Compositions, products comprising the same and uses thereof

Abstract

The present invention relates to a composition, products comprising the same and uses thereof. The composition comprises Bifidobacterium animalis lactis HN019 and medium long chain triglycerides (MLCT), wherein the number of viable cells of the Bifidobacterium animalis lactis HN019 is 1.44 x 10 7 to 5.77 x 10 9 CFU per gram (g) of the MLCT. The composition has a synergistic effect in improving intestinal colic and / or abdominal distension.

Description

Technical Field

[0001] This invention relates to compositions, products comprising the same, and their uses, particularly to the use of the compositions in the preparation of products for improving intestinal colic and / or bloating. Background Technology

[0002] Colic and bloating are common functional gastrointestinal symptoms with a high incidence in both infants and adults. Infantile colic is mainly characterized by paroxysmal, persistent crying without obvious cause, often accompanied by abdominal distension, increased flatulence, and other signs of discomfort, with an incidence of approximately 5%–25%. It may be related to factors such as immature gastrointestinal development, food intolerance, food allergies, or intestinal gas accumulation, but currently there is no clear cause or universally accepted effective prevention or treatment method, causing significant distress to infants and their caregivers.

[0003] In adults, symptoms such as bloating and flatulence are also common and closely related to lifestyle, psychological stress, and functional gastrointestinal disorders. Related studies show that more than half of the population has experienced varying degrees of gastrointestinal discomfort, with bloating being one of the most common symptoms, suggesting that such problems have a wide range of health impacts.

[0004] Currently, interventions for colic and bloating mainly include behavioral adjustments, medication, or medical nutrition support. However, caution is needed when administering medication to infants and young children, and long-term medication use in adults may have side effects or compliance issues. Existing nutritional intervention programs are mostly compound formulas, and their mechanisms of action are not yet clear, leaving room for improvement.

[0005] Intervening in gastrointestinal function through food intake has advantages such as high safety, direct action, and ease of long-term adherence, making it especially suitable for infants and young children and people with limited drug tolerance.

[0006] Medium and long-chain triacylglycerols (MLCTs) have attracted widespread attention as a novel type of structured lipid. Patent CN121795626A discloses a composition containing a specific ratio of partially hydrolyzed protein and medium and long-chain triacylglycerols (MLCTs), which has a synergistic effect in improving intestinal colic and / or bloating.

[0007] Patent CN111227044B discloses a dairy product composed of whole milk powder, skim milk powder, lactose, whey protein powder, demineralized whey powder, α-lactalbumin powder, OPO, vegetable oil, prebiotics, vitamins, and bifidobacteria, which is effective in preventing, relieving, or treating abdominal discomfort symptoms in infants and young children (such as colic). Currently, there are no literature reports on whether Bifidobacterium lactis HN019 alone has the effect of improving colic.

[0008] No literature or patents, either domestically or internationally, have disclosed compositions containing both MLCT and HN019, or their effects on improving intestinal colic and bloating. Researching the effects of MLCT, HN019, and their combinations is of great significance for developing products and / or methods to improve intestinal colic and / or bloating through dietary intake.

[0009] There is still a need in the field for methods that can address gastrointestinal functional discomfort symptoms such as intestinal colic and / or bloating. Summary of the Invention

[0010] To address the above and other technical issues, the inventors conducted in-depth research and discovered that Bifidobacterium animalis subsp. lactis HN019 and medium-long chain triglycerides (MLCT) have a synergistic effect in improving gastrointestinal functional discomfort symptoms such as colic and / or bloating.

[0011] In a first aspect, the present invention provides a composition comprising Bifidobacterium animalis subsp. lactis HN019 and medium- and long-chain triglycerides (MLCT), wherein the viable count of Bifidobacterium animalis subsp. lactis HN019 is 1.44 x 10⁻⁶ per gram (g) of the MLCT. 7 CFU up to 5.77x10 9 CFU.

[0012] In some preferred embodiments, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 1.44 x 10⁻⁶ per g of the MLCT. 7 CFU up to 2.31x10 9 CFU. In some preferred embodiments, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 5.77 x 10⁻⁶ per g of MLCT. 7 CFU up to 2.31x10 9 CFU. In some preferred embodiments, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 5.77 x 10⁻⁶ per g of MLCT. 7 CFU up to 5.77x10 8 CFU.

[0013] In some embodiments, the composition is used to improve intestinal colic and / or bloating.

[0014] In the second aspect, a product is provided which comprises the composition according to the first aspect.

[0015] In some implementations, the product is used to improve intestinal colic and / or bloating.

[0016] In some embodiments, the content of medium- and long-chain triglycerides (MLCT) is from 0.1% to 30.0% by dry weight, based on the total dry weight of the product.

[0017] In a third aspect, the use of the composition according to the first aspect in the preparation of a product for improving intestinal colic and / or bloating is provided.

[0018] In the fourth aspect, the composition according to the first aspect is provided for use for non-therapeutic purposes in improving intestinal colic and / or bloating.

[0019] The beneficial effects of this invention include: when medium- and long-chain triglycerides (MLCT) are used in combination with Bifidobacterium animalis subsp. lactis HN019, the two can synergistically regulate gastrointestinal function, thereby improving gastrointestinal functional discomfort symptoms such as intestinal colic and / or bloating. For example, based on each gram (g) of the MLCT, the viable count of Bifidobacterium animalis subsp. lactis HN019 is 1.44 x 10⁻⁶. 7 CFU up to 5.77x10 9 CFU. Attached Figure Description

[0020] Figure 1 This is a statistical graph showing the total movement distance of zebrafish after treatment with the composition of this application. Note: Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001; compared with Comparative Example 1, !!p < 0.01, !!!p < 0.001; compared with Comparative Example 2, ###p < 0.001; compared with Comparative Example 3, @p < 0.05, @@p < 0.01; compared with Comparative Example 4, $p < 0.05, $$p < 0.01; compared with Comparative Example 5, &p < 0.05, &&p < 0.01; compared with Comparative Example 6, ++p < 0.01, +++p < 0.001.

[0021] Figure 2 The image shows typical behavior of zebrafish after treatment with the composition of this application, where the black line represents the distance of slow movement, the green line represents the distance of medium movement, and the red line represents the distance of fast movement.

[0022] Figure 3This is a statistical graph showing the intestinal lumen area of ​​zebrafish after treatment with the composition of this application. Note: Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001; compared with Comparative Example 1, !p < 0.05, !!!p < 0.001; compared with Comparative Example 2, ###p < 0.001; compared with Comparative Example 3, @p < 0.05, @@@p < 0.001; compared with Comparative Example 4, $p < 0.05, $$p < 0.01, $$$p < 0.001; compared with Comparative Example 5, &p < 0.05, &&p < 0.01; compared with Comparative Example 6, +p < 0.05, ++p < 0.01.

[0023] Figure 4 A graph showing the measurement of the intestinal lumen area of ​​zebrafish after treatment with the composition according to this application. Detailed Implementation

[0024] The following definitions are provided to enable those skilled in the art to understand the invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Preferred materials and methods are described herein, but any methods and materials similar to or equivalent to those described herein may be used in the practice of testing the invention. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0025] definition

[0026] As used herein, the term "medium-long chain triglyceride (MLCT)" refers to a class of triglycerides that contain both medium-chain and long-chain fatty acid residues in their molecular structure. MLCTs possess different physicochemical properties, metabolic characteristics, and nutritional value from long-chain triglycerides (LCT) or medium-chain triglycerides (MCT). Studies (see, for example, Yuan Tinglan, Composition and Metabolic Characteristics of Medium- and Long-Chain Triglycerides in Breast Milk Fat [D], Jiangnan University, 2021) show that medium- and long-chain triglycerides (MLCT) are not simply equivalent to a physical mixture of long-chain triglycerides (LCT) and medium-chain triglycerides (MCT) (nor are they simply equivalent to a mixture of long-chain and medium-chain fatty acids), and the latter does not possess the physicochemical properties, metabolic characteristics, and nutritional value of the former.

[0027] According to the relevant provisions of the "Administrative Measures for New Resource Foods", MLCT is made from edible vegetable oil and medium-chain triglycerides as raw materials, through lipase transesterification reaction, and then through processes such as distillation separation, decolorization, and deodorization.

[0028] This invention does not have any particular requirements for the MLCT used; any MLCT commonly used in the art can be used. MLCT can be used in pure form or in an impure form rich in MLCT. The MLCT component contained in the composition can be a single-component MLCT (i.e., each molecular chain has the same long-chain fatty acid residues and the same medium-chain fatty acid residues) or a mixture of two or more different-component MLCTs (i.e., each molecular chain has different long-chain fatty acid residues and / or different medium-chain fatty acid residues).

[0029] In some embodiments, the MLCT used in this invention comprises C6-C 12 (e.g., C6, C7, C8, C9, C) 10 C 11 C 12 (or the range defined by either or both) fatty acid residues and C 14 -C 30 (e.g., C) 14 C 15 C 16 C 17 C 18 C 19 C 20 C 21 C 22 C 23 C 24 C 25 C 26 C 27 C 28 C 29 C 30 (or the range defined by either or both) fatty acid residues. The C6-C 12 The fatty acid residues may be derived from one or more of the following: hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, etc., preferably hexanoic acid, octanoic acid, decanoic acid, and lauric acid. The C 14 -C 30 The fatty acid residues may be derived from one or more of the following: myristic acid, palmitic acid, heptadecanic acid, oleic acid, linoleic acid, stearic acid, nonadecanic acid, eicosapentaenoic acid, docosahexaenoic acid, docosapatraenoic acid, docosahexaenoic acid, tricarboxylic acid, docosapetraenoic acid, docosapatraenoic acid, docosahexaenoic acid, arachidonic acid, etc., preferably myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapatraenoic acid and docosahexaenoic acid.

[0030] In some embodiments, the C6-C in the MLCT 12 Fatty acid residues on the C14 -C 30 The weight ratio of fatty acid residues is from 0.124 to 2.000. As an example, the C6-C... 12 Fatty acid residues on the C 14 -C 30 The weight ratio of fatty acid residues may be 0.124, 0.125, 0.130, 0.140, 0.150, 0.160, 0.170, 0.180, 0.190, 0.200, 0.300, 0.400, 0.500, 0.600, 0.700, 0.800, 0.900, 1.000, 1.100, 1.200, 1.300, 1.400, 1.500, 1.600, 1.700, 1.800, 1.900, 2.000, or within the range defined by any two of these.

[0031] As used herein, the terms "Bifidobacterium animalis subsp. Lactis HN019", "Bifidobacterium lactis HN019", "Bifidobacterium HN019", or "HN019" are used interchangeably. HN019 is a Gram-positive, anaerobic, non-spore-forming bacterium of the genus *Bifidobacterium*, originally isolated from yogurt.

[0032] As used herein, the phrase “the number of viable Bifidobacterium animalis subsp. lactis HN019 per gram (g) of MLCT” refers to the ratio of the number of viable Bifidobacterium animalis subsp. lactis HN019 (colony forming units, CFU) to the mass (g) of medium- and long-chain triglycerides (MLCT). In this document, this ratio may also be expressed as the HN019 / MLCT ratio in CFU / g.

[0033] As used herein, the terms “individual” or “subject” refer to any animal, such as a mammal, preferably a human.

[0034] As used in this article, the term “infant” refers to a person aged from birth to 12 months; the term “toddler” refers to a person aged 1 to 3 years; the term “child” refers to a person aged 3 to 7 years; the term “teenager” refers to a person aged 7 to 17 years; the term “adult” refers to a person aged 18 years or older; the term “young adult” refers to a person aged 18 to 40 years; the term “middle-aged” refers to a person aged 41 to 65 years; and the term “elderly” or “senior” refers to a person aged 65 years or older.

[0035] Composition

[0036] A first aspect of the present invention provides a composition comprising Bifidobacterium animalis subsp. lactis HN019 and medium- and long-chain triglycerides (MLCT). The inventors have surprisingly discovered that when HN019 and MLCT are used in combination at a defined dosage ratio, there is a synergistic effect between them, which can synergistically improve intestinal colic and bloating.

[0037] In some embodiments of the composition described in this application, the viable count of Bifidobacterium animalis subsp. lactis HN019 is 1 x 10⁻⁶ per gram (g) of the MLCT. 7 CFU up to 6x10 9 CFU, for example 1.44x10 7 CFU up to 5.77x10 9 CFU. In some embodiments of the compositions described in this application, the viable count of the *Bifidobacterium animalis* subsp. *lactamase* HN019 is 1 x 10⁻⁶ per gram (g) of the MLCT. 7 CFU, 2x10 7 CFU, 4x10 7 CFU, 6x10 7 CFU, 8x10 7 CFU, 1x10 8 CFU, 2x10 8 CFU, 4x10 8 CFU, 6x10 8 CFU, 8x10 8 CFU, 1x10 9 CFU, 2x10 9 CFU, 4x10 9 CFU or 6x10 9 CFU, or within the range defined by any two of the above. In some example embodiments of the compositions described in this application, the viable count of Bifidobacterium animalis subsp. lactis HN019 is 1.44 x 10⁻⁶ per gram (g) of the MLCT. 7 CFU, 5.77x10 7 CFU, 1.44x10 8 CFU, 2.31x10 8 CFU, 5.77x10 8 CFU, 1.44x10 9 CFU, 2.31x10 9 CFU or 5.77x10 9 CFU, or within the range defined by either of the above.

[0038] In some preferred embodiments, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 1.44 x 10⁻⁶ per gram (g) of the MLCT. 7 CFU up to 2.31x10 9 Within the range of CFU, the composition can exhibit a more significant synergistic effect, particularly in synergistically improving intestinal colic and / or bloating.

[0039] In some preferred embodiments, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 5.77 x 10⁻⁶ per gram (g) of the MLCT. 7 CFU up to 2.31x10 9 Within the range of CFU, the composition can exhibit a more significant synergistic effect, particularly in synergistically improving intestinal colic and / or bloating.

[0040] In some further preferred embodiments, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 5.77 x 10⁻⁶ per gram (g) of the MLCT. 7 CFU up to 5.77x10 8 Within the range of CFU, the composition can exhibit a more significant synergistic effect, particularly in synergistically improving intestinal colic and / or bloating.

[0041] In some embodiments, the composition is used to improve intestinal colic. In some embodiments, the composition is used to improve bloating. In some embodiments, the composition is used to improve both intestinal colic and bloating.

[0042] product

[0043] A second aspect of the invention provides a product comprising the composition according to the first aspect described above.

[0044] All descriptions of the composition above apply here and will not be repeated here.

[0045] In some embodiments, the composition according to the first aspect of this application may be added to a product as a beneficial ingredient.

[0046] In some embodiments, the product may include a drug.

[0047] In some embodiments, the dosage form of the drug is selected from powders, tablets, and liquids. This invention does not place particular requirements on the dosage form of the drug, and commonly used drug dosage forms in the art can be used. As examples, the drug of this invention can be a solid dosage form, such as powders, granules, tablets, capsules, drop pills, pills, etc.; a semi-solid dosage form, such as ointments, creams, eye ointments, gels, etc.; or a liquid dosage form, such as oral liquid preparations, such as syrups, mixtures, suspensions, emulsions, etc. Those skilled in the art will readily understand that, depending on the dosage form and application scenario, the drug may also include various pharmaceutically permissible excipients. Furthermore, the drug of this invention can be prepared using methods commonly employed in the art, which will not be elaborated upon here.

[0048] As an example, based on the weight of the product, the content of medium- and long-chain fatty acid triglycerides may be 0.1, 0.2, 0.5, 0.8, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, 12.0, 13.0, 14.0, 15.0, 16.0, 17.0, 18.0, 19.0, 20.0, 21.0, 22.0, 23.0, 24.0, 25.0, 26.0, 27.0, 28.0, 29.0, or 30.0% by dry basis, or within the range defined by any two of the above.

[0049] The product can be prepared by methods commonly used in the art, which will not be described in detail here.

[0050] use

[0051] The present invention also relates to the use of the composition of the first aspect of the present invention.

[0052] A third aspect of the invention provides the use of the composition according to the first aspect in the preparation of a product for improving intestinal colic and / or bloating.

[0053] A fourth aspect of the invention provides the use of the composition according to the first aspect for non-therapeutic purposes of improving intestinal colic and / or bloating.

[0054] All descriptions of the compositions and products described in this application above are applicable here and will not be repeated here.

[0055] In some embodiments, a method for improving intestinal colic and / or bloating in a subject is provided, comprising administering to the subject a composition according to the first aspect or a product according to the second aspect.

[0056] In some embodiments, the subject for improving intestinal colic and / or abdominal distension is a human. In some embodiments, the individual / subject for improving intestinal colic and / or abdominal distension is not particularly limited and may include, for example, infants, toddlers, children, juveniles, adults, young people, middle-aged people and / or elderly people.

[0057] Examples

[0058] The present invention will be more readily understood with reference to the following examples, which are only intended to illustrate some aspects and embodiments of the present invention, and are not intended to limit the present invention.

[0059] Unless otherwise stated, the materials used in the examples are all commercially available materials or conventional materials.

[0060] laboratory animals

[0061] All zebrafish are raised in fish culture water at 28°C (water quality: 200 mg of instant sea salt is added to every 1 L of reverse osmosis water, conductivity is 450~550 μS / cm; pH is 6.5~8.5; hardness is 50~100 mg / L CaCO3), and are bred and provided by the fish culture center of Quantao Biotechnology Co., Ltd. The license number for the use of experimental animals is: SYXK (Zhejiang) 2022-0004, the feeding management meets the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethical review number is: IACUC-2025-202510290009-01.

[0062] Instruments, consumables and reagents

[0063] Dissecting microscope (SZX7, OLYMPUS, Japan);

[0064] Zebrafish behavior analysis system (Zebra Lab 3.22.3.31, Viewpoint, France);

[0065] 150 mm*25 mm petri dish (Orange Scientific, Belgium);

[0066] 6-well plate (Zhejiang Bellan Biotechnology Co., Ltd., China);

[0067] 96-well plate (Nest Biotech, China);

[0068] Trinitrobenzenesulfonic acid (TNBS, batch number SLCN9362, Sigma, USA);

[0069] Dimethyl sulfoxide (DMSO, batch number BCCD8942, Sigma, Switzerland);

[0070] The medium- and long-chain fatty acid triglycerides were derived from medium- and long-chain fatty acid edible oil (production batch number 20250804) supplied by Yihai Kerry, with a purity of 61%.

[0071] HN019 is Bifidobacterium lactis HN019, which is derived from Danisco Nutritional Foods Co., Ltd. (production batch number 7084874495).

[0072] Prednisolone, white powder, batch number C10016501, purchased from Shanghai Maclean Biochemical Technology Co., Ltd., solvent is DMSO.

[0073] Evaluation of the efficacy of the composition in improving intestinal colic

[0074] Wild-type AB strain zebrafish (3 dpf) were randomly selected and placed in culture dishes. Except for the normal control group, all experimental groups were treated with 2,4,6-trinitrobenzenesulfonic acid (TNBS) in water to establish a zebrafish colic model. After treatment at 28℃ for 2 days, the TNBS was removed and the zebrafish were distributed into 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples were administered in water (concentrations shown in Table 1). The positive control group received prednisone at a concentration of 15.0 μg / mL. A normal control group and a model control group were also included, with a volume of 3 mL per well. After another 2 days of treatment at 28℃, 10 zebrafish were randomly selected from each group and placed in a behavioral analyzer for 1 h. The total movement distance of the zebrafish was analyzed, and the statistical analysis results of this index were used to evaluate the efficacy of the samples in improving colic. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS software; p < 0.05 indicated statistical significance.

[0075] Then, for each embodiment and comparative example, the increase in total movement distance (mm) compared to the model control group was calculated (denoted as A), where A = total movement distance of the embodiment or comparative example minus the total movement distance of the model control group. For each embodiment and comparative example 7, the sum of the increase in total movement distance (mm) of the corresponding comparative example compared to the model control group was calculated (denoted as B), where B = the sum of A for the single-component comparative examples corresponding to embodiment or comparative example 7. For example, for embodiment 1 (MLCT concentration 39 μg / mL + HN019 concentration 9000 CFU / mL), the "sum of the increase in total movement distance (B) of the corresponding comparative examples compared to the model control group" is the sum of the "increase in total movement distance compared to the model control group" of comparative example 1 (MLCT concentration 39 μg / mL) and comparative example 4 (HN019 concentration 9000 CFU / mL). Finally, for each embodiment and comparative example 7, the synergy coefficient (C = A / B) was calculated. When the synergy coefficient > 1, a synergistic effect exists; the larger the value, the stronger the synergistic effect.

[0076] Following the above method, zebrafish were administered HN019 and MLCT at different ratios to evaluate their efficacy in improving intestinal colic. The dosages are shown in Table 1. The results are also shown in Table 1. Figure 1 and Figure 2 As shown.

[0077] Table 1. Experimental results evaluating the efficacy of the samples in improving intestinal colic.

[0078]

[0079] Note: Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001;

[0080] Compared with Comparative Example 1, !!p < 0.01, !!!p < 0.001;

[0081] Compared with Comparative Example 2, p < 0.001;

[0082] Compared with Comparative Example 3, @p < 0.05, @@p < 0.01;

[0083] Compared with Comparative Example 4, $p < 0.05, $$p < 0.01;

[0084] Compared with Comparative Example 5, &p < 0.05, &&p < 0.01;

[0085] Compared with Comparative Example 6, ++p < 0.01, +++p < 0.001.

[0086] From Table 1, Figure 1 and Figure 2 The results showed that, compared with the normal control group, the total movement distance of wild-type AB strain zebrafish in the model control group induced by TNBS water-soluble administration was significantly reduced (P < 0.05), indicating that the TNBS-induced intestinal colic model was successfully established. The positive control drug (prednisone), single-component MLCT (625 μg / mL), single-component HN019 (9000 CFU / mL, 90000 CFU / mL, and 900000 CFU / mL), Examples 1-8, and Comparative Example 7 all showed a significantly increased total movement distance compared with the model control group (P < 0.05), indicating a significant improvement in intestinal colic symptoms.

[0087] Furthermore, compared to the single-component comparative examples using MLCT or HN019 alone, Examples 1-8 exhibited a significant synergistic effect when used in combination. For example, as shown in Table 1, Example 1 (HN019 / MLCT ratio was 2.31 x 10⁻⁶) showed a significant synergistic effect. 8The CFU / g ratio was equivalent to a combination of Comparative Example 1 (single-component MLCT, concentration 39 µg / mL) and Comparative Example 4 (single-component HN019, concentration 9000 CFU / mL). Compared with the model control group, the increase in total movement distance in Example 1 was 1373 mm, while the increases in total movement distance in Comparative Examples 1 and 4 were 211 mm and 696 mm, respectively. The former was greater than the sum of the latter two (907 mm), with a synergy coefficient of 1.51, indicating a synergistic effect of Example 1 in improving intestinal colic. Similarly, Examples 2-8 also showed a synergistic effect in improving intestinal colic, while Comparative Example 7 did not show a synergistic effect in improving intestinal colic. These results indicate that the HN019 / MLCT ratio of 1.44 x 10⁻⁶ is suitable for this condition. 7 CFU / g up to 5.77x10 9 At CFU / g, the two have a synergistic effect in improving intestinal colic (synergistic coefficient > 1).

[0088] Furthermore, in Examples 3 and 4, the HN019 / MLCT dosage ratio was 5.77 x 10⁻⁶. 7 CFU / g and 5.77x10 8 The synergy coefficient (CFU / g) was higher than that of other embodiments (synergy coefficient > 1.7), indicating that the two had a more significant synergistic effect in improving intestinal colic at this dosage ratio.

[0089] Evaluation of the efficacy of the composition in improving bloating

[0090] Wild-type AB strain zebrafish (3 dpf) were randomly selected and placed in culture dishes. Except for the normal control group, all experimental groups were treated with TNBS dissolved in water. After treatment at 28℃ for 2 days, the TNBS was removed and the zebrafish were distributed into 6-well plates, with 30 zebrafish treated in each well (experimental group). Water-dissolved samples (concentrations shown in Table 2) were used as examples / comparative examples, and prednisone 15.0 μg / mL was used as a positive control. A normal control group and a model control group were also set up, with a volume of 3 mL per well. After further treatment at 28℃ for 2 days, 10 zebrafish from each group were randomly selected, photographed under a dissecting microscope, and the images were saved. Data were collected using NIS-Elements D 3.20 advanced image processing software, and the intestinal lumen area of ​​the zebrafish was analyzed. The statistical analysis results of this index were used to evaluate the efficacy of the samples in improving abdominal distension in zebrafish. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS software; p < 0.05 indicated statistical significance.

[0091] Then, the reduction in intestinal lumen area (pixels) compared to the model control group (denoted as A) is calculated, where A = intestinal lumen area of ​​the embodiment or comparative example minus intestinal lumen area of ​​the model control group; and for each embodiment and comparative example 7, the sum of the reduction in intestinal lumen area (pixels) of the corresponding comparative example compared to the model control group is calculated (denoted as B), where B = the sum of A of the single-component comparative examples corresponding to embodiment or comparative example 7; subsequently, the synergy coefficient (C) is calculated as described above.

[0092] Following the above method, zebrafish were administered HN019 and MLCT at different ratios to evaluate their efficacy in improving abdominal distension. The dosages are shown in Table 2. The results are also shown in Table 2. Figure 3 and Figure 4 As shown.

[0093] Table 2. Experimental results evaluating the efficacy of the samples in improving abdominal distension.

[0094]

[0095] Note: Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001;

[0096] Compared with Comparative Example 1, !p < 0.05;

[0097] Compared with Comparative Example 2, ##p < 0.01, ###p < 0.001;

[0098] Compared with Comparative Example 3, p < 0.001;

[0099] Compared with Comparative Example 4, $p < 0.05, $$p < 0.01;

[0100] Compared with Comparative Example 5, p < 0.01;

[0101] Compared with Comparative Example 6, +p < 0.05.

[0102] From Table 2, Figure 3 and Figure 4 The results showed that, compared with the normal control group, the intestinal lumen area of ​​wild-type AB strain zebrafish induced by TNBS water-soluble administration at 3 dpf was significantly increased (P < 0.05), indicating successful modeling. The positive control drug (prednisone), single-component MLCT (625 μg / mL), single-component HN019 (9000 CFU / mL, 90000 CFU / mL, and 900000 CFU / mL), Examples 1-6, and Comparative Example 7 all showed significantly reduced intestinal lumen area (pixels) compared with the model control (P < 0.05), indicating a significant improvement in abdominal distension symptoms.

[0103] Furthermore, compared to the single-component comparative examples using MLCT or HN019 alone, Examples 1-6 exhibited a significant synergistic effect when used in combination. For example, as shown in Table 2, Example 1 (HN019 / MLCT ratio was 2.31 x 10⁻⁶) showed a significant synergistic effect. 8 The CFU / g ratio corresponds to a combination of Comparative Example 1 (single-component MLCT, concentration 39 µg / mL) and Comparative Example 4 (single-component HN019, concentration 9000 CFU / mL). Compared to the model control group, Example 1 showed a reduction of 9995 pixels in intestinal lumen area (pixels), while Comparative Examples 1 and 4 showed reductions of 2844 pixels and 5519 pixels, respectively. The former was greater than the sum of the latter two (8363 pixels), with a synergy coefficient of 1.2, indicating a synergistic effect of Example 1 in improving abdominal distension. Similarly, Examples 2-6 also showed a synergistic effect in improving abdominal distension, while Comparative Example 7 did not show a synergistic effect. These results indicate that the HN019 / MLCT ratio of 1.44 x 10⁻⁶ is suitable for this purpose. 7 CFU / g up to 5.77x10 9 At CFU / g, the two have a synergistic effect in improving abdominal distension (synergistic coefficient > 1).

[0104] Furthermore, Examples 1-4 and 6 (HN019 / MLCT ratio is 1.44 x 10⁻⁶) 7 CFU / g up to 2.31x10 9 Compared to Example 5, the HN019 / MLCT ratio (CFU / g) has a higher synergistic coefficient (synergistic coefficient > 1.05), indicating that the synergistic effect of the two in improving abdominal distension is more significant at this dosage ratio. Specifically, the preferred HN019 / MLCT dosage ratio is 1.44 x 10⁻⁶. 7 CFU / g up to 2.31x10 9 At CFU / g, the synergistic effect of the two is more significant, especially in synergistically improving abdominal distension.

[0105] Furthermore, in Examples 1-4 (HN019 / MLCT ratio was 5.77 x 10⁻⁶), the HN019 / MLCT dosage ratio was 5.77 x 10⁻⁶. 7 CFU / g up to 2.31x10 9 Compared to Examples 5 and 6, the HN019 / MLCT ratio (CFU / g) exhibits a higher synergistic coefficient (synergistic coefficient > 1.06), indicating a more significant synergistic effect in improving abdominal distension at this dosage ratio. More preferably, the HN019 / MLCT dosage ratio is 5.77 x 10⁻⁶. 7 CFU / g up to 2.31x10 9 Within the range of CFU / g, the synergistic effect is more significant, especially in synergistically improving bloating.

[0106] Furthermore, in Examples 1, 3, and 4 (HN019 / MLCT ratio of 5.77 x 10⁻⁶), the... 7 CFU / g up to 5.77x10 8 The HN019 / MLCT ratio (CFU / g) exhibits a higher synergistic coefficient (synergistic coefficient > 1.09) compared to other embodiments, indicating a more significant synergistic effect in improving abdominal distension at this dosage ratio. More preferably, the HN019 / MLCT dosage ratio is 5.77 x 10⁻⁶. 7 CFU / g up to 5.77x10 8 Within the range of CFU / g, the synergistic effect is more significant, especially in synergistically improving bloating.

[0107] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition, characterized in that, The composition comprises *Bifidobacterium animalis* subsp. *lactamase* HN019 and medium- and long-chain triglycerides (MLCT), wherein the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 1.44 x 10⁻⁶ per g of MLCT. 7 CFU up to 5.77x10 9 CFU.

2. The composition according to claim 1, characterized in that, Based on the MLCT per gram, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 1.44 x 10⁻⁶. 7 CFU up to 2.31x10 9 CFU.

3. The composition according to claim 1, characterized in that, Based on the MLCT per gram, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 5.77 x 10⁻⁶. 7 CFU up to 2.31x10 9 CFU.

4. The composition according to claim 1, characterized in that, Based on the MLCT per gram, the viable count of *Bifidobacterium animalis* subsp. *lactamase* HN019 is 5.77 x 10⁻⁶. 7 CFU up to 5.77x10 8 CFU.

5. The composition according to any one of claims 1-4, characterized in that, The composition is used to improve intestinal colic and / or bloating.

6. The product, characterized in that, The product comprises the composition according to any one of claims 1-4.

7. The product according to claim 6, characterized in that, The product is used to improve intestinal colic and / or bloating.

8. Use of the composition according to any one of claims 1-5 in the preparation of a product for improving intestinal colic and / or bloating.

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