Food composition for reducing amount of mycotoxin absorbed into body

Abstract

Provided is a food composition that contains lactic acid bacteria as an active ingredient and reduces the amount of mycotoxins absorbed into a body.

Description

Food composition that reduces mycotoxins absorbed into the body

[0001] The present invention relates to a food composition that reduces mycotoxins absorbed into the body.

[0002] Mycotoxins are chemical substances produced by molds that contaminate food or feed. Ingestion of food or feed contaminated with mycotoxins can cause serious health problems for humans and livestock.

[0003] For example, aflatoxin, produced by certain Aspergillus molds, is a toxic substance with strong carcinogenic properties. Because aflatoxin is colorless, odorless, and tasteless, humans and livestock cannot detect it with their sensory organs, and there is a risk of unknowingly being exposed to large amounts. Furthermore, aflatoxin is physically and chemically stable, and remains in food and feed even after the mold has died, making it difficult to remove by approaches such as heating or washing.

[0004] Under these circumstances, attempts have been made in recent years to remove mycotoxins such as aflatoxins by biocontrol using beneficial microorganisms such as beneficial bacteria. Non-Patent Document 1 discloses that in an in vitro test, the adsorption capacity of aflatoxin was improved by heat inactivating a Lacticaseibacillus rhamnosus strain.

[0005] Effect of digestion and thermal processing on the stability of microbial cell-aflatoxin B1 complex

[0006] However, there was still no knowledge about foods and beverages that could reduce mycotoxins once they had been absorbed into the body, and there was still room for research and development.

[0007] Therefore, an object of the present invention is to provide a food composition that reduces mycotoxins absorbed into the body.

[0008] As a result of extensive research, the present inventors have discovered that a food composition containing lactic acid bacteria is effective in reducing mycotoxins absorbed into the body, and have thus completed the present invention.

[0009] [1] A food composition containing lactic acid bacteria as an active ingredient, which reduces mycotoxins absorbed into the body. [2] The food composition according to [1], wherein the mycotoxin is aflatoxin. [3] The food composition according to [2], which is a food composition that reduces aflatoxins in the blood or urine of the body. [4] The food composition according to [3], wherein the lactic acid bacteria are ingested by a subject at a dose of 5 to 200 billion bacteria per day for at least 6 weeks. [5] The substance to be detected as aflatoxin in urine is aflatoxin M １ and the substance to be detected for aflatoxin in the blood is aflatoxin B １ The food composition according to [3], wherein the aflatoxin concentration is 0.01% or more. [6] The food composition according to [3], wherein the subject for reducing aflatoxin in urine or blood is a human or a livestock. [7] The food composition according to any of [1] to [6], wherein the lactic acid bacteria is administered for at least 10 weeks. [8] The food composition according to any of [1] to [6], wherein the lactic acid bacteria is administered for at least 12 weeks. [9] The food composition according to any of [1] to [6], wherein the food composition is a beverage further comprising a carbohydrate and a milk protein.

[10] The food composition according to any of [1] to [6], wherein the lactic acid bacteria comprises the genus Lacticaseibacillus.

[11] The food composition according to

[10] , wherein the lactic acid bacteria is Lacticaseibacillus paracasei YIT 9029 (FERM BP-1366).

[12] A mycotoxin reducer containing Lacticase Bacillus paracasei as an active ingredient, which reduces mycotoxins absorbed into the body.

[13] The mycotoxin reducer according to

[12] , wherein the Lacticase Bacillus paracasei is Lacticase Bacillus paracasei YIT 9029 (FERM BP-1366).

[0010] According to the present invention, a food composition containing lactic acid bacteria as an active ingredient and capable of reducing mycotoxins absorbed into the body can be provided.

[0011] Figure 1 is a diagram illustrating a test schedule in an example of the present invention. Figure 2 is a table illustrating the amount of aflatoxin in urine in an example of the present invention. Figure 3 is a table illustrating the amount of aflatoxin in blood in an example of the present invention. Figure 4 is a table showing the relationship between the amount of aflatoxin in urine or blood and the foods ingested by the subject in an example of the present invention.

[0012] The present invention will be described in detail below. The following description of the constituent elements is an example of one embodiment of the present invention, and the present invention is not limited to these contents.

[0013] The present invention has the effect of reducing mycotoxins absorbed into the body by having a subject ingest a predetermined dose of a food composition containing lactic acid bacteria as an active ingredient for a predetermined period of time.

[0014] The food composition of the present invention has applications as a mycotoxin-reducing food composition, a mycotoxin-removing food composition, or a mycotoxin-absorption-inhibiting food composition, which reduce mycotoxins absorbed into the body. The present invention is also intended for use in humans or livestock. Furthermore, the present invention has applications as a mycotoxin-reducing agent, mycotoxin-removing agent, or mycotoxin-absorption inhibitor, which reduce mycotoxins absorbed into the body.

[0015] Here, "mycotoxins absorbed into the body" refers to mycotoxins or their metabolites that are absorbed into the body through cells in the digestive tract via oral ingestion. Mycotoxins absorbed into the body can be detected in blood or urine samples taken from a subject.

[0016] "Mycotoxins" include, for example, aflatoxin, ochratoxin A, deoxynivalenol, nivalenol, and batulin, with aflatoxin being particularly preferred. Aflatoxins detected in urine or blood include aflatoxin M, １ (aflatoxin M １ or AFM １ ) or aflatoxin B １ (aflatoxin B１ or AFB １ ) Aflatoxin M １ is aflatoxin B that has been absorbed into the body. １ is a metabolite broken down in the liver.

[0017] Generally, AFM １ is known as a marker that reflects aflatoxin intake in the last few days, and is used as a marker detected in urine. １ is known as a marker that reflects aflatoxin intake over the past two to three months, and is used as a marker that can be detected in the blood.

[0018] From the viewpoint of the effectiveness of mycotoxin reduction, it is preferable to ingest the food composition containing lactic acid bacteria of the present invention at a dose of 5 to 200 billion lactic acid bacteria per day. Also, from the viewpoint of the effectiveness of mycotoxin reduction, it is preferable to ingest the food composition containing lactic acid bacteria for at least 6 weeks.

[0019] The food composition of the present invention contains lactic acid bacteria as an active ingredient. Furthermore, from the viewpoint of demonstrating the effects of the present invention, it is preferable that the lactic acid bacteria be viable. Examples of lactic acid bacteria include Lacticaceibacillus bacteria such as Lacticaceibacillus paracasei, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus mali, Lactobacillus helveticus, Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus salivarius, Lactobacillus eugleti, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus johnsonii, Streptococcus bacteria such as Streptococcus thermophilus, and Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. Examples of bacteria that can be used include bacteria of the genus Lactococcus such as Lactococcus cremoris, Lactococcus plantarum, and Lactococcus raffinolactis, and bacteria of the genus Enterococcus such as Enterococcus faecalis and Enterococcus faecium.

[0020] Of the above Lacticase Bacillus paracasei, Lacticase Bacillus paracasei YIT 9029 (FERM BP-1366, date of deposit: May 1, 1981) has been internationally deposited with the International Patent Organism Depositary of the National Institute of Technology and Evaluation (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan), which is the international depositary authority of the Budapest Treaty. Lacticaseibacillus paracasei YIT 9029 was previously classified as Lactobacillus casei, but in recent years the genus Lactobacillus has been reclassified and it has been classified as Lacticaseibacillus paracasei (Zheng et al., A taxonomic note on the genus Lactobacillus: Description of 23 novel genera, emended description of the genus Lactobacillus). Beijerinck 1901, and union of Lactobacillaceae and Leuconostoceae. Int. J. Syst. Evol. Microbiol. 2020 Apr; 70(4):2782-2858 DOI 10.1099 / ijsem. 0.004107). In this specification, the classification of lactic acid bacteria is described using a new classification based on the above literature.

[0021] Among these, from the viewpoint of effectiveness in reducing mycotoxins, lactic acid bacteria of the genus Lacticaseibacillus are preferred, and Lacticaseibacillus paracasei YIT 9029 (FERM BP-1366) is more preferred.

[0022] The food composition may be in any form, for example, any food or drink in a liquid, semi-solid, solid, etc., as long as it effectively reduces mycotoxins. Specific examples include fermented food or drink, dried bacterial powder, etc., and food or drink containing fermented milk is particularly preferred.

[0023] The food composition may contain, in addition to the above-mentioned lactic acid bacteria, various carbohydrates and milk proteins, as well as dietary fiber, flavorings, etc., as needed.

[0024] The various carbohydrates may include sugars such as fructose, maltitol, glucose, lactose, sucrose, isomerized sugar, palatinose, trehalose, and xylose; sugar alcohols such as sorbitol, xylitol, erythritol, lactitol, palatinit, reduced starch syrup, and reduced maltose syrup; and high-intensity sweeteners such as aspartame, sucralose, stevia, and alitame.

[0025] Milk proteins include casein, whey protein, and the like, and milk proteins may include skim milk, whole milk powder, milk protein concentrate, whey protein concentrate, whey protein isolate, caseinate, and the like, which contain these components.

[0026] Dietary fiber may include polydextrose, dextrin, starch, pullulan, mannan, glucomannan, alginic acid, xanthan gum, guar gum, tamarind gum, chitin, chitosan, glycogen, inulin, pectin, agar, and the like.

[0027] The flavoring may include yogurt, berry, citrus, quince, perilla, apple, mint, grape, apricot, and the like.

[0028] Furthermore, the food composition may contain appropriate additives such as excipients, stabilizers, preservatives, humectants, emulsifiers, lubricants, sweeteners, colorants, flavorings, buffers, antioxidants, and pH adjusters, as needed.

[0029] From the viewpoint of the effectiveness of reducing mycotoxins, the intake amount of the food composition of the present invention is preferably 5 to 200 billion lactic acid bacteria per day, more preferably 10 to 200 billion lactic acid bacteria per day, and particularly preferably 20 to 200 billion lactic acid bacteria per day.

[0030] If the concentration is below the lower limit, it may not be possible to confirm the effectiveness of mycotoxin reduction, while if the concentration is above the upper limit, in the case of beverages, sedimentation may occur in the container, which may result in a poor appearance of the product.

[0031] The ingestion period of the food composition of the present invention is preferably at least 6 weeks, more preferably at least 10 weeks, and most preferably at least 12 weeks. If the ingestion period is below the lower limit, the effectiveness of mycotoxin reduction may not be confirmed.

[0032] EXAMPLES The present invention will be specifically described based on the following examples, but the present invention is not limited to these examples.

[0033] (Example 1) <Test Food and Drink> A lactic acid bacteria drink and a placebo drink were prepared in bottles, each 80 mL, as test drinks. The placebo drink had the same taste, color, appearance, and nutrients as the lactic acid bacteria drink, but did not contain lactic acid bacteria. The ingredients of the lactic acid bacteria drink and the placebo drink are shown below.

[0034] <Ingredients of lactic acid bacteria drink> Fructose, maltitol, skim milk, polydextrose, glucose, live bacteria Lacticase Bacillus paracasei YIT 9029 (FERM BP-1366, 3 x 1010 CFU / 80 mL), flavoring <Ingredients of placebo drink> Fructose, maltitol, skim milk, polydextrose, glucose, flavoring

[0035] (Example 2) <Subjects> The subjects were 174 healthy adults aged 20 to 60 years old, whose blood and urine samples contained detectable aflatoxins in a preliminary survey. A total of 174 subjects were tested, including 52 Malays, 96 Chinese, and 26 Indians. Of the 174 subjects, 87 were assigned to a group that consumed a lactic acid bacteria drink (hereinafter referred to as the lactic acid bacteria intake group) and a group that consumed a placebo drink (hereinafter referred to as the placebo intake group). After the start of the study, the subjects consumed the test drink twice a day for 12 weeks, within one hour after lunch and one hour after dinner. Furthermore, the subjects did not consume the test drink during the follow-up period after 12 weeks.

[0036] (Example 3) Evaluation Items Urine samples and blood samples were collected from subjects, and urinary aflatoxin (AFM) was measured. １ ) levels and blood aflatoxin (AFB １ The amount of omega-3 fatty acids was evaluated. In addition, a dietary survey was conducted on the subjects, and AFM １ and AFB１ The correlation with

[0037] <Test Method> A generalized estimating equation (GEE) model was used for statistical analysis in the examples of the present application. Statistical significance was determined using known testing methods. Specifically, to compare continuous variables between the lactic acid bacteria intake group and the placebo intake group, or to determine the baseline association between categorical variables, the Mann-Whitney U test or Kruskal-Wallis H test was used to determine the presence or absence of significant differences between two or more groups. Furthermore, Spearman's correlation coefficient was used to determine the association between continuous variables. A p-value was used as an indicator of the test results, and a statistically significant relationship was defined as a p-value of less than 0.05. Note that the implementation of the present invention is not limited to the above methods, as long as they are capable of determining statistical significance.

[0038] (Example 4) <Test Schedule> Figure 1 is a diagram illustrating the test schedule of the present invention. The horizontal axis indicates the time of sampling, the upper row indicates the lactic acid bacteria intake group, and the lower row indicates the placebo intake group.

[0039] In both the lactobacillus intake group and the placebo intake group, "△" indicates the timing of urine collection, and "▲" indicates the timing of blood collection. Urine was collected a total of nine times, and blood was collected a total of five times. In addition, intake of the lactobacillus drink and placebo drink was continued for 12 weeks from the start of intake. Furthermore, sampling of urine and blood continued for four weeks after the end of the intake period as a follow-up period.

[0040] AFM １ Quantitation of aflatoxin in urine (AFM) １ The amount of α-amyloid was determined using a commercially available kit (Aflatoxin M １The ELISA quantitative test kit, catalog number 991AFLM01U-96, Helica Biosystems Inc., was used to measure the IgG antibody titer according to the method of Mold et al. (Association between aflatoxin M １ The assay was performed using a modified version of the method described in Bulletin of Environmental Contamination and Toxicology, 89(6), 1115-1119 (extracted from human urine samples with the consumption of milk and dairy products). Specifically, urine samples were centrifuged at 3000 x g for 5 minutes to remove sediment. The supernatant and urine standards (0-4 ng / mL) were then diluted 1:20 with distilled water. The diluted standard or sample (100 μL) was mixed with assay buffer (200 μL), and 100 μL of the mixture was transferred to an antibody-precoated microwell. The microwells were incubated for 1 hour to measure the AFM in the standard or sample. １ After the conjugate was bound to the coated antibody, the contents were decanted from the microwells and washed three times with PBS-Tween wash buffer using a microplate washer. Approximately 100 μL of conjugate was added and incubated for 15 minutes to allow the remaining antibody coated on the wells to bind. This was followed by three rounds of decantation and washing, followed by the addition of substrate reagent (100 μL) and incubation for 15 minutes at room temperature in the dark. The color development was then immediately measured at absorbance 450 nm and compared with a standard curve using AFM. １ was quantified.

[0041] <AFB １ Quantitative determination of aflatoxin (AFB) in blood １ The amount of serum AFB was determined by the following method. First, the method of Mold et al. (Detection of serum AFB) １Serum was separated using a PBS-lysine adduct in Malaysia and its association with liver and kidney functions. International Journal of Hygiene and Environmental Health, 217(4-5), 443-451), and 250 μL of the resulting serum samples were transferred into capped vials and stored frozen at -80°C until analysis. Subsequently, the method of Andrews et al. R., Pokharel, A., & Acharya, S. (2021). growth over time in Nepal. Clinical Nutrition, 113(4), 874-883) and AFB was analyzed by high performance liquid chromatography with fluorescence detection. １ was quantified.

[0042] (Example 5) <Amount of aflatoxin in urine> Figure 2 is a table illustrating the amount of aflatoxin in urine in an example of the present invention. In the table, the first column shows the sampling time, and the second column shows the AFM in the lactic acid bacteria intake group. １ The geometric mean of the amount (ng / ml), the third column is AFM in the placebo group １ The fourth column shows the p-value calculated by the Mann-Whitney U test / Kruskal-Wallis H test. In the following paragraphs, cells in the figures that show significant differences are shown in gray.

[0043] As can be seen from Figure 2, the AFM of the lactobacillus intake group was １It was confirmed that the geometric mean of the amount was lower than that of the placebo intake group. This state continued not only during the intake period but also until the follow-up period four weeks after the end of intake. In addition, from the sixth week of intake onwards, the AFM of the lactic acid bacteria intake group was significantly lower than that of the first four weeks of intake. １ It was confirmed that the difference between the amount of lactic acid bacteria intake and that of the placebo intake group became larger. １ The amount of AFM continued to decrease compared to that of the placebo group, and the AFM at 12 weeks of intake １ It was confirmed that the amount was significantly reduced.

[0044] (Example 6) <Aflatoxin Levels in Blood> Figure 3 is a table illustrating the aflatoxin levels in blood in an example of the present invention. In the table, the first column shows the sampling time, and the second column shows the AFB levels in the lactic acid bacteria intake group. １ The geometric mean of the amount (pg / mg alb), the third column is the AFB in the placebo group １ The fourth column shows the p-value calculated by the Mann-Whitney U test / Kruskal-Wallis H test. １ AFB １ As shown in Figure 3, the AFB of the lactobacillus intake group was significantly higher than that of the control group at all time points after the start of intake. １ The geometric mean of the amount was confirmed to be lower than that of the placebo group. This state continued not only during the intake period but also until the follow-up period four weeks after the end of intake.

[0045] (Example 7) <Food Characteristics> Figure 4 is a table showing the relationship between the amount of aflatoxin in urine or blood during the sampling period and the food ingested by the subject in an example of the present invention. In the table, the first column shows the type of food, and the second column shows the amount of aflatoxin in urine (AFM). １ The third column shows the correlation coefficient (rho, ng / ml) and p-value of the blood aflatoxin level (AFB １The correlation coefficient and p-value of the AFB (amount) are shown. Spearman's correlation coefficient was used as the correlation coefficient. １ The amount is AFB １ -lysine adducts, the unit is "pg / mg alb." Figure 4 shows the relationship between grain intake and AFM １ While a significant correlation was found between the intake of protein (meat, fish, beans, eggs, dairy products) and AFM, １ It was found that there was no significant correlation between the amount of aflatoxin in the body. Note that a negative correlation coefficient indicates that consuming the corresponding type of food reduced the amount of aflatoxin in the body.

[0046] (Summary) From the above, the following points became clear: ・From Figure 2, it can be seen that the AFM of the lactobacillus intake group was １ It was confirmed that the geometric mean of the amount of aflatoxin in urine (i.e., the amount of aflatoxin in urine) was lower than that of the placebo group. This state continued not only during the intake period but also until the follow-up period four weeks after the end of intake. In addition, from the sixth week of intake onwards, the AFM of the lactobacillus intake group was significantly lower than that of the first four weeks of intake. １ It was confirmed that the difference between the amount of lactic acid bacteria intake and that of the placebo intake group became larger. １ The amount of AFB in the lactobacillus intake group continued to decrease compared to that in the placebo intake group, and it was confirmed that it had significantly decreased at the 12th week of intake. １ It was confirmed that the geometric mean of the amount was lower than that of the placebo group. This state continued not only during the intake period but also until the follow-up period four weeks after the end of intake. - Figure 4 reveals that grains are susceptible to aflatoxin contamination. Therefore, it can be said that subjects with a lifestyle that involves consuming grains are more likely to benefit from the effects of the food composition of the present application, which reduces mycotoxins absorbed into the body.

Claims

1. A food composition containing lactic acid bacteria as an active ingredient that reduces mycotoxins absorbed into the body.

2. The food composition of claim 1, wherein the mycotoxin is an aflatoxin.

3. The food composition according to claim 2, which is a food composition that reduces aflatoxin in the blood or urine of the body.

4. The food composition according to claim 3, wherein the lactic acid bacteria is administered to a subject at a dose of 5 to 200 billion cells per day for at least six weeks.

5. The substance to be detected for aflatoxin in the urine is aflatoxin M １ and the substance to be detected for aflatoxin in the blood is aflatoxin B １ The food composition according to claim 3, wherein 6. The food composition according to claim 3, wherein the subject for which aflatoxin in urine or blood is to be reduced is a human or a livestock.

7. A food composition according to any one of claims 1 to 6, wherein the lactic acid bacteria are to be ingested for at least 10 weeks.

8. A food composition according to any one of claims 1 to 6, wherein the lactic acid bacteria are to be ingested for at least 12 weeks.

9. The food composition according to any one of claims 1 to 6, which is a beverage product further comprising carbohydrates and milk proteins.

10. A food composition according to any one of claims 1 to 6, wherein the lactic acid bacteria include the genus Lacticaseibacillus.

11. The food composition according to claim 10, wherein the lactic acid bacterium of the genus Lacticaseibacillus is Lacticaseibacillus paracasei YIT 9029 (FERM BP-1366).

12. A mycotoxin reducer containing Bacillus paracasei as its active ingredient, which reduces mycotoxins absorbed into the body.

13. The mycotoxin-reducing agent according to claim 12, wherein the Lacticase Bacillus paracasei is Lacticase Bacillus paracasei YIT 9029 (FERM BP-1366).

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