Nutritional composition and use of the nutritional composition

Abstract

The present invention provides a nutritional composition and the use of the nutritional composition. The nutritional composition comprises medium- and long-chain triglycerides (MLCT) and milk fat globule membrane (MFGM) in a mass ratio of 2:1 to 80:1, and the combination of the two has a synergistic effect of improving diarrhea, inhibiting Staphylococcus aureus in vivo or in vitro, and / or regulating the intestinal flora.

Description

Technical Field

[0001] The present disclosure relates to a nutritional composition for inhibiting Staphylococcus aureus and / or improving diarrhea, and uses 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 diarrhea diseases in developing countries are serious, so it remains a global public health problem worthy of attention. The World Health Organization (WHO) estimates that tens of millions of people suffer from diarrhea diseases every day globally, with 1.7 billion cases of diarrhea occurring annually, and 2.2 million patients die due to severe diarrhea (Miao Xiaohui et al., Chinese Journal of Digestion, 2013, 33(12): 793 - 802). In addition, infectious diarrhea diseases have a high incidence rate and are widespread among children, seriously endangering children's health and being one of the main causes of childhood malnutrition (Ni Xin et al., China Medicine and Pharmacy, 2020, 10(21): 249 - 256). Infectious diarrhea is an intestinal infectious disease caused by bacteria, fungi, viruses or parasites, with diarrhea as the main clinical manifestation.

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

[0004] Medium and long-chain triacylglycerols (MLCT), as a star product among new structural lipids, have received extensive attention. The main structural feature of MLCT is that both medium-chain fatty acids and long-chain fatty acids are bound to a glycerol backbone. Early studies have shown that MLCT has many health functions such as reducing blood lipids, inhibiting obesity, reducing diabetes and cardiovascular diseases, and reducing the risk of cancer (Lai Yundong et al., Chinese Journal of Oil Crop Sciences, 2024, 46(04): 719-727.). In addition, the fatty acid composition of MLCT is closer to breast milk, which is beneficial to fat digestion and absorption, improves the absorption of lipid nutrients, inhibits the accumulation of body fat, and can provide energy quickly and stably (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 Sci Technol. 2024 Jun;15(1):381-408.). Currently, it is 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 Care, 2016, 24(01): 43-47.).

[0005] There have been no reports at home and 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 drugs with anti-infective diarrhea efficacy.

[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 the intestinal flora. Summary of the Invention

[0007] The present invention is made in view of the above problems in the art.

[0008] In a first aspect of the present invention, a nutritional composition is provided, which contains medium and long-chain triacylglycerols and milk fat globule membrane, wherein the mass ratio of the medium and long-chain triacylglycerols to the milk fat globule membrane is 2:1 to 80:1.

[0009] In a third aspect of the present invention, there is provided the use of the nutritional composition according to the first aspect of the present invention in the preparation of a drug for improving (such as alleviating or treating) diarrhea.

[0010] The fourth aspect of the present invention provides the use of the nutritional composition described in the first aspect of the present invention in the preparation of a medicament for inhibiting Staphylococcus aureus infection.

[0011] The fifth aspect of the present invention provides the use of the nutritional composition described in the first aspect of the present invention in the preparation of a product for regulating the intestinal flora.

[0012] The sixth aspect of the present invention provides the use of the nutritional composition described in the first aspect of the present invention for the non-therapeutic purpose of improving diarrhea.

[0013] The seventh aspect of the present invention provides the use of the nutritional composition described in the first aspect of the present invention for the non-therapeutic purpose of inhibiting Staphylococcus aureus infection.

[0014] The present invention discovers that when medium- and long-chain fatty acid triglycerides and milk fat globule membrane are used in combination at the defined mass ratio, there is a synergistic effect between them, which can synergistically improve diarrhea, especially improve (such as relieve or treat) diarrhea caused by Staphylococcus aureus, and inhibit Staphylococcus aureus (in vivo or in vitro). Description of the Drawings

[0015] Figure 1 Showing the fluorescence intensity of the zebrafish intestine after sample treatment. The P value is the significance analysis of the difference between each example group and the model control group. Indicating P < 0.01, Indicating P < 0.001.

[0016] Figure 2 Showing the fluorescence intensity of the zebrafish intestine after treatment with the composition sample. The P value is the significance analysis of the difference between each example group and the model control group, Indicating P < 0.01, Indicating P < 0.001.

[0017] Figure 3 Showing a typical image of the intestine of zebrafish treated with the sample.

[0018] Figure 4 Showing the fluorescence intensity of Staphylococcus aureus in the zebrafish intestine after sample treatment. The P value is the significance analysis of the difference between each example group and the model control group, Indicating P < 0.05, Indicating P < 0.01, Indicating P < 0.001.

[0019] Figure 5 Showing a typical image of the fluorescence intensity of Staphylococcus aureus in the zebrafish intestine after sample treatment. Detailed Embodiments

[0020] 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 meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Preferred materials and methods are described herein, but any methods and materials similar to or equivalent to those described herein can be used in the practice of testing the present invention. It should also be understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting.

[0021] Term Definitions

[0022] Unless otherwise indicated or defined, all terms used have their ordinary meanings in the art, which will be understood by those skilled in the art. In addition, unless otherwise stated, all methods, steps, techniques, and operations not specifically detailed can and have been carried out in a manner known per se, which will be understood by those skilled in the art.

[0023] As used herein, the term "medium-chain triglyceride" or "MLCT" is a special triglyceride in which both medium-chain fatty acids and long-chain fatty acids are present on the triglyceride backbone. As used herein, the term "medium-chain triglyceride" or "MLCT" is a special triglyceride in which both 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 oils and medium-chain triglycerides through a transesterification reaction by lipase, and through processes such as distillation separation, decolorization, and deodorization. Medium-chain triglycerides can be derived from medium-chain fatty acid edible oils.

[0024] As used herein, the term "medium-chain fatty acid" refers to fatty acids having 6-12 carbon atoms in the carbon chain, such as caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, etc.; the term "long-chain fatty acid" refers to fatty acids 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 triglyceride" and "long-chain triglyceride" respectively refer to the esterification reaction products of "medium-chain fatty acid" and "long-chain fatty acid" with glycerol; the term "medium-chain 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-chain triglycerides used, and the medium-chain triglycerides commonly used in the art can be used. Medium-chain triglycerides can be used in pure form or in non-pure form rich in medium-chain triglycerides. For example, the medium-chain 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: caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, etc. C 14 -C 30 The fatty acid residue may be derived from one or more of the following: myristic acid, palmitic acid, margaric acid, oleic acid, linoleic acid, stearic acid, nonadecanoic acid, eicosapentaenoic acid, heneicosanoic acid, docosatetraenoic acid, docosahexaenoic acid, tricosanoic acid, tetracosenoic acid, pentacosapentaenoic acid, docosahexaenoic 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. In some embodiments, C6-C 12 The weight ratio of the fatty acid residue to the C 14 -C 30 The weight ratio of the fatty acid residue is from 0.124 to 2.000. In certain embodiments, C6-C 12 The weight ratio of the fatty acid residue to the C 14 -C 30 The weight ratio of the fatty acid residue 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 them. In some embodiments, the "medium-chain and long-chain fatty acid triglyceride" may be prepared by referring to the method of Patent CN115369132A.

[0025] As used herein, the term "milk fat globule membrane" or "MFGM", which is Milk Fat Globule Membrane in English, is a complex biological membrane structure present in breast milk. The milk fat globule membrane is a complex three-layer phospholipoprotein membrane that envelopes the surface of milk fat droplets and is composed of polar lipids, cholesterol, proteins, etc. In principle, there is no particular limitation on the source of the milk fat globule membrane of the present invention. Generally, it can be obtained by extraction from animal milk or its products. In some preferred embodiments, such animal milk or its products can be cow milk, goat milk, camel milk, horse milk, or dairy products based on them (such as cheese), etc. More preferably, it can be extracted from cow milk, including colostrum or regular cow milk. The present invention does not particularly limit the method for extracting MFGM from the above-mentioned animal milk or its products. For example, the method of acid precipitation - centrifugation - isoelectric point enrichment - drying with acidification can be used, or the existing membrane filtration method can also be used to separate 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, etc. Milk fat globule membrane whey protein powder refers to a powdery product containing milk fat globule membrane, which is made from raw milk or whey through processes such as separation, concentration, and drying. Milk fat globule membrane milk protein powder refers to a powdery product containing milk fat globule membrane, which is made from raw milk or cream through processes such as separation, concentration, and drying. Commercially available sources of MFGM as the present invention include LacprodanⓇ MFGM-10, LacprodanⓇ PL-20, Cor-PowerⓇ SM2, lipid-rich MFGM fraction, or buttermilk powder concentrates BPC50, BPC60, G600, PC700, Hilmar Cor-PowerⓇ WPC7500MEGM ENRICHED WPC, etc.

[0026] As used herein, "Staphylococcus aureus" is also called "S. aureus" and belongs to the genus Staphylococcus. It is a representative of Gram-positive bacteria and is a common foodborne pathogenic microorganism widely present in the natural environment. Staphylococcus aureus can produce enterotoxin under appropriate conditions, causing food poisoning. Enterotoxin is a single-chain small molecule protein with a molecular weight of about 26 - 29 kDa, a relatively low molecular weight, heat stability, and can damage the human intestine, resulting in symptoms such as vomiting and diarrhea.

[0027] As used herein, a "kit" refers to a set 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 and long-chain fatty acid triglycerides and / or milk fat globule membrane. 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

[0029] Nutritional composition

[0030] In a first aspect of the present invention, there is provided a nutritional composition comprising medium-chain and long-chain triglycerides (MLCT) and milk fat globule membrane (MFGM), wherein the mass ratio of the medium-chain and long-chain triglycerides to the milk fat globule membrane is from 2:1 to 80:1.

[0031] The present invention has found that when medium-chain and long-chain triglycerides and milk fat globule membrane are used in combination at the defined mass ratio, there is a synergistic effect between them, which can synergistically improve diarrhea, especially improve (such as relieve or treat) diarrhea caused by Staphylococcus aureus, and / or inhibit Staphylococcus aureus (in vivo or in vitro).

[0032] In some embodiments, the nutritional composition consists of medium-chain and long-chain triglycerides (MLCT) and milk fat globule membrane (MFGM).

[0033] In an embodiment, the mass ratio of the medium-chain and long-chain triglycerides to the milk fat globule membrane in the nutritional composition is from 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 and long-chain triglycerides to the milk fat globule membrane is from 2:1 to 40:1, within this range, the synergistic effect is more significant, especially it can synergistically improve diarrhea, especially synergistically improve (such as relieve or treat) diarrhea caused by Staphylococcus aureus, and synergistically inhibit Staphylococcus aureus (in vivo or in vitro). More preferably, the mass ratio of the medium-chain and long-chain triglycerides to the milk fat globule membrane is from 3:1 to 40:1, within this range, the synergistic effect in improving diarrhea, especially synergistically improving (such as relieve or treat) diarrhea caused by Staphylococcus aureus is more significant. Further preferably, the mass ratio of the medium-chain and long-chain triglycerides to the milk fat globule membrane is from 3:1 to 20:1, preferably from 3:1 to 10:1, within this range, the synergistic effect in inhibiting Staphylococcus aureus (in vivo or in vitro) is more significant.

[0034] Product

[0035] In a second aspect of the present invention, there is provided a product comprising the nutritional composition according to the first aspect of the present invention.

[0036] As an example, based on the weight of the product, the content of medium and long-chain fatty acid triglycerides can 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 basis 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.

[0037] In some embodiments, based on the dry basis weight of the product, the content of milk fat globule membrane can be from 0.0005% to 30%. As an example, based on the weight of the product, the content of milk fat globule membrane can 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 basis weight, or within the range defined by any two of them.

[0038] Application

[0039] The present invention also relates to the application of the nutritional composition of the first aspect of the present invention.

[0040] In a third aspect, the application is in the preparation of a medicament for improving (such as alleviating or treating) diarrhea.

[0041] In some embodiments, the diarrhea is bacterial diarrhea. The bacterial diarrhea can be caused, for example, by Staphylococcus aureus infection.

[0042] In a fourth aspect, the application is in the preparation of a medicament for inhibiting Staphylococcus aureus infection.

[0043] In some embodiments, the inhibition is in vivo inhibition or in vitro inhibition.

[0044] In some embodiments, the inhibition includes inhibiting the growth, proliferation or metabolite production of Staphylococcus aureus.

[0045] In a fifth aspect, the application is in the preparation of a product for regulating the intestinal flora. In some embodiments, the intestinal flora includes Staphylococcus aureus.

[0046] In the sixth aspect, the application is for non-therapeutic purposes of improving diarrhea.

[0047] In some embodiments, the diarrhea is bacterial diarrhea. The bacterial diarrhea can be caused, for example, by Staphylococcus aureus infection.

[0048] In the seventh aspect, the application is for non-therapeutic purposes of inhibiting Staphylococcus aureus infection.

[0049] In some embodiments, the inhibition is in vivo inhibition or in vitro inhibition.

[0050] In some embodiments, the inhibition includes inhibiting the growth, proliferation, or metabolite production of Staphylococcus aureus.

[0051] All the descriptions regarding the first aspect of the present invention above are applicable here and will not be elaborated further.

[0052] The product in the fifth aspect of the present invention can be, for example, a drug.

[0053] For the drug, in some embodiments, its dosage forms include but are not limited to tablets, granule powders, capsules, and liquids.

[0054] In some embodiments, when the composition or drug is orally administered to a human, the dosage of medium- and long-chain fatty acid triglycerides is 0.1 - 30 g / day, such as 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 to the medium- and long-chain fatty acid triglycerides.

[0055] In some embodiments, the dosage of the milk fat globule membrane is 0.0005 to 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, 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-chain and long-chain triglycerides and milk fat globule membrane can be formulated into a composition or preparation for easy administration to the subject. Alternatively, medium-chain and long-chain triglycerides and milk fat globule membrane can be administered separately without being formulated into the same composition, for example, simultaneously or separately. In the case of separate administration, medium-chain and long-chain triglycerides and milk fat globule membrane can be administered at intervals within a day. Long-chain triglycerides and milk fat globule membrane can also be administered in several doses within a day. The time interval between the administration of the two components or the number of separate administrations can be easily determined by those skilled in the art.

[0056] Examples

[0057] The present invention will be more easily understood with reference to the following examples, which are only used to illustrate certain aspects and embodiments of the present invention and are not intended to limit the present invention.

[0058] Unless otherwise specified, the reagents used in this example are all commercially available materials or conventional materials. In all the following examples, medium-chain and long-chain triglycerides (MLCT) are derived from medium-chain and long-chain fatty acid edible oil (purity 69 ± 10%), produced by Qingdao Haizhiyuan Life Science Co., Ltd., and the production batch number is Y1505 - 22120101.

[0059] Milk fat globule membrane (MFGM), purchased from Arla, with the product name Lacprodan® MFGM-10, batch number P510216. The phospholipid content in this raw material is 8 ± 2 %, the protein content is 71.5 ± 5 wt%, and the sphingomyelin per 100 g of phospholipid is 25 ± 10 g.

[0060] Vancomycin is sourced from Shanghai Macklin Biochemical Co., Ltd., in the form of white powder, with the batch number C12976208.

[0061] Example 1 Evaluation of the Efficacy of MLCT or MFGM in Improving Diarrhea

[0062] Wild-type AB strain zebrafish at 4 days post-fertilization (4 dpf) were randomly selected and placed in 6-well plates, with 30 zebrafish in each well (experimental group). The samples were administered in aqueous solution (concentrations are shown in Table 1), and the positive control was vancomycin at a concentration of 1000 μg / mL. At the same time, a normal control group and a model control group were set up, with a volume of 3 mL per well. After treatment at 28 °C for 8 h, each experimental group was administered Nile red as a fluorescent indicator of intestinal contents in aqueous solution and fed overnight. The next day, the samples and Nile red were washed away, and the samples were continued to be administered in aqueous solution (concentrations are shown in Table 1, Figure 1 ). Except for the normal control group, the rest of the experimental groups were administered Staphylococcus aureus in aqueous solution to establish a zebrafish diarrhea model. After treatment at 28 °C for 30 h, 10 zebrafish were randomly selected from each experimental group and placed under a fluorescence microscope for photographing. The NIS-Elements D 3.20 advanced image processing software was used to analyze and collect data, and the fluorescence signal intensity of the zebrafish intestine was analyzed. The efficacy of the samples in improving diarrhea induced by Staphylococcus aureus was evaluated based on the statistical analysis results of this index. The statistical treatment 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.

[0063] Following the above method, MLCT and MFGM were respectively administered to zebrafish at different doses to evaluate the efficacy (intestinal fluorescence intensity) of the two single components at different concentrations in improving diarrhea. The dosing doses are shown in Table 1, and the dosing results are shown in Table 1 and Figure 1 as shown.

[0064] Table 1 Experimental Results of the Efficacy Evaluation of Single Components in Improving Diarrhea (Intestinal Fluorescence Intensity)

[0065]

[0066] Note: The P value is the result of the analysis of the difference in intestinal fluorescence intensity between the medium-chain and long-chain fatty acid triglyceride group or the MFGM group and the model control group.

[0067] From Table 1 and Figure 1The results showed that after induction with Staphylococcus aureus, the intestinal fluorescence intensity of 4-dpf wild-type AB strain zebrafish was significantly decreased 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), monomer medium- and long-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, 41.6 μg / mL) all had significant effects on improving diarrhea. Specifically, compared with the model control group, the intestinal fluorescence intensity of zebrafish became stronger (P < 0.05). Monomer medium- and long-chain fatty acid triglycerides had the strongest effect on improving diarrhea at a concentration of 250 μg / mL. When its concentration was further increased (500 μg / mL, 750 μg / mL), its effect on improving diarrhea became weaker or similar. Therefore, a concentration of 250 μg / mL was the optimal concentration for medium- and long-chain fatty acid triglycerides to improve diarrhea. Monomer MFGM had the strongest effect on improving diarrhea at a concentration of 31.2 μg / mL. When its concentration was further increased (41.6 μg / mL), the effect of zebrafish on improving diarrhea became weaker. When the concentration increased to 62.5 μg / mL, there was no effect on improving diarrhea. Therefore, 31.2 μg / mL was the optimal concentration for MFGM to improve diarrhea.

[0068] Example 2 Evaluation of the Efficacy of the Composition of MLCT and MFGM in Improving Diarrhea

[0069] Randomly select 4-dpf wild-type AB strain zebrafish into 6-well plates, with 30 zebrafish in each well (experimental group). Administer the samples in water (concentrations are shown in Table 2, Figure 2 , Figure 3 ), the positive control vancomycin at a concentration of 1000 μg / mL. At the same time, set up a normal control group and a model control group, with a volume of 3 mL in each well. After treatment at 28°C for 8 h, each experimental group was administered Nile red as a fluorescent indicator of intestinal contents in water and fed overnight. The next day, the samples and Nile red were washed off, and the samples were continued to be administered in water (concentrations are shown in Table 2, Figure 2 , Figure 3 ). Except for the normal control group, the rest of the experimental groups were administered Staphylococcus aureus in water to establish a zebrafish diarrhea model. After treatment at 28°C for 30 h, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The NIS-Elements D 3.20 advanced image processing software was used to analyze and collect data, and the intestinal fluorescence signal intensity of zebrafish was analyzed. The efficacy of the samples in improving diarrhea induced by Staphylococcus aureus was evaluated based on the statistical analysis results of this index.

[0070] The administration doses and results are shown in Table 2. The administration results are shown in Table 2,Figure 2 and Figure 3 as shown

[0071] Table 2 Results of the efficacy evaluation experiment on the effect of the composition in improving diarrhea (intestinal fluorescence intensity)

[0072]

[0073] Note: The P value is the result of the analysis of the significance of the difference in intestinal fluorescence intensity between the medium and long-chain fatty acid triglyceride group or the MFGM group and the model control group.

[0074] From Table 2 Figure 2 and Figure 3 It can be seen from the results that compared with the model control group, the compositions of Examples 3-10 all have the effect of improving diarrhea, specifically manifested as an increase in intestinal fluorescence intensity (P < 0.05). Compared with the effects of individual MLCT (concentration 250 μg / mL) and individual MFGM (concentration 31.2 μg / mL) at the optimal concentration, the intestinal fluorescence intensity values of zebrafish in Examples 3-9 with the ratio of MLCT to MFGM being (2-80):1 are all significantly increased, indicating that there is a synergistic effect between MLCT and MFGM in improving diarrhea; the intestinal fluorescence intensity values of zebrafish in Examples 4-8 with the ratio of MLCT to MFGM being (3-40):1 are further increased, indicating that the synergistic effect between MLCT and MFGM in improving diarrhea is more significant.

[0075] Efficacy experiment of the composition of MLCT and MFGM in Example 3 against Staphylococcus aureus

[0076] Randomly select 4 dpf wild-type AB strain zebrafish into 6-well plates, with 30 zebrafish in each well (experimental group). Administer the sample in water (concentration as shown in Table 3), the positive control vancomycin at a concentration of 1000 μg / mL, and at the same time set up a model control group, with a volume of 3 mL per well. After treatment at 28°C for 24 h, all experimental groups were administered Staphylococcus aureus in water to establish a zebrafish Staphylococcus aureus infection model. After labeling Staphylococcus aureus with CM-DiI at 28°C for 6 h, randomly select 10 zebrafish from each experimental group and place them under a fluorescence microscope for photographing. Use the NIS-Elements D 3.20 advanced image processing software to analyze and collect data, analyze the fluorescence intensity of Staphylococcus aureus in the zebrafish intestine, and evaluate the antibacterial efficacy of the sample based on the statistical analysis results of this index. The statistical processing results are expressed as mean ± SE. Use SPSS 26.0 software for statistical analysis, and p < 0.05 indicates that the difference is statistically significant.

[0077] The administration dose is as shown in Table 3. The administration results are as shown in Table 3 Figure 4 and Figure 5 as shown

[0078] Table 3 Evaluation of the efficacy against Staphylococcus aureus

[0079]

[0080] Note: The P value is the result of the significance analysis of the difference in the fluorescence intensity of Staphylococcus aureus between the medium- and long-chain fatty acid triglyceride group or the MFGM group and the model control group.

[0081] From Table 3, Figure 4 , Figure 5 it can be seen that compared with the model control group induced by Staphylococcus aureus, the positive control (vancomycin, 1000 µg / mL), the monomer medium- and long-chain fatty acid triglyceride (250 µg / mL), MFGM (31.2 µg / mL), and Examples 2-10 all have significant efficacy against Staphylococcus aureus. Specifically, compared with the model control group, the fluorescence intensity of Staphylococcus aureus in zebrafish becomes weaker (P<0.05). Compared with the monomer medium- and long-chain fatty acid triglyceride (250 µg / mL) and MFGM (31.2 µg / mL), the fluorescence intensity of Staphylococcus aureus becomes weaker in Examples 3-8 where the ratio of MLCT to MFGM is (2-40):1, indicating a synergistic effect between MLCT and MFGM in terms of anti-Staphylococcus aureus efficacy; the fluorescence intensity of Staphylococcus aureus is even lower in Examples 4-7 where the ratio of MLCT to MFGM is (3-20):1, indicating a stronger synergistic effect between MLCT and MFGM in terms of anti-Staphylococcus aureus efficacy; the fluorescence intensity of Staphylococcus aureus is further reduced in Examples 4-6 where the ratio of MLCT to MFGM is (3-10):1, indicating a further enhanced synergistic effect between MLCT and MFGM in terms of anti-Staphylococcus aureus efficacy.

[0082] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A nutritional composition, characterized in that, Containing medium- and long-chain triglycerides and milk fat globule membrane, wherein the mass ratio of the medium- and long-chain triglycerides to the milk fat globule membrane is from 2:1 to 80:

1.

2. The nutritional composition according to claim 1, wherein The mass ratio of the medium- and long-chain triglycerides to the milk fat globule membrane is from 2:1 to 40:

1.

3. The nutritional composition according to claim 2, wherein The mass ratio of the medium- and long-chain triglycerides to the milk fat globule membrane is from 3:1 to 40:

1.

4. The nutritional composition according to claim 3, wherein The mass ratio of the medium- and long-chain triglycerides to the milk fat globule membrane is from 3:1 to 20:

1.

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

1.

6. Product, characterized in that, Containing the nutritional composition according to any one of claims 1-5.

7. Use of the nutritional composition according to any one of claims 1-5 in the preparation of a product for improving bacterial diarrhea caused by Staphylococcus aureus infection.

8. The application according to claim 7, wherein The product is a drug.

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

10. The application according to claim 9, wherein The product is a drug.

11. Use of the nutritional composition according to any one of claims 1-5 in the preparation of a product for regulating the intestinal flora of a subject with bacterial diarrhea caused by Staphylococcus aureus infection.

12. The application according to claim 11, wherein The product is a drug.

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