed vinegar to make 100 parts by weight of brewed vinegar, which is sucrose fatty acid ester, glycerin fatty acid A biofilm control agent having a function of controlling airborne bacteria in foods containing 1 to 50 parts by weight of one of the esters. One to seven parts by weight of one or more animal calcium ions such as shells, eggshells, fish bones, and livestock skeletons are dissolved in brewed vinegar to make 100 parts by weight of brewed vinegar. A biofilm control agent having a function of controlling floating bacteria in food, comprising 10 to 90 parts by weight of seeds or two kinds. The biofilm control agent according to any one of claims 1 to 4, which has a function of controlling bacteria inside and outside the cuticle layer of a plant. The foodstuff which added the biofilm control agent in any one of Claims 1-4. An apparatus and method for measuring the number of bacteria in an object in a container and the number of bacteria in a biofilm by attaching to metal, plastics, rubber, ceramics, wood, paper, glass, or fiber installed in the container.