Nutritional composition and application thereof

Abstract

The invention provides a nutritional composition and application of the nutritional composition. The nutritional composition comprises medium-chain and long-chain fatty acid triglyceride (MLCT) and milk fat globule membrane (MFGM) in a mass ratio of 2: 1 to 80: 1, and the combination of the two has the effects of synergistically improving diarrhea, inhibiting staphylococcus aureus in vivo or in vitro and / or regulating intestinal flora.

Description

Technical Field

[0001] The present disclosure relates to the field of food technology, and in particular to a nutritional composition for inhibiting Staphylococcus aureus and / or improving diarrhea, a food containing the nutritional composition, and a use of the nutritional composition. Background Art

[0002] Infectious diarrhea is highly prevalent and has a high incidence rate. The medical burden and economic burden caused by diarrheal diseases in developing countries are serious, so it is still a global public health problem worthy of attention. The World Health Organization (WHO) estimates that tens of millions of people suffer from diarrheal diseases every day in the world, and 1.7 billion cases of diarrheal diseases occur each year, of which 2.2 million patients die from severe diarrhea (Miao Xiaohui et al., Chinese Journal of Digestion, 2013, 33 (12): 793-802.). In addition, infectious diarrheal diseases have a high incidence rate and are widely prevalent in children, which seriously endangers children's health and is also the main cause of malnutrition in children (Ni Xin et al., Chinese Medical Science, 2020, 10 (21): 249-256). Infectious diarrhea is an intestinal infectious disease with diarrhea as the main clinical manifestation caused by bacteria, fungi, viruses or parasites.

[0003] Staphylococcus aureus is a zoonotic Gram-positive bacterium that is detected in 30% to 50% of healthy adults, and about 20% of infected people can continue to carry the bacteria; it can cause chronic infections in cattle, sheep, pigs, chickens and other animals (Wu Yunpu et al., Laboratory Animal Science, 2024, 41(03):85-89). The severity of diseases caused by Staphylococcus aureus varies, from moderate infections such as skin infections to fatal diseases such as severe pneumonia and sepsis. The treatment process of the disease can be complicated by antibiotic resistance, and there is no effective vaccine to date. It is one of the most common causes of morbidity and mortality caused by infectious pathogens worldwide. Chronic infections and antibiotic resistance caused by Staphylococcus aureus pose serious challenges to the public health field. In addition, Staphylococcus aureus is also an important pathogen that induces infectious diarrhea.

[0004] Medium and long-chain triacylglycerols (MLCT) have attracted widespread attention as a star product among new structural lipids. The main structural feature of MLCT is that medium-chain fatty acids and long-chain fatty acids are simultaneously combined on a glycerol backbone. Early studies have shown that MLCT has many health functions such as lowering blood lipids, inhibiting obesity, reducing diabetes and cardiovascular diseases, and reducing the risk of cancer (Lai Yundong et al., Journal of Oil Crops, 2024, 46(04):719-727.). In addition, the fatty acid composition of MLCT is closer to breast milk, which is conducive to fat digestion and absorption, improves the absorption of lipid nutrients, inhibits the accumulation of body fat, and can quickly and stably supply energy (Cheng X, Jiang C, Jin J, Jin Q, Akoh CC, Wei W, Wang X. Medium- and Long-Chain Triacylglycerol: Preparation, Health Benefits, and Food Utilization. Annu Rev Food SciTechnol. 2024 Jun;15(1):381-408.). It is currently known that milk fat globule membrane (MFGM) plays an important role in improving cognitive ability, improving metabolism, and reducing the incidence of infectious diseases (Zhang Bo et al., Chinese Journal of Child Health, 2016, 24(01):43-47).

[0005] There have been no reports at home or abroad on the effects of MLCT and MFGM on Staphylococcus aureus infection and diarrhea. Studying the effects of MLCT, MFGM and their combination is of great significance for developing foods and drugs with anti-infective diarrhea effects.

[0006] There is still a need in the art for methods that can improve diarrhea, inhibit Staphylococcus aureus in vivo or in vitro, and / or regulate intestinal flora. Summary of the invention

[0007] The present invention has been made in view of the above-mentioned problems in the art.

[0008] A first aspect of the present invention provides a nutritional composition, comprising medium-chain triglycerides and milk fat globule membrane, wherein the mass ratio of the medium-chain triglycerides to the milk fat globule membrane is 2:1 to 80:1.

[0009] The second aspect of the present invention provides a food comprising the nutritional composition according to the first aspect of the present invention.

[0010] The third aspect of the present invention provides use of the nutritional composition according to the first aspect of the present invention in preparing a product for improving (eg, alleviating or treating) diarrhea.

[0011] The fourth aspect of the present invention provides use of the nutritional composition according to the first aspect of the present invention in preparing a product for inhibiting Staphylococcus aureus infection.

[0012] The fifth aspect of the present invention provides use of the nutritional composition according to the first aspect of the present invention in the preparation of a product for regulating intestinal flora (eg, intestinal flora in a subject with diarrhea).

[0013] The sixth aspect of the present invention provides use of the nutritional composition of the first aspect of the present invention for the non-therapeutic purpose of improving diarrhea.

[0014] The seventh aspect of the present invention provides use of the nutritional composition of the first aspect of the present invention for non-therapeutic purposes of inhibiting Staphylococcus aureus infection.

[0015] The present invention finds that when medium-chain fatty acid triglycerides and milk fat globule membrane are used in combination at a defined mass ratio, there is a synergistic effect between the two, which can synergistically improve diarrhea, especially improve (e.g., alleviate or treat) diarrhea caused by Staphylococcus aureus, and inhibit Staphylococcus aureus (in vivo or in vitro). BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The fluorescence intensity of zebrafish intestine after sample treatment is shown. The P value is the significant difference analysis between each example group and the model control group. ** indicates P < 0.01, *** indicates P < 0.001.

[0017] Figure 2 The fluorescence intensity of zebrafish intestine after treatment with the composition sample is shown. The P value is the difference between each example group and the model control group, ** indicates P < 0.01, *** indicates P < 0.001.

[0018] Figure 3 Typical images of the intestine of zebrafish showing sample treatments.

[0019] Figure 4 The fluorescence intensity of Staphylococcus aureus in zebrafish after sample treatment is shown. The P value is the significant difference analysis between each example group and the model control group, * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0020] Figure 5 Typical graph showing the fluorescence intensity of Staphylococcus aureus in zebrafish after sample treatment. DETAILED DESCRIPTION

[0021] The following definitions are provided to facilitate understanding of the present invention by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs. 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 the present invention's tests. It should also be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to be limiting.

[0022] Definition of terms

[0023] Unless otherwise indicated or defined, all terms used have the usual meaning in the art, which will be understood by those skilled in the art. In addition, unless otherwise specified, all methods, steps, techniques and operations not specifically described in detail can and have been performed in a manner known per se, which will be understood by those skilled in the art.

[0024] As used herein, the term "medium-chain triglyceride of fatty acids" or "MLCT" is a special triglyceride in which medium-chain fatty acids and long-chain fatty acids are present on the triglyceride backbone. As used herein, the term "medium-chain triglyceride of fatty acids" or "MLCT" is a special triglyceride in which medium-chain fatty acids and long-chain fatty acids are present on the triglyceride backbone. It is well known in the art that MLCT is made from edible vegetable oil and medium-chain triglycerides, through ester exchange reaction by lipase, and through distillation separation, decolorization, deodorization and other processes. Medium-chain triglycerides of fatty acids can be derived from medium-chain edible oil of fatty acids.

[0025] As used herein, the term "medium-chain fatty acid" refers to a fatty acid having 6-12 carbon atoms in the carbon chain, such as caproic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, etc.; the term "long-chain fatty acid" refers to a fatty acid having more than 14 carbon atoms, generally 14-30 carbon atoms, in the carbon chain, such as myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid, etc. The terms "medium-chain fatty acid triglyceride" and "long-chain fatty acid triglyceride" refer to the esterification reaction products of "medium-chain fatty acid" and "long-chain fatty acid" with glycerol, respectively; the term "medium-long chain fatty acid triglyceride" refers to triglycerides containing both medium-chain fatty acid residues and long-chain fatty acid residues in the molecular structure. The present invention has no special requirements for the medium-long chain fatty acid triglyceride used, and the medium-long chain fatty acid triglyceride commonly used in the art can be used. The medium-chain fatty acid triglycerides can be used in pure form or in an impure form rich in medium-chain fatty acid triglycerides. For example, the medium-chain fatty acid triglycerides used in the present invention can contain C6-C 12 Fatty acid residues and C14 -C 30 Fatty acid residue. C6-C 12 The fatty acid residue 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. 14 -C 30 The fatty acid residue may be derived from one or more of the following: myristic acid, palmitic acid, heptadecanoic acid, oleic acid, linoleic acid, stearic acid, nonadecanoic acid, eicosapentaenoic acid, heneicosanoic acid, docosatetraenoic acid, docosahexaenoic acid, tricosanoic acid, tetracosenoic acid, eicosapentaenoic acid, hexacosahexenoic acid, arachidonic acid, etc. In some embodiments, the C6-C 12 The fatty acid residue is derived from one or more of the following: caproic acid, caprylic acid, capric acid and lauric acid; and / or the C 14 -C 30 The fatty acid residue is derived from one or more of the following: myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid. 12 The fatty acid residues on the C 14 -C 30 The weight ratio of fatty acid residues is 0.124 to 2.000. In certain embodiments, C6-C 12 The fatty acid residues on the C 14 -C 30 The weight ratio of fatty acid residues can 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 any two thereof. In some embodiments, the "medium and long chain fatty acid triglyceride" can be prepared with reference to the method of patent CN115369132A.

[0026] As used herein, the term "milk fat globule membrane" or "MFGM" in English is a complex biological membrane structure present in breast milk. The milk fat globule membrane is a complex three-layer phospholipid protein membrane composed of polar lipids, cholesterol and proteins, etc., which is encapsulated on the surface of milk fat droplets. In principle, there is no particular restriction on the source of the milk fat globule membrane of the present invention. It can usually be obtained by extraction from animal milk or its products. In some preferred embodiments, such animal milk or its products can be cow's milk, sheep's milk, camel's milk, horse's milk or dairy products based on them (such as cheese), etc. More preferably, it can be extracted from cow's milk, including bovine colostrum or bovine milk. The present invention does not specifically limit the method for extracting MFGM from the above-mentioned animal milk or its products. For example, it can be carried out by acidified acid precipitation-centrifugation-isoelectric point enrichment-drying method, or it can also be separated by existing membrane filtration methods to obtain MFGM. In addition, MFGM can also be obtained through commercially available products, including milk fat globule membrane whey protein powder and milk fat globule membrane milk protein powder. Milk fat globule membrane whey protein powder refers to a powdered product containing milk fat globule membrane made from raw milk or whey through separation, concentration, drying and other processes. Milk fat globule membrane milk protein powder refers to a powdered product containing milk fat globule membrane made from raw milk or cream through separation, concentration, drying and other processes. The commercial sources of MFGM in the present invention include Lacprodan® MFGM-10, Lacprodan® PL-20, Cor-Power® SM2, lipid-rich MFGM fraction or buttermilk powder concentrate BPC50, BPC60, G600, PC700, Hilmar Cor-Power® WPC7500MEGM ENRICHED WPC, etc.

[0027] As used herein, "Staphylococcus aureus" is also called "golden Staphylococcus", which belongs to the genus Staphylococcus, is a representative of Gram-positive bacteria, is a common foodborne pathogenic microorganism, and is widely present in the natural environment. Under appropriate conditions, Staphylococcus aureus can produce enterotoxins and cause food poisoning. Enterotoxins are single-chain small molecule proteins with a molecular weight of about 26-29k Da, a relatively low molecular mass, and thermal stability. They damage the human intestine and cause symptoms such as vomiting and diarrhea.

[0028] As used herein, "kit" refers to a group of packaged related components, such as one or more compounds or compositions and one or more related materials, such as solvents, solutions, buffers, instructions or desiccants. The "kit" contains medium-chain fatty acid triglycerides and / or milk fat globule membranes. The "kit" is used to inhibit Staphylococcus aureus in vitro, including inhibiting the growth, proliferation and / or production of metabolites of Staphylococcus aureus. DETAILED DESCRIPTION OF THE INVENTION

[0030] Nutritional composition

[0031] A first aspect of the present invention provides a nutritional composition, comprising medium-chain triglycerides (MLCT) and milk fat globule membrane (MFGM), wherein the mass ratio of the medium-chain triglycerides to the milk fat globule membrane is 2:1 to 80:1.

[0032] The present invention finds that when medium- and long-chain fatty acid triglycerides and milk fat globule membrane are used in combination at a defined mass ratio, there is a synergistic effect between the two, which can synergistically improve diarrhea, especially improve (e.g., alleviate or treat) diarrhea caused by Staphylococcus aureus, and / or inhibit Staphylococcus aureus (in vivo or in vitro).

[0033] In some embodiments, the nutritional composition consists of medium-chain triglycerides (MLCT) and milk fat globule membrane (MFGM).

[0034] In an embodiment, the mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane in the nutritional composition is 2:1 to 80:1, such as 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1 or within the range defined by any two of them. Preferably, the mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane is 2:1 to 40:1, within the range, the synergistic effect is more significant, in particular, it can synergistically improve diarrhea, in particular synergistically improve (e.g., alleviate or treat) diarrhea caused by Staphylococcus aureus, and synergistically (in vivo or in vitro) inhibit Staphylococcus aureus. More preferably, the mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane is 3:1 to 40:1, within which the synergistic effect in improving diarrhea, especially synergistically improving (e.g., alleviating or treating) diarrhea caused by Staphylococcus aureus is more significant. Further preferably, the mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane is 3:1 to 20:1, preferably 3:1 to 10:1, within which the synergistic effect in inhibiting Staphylococcus aureus (in vivo or in vitro) is more significant.

[0035] product

[0036] The second aspect of the present invention relates to a food, which comprises the nutritional composition according to the first aspect of the present invention.

[0037] All the above descriptions about the first aspect of the present invention are applicable here and will not be repeated here.

[0038] In some embodiments, the food may be, for example, a functional food, a health food, or a health product.

[0039] In some embodiments, the food is a common food or a special food. As an example, the food can be selected from dairy products, candies, beverages, bread and biscuits, for example, the food can be selected from milk powder or fermented food. In some embodiments, the milk powder is infant milk powder, such as infant formula milk powder.

[0040] In some embodiments, the food is an infant food such as an infant formula.

[0041] In some embodiments, the food product may be in the form of a powder, a tablet, or a liquid.

[0042] In some embodiments, based on the dry weight of the food, the content of medium-chain fatty acid triglycerides may be 0.1% to 30.0%. As an example, based on the weight of the product, the content of medium-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, 30.0 dry weight%, or within the range defined by any two of them. The content of milk fat globule membrane can be calculated according to the ratio of the two.

[0043] In some embodiments, the content of milk fat globule membrane may be 0.0005% to 30% based on the dry weight of the product. As an example, the content of milk fat globule membrane may be 0.0005, 0.001, 0.005, 0.01, 0.05, 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, 30.0 dry weight%, or within the range defined by any two of them, based on the weight of the product.

[0044] In addition, it is easy for those skilled in the art to understand that, in addition to the nutritional composition of the first aspect of the present invention, the food also includes one or more selected from the following: raw milk, desalted whey powder, concentrated whey protein, lactose, edible vegetable blending oil, oligofructose, oligogalactose, nucleotides, choline, vitamins, minerals, DHA and taurine.

[0045] For example, when the food is milk powder, in addition to the nutritional composition described in the first aspect of the present invention, the milk powder may also include proteins such as α-lactalbumin and milk fat globule membrane protein; carbohydrates such as lactose; lipids; minerals such as calcium, iron, phosphorus, etc.; vitamins; other additives such as whey powder, choline bitartrate, docosahexaenoic acid, arachidonic acid, walnut oil, etc.

[0046] The food can be produced by a preparation method commonly used in the art, which will not be described in detail here.

[0047] application

[0048] The present invention also relates to the use of the nutritional composition according to the first aspect of the present invention.

[0049] In the third aspect, the use is for preparing a product for improving (eg, alleviating or treating) diarrhea.

[0050] In some embodiments, the diarrhea is bacterial diarrhea. The bacterial diarrhea may be caused, for example, by Staphylococcus aureus infection.

[0051] In a fourth aspect, the use is for preparing a product for inhibiting Staphylococcus aureus infection.

[0052] In some embodiments, the inhibition is in vivo inhibition or in vitro inhibition.

[0053] In some embodiments, the inhibition comprises inhibiting the growth, proliferation, or metabolite production of S. aureus.

[0054] In a fifth aspect, the use is for preparing a product for regulating intestinal flora (eg, intestinal flora in a subject with diarrhea). In some embodiments, the intestinal flora includes Staphylococcus aureus.

[0055] In the sixth aspect, the use is for the non-therapeutic purpose of improving diarrhea.

[0056] In some embodiments, the diarrhea is bacterial diarrhea. The bacterial diarrhea may be caused, for example, by Staphylococcus aureus infection.

[0057] In a seventh aspect, the use is for non-therapeutic purposes of inhibiting Staphylococcus aureus infection.

[0058] In some embodiments, the inhibition is in vivo inhibition or in vitro inhibition.

[0059] In some embodiments, the inhibition comprises inhibiting the growth, proliferation, or metabolite production of S. aureus.

[0060] All the above descriptions about the first aspect of the present invention are applicable here and will not be repeated here.

[0061] The product of the third to fifth aspects of the present invention may be, for example, a food or a medicine.

[0062] For the food, the above description about the second aspect of the present invention is applicable here and will not be repeated here.

[0063] Regarding the drug, in some embodiments, its dosage form includes but is not limited to tablets, granular powders, capsules and liquids.

[0064] In some embodiments, when the composition, food or medicine is orally administered to a human body, the dosage of medium and long chain fatty acid triglycerides is 0.1 to 30 g / day, for example, 0.1 g / day, 0.2 g / day, 0.3 g / day, 0.4 g / day, 0.5 g / day, 0.6 g / day, 0.7 g / day, 0.8 g / day, 0.9 g / day, 1 g / day, 2 g / day, 3 g / day, 4 g / day, 5 g / day, 6 g / day, 7 g / day, 8 g / day, 9 g / day, 10 g / day, 11 g / day, 12 g / day, 13 g / day, 14 g / day, 15 g / day, 16 g / day, 17 g / day, 18 g / day, 19 g / day, 20 g / day, 21 g / day, 22 g / day, 23 g / day, 24 g / day, 25 g / day, 26 g / day, 27 g / day, 28 g / day, 29 g / day, or 30 g / day or any range therebetween. The dosage of the milk fat globule membrane can be converted according to the ratio with medium and long chain fatty acid triglycerides.

[0065] In some embodiments, the dosage of the milk fat globule membrane is 0.0005-30 g / day, for example, 0.0005 g / day, 0.0006 g / day, 0.0007 g / day, 0.0008 g / day, 0.0009 g / day, 0.001 g / day, 0.002 g / day, 0.003 g / day, 0.004 g / day, 0.005 g / day, 0.006 g / day, 0.007 g / day, 0.008 g / day, 0.009 g / day, 0.01 g / day, 0.02 g / day, 0.03 g / day, 0.04 g / day, 0.05 g / day, 0.06 g / day, 0.07 g / day, 0.08 g / day, 0.09 g / day, 1 g / day, 2 g / day, 3 g / day, 4 g / day, 5 g / day, 6 g / day, 7 g / day, 8 g / day, 9 g / day, 10 g / day, 11 g / day, 12 g / day, 13 g / day, 14 g / day, 15 g / day, 16 g / day, 17 g / day, 18 g / day, 19 g / day, 20 g / day, 21 g / day, 22 g / day g / day, 4 g / day, 5 g / day, 6 g / day, 7 g / day, 8 g / day, 9 g / day, 10 g / day, 11 g / day, 12 g / day, 13 g / day, 14 g / day, 15 g / day, 16 g / day, 17 g / day, 18 g / day, 19 g / day, 20 g / day, 21 g / day, 22 g / day, 23 g / day, 24 g / day, 25 g / day, 26 g / day, 27 g / day, 28 g / day, 29 g / day, or 30 g / day or any range therebetween. Medium and long chain fatty acid triglycerides and milk fat globule membranes can be formulated into compositions or preparations for the convenience of subject taking. Alternatively, medium and long chain fatty acid triglycerides and milk fat globule membranes can be taken separately without being formulated into the same composition, for example, taken simultaneously or taken separately. In the case of separate taking, medium and long chain fatty acid triglycerides and milk fat globule membranes can be taken at intervals of a certain time within a day. Long-chain fatty acid triglycerides and milk fat globule membrane can also be taken several times a day. The interval between the administration of the two components or the number of separate administrations can be easily determined by those skilled in the art.

[0066] Example

[0067] The present invention will be more easily understood with reference to the following examples, which are merely provided to illustrate certain aspects and embodiments of the present invention and are not intended to limit the present invention.

[0068] Unless otherwise stated, the reagents used in this embodiment are all commercially available materials or conventional materials. In all the following embodiments, medium-chain fatty acid triglycerides (MLCT) are derived from medium-chain fatty acid edible oil (purity of 69±10%), produced by Qingdao Haizhiyuan Life Science Technology Co., Ltd., with a production batch number of Y1505-22120101.

[0069] Milk fat globule membrane (MFGM) was purchased from Arla under the product name Lacprodan® MFGM-10, batch number P510216. The raw material had a phospholipid content of 8±2%, a protein content of 71.5±5% by weight, and a sphingomyelin g / 100g phospholipid content of 25±10g.

[0070] Vancomycin was obtained from Shanghai MacLean Biochemical Technology Co., Ltd., white powder, batch number C12976208.

[0071] Example 1 Evaluation of the efficacy of MLCT or MFGM in improving diarrhea

[0072] Wild-type AB strain zebrafish at 4 days post-fertilization (4 dpf) were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations are shown in Table 1), and the positive control vancomycin concentration was 1000 μg / mL. A normal control group and a model control group were also set up, with a volume of 3 mL per well. After treatment at 28℃ for 8 h, each experimental group was given water-soluble Nile red as a fluorescent indicator of intestinal contents, and fed overnight. The next day, the samples and Nile red were washed off, and the samples were given water-soluble (concentrations are shown in Table 1, Figure 1 ). Except for the normal control group, all other experimental groups were given water-soluble Staphylococcus aureus to establish a zebrafish diarrhea model. After treatment at 28℃ for 30 h, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The intensity of zebrafish intestinal fluorescence signals was analyzed, and the results of statistical analysis were used to evaluate the efficacy of the samples in improving diarrhea induced by Staphylococcus aureus. The statistical processing results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0073] According to the above method, different doses of MLCT and MFGM were set for zebrafish administration to evaluate the efficacy of different concentrations of the two single components in improving diarrhea (intestinal fluorescence intensity). The dosages are shown in Table 1, and the results of the administration are shown in Tables 1 and Figure 1 shown.

[0074] Table 1 Results of the efficacy evaluation experiment on the efficacy of single component in improving diarrhea (intestinal fluorescence intensity)

[0075]

[0076] Note: P value is the result of the significant difference analysis of intestinal fluorescence intensity between the medium-chain fatty acid triglyceride group or MFGM group and the model control group.

[0077] From Table 1 and Figure 1The results showed that the intestinal fluorescence intensity of 4dpf wild-type AB strain zebrafish significantly decreased after induction with Staphylococcus aureus compared with the normal control group (P < 0.05), indicating that the diarrhea model induced by Staphylococcus aureus was successfully established. The positive control (vancomycin 1000 μg / mL), monomeric medium-chain fatty acid triglycerides (125 µg / mL, 250 µg / mL, 500 µg / mL, 750 µg / mL) and MFGM (15.6 µg / mL, 31.2 µg / mL, 41.6µg / mL) all had the effect of significantly improving diarrhea, as shown by: the intestinal fluorescence intensity of zebrafish became stronger compared with the model control group (P < 0.05). The effect of monomeric medium-chain fatty acid triglycerides on improving diarrhea was the strongest at a concentration of 250 µg / mL. When the concentration was further increased (500 µg / mL, 750 µg / mL), the effect of improving diarrhea became weaker or similar. Therefore, 250 µg / mL was the optimal concentration of medium-chain fatty acid triglycerides for improving diarrhea. The effect of monomeric MFGM on improving diarrhea was the strongest at a concentration of 31.2 µg / mL. When the concentration was further increased (41.6 µg / mL), the effect of improving diarrhea in zebrafish became weaker. When the concentration increased to 62.5 µg / mL, it had no effect on improving diarrhea. Therefore, 31.2 µg / mL was the optimal concentration of MFGM for improving diarrhea.

[0078] Example 2 Evaluation of the efficacy of the combination of MLCT and MFGM in improving diarrhea

[0079] 4 dpf wild-type AB strain zebrafish were randomly selected and placed in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). Figure 2 , Figure 3 ), positive control vancomycin concentration of 1000 μg / mL, normal control group and model control group were set up at the same time, and the volume of each well was 3 mL. After 8 h of treatment at 28℃, each experimental group was given Nile red as a fluorescent indicator of intestinal contents in water, fed overnight, and the next day, the samples and Nile red were washed off, and the samples were given water again (concentrations are shown in Table 2, Figure 2 , Figure 3 ). Except for the normal control group, all other experimental groups were given water-soluble Staphylococcus aureus to establish a zebrafish diarrhea model. After treatment at 28℃ for 30 h, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The intensity of zebrafish intestinal fluorescence signals was analyzed, and the results of statistical analysis of this index were used to evaluate the efficacy of the samples in improving diarrhea induced by Staphylococcus aureus.

[0080] The dosage and results are shown in Table 2. The results of the administration are shown in Table 2. Figure 2 and Figure 3 shown.

[0081] Table 2 Results of efficacy evaluation experiment on the efficacy of the composition in improving diarrhea (intestinal fluorescence intensity)

[0082]

[0083] Note: P value is the result of the significant difference analysis of intestinal fluorescence intensity between the medium-chain fatty acid triglyceride group or MFGM group and the model control group.

[0084] From Table 2, Figure 2 and Figure 3 The results show that compared with the model control group, the compositions of Examples 3 to 10 all have the effect of improving diarrhea, which is specifically manifested by the enhancement of intestinal fluorescence intensity (P<0.05). Compared with the effects of MLCT alone (concentration 250 µg / mL) and MFGM alone (concentration 31.2 µg / mL) at the optimal concentration, the values ​​of zebrafish intestinal fluorescence intensity in Examples 3 to 9 with a ratio of MLCT to MFGM of (2-80):1 were significantly enhanced, indicating that there is a synergistic effect between MLCT and MFGM in improving diarrhea; the values ​​of zebrafish intestinal fluorescence intensity in Examples 4 to 8 with a ratio of MLCT to MFGM of (3-40):1 were further enhanced, indicating that the synergistic effect between MLCT and MFGM in improving diarrhea was more significant.

[0085] Example 3 Experiment on the efficacy of the combination of MLCT and MFGM in inhibiting Staphylococcus aureus

[0086] 4 dpf wild-type AB strain zebrafish were randomly selected in 6-well plates, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations are shown in Table 3), the positive control vancomycin concentration was 1000 μg / mL, and the model control group was set up at the same time, with a volume of 3 mL per well. After 24 h of treatment at 28℃, all experimental groups were given water-soluble Staphylococcus aureus to establish the zebrafish Staphylococcus aureus infection model. After CM-DiI was used to label Staphylococcus aureus for 6 h at 28℃, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software, and the fluorescence intensity of Staphylococcus aureus in the zebrafish intestine was analyzed. The antibacterial efficacy of the samples was evaluated by statistical analysis of the index. The statistical analysis results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0087] The dosage is shown in Table 3. The results of the administration are shown in Table 3. Figure 4 and Figure 5 shown.

[0088] Table 3 Anti-Staphylococcus aureus efficacy evaluation experiment

[0089]

[0090] Note: P value is the result of the significant difference analysis of the fluorescence intensity of Staphylococcus aureus between the medium-chain fatty acid triglyceride group or MFGM group and the model control group.

[0091] From Table 3, Figure 4 , Figure 5 The results show that compared with the model control group induced by Staphylococcus aureus, the positive control (vancomycin, 1000 µg / mL), monomeric medium-chain fatty acid triglycerides (250 µg / mL), MFGM (31.2 µg / mL) and Example 2-10 all have significant anti-Staphylococcus aureus efficacy, which is specifically manifested in that the fluorescence intensity of Staphylococcus aureus in zebrafish is weakened compared with the model control group (P<0.05). Compared with monomeric medium-chain fatty acid triglycerides (250 µg / mL) and MFGM (31.2 µg / mL), the fluorescence intensity of Staphylococcus aureus in Example 3-8, in which the ratio between MLCT and MFGM was (2-40:1), weakened, indicating that there was a synergistic effect between MLCT and MFGM in anti-Staphylococcus aureus; the fluorescence intensity of Staphylococcus aureus in Example 4-7, in which the ratio between MLCT and MFGM was (3-20:1), was even lower, indicating that the synergistic effect between MLCT and MFGM in anti-Staphylococcus aureus was stronger; the fluorescence intensity of Staphylococcus aureus in Example 4-6, in which the ratio between MLCT and MFGM was (3-10:1), further decreased, indicating that the synergistic effect between MLCT and MFGM in anti-Staphylococcus aureus was further enhanced.

[0092] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A nutritional composition, characterized in that The invention comprises medium-chain triglycerides of fatty acids and milk fat globule membrane, wherein the mass ratio of the medium-chain triglycerides of fatty acids to the milk fat globule membrane is 2:1 to 80:

1.

2. The nutritional composition according to claim 1, characterized in that The mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane is 2:1 to 40:

1.

3. The nutritional composition according to claim 2, characterized in that The mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane is 3:1 to 40:

1.

4. The nutritional composition according to claim 3, characterized in that The mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane is 3:1 to 20:

1.

5. The nutritional composition according to claim 4, characterized in that The mass ratio of the medium-chain fatty acid triglyceride to the milk fat globule membrane is 3:1 to 10:

1.

6. Food, characterized in that A nutritional composition comprising any one of claims 1 to 5.

7. The food according to claim 6, characterized in that The food is selected from one or more of dairy products, candies, beverages, bread and biscuits.

8. The food according to claim 6, characterized in that The food is milk powder or fermented food.

9. The food according to claim 8, characterized in that The milk powder is infant formula milk powder.

10. The food according to claim 6, characterized in that The food is infant food.

11. Use of the nutritional composition according to any one of claims 1 to 5 in preparing a product for improving diarrhea.

12. The use according to claim 11, characterized in that: The diarrhea is bacterial diarrhea.

13. The use according to claim 12, characterized in that: The bacterial diarrhea is caused by Staphylococcus aureus infection.

14. The use according to claim 11, characterized in that: The product is a food or a medicine.

15. The use according to claim 11, characterized in that: The product is a health food.

16. Use of the nutritional composition according to any one of claims 1 to 5 in the preparation of a product for inhibiting Staphylococcus aureus infection, wherein the inhibition is in vivo inhibition or in vitro inhibition.

17. The use according to claim 16, characterized in that The product is a food or a medicine.

18. The use according to claim 16, characterized in that The product is a health food.

19. Use of the nutritional composition according to any one of claims 1 to 5 in the preparation of a product for regulating intestinal flora in a subject with diarrhea.

20. The use according to claim 19, characterized in that The product is a food or a medicine.

21. The use according to claim 19, characterized in that The product is a health food.

22. Use of the nutritional composition according to any one of claims 1 to 5 for the non-therapeutic purpose of improving diarrhea.

23. Use of the nutritional composition according to any one of claims 1 to 5 for non-therapeutic purposes of inhibiting Staphylococcus aureus infection.

Images