Food composition containing soy food and food composition containing solanaceous vegetable, and method for improving stress condition

Abstract

To provide a method for evaluating whether a specific diet is suitable for an object person or an object animal by clarifying the relationship between intestinal bacteria and stress symptoms.SOLUTION: The present invention provides a processed soybean food-containing food composition and / or a solanaceous vegetable-containing food composition, and a method for evaluating whether or not ingestion of a processed soybean food or a solanaceous vegetable is suitable, based on the ratio of the processed soybean food and the solanaceous vegetable to the genus Lachnospira and the causal relationship between the genus Lachnospira and stress symptoms.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a soybean food-containing food composition, a solanaceous vegetable-containing food composition, and a method for improving stress conditions. [Background technology]

[0002] We are exposed to various stresses throughout our lives. When the body is exposed to stress, it triggers a stress response through reactions in the endocrine, autonomic, and immune systems. These include increased cortisol secretion via the hypothalamic-pituitary-adrenal axis, increased granulocytes and monocytes via activation of the sympathetic nervous system, and increased production of inflammatory cytokines. Long-term exposure to stress can have a variety of adverse effects on the mind and body, leading to physical symptoms such as gastric and duodenal ulcers, migraines, neurosis, and autonomic dysfunction, as well as psychiatric disorders such as depression and anxiety disorders (Xavier CH et al., Front Physiol. 2022;13:896-842; Kanno T et al., J Gastroenterol. 2013;48:483-90; Margolis KG et al., Gastroenterology. 2021;160:1486-501). In the Basic Survey on National Life conducted in 2019, the percentage of people who responded that they experienced anxiety or stress in their daily lives rose to 47.9%. Furthermore, according to the World Health Organization, the number of people suffering from depression worldwide has reached 280 million (Reference URL: https: / / www.who.int / publications / i / item / depression-global-health-estimates), and nearly 800,000 people die by suicide each year (Reference URL: https: / / www.who.int / publications / i / item / suicide-in-the-world). Therefore, how to control stress, which can be a cause of depression, is an extremely important issue.

[0003] In recent years, it has become clear that the intestinal microbiota is closely related to stress. It has long been reported that stress alters the intestinal microbiota. It has been reported that overcrowding in rats increases the number of Staphylococcus and Corynebacterium species and decreases the number of Bifidobacterium species (Non-Patent Document 1). Separation of mother and child monkeys reduces the diversity of the infant monkey's microbiota, with a particularly significant decrease in the Lactobacillus species (Non-Patent Document 2). It has also been reported that acute psychosocial stress testing in humans alters the composition of the intestinal microbiota (Non-Patent Document 3). It has also been reported that psychological stress is negatively correlated with the number of Lachnospira, Lachnospiraceae, Phascolarctobacterium, Stellera, and Veillonella species in the intestinal microbiota (Non-Patent Document 4). On the other hand, it is also known that the intestinal microbiota influences stress responses. Germ-free mice, which have no intestinal microbiota at all, are known to have increased reactivity to stress and reduced levels of brain-derived neurotrophic factor (BDNF), which promotes the development of the nervous system in the brain, compared to normal mice (Non-Patent Document 5). It has also been reported that the intake of probiotics such as lactic acid bacteria and bifidobacteria reduces scores on anxiety and depression tests (Non-Patent Document 6). Thus, controlling the intestinal microbiota can change responses to stress, suggesting that diet may be able to alter the intestinal microbiota and influence stress responses.

[0004] In fact, dietary interventions designed by registered dietitians that include high amounts of indigestible foods such as onions, cabbage, and bananas have been shown to reduce stress scores in questionnaires and significantly improve sleep quality (Non-Patent Document 7). It is also known that the ingestion of soy polyphenols and isoflavones can alter the intestinal microflora (Patent Documents 1 and 2). Furthermore, feeding mice with genistein (400 mg / kg), a type of isoflavone found in soybeans, has been reported to increase the number of Lachnospira bacteria in their intestines (Non-Patent Document 8). However, the median soy isoflavone intake of Japanese people aged 15 years and older is reported to be approximately 20 mg / day (Food Safety Commission, Basic Concepts for Safety Assessment of Specified Health Foods Containing Soy Isoflavones, 2006), making 400 mg / kg of genistein an unrealistic amount to ingest as a dietary source. On the other hand, α-linolenic acid, eicosapentaenoic acid, and docosahexaenoic acid, which are widely associated with mental illness, have been reported to not contribute to the improvement of depression on their own, suggesting that a combination of multiple nutrients may be effective (Sarris J et al., Nutr Rev. 2009;67:125-31). This suggests that various dietary and nutrient sources affect stress, and that their intake may lead to the prevention of mental illnesses such as depression and anxiety disorders. Therefore, when investigating the relationship between dietary and nutrient sources and stress, it is necessary to consider multiple nutrients, not just a single nutrient.

[0005] It is known that the effectiveness of ingesting a particular food depends on the proportion of bacteria in the intestinal flora. For example, a method is known for determining whether a patient with dyslipidemia or hypertension will become healthy by ingesting barley, based on the proportion of intestinal normal bacteria or the taxonomic group of intestinal normal bacteria (Patent Document 3).

[0006] The genus Lachnospira has been reported as an anti-inflammatory bacterium (Jayasudha R et al., J Biosci. 2018;43(5):835-856.), and a low proportion of Lachnospira in the intestine has been reported to be a risk factor for systemic lupus erythematosus (SLE) and allergies (Xiang K et al., Front Immunol. 2021;12:667097.; Mousavian AH et al., J Asthma. 2024:1-15.). [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Patent No. 7437216 [Patent Document 2] Patent No. 2023-080372 [Patent Document 3] Patent Publication No. 2022-166730 [Non-patent literature]

[0008] [Non-Patent Document 1] Suzuki K et al., Nihon Juigaku Zasshi. 1983;45:331-8. [Non-patent document 2] Bailey MT et al., Dev Psychobiol. 1999;35:146-55. [Non-patent document 3] Yamaoka K et al.,Neurobiol Stress. 2022;20:100479. [Non-patent document 4] Lu Ma et al., Neuropsychobiology. 2023;82:247-262. [Non-patent document 5] Sudo N et al., J Physiol. 2004;558:263-75. [Non-patent document 6] Messaoudi M et al.,Gut Microbes. 2011;2:256-61. [Non-Patent Document 7] Berding K et al.,Mol Psychiatry. 2023;28:601-10. [Non-patent document 8] Hou Q et al.,Pharmacological Research. 2023;188:106676 Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention aims to increase the proportion of Lachnospira in intestinal bacteria in order to improve stress conditions. [Means for solving the problem]

[0010] The present inventors attempted to clarify the relationship between intestinal bacteria, diet, and stress using various statistical analysis methods from a database of human intestinal microbiota, dietary habits, and stress states, and after considerable ingenuity and investigation, discovered that groups with a low proportion of Lachnospira in their intestinal bacteria are in a state of high stress, and that the intake of beans and vegetables increases the proportion of Lachnospira and improves stress states. Based on these findings, the present inventors have completed the present invention.

[0011] The present invention includes, for example, the inventions shown in [1] to [9] below. [1] A food composition containing a processed soybean food and / or a food composition containing a Solanaceae vegetable for enhancing the proportion of Lachnospira in intestinal bacteria. [2] The food or food composition according to [1], wherein the soybean processed food is a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, and natto. [3] The food or food composition according to [1], wherein the solanaceous vegetables are foods selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato. [4] A method for evaluating whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state in a subject or a target animal, comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that intake of processed soybean foods or Solanaceae vegetables is suitable for the subject or target animal when the proportion of Lachnospira in the subject or target animal is relatively low. A method comprising: [5] The method according to [4], wherein the soybean processed food is a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, and natto. [6] The method according to [4], wherein the solanaceous vegetables are foods selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato. [7] A program that outputs an evaluation of whether intake of processed soybean foods or nightshade vegetables is suitable for improving a stress state of a subject or a subject animal, the program comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results. A program that outputs an evaluation that the intake of soybean processed foods or Solanaceae vegetables is suitable for the subject or target animal when the proportion of Lachnospira genus in the subject or target animal is relatively low, based on the data obtained by the above. [8] A program for transmitting to a client an evaluation of whether intake of processed soybean foods or nightshade vegetables is suitable for improving a stress state of a subject or a target animal, the program being executed on a server and comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results. and transmits to a client via a communication network an assessment that the intake of processed soybean foods or Solanaceae vegetables is suitable for the subject or target animal if the proportion of Lachnospira genus in the subject or target animal is relatively low, based on the data obtained by the method. [9] A kit for evaluating whether the intake of processed soybean foods or nightshade vegetables is suitable for improving the stress state of a subject or a subject animal, comprising a detection reagent for detecting the genus Lachnospira. [Effects of the Invention]

[0012] The present invention may be able to increase the proportion of Lachnospira in intestinal bacteria in order to improve stress conditions. [Brief explanation of the drawings]

[0013] [Figure 1] (Top): Subjects were clustered using Ward's method based on their mental and physical stress response scores. The horizontal dotted line indicates the number of clusters determined this time, and the numbers on the line are the cluster numbers. (Bottom): The mental and physical stress response scores were tallied for each cluster. The results are shown as the mean ± standard error. Note that significant differences are considered to be between different letters, and the significance level is set at 0.05. [Figure 2]For each cluster, the amount of food intake calculated using the Nutrient Rich Food Index 9.3 (NRF9.3) and the Food Frequency Questionnaire (FFQ) was compiled. Results are shown as mean ± standard error. Significant differences are indicated by different letters, with a significance level of 0.05. [Figure 3] The results for the abundance of gut bacteria and the subjects' physical characteristics were summarized for each cluster. The results are shown as the mean ± standard error. Note that significant differences are indicated by different letters, and the significance level is set at 0.05. [Figure 4] The results of a Bayesian network-based causal search for mental and physical stress response scores, intestinal bacteria, food groups, and physical characteristics are shown below. Links with a rate of appearance greater than 0.5 were deemed significant, and this value was set as the threshold to create a network model. The thickness of the lines indicates the strength of the links. [Figure 5] The intake of foods included in the legume and vegetable categories in the FFQ was calculated for each cluster. The results are shown as mean ± standard error. Note that significant differences are indicated by different letters, and the significance level is set at 0.05. [Figure 6] The results of a Bayesian network-based causal search for mental and physical stress response scores, intestinal bacteria, food, and physical characteristics are shown below. Links with a rate of appearance greater than 0.5 were deemed significant, and this value was set as the threshold to create a network model. The thickness of the lines indicates the strength of the links. DETAILED DESCRIPTION OF THE INVENTION

[0014] The present invention will be described in detail below.

[0015] Intestinal bacteria are bacteria that inhabit the digestive tract. Intestinal flora is a group formed by inhabiting separate regions of the digestive tract by individual intestinal bacteria. Intestinal flora may also be called intestinal bacterial flora, intestinal microbiota, intestinal microbiome, etc. Intestinal flora specimens are specimens derived from intestinal flora. In one embodiment, the specimen derived from intestinal flora in the present invention is feces.

[0016] Methods for analyzing the bacterial composition of samples derived from intestinal microbiota used in the present invention include, for example, culture in selective media, metabolomic analysis, proteomic analysis, 16s rRNA analysis, shotgun metagenomic analysis, and whole genome analysis, but are not limited to these, as long as the proportion of any bacteria can be compared with the proportion in any population.

[0017] In one embodiment, the analysis of the present invention is performed by analyzing a shotgun library prepared from metagenomic DNA of bacteria contained in the intestinal microbiota. In one embodiment, the analysis is performed using a next-generation sequencer.

[0018] In one embodiment, the analysis of the present invention is performed by analyzing a library prepared by amplifying a region containing the variable region of 16s rRNA of bacteria contained in the intestinal microbiota. In another embodiment, the variable region is a region containing the V3-V4 variable region. In one embodiment, the analysis is performed using a next-generation sequencer.

[0019] Examples of next-generation sequencers that can be used in the present invention include GS Junior (Roche), Genome Sequencer FLX System (454 Life Sciences), MiSeq (Illumina), NovaSeq (Illumina), and Ion Proton System (Thermo Fisher Scientific), but are not limited to these as long as they are capable of analyzing the composition of the intestinal microbiota or a 16s rRNA library.

[0020] The genus Lachnospira (Lachnospira / NCBI: txid28050) is a bacterium belonging to the family Lachnospiraceae (Lachnospiraceae / NCBI: txid186803) that is generally known to be present in the human intestine. Examples of bacteria of the genus Lachnospira include Lachnospira pectinoschiza, Lachnospira multipara, Lachnospira hominis, and Lachnospira eligens, but the Lachnospira genus targeted by the present invention is not limited to these as long as it is a bacterium belonging to the genus Lachnospira.

[0021] Any detection reagent can be used in the analysis of the composition of the genus Lachnospira. The detection reagent of the present invention is not particularly limited as long as it can detect the genus Lachnospira. In one embodiment, the detection reagent of the present invention is a selective medium capable of selectively culturing the genus Lachnospira. In another embodiment, the detection reagent of the present invention is a detection agent for bacterial metabolites of the genus Lachnospira. In another embodiment, the detection reagent of the present invention is an antibody that selectively binds to a protein of the genus Lachnospira. In another embodiment, the detection reagent of the present invention is a nucleic acid that specifically hybridizes with a nucleic acid sequence of the genus Lachnospira. The nucleic acid sequence of the genus Lachnospira can be a sequence known to those skilled in the art.

[0022] In one embodiment, the nucleic acid that specifically hybridizes to a nucleic acid sequence of the genus Lachnospira is a nucleic acid that hybridizes to a region of the 16s rRNA of the genus Lachnospira, preferably a nucleic acid that hybridizes to a region including the variable region of the 16s rRNA, and more preferably a nucleic acid that hybridizes to a region including the V3-V4 region of the 16s rRNA.

[0023] In one embodiment, analysis of the composition of the genus Lachnospira can be performed using quantitative PCR using nucleic acids that specifically hybridize to nucleic acid sequences of the genus Lachnospira.

[0024] In one embodiment, in the present invention, in addition to the genus Lachnospira, any other bacteria can also be simultaneously targeted for analysis.

[0025] In one embodiment, the arbitrary population to which the proportion of any bacteria in the intestinal bacteria of a subject is compared is a population including the subject from whom the sample was derived. In another embodiment, the arbitrary population to which the proportion of bacteria is compared is a group similar to the subject in terms of race, residential area, food culture, physical characteristics, and / or health condition. In another embodiment, the arbitrary population to which the proportion of bacteria is compared is a group of 10 or more individuals, preferably a group of 100 or more individuals, and more preferably a group of 500 or more individuals.

[0026] In one embodiment, the step of comparing the proportion of Lachnospira in enterobacteria with the proportion of the bacteria in an arbitrary population refers to a step of comparing the proportion of Lachnospira in enterobacteria with the average proportion of the bacteria in an arbitrary population.

[0027] In one embodiment, the significance level for determining whether the proportion of any bacteria in a subject's intestinal bacteria is high or low compared to any population is 10%, preferably 5%.

[0028] In one embodiment, if the proportion of Lachnospira in a gut microbiota sample is low compared to any population, it can be determined that the subject of the sample is suitable for consuming a processed soybean food or a solanaceous vegetable, as well as a food composition containing a processed soybean food or a food composition containing a solanaceous vegetable, in order to improve stress conditions.

[0029] Soybean processed foods are foods produced using soybeans as one of the ingredients. Examples of soybean processed foods used in the present invention include roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, natto, etc., but are not limited to these as long as they are processed foods using soybeans as one of the ingredients. In this specification, processed soybeans also include processed foods using roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, natto, etc. as one of the ingredients. In one embodiment, the soybean processed food is a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, and natto, and is preferably natto.

[0030] Solanaceous vegetables are edible plants belonging to the Solanaceae family. Examples of solanaceous vegetables used in the present invention include eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, potato, etc., but are not limited to these as long as they are edible plants belonging to the Solanaceae family. Solanaceous fruit vegetables also fall under the category of solanaceous vegetables. Examples of solanaceous fruit vegetables used in the present invention include eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, etc. In one embodiment, the solanaceous vegetable is a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato, and is preferably tomato or bell pepper.

[0031] As used herein, a food composition is an edible composition that contains at least one food ingredient.

[0032] A stress state is a state in which mental and physical symptoms are manifested by stress stimuli. Examples of mental symptoms include depression, irritability, lack of motivation, decreased concentration, distractibility, and decreased interest. Examples of physical symptoms include insomnia, hypersomnia, fatigue, lethargy, loss of appetite, and headaches. A stress state can also be assessed by questionnaires corresponding to stress checks.

[0033] In one embodiment, the subject is a human. In another embodiment, the subject is a non-human mammal. The present invention can be applied to mammals, including humans, humans, or non-human mammals. The present invention can also be applied to mammals, including humans, humans, or non-human mammals, that are in a stressed state or are at risk of becoming stressed.

[0034] A storage medium is a medium that stores computer programs and data, and includes, but is not limited to, hard disk drives (HDDs), solid-state drives (SSDs), magnetic tape drives, optical disk drives, flash memory, etc., as long as the contents can be read by a computer.

[0035] A server is a computer that provides various functions and services to other computers over a network. A client is a computer that uses the server's functions and services by communicating with the server. In one embodiment, the client is a smartphone.

[0036] In the present invention, a computer that executes a program is configured to have a CPU, RAM (Random Access Memory), ROM (Read Only Memory), storage, a user interface as needed, and a communication interface as needed. The components of the computer are connected to each other via a bus so that they can communicate with each other.

[0037] The CPU is a central processing unit that executes various programs and controls each part. That is, the CPU reads the program from ROM or storage and executes the program using RAM as a work area. The CPU controls each component of the computer and performs various arithmetic processing according to the program recorded in ROM or storage. In one embodiment of the present invention, the program of the present invention is stored in storage or an additional storage medium.

[0038] The ROM stores various programs and various data. The RAM temporarily stores programs or data as a working area. The storage is composed of an HDD or SSD and stores various programs including the operating system and various data. The user interface is configured to include, for example, at least one of an LCD display with a touch panel that allows the user to perform touch operations, a voice input reception unit that receives voice input from the user, and a button that the user can press. The communication interface is an interface that allows the computer to communicate with other devices, and uses standards such as Ethernet (registered trademark), FDDI, and Wi-Fi (registered trademark).

[0039] Output from a program refers to transmitting the results of processing by the program to an external device. In one embodiment, output from a program includes displaying the results as an image so that humans can visually perceive them, playing the results as sound so that humans can audibly perceive them, saving the results as readable data in a memory area, or transmitting the results over a network, etc.

[0040] In one embodiment, the computer uses a program recorded on a storage medium to process data comparing the proportion of Lachnospira in the intestinal bacteria of a subject or a target animal with the proportion of said bacteria in an arbitrary population, and can output an assessment that the intake of processed soybean foods or Solanaceae vegetables is suitable for said subject or target animal if the proportion of Lachnospira in the intestinal bacteria of said subject or target animal is relatively low. Preferably, the output is configured to be able to be transmitted to a client via a communication network.

[0041] <Food and food composition of the present invention> The present invention includes a food composition containing a processed soybean food and a food composition containing a Solanaceae vegetable for enhancing the proportion of Lachnospira in intestinal bacteria.

[0042] In one embodiment, the present invention is a food composition containing a processed soybean food or a food composition containing a solanaceous vegetable for enhancing the proportion of Lachnospira genus in intestinal bacteria.

[0043] In one embodiment, the present invention relates to a food composition containing a processed soybean food for enhancing the proportion of Lachnospira in intestinal bacteria, wherein the processed soybean food is a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba (tofu skin), tofu, fried tofu, miso, and natto.

[0044] In one embodiment, the present invention is a natto-containing food composition for enhancing the proportion of Lachnospira genus in intestinal bacteria.

[0045] In one embodiment, the present invention relates to a food composition containing a solanaceous vegetable for enhancing the proportion of Lachnospira in intestinal bacteria, wherein the solanaceous vegetable is a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato.

[0046] In one embodiment, the present invention relates to a food composition containing Solanaceae fruit vegetables for enhancing the proportion of Lachnospira in intestinal bacteria, wherein the Solanaceae fruit vegetables are foods selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, and chili pepper.

[0047] In one embodiment, the present invention is a tomato and / or bell pepper-containing food composition for enhancing the proportion of Lachnospira genus in intestinal bacteria.

[0048] In one embodiment, the present invention is a food composition containing natto, tomato, and / or bell pepper for enhancing the proportion of Lachnospira genus in intestinal bacteria.

[0049] Increasing the proportion of Lachnospira in intestinal bacteria can be expected to improve, alleviate, and prevent stress conditions, or to have anti-inflammatory effects, more specifically, to reduce the risk of SLE and allergies.

[0050] <Evaluation method of the present invention> The present invention includes a method for evaluating whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state in a subject or a subject animal.

[0051] In one embodiment, the present invention provides a method for evaluating whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state in a subject or a subject animal, the method comprising the steps of: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that intake of processed soybean foods or Solanaceae vegetables is suitable for the subject or target animal when the proportion of Lachnospira in the subject or target animal is relatively low. The method includes:

[0052] In one embodiment, the present invention provides a method for evaluating whether ingestion of a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, and natto is suitable for improving a stress state in a subject or a subject animal, the method comprising the steps of: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that the intake of a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba (dried soybeans skin), tofu, fried tofu, miso, and natto is suitable for the subject or the target animal when the proportion of Lachnospira in the subject or the target animal is relatively low. The method includes:

[0053] In one embodiment, the present invention provides a method for evaluating whether ingestion of natto is suitable for improving a stress state in a subject or a target animal, the method comprising the steps of: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that the intake of natto is suitable for the subject or the target animal when the proportion of Lachnospira in the subject or the target animal is relatively low; The method includes:

[0054] In one embodiment, the present invention provides a method for evaluating whether ingestion of a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato is suitable for improving a stress state in a subject or a subject animal, the method comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that the intake of a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato is suitable for the subject or target animal if the proportion of Lachnospira in the subject or target animal is relatively low. The method includes:

[0055] In one embodiment, the present invention provides a method for evaluating whether ingestion of a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, and chili pepper is suitable for improving a stress state in a subject or a target animal, the method comprising the steps of: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that the intake of a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, and chili pepper is suitable for the subject or target animal when the proportion of Lachnospira in the subject or target animal is relatively low. The method includes:

[0056] In one embodiment, the present invention provides a method for evaluating whether ingestion of tomatoes or bell peppers is suitable for improving a stress state in a subject or a subject animal, the method comprising the steps of: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that the subject or target animal is suitable for consumption of tomatoes or bell peppers if the proportion of Lachnospira in the subject or target animal is relatively low. The method includes:

[0057] In one embodiment, the present invention provides a method for evaluating whether ingestion of natto, tomatoes, or bell peppers is suitable for improving a stress state in a subject or a subject animal, the method comprising the steps of: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that the intake of natto, tomatoes, or bell peppers is suitable for the subject or the target animal when the proportion of Lachnospira in the subject or the target animal is relatively low. The method includes:

[0058] <Kit of the Present Invention> The present invention includes a kit for evaluating whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state in a subject or a subject animal.

[0059] In one embodiment, the present invention is a kit for evaluating whether the intake of processed soybean foods or solanaceous vegetables is suitable for improving a stress state in a subject or subject animal, comprising a detection reagent for detecting the genus Lachnospira.

[0060] In one embodiment, the kit of the present invention is a kit used in the method of the present invention.

[0061] In one embodiment, the present invention is a kit for evaluating whether the intake of processed soybean foods or solanaceous vegetables is suitable for improving a stress state in a subject or animal, the kit comprising a nucleic acid that hybridizes to a nucleic acid sequence of the genus Lachnospira.

[0062] In one embodiment, the present invention is a kit for evaluating whether the intake of processed soybean foods or solanaceous vegetables is suitable for improving a stress state in a subject or animal, the kit comprising a nucleic acid that hybridizes to a region of 16s rRNA of a nucleic acid sequence of the Lachnospira genus.

[0063] <Program of the present invention> The present invention includes a program for evaluating whether intake of processed soybean foods or solanaceous vegetables is suitable for improving the stress state of a subject or subject animal.

[0064] In one embodiment, the present invention provides a program for outputting an evaluation of whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state of a subject or a subject animal, the program comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results. A program that outputs an evaluation that the intake of soybean processed foods or Solanaceae vegetables is suitable for the subject or target animal when the proportion of Lachnospira genus in the subject or target animal is relatively low, based on the data obtained by the above.

[0065] In one embodiment, the present invention provides a program for transmitting to a client an evaluation of whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state of a subject or a target animal, the program being executed on a server and comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results. and transmits to a client via a communication network an assessment that the intake of processed soybean foods or Solanaceae vegetables is suitable for the subject or target animal if the proportion of Lachnospira genus in the subject or target animal is relatively low, based on the data obtained by the method.

[0066] In one embodiment, the present invention provides a storage medium storing a program for outputting an evaluation of whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state of a subject or a subject animal, the program comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results. A storage medium that stores a program that outputs an evaluation that the intake of soybean processed foods or solanaceous vegetables is suitable for the subject or target animal when the proportion of Lachnospira genus in the subject or target animal is relatively low based on the data obtained by the method.

[0067] In one embodiment, the present invention provides a storage medium storing a program for transmitting to a client an evaluation of whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state of a subject or a target animal, the storage medium being installed in a server and comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results. A storage medium storing a program for transmitting to a client via a communication network an assessment that the intake of processed soybean foods or Solanaceae vegetables is suitable for the subject or target animal if the proportion of Lachnospira genus in the subject or target animal is relatively low based on the data obtained by the method.

[0068] For a better understanding of the present invention, reference is made to specific examples which are provided herein for purposes of illustration and not limitation. [Example]

[0069] <Means> Survey target This study was conducted using data from the "Comprehensive Research Survey for the Construction of an Integrated Database of Food, Gut Microbiome, and Health Information (Healthy Health Survey)," conducted by Hokkaido Information University, University of Miyazaki, Kyoto University, and University of Nagasaki. The Healthy Health Survey is a survey of healthy Japanese men and women aged 20 to 80, excluding those with serious cerebrovascular disease, heart disease, liver disease, kidney disease, gastrointestinal disease, or notifiable infectious diseases. It is conducted twice a year, in summer and winter, per subject. The Healthy Health Survey was conducted in fiscal years 2019 and 2020 as part of the Strategic Innovation Promotion Program (SIP). This study used data from the summer of fiscal years 2019 and 2020. Of these data, analysis was conducted using data from 1,268 people, including "subject attributes," "body composition measurements," "questionnaire survey," "microbial composition," and "dietary survey using FFQ (Food Frequency Questionnaire)" (Reference URL: https: / / humandbs.dbcls.jp / data-use / all-researches). ·Mental and physical stress response score The mental and physical stress response score was calculated using the Simple Occupational Stress Questionnaire included in the questionnaire survey. The Simple Occupational Stress Questionnaire is a questionnaire recommended by the Ministry of Health, Labor and Welfare for the implementation of the stress check system, and is a questionnaire survey consisting of 57 items created by research by the Stress Measurement Group of the Ministry of Labor's commissioned research "Research on the Prevention of Work-Related Illnesses" from 1995 to 1999. Of the 57 items, the survey results for 29 items that correspond to "mental and physical stress response" were used. For each item, high stress was assigned 4 points and low stress was assigned 1 point, and the total score was defined as the mental and physical stress response score. ·Nutrient rich food index 9.3(NRF9.3) The NRF9.3 was used to evaluate overall dietary quality based on the nutritional intake results obtained from the FFQ. To calculate the NRF9.3, various reference values ​​from the 2020 Dietary Reference Intakes for Japanese were used (recommended intakes for protein, vitamin A, vitamin C, calcium, iron, and magnesium, estimated intakes for vitamin D, and target intakes for saturated fatty acids, dietary fiber, potassium, and sodium). Regarding added sugars, because there are no recommended intake values ​​for added sugars in Japan and intakes are low, the value proposed by the WHO (5% of the estimated energy requirement) was used. The results of the "Sugars and Sweeteners" section of the FFQ were used to determine added sugar intake. The NRF9.3 score was calculated by subtracting the sum of the points calculated from the proportions of the reference intakes of the three undesirable nutrients (added sugars, saturated fatty acids, and sodium) from the sum of the points calculated from the proportions of the reference intakes of the nine desirable nutrients (protein, dietary fiber, vitamin A, vitamin C, vitamin D, iron, calcium, potassium, and magnesium). The reference values ​​for each nutrient are shown in Table 1. For nutrients with desirable intakes, the percentage of the recommended intake amount is capped at 100, and any amount exceeding 100% is rounded down to prevent a high intake of one nutrient from compensating for a low intake of another. In other words, if all nine nutrients exceed the recommended intake amount, the score is calculated as a full 900 points. Furthermore, for nutrients with undesirable intakes, the score is calculated based on the percentage that exceeds the recommended amount. In other words, if all three nutrients are below the recommended intake amount, the score is 0, and no deductions are made from the score of the desirable nutrients.

[0070] [Table 1]

[0071] ·Statistical analysis methods Statistical analysis was performed using GraphPad Prism (version 9.5.1, GraphPad Software Inc., USA) and the free software "R" (version 4.3.1, CRAN). Exclusion criteria for analysis Those with any missing values ​​in any of the aforementioned datasets were excluded from the analysis. Based on this exclusion criterion, data from 1,058 individuals were used for the analysis. The average values ​​of the physical characteristics of the 1,058 individuals are shown in Table 2.

[0072] [Table 2]

[0073] Hierarchical Cluster Analysis Hierarchical cluster analysis was performed using R. To perform the hierarchical cluster analysis, the mental and physical stress response scores were used as a dataset. The dataset was loaded as a CSV file and converted into a data frame. This data frame was then standardized using the scale function. The standardized data frame was then used for the hierarchical cluster analysis. First, the distance between samples was calculated using the dist function in the stats package. The euclidean distance calculation method was used. Next, the hclust function in the stats package was used, specifying ward.D as the cluster merging algorithm. A dendrogram of the subjects was created using the plot function in the gplots package. The dendrogram and heatmap results of the subjects created by the hierarchical cluster analysis were confirmed, and the subjects were classified into three clusters using the cutree function in the stats package. The cluster results separated by the cutree function were converted to factor types using the facter function and then combined with the mental and physical stress response score data frame using the cbind function. Finally, the write.csv function was used to output the data to CSV format, and the resulting CSV file was used for analysis of variance.

[0074] One-way analysis of variance and nonparametric tests Based on the cluster results obtained by hierarchical cluster analysis, the mean values ​​for each item of gut microbiota, dietary records, and body composition were calculated for each cluster, and significant changes between clusters were examined using Prism. For each item, the data were first examined for normal distribution using the D'Agostino-Pearson test, and then for variance using the Bartlett's test. If the data were normally distributed and had equal variances, a one-way analysis of variance was used, followed by Tukey's multiple comparison test as a post-hoc test. If the data were non-normally distributed or had unequal variances, a nonparametric test, the Kruskal-Wallis test, was used, followed by Dunn's multiple comparison test as a post-hoc analysis. The p-values ​​for each test were calculated, with p<0.05 considered significant, to identify the characteristics of each cluster.

[0075] Chi-square test To test whether the gender ratio differed between groups, a chi-square test included in Prism was used.

[0076] ·Causal search The dataset used consisted of items that showed significant differences between the mental and physical stress response scores and the one-way analysis of variance and nonparametric tests. This dataset was standardized as described above and exported to a CSV file. First, because it was unlikely that gut bacteria affect age, a constraint was added to prevent the emergence of a link between gut bacteria and age. The constraints were implemented using the tiers2dblacklist function in the bnlearn package. Subdatasets were then created from the dataset using the bootstrap method. A hill-climbing structural learning algorithm was then applied to each subdataset to learn a directed acyclic graph (DAG). The number and direction of links in the DAG were recorded, and the proportion of links relative to the entire subdataset was calculated. Significant proportions were then used as a reliability index to model the causal relationships. The structural learning process described above was performed using the boot.strength function in the bnlearn package, and graphs were created using the strength.plot function. Next, parameter estimation was performed using the bn.fit function in the bnlearn package. The estimation method used was maximum likelihood estimation.

[0077] <Result> The subjects were classified into clusters based on their mental and physical stress response scores. The results are shown in Figure 1 (top). From the dendrogram created, the subjects were classified into three major clusters, and cluster classification was performed. The positions of the clusters are indicated by the dotted lines in Figure 1 (top).

[0078] The mental and physical stress response scores for each cluster were aggregated and averaged, as shown in Figure 1 (bottom). Cluster 2 had significantly higher mental and physical stress response scores than the other clusters. On the other hand, Cluster 3 had significantly lower mental and physical stress response scores than the other clusters. In other words, Cluster 2 can be considered a high-stress group, Cluster 3 a low-stress group, and Cluster 1 a medium-stress group.

[0079] Next, the NRF9.3 scores were aggregated across clusters. The results are shown in Figure 2. Cluster 2 had significantly lower NRF9.3 scores than Clusters 1 and 3. This suggests that the high-stress group may have poorer dietary quality. The amount of food intake for each food group obtained from the FFQ dietary survey was also aggregated for each cluster to determine which food groups differed in intake between clusters. Figure 2 shows the results of aggregating the 14 food groups by cluster. Significant differences were observed between clusters for legumes and vegetables, with Cluster 2 having significantly lower intakes of both foods than the other clusters. This suggests that Cluster 2's low intake of legumes and vegetables may have contributed to its lower NRF9.3 score.

[0080] Next, we compared the intestinal bacteria between the clusters. As a result, significant differences were observed between the clusters for the genera Lachnospira, Ruminococcus_E, and Collinsella. The results are shown in Figure 3. For the genera Lachnospira and Ruminococcus_E, the abundance ratio was significantly lower in Cluster 2 than in Cluster 3, and for the genus Collinsella, the abundance ratio was significantly lower in Cluster 2 than in Cluster 1.

[0081] Next, we checked whether there were differences in physical characteristics such as age and gender within each cluster. Table 3 shows the male / female ratio for each cluster, and Figure 3 shows age, BMI, systolic blood pressure, and diastolic blood pressure. As a result, no significant differences were found in the male / female ratio for each cluster (p=0.3984), but significant differences were found between clusters in age, systolic blood pressure, and diastolic blood pressure. Significant differences were found between all clusters in age, and cluster 2 had significantly lower blood pressure than cluster 3 in systolic blood pressure and diastolic blood pressure.

[0082] [Table 3]

[0083] These findings suggest that physical and mental stress response scores are influenced by diet quality, food groups consumed, intestinal bacteria, and physical characteristics. Therefore, we next sought to estimate causal relationships among these factors. Using the items that previously showed significant differences, we conducted a causal search using Gaussian Bayesian networks. We also added ethically unrelated causal relationships as constraints. The results are shown in Table 4. Figure 4 shows the structural model created after the causal search. As a result, a causal relationship was identified between NRF9.3, Lachnospira genus, and physical and mental stress response scores. Furthermore, food groups such as beans and vegetables appeared upstream of NRF9.3. Furthermore, the results of parameter estimation using regression analysis of the relationships between each link are shown in Table 5. The coefficient indicating the relationship between NRF9.3 and Lachnospira genus was 0.1026, and the coefficient indicating the relationship between Lachnospira genus and physical and mental stress response scores was -0.0841. Furthermore, the coefficients showing the relationship between the links "legumes → NRF9.3" and "vegetables → NRF9.3" were 0.2281 and 0.4981, respectively. From the above, it was inferred that a causal relationship exists in which a high NRF9.3 increases the abundance of the genus Lachnospira and also reduces the mental and physical stress response score. In particular, it was suggested that the intake of legumes and vegetables may improve dietary quality, increase the abundance of the genus Lachnospira, and thereby reduce mental and physical stress responses.

[0084] [Table 4]

[0085] [Table 5]

[0086] To investigate how specific bean and vegetable intakes affect NRF9.3 scores, we aggregated the intakes of four foods included in the bean category (soybeans and soybean products, tofu, fried tofu, and natto) and nine foods included in the vegetable category (tomatoes, carrots, spinach, bell peppers, cabbage, cucumbers, daikon radishes, onions, and Chinese cabbage) by cluster in the FFQ and examined which foods differed in intake between clusters. Figure 5 shows the results for all 13 foods aggregated by cluster. Significant differences were observed between clusters for natto, tomatoes, and bell peppers, with Cluster 2 having significantly lower intakes for each of these foods compared to the other clusters. This suggests that Cluster 2's lower intake of natto, tomatoes, and bell peppers may have contributed to the lower NRF9.3 scores. Furthermore, we created a structural model for specific foods, such as natto, tomatoes, and bell peppers, based on the structural model created for beans and vegetables. The constraints used in this study are shown in Table 6, and the structural model created through causal analysis is shown in Figure 6. As a result, the causal relationship "NRF9.3 → Lachnospira → mental and physical stress response score" was observed, as before, with foods such as natto, tomatoes, and green peppers appearing upstream of NRF9.3. Table 7 shows the results of parameter estimation using regression analysis of the relationships between each link. The coefficient representing the relationship between "NRF9.3 → Lachnospira" was 0.1466, and the coefficient representing the relationship between "Lachnospira → mental and physical stress response score" was -0.0841. Furthermore, the coefficients representing the relationships between "natto → NRF9.3" and "tomato → NRF9.3" and "green pepper → NRF9.3" were 0.2446, 0.1892, and 0.1230, respectively. These findings suggest that, among legumes and vegetables, consuming natto, tomatoes, and bell peppers in particular may improve dietary quality and increase the number of Lachnospira species, potentially reducing physical and mental stress responses.

[0087] [Table 6]

[0088] [Table 7] [Industrial Applicability]

[0089] The foods and food compositions of the present invention are expected to be useful for increasing the proportion of Lachnospira in intestinal bacteria to ameliorate stress conditions.Furthermore, the evaluation methods, kits, and programs of the present invention are expected to be useful for evaluating whether the intake of processed soybean foods or solanaceous vegetables is suitable for ameliorating stress conditions.

Claims

1. A food composition containing a processed soybean food and / or a food composition containing a soybean vegetable for enhancing the proportion of Lachnospira in intestinal bacteria.

2. 2. The food or food composition according to claim 1, wherein the soybean processed food is a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, and natto.

3. 2. The food or food composition of claim 1, wherein the solanaceous vegetable is a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato.

4. A method for evaluating whether intake of a processed soybean food or a solanaceous vegetable is suitable for improving a stress state in a subject or a subject animal, comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and (c) determining, based on the comparison, that intake of processed soybean foods or Solanaceae vegetables is suitable for the subject or the target animal when the proportion of Lachnospira in the subject or the target animal is relatively low. A method comprising:

5. 5. The method according to claim 4, wherein the soybean processed food is a food selected from the group consisting of roasted soybeans, soybean flour, soy milk, yuba, tofu, fried tofu, miso, and natto.

6. 5. The method of claim 4, wherein the solanaceous vegetable is a food selected from the group consisting of eggplant, tomato, bell pepper, shishito pepper, paprika, chili pepper, and potato.

7. A program that outputs an evaluation of whether intake of processed soybean foods or nightshade vegetables is suitable for improving a stress state of a subject or a subject animal, the program comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; A program that outputs an evaluation that the intake of soybean processed foods or Solanaceae vegetables is suitable for the subject or target animal when the proportion of Lachnospira genus in the subject or target animal is relatively low, based on the data obtained by the above.

8. A program for transmitting to a client an evaluation of whether intake of processed soybean foods or nightshade vegetables is suitable for improving a stress state of a subject or a target animal, the program being executed on a server and comprising the following steps: (a) analyzing the bacterial composition of an intestinal microbiota sample derived from the subject or subject animal; and (b) comparing the proportion of Lachnospira in the intestinal bacteria of the subject or subject animal with the proportion of the bacteria in an arbitrary population based on the analysis results; and transmits to a client via a communication network an assessment that the intake of processed soybean foods or Solanaceae vegetables is suitable for the subject or target animal if the proportion of Lachnospira genus in the subject or target animal is relatively low, based on the data obtained by the method.

9. A kit for evaluating whether the intake of processed soybean foods or nightshade vegetables is suitable for improving the stress state of a subject or a subject animal, comprising a detection reagent for detecting the genus Lachnospira.

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