A bifidobacterium-lactobacillus plantarum complex composition for regulating intestinal flora

Abstract

The present invention discloses a Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora, relating to the technical fields of microbial technology, functional foods, and biomedicine. The present invention combines three active probiotics, namely Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum, and prepares them into single-strain powders through a specific ratio and vacuum freeze-drying technology. Freeze-drying protectants and prebiotics are further added. Through the synergistic action of these components, the composition not only effectively regulates the balance of intestinal flora, inhibits the growth of harmful bacteria, and promotes the proliferation of beneficial bacteria, but also significantly enhances the survival rate and stability of the probiotics during storage, transportation, and within the intestinal tract, thereby ensuring that the probiotics maintain high activity in the intestine and comprehensively maintaining and improving intestinal health.

Description

[0001] DESCRIPTION

[0002] A Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora

[0003] TECHNICAL FIELD

[0004] The present invention relates to the technical fields of microbial technology, functional foods, and biomedicine, and in particular to a Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora.

[0005] BACKGROUND ART

[0006] With the accelerating pace of modern life and changes in dietary structure, intestinal health issues have received increasing attention. As the largest digestive and immune organ in the human body, the balance of the microbial community in the intestine is of great importance to human health. Imbalance of intestinal flora not only leads to common problems such as dyspepsia, constipation, and diarrhea, but is also closely related to the occurrence and development of a variety of chronic diseases such as obesity, diabetes, and autoimmune diseases.

[0007] Traditional probiotic products often suffer from problems such as a single strain, an unreasonable ratio, and a low survival rate. Probiotic products containing only a single strain are unable to comprehensively regulate the balance of intestinal flora, and an unreasonable ratio among different strains also affects their synergistic performance. In addition, traditional probiotic products are readily affected by environmental factors such as temperature and humidity during storage and transportation, resulting in a reduction in probiotic survival rate and thus affecting their use effect. Meanwhile, some probiotic products lack necessary auxiliary components such as prebiotics and therefore cannot provide sufficient nutritional support for probiotics, further limiting their growth and activity in the intestine.

[0008] In view of the problems of traditional probiotic products, such as a single strain, an unreasonable ratio, and a low survival rate, it is particularly DESCRIPTION important to provide a Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora.

[0009] SUMMARY OF THE INVENTION

[0010] An object of the present invention is to overcome the shortcomings of the prior art by providing a Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora. By combining three active probiotics, namely Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum, preparing them into single-strain powders through a specific ratio and vacuum freeze-drying technology, and further adding auxiliary components such as a freeze-drying protectant and a prebiotic, the present invention effectively overcomes the defects of traditional probiotic products. The complex composition of the present invention not only comprehensively regulates the balance of intestinal flora, inhibits the growth of harmful bacteria, and promotes the proliferation of beneficial bacteria, but also significantly enhances the survival rate and stability of probiotics during storage, transportation, and in the intestinal tract, thereby providing a new solution for maintaining intestinal health.

[0011] To solve the above technical problems, the present invention provides the following technical solution. A Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora is prepared from the following raw materials in parts by weight:

[0012] The active probiotic component of the complex composition consists of Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum.

[0013] Further, the viable-count ratio of Bifidobacterium longum, Bifidobacterium DESCRIPTION adolescentis, and Lactobacillus plantarum is (1 -3):(1 -3):(l-2).

[0014] Furthermore, the viable-count ratio of Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum is 1:1 :1.

[0015] Furthermore, per gram of the complex composition, each of Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum has a viable count of 1.0x10A9-5.0xl0A10 CFU.

[0016] Furthermore, the active probiotic components are all single-strain powders prepared by vacuum freeze-drying.

[0017] Furthermore, the complex composition further comprises a prebiotic, and the prebiotic is one or more selected from fructooligosaccharide, galactooligosaccharide, inulin, isomaltooligosaccharide, and stachyose.

[0018] Furthermore, the complex composition further comprises a freeze-drying protectant, and the freeze-drying protectant is one or more selected from skimmed milk powder, trehalose, mannitol, and glycerol.

[0019] Furthermore, the complex composition further comprises a food- grade or pharmaceutically acceptable filler, and the filler is one or more selected from maltodextrin, com starch, and microcrystalline cellulose.

[0020] Furthermore, the complex composition further comprises a food- grade or pharmaceutically acceptable glidant, and the glidant is one or more selected from magnesium stearate, silicon dioxide, and micronized silica gel.

[0021] Furthermore, the dosage form of the complex composition is any one of powder, capsule, tablet, granule, and oral liquid.

[0022] Compared with the prior art, the Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora provided by the present DESCRIPTION invention has the following beneficial effects:

[0023] I. By combining three active probiotics, namely Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum, preparing them into single-strain powders through a specific ratio and vacuum freeze-drying technology, and further adding a freeze-drying protectant and a prebiotic, this complex composition achieves synergistic effects among these components. It not only effectively regulates the balance of intestinal flora, inhibits the growth of harmful bacteria, and promotes the proliferation of beneficial bacteria, but also significantly enhances the survival rate and stability of probiotics during storage, transportation, and in the intestinal tract, thereby ensuring that the probiotics maintain high activity in the intestine and comprehensively maintaining and improving intestinal health.

[0024] Other advantages, objectives, and features of the present invention will be set forth in part in the following description, and in part will become apparent to those skilled in the art upon examination of the following disclosure, or may be learned from practice of the invention.

[0025] BRIEF DESCRIPTION OF THE DRAWINGS

[0026] To describe the technical solutions in the embodiments of the present invention or in the prior art more clearly, the drawings required for describing the embodiments or the prior art are briefly introduced below. Obviously, the drawings in the following description merely illustrate certain embodiments of the present invention, and those of ordinary skill in the art can derive other drawings therefrom without inventive effort.

[0027] Figure 1 is a flow chart of the Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora. DESCRIPTION

[0028] DETAILED DESCRIPTION

[0029] In order to further illustrate the technical means and effects adopted by the present invention to achieve the intended objectives, the specific implementation, structure, features, and effects of the present invention are described in detail below in conjunction with the accompanying drawings and preferred embodiments.

[0030] Embodiments 1-3 differ only in the amount of Bifidobacterium longum.

[0031] Embodiment 1

[0032] I. Formulation composition

[0033] Active probiotic component

[0034] Bifidobacterium longum lyophilized powder: 15 parts, viable count in powder >1. Ox 10A12 CFU / g

[0035] Bifidobacterium adolescentis lyophilized powder: 10 parts, viable count in powder >1.0xl0A12 CFU / g

[0036] Lactobacillus plantarum lyophilized powder: 10 parts, viable count in powder >1.0xl0A12 CFU / g

[0037] The viable-count ratio of the above three strains is 1.5: 1:1. Per gram of the final complex composition, the viable count of Bifidobacterium longum is 1.5xl0A10 CFU, and the viable counts of Bifidobacterium adolescentis and Lactobacillus plantarum are both 1.0xl0A10 CFU.

[0038] Prebiotic component

[0039] Fructooligosaccharide: 180 parts

[0040] Stachyose: 120 parts

[0041] Filler

[0042] Maltodextrin: 645 parts

[0043] Glidant

[0044] Silicon dioxide: 20 parts

[0045] II. Preparation method

[0046] 1. Preparation of single-strain lyophilized powders DESCRIPTION

[0047] (1) Strain activation: glycerol-preserved tubes of the corresponding strains were taken and streaked onto MRS solid medium. Bifidobacterium longum and Bifidobacterium adolescentis were cultured anaerobically at 37 °C for 48 h, and Lactobacillus plantarum was cultured at 37 °C for 24 h. Single colonies with full morphology were picked and inoculated into MRS liquid medium, and then subcultured for two generations under the same culture conditions to obtain seed cultures with stable activity.

[0048] (2) Fermentation culture: the seed cultures were inoculated into optimized MRS fermentation medium at an inoculation amount of 3% (v / v). Bifidobacterium longum and Bifidobacterium adolescentis were anaerobically fermented at 37 °C for 24 h, and Lactobacillus plantarum was fermented at 37 °C for 18 h. After fermentation, the cultures were centrifuged at 8000 r / min for 15 min using a disc centrifuge, the supernatant was discarded, and highly active bacterial sludge was collected.

[0049] (3) Vacuum freeze-drying: a sterile freeze-drying protectant was added to the bacterial sludge, and the mass ratio of bacterial sludge to freeze-drying protectant was 1:3. After thorough mixing, a uniform bacterial suspension was obtained. The suspension was pre-frozen in a -40 °C low-temperature freezer for 8 h and then transferred to a vacuum freeze-dryer for drying for 24 h under conditions of a cold-trap temperature of -55 °C and a vacuum degree of 10 Pa, thereby obtaining the corresponding single-strain lyophilized powder, which was stored at 4 °C in a sealed light-proof state for later use.

[0050] 2. Preparation of the powder form of the complex composition

[0051] (1) Sieving pretreatment: the three single-strain lyophilized powders prepared above, together with fructooligosaccharide, stachyose, maltodextrin, and silicon dioxide, were separately passed through an 80-mesh sieve to remove agglomerates for later use.

[0052] (2) Equal-increment mixing: according to the formulation in parts by weight, the three strain powders and an equal mass of maltodextrin were first DESCRIPTION pre-mixed in a three-dimensional mixer for 10 min. The remaining maltodextrin, fructooligosaccharide, and stachyose were then added successively by the equal-increment method and mixed for 20 min, after which silicon dioxide was added and mixing was continued for another 10 min. The final material-mixing uniformity had an RSD of <5%.

[0053] (3) Packaging and storage: the uniformly mixed material was aseptically packed into aluminum foil bags in a Class 10,000 clean environment at 1 g per bag, sealed, and stored in a cool and dry place, thereby obtaining the Bifidobacterium-Lactobacillus plantarum complex powder for regulating intestinal flora.

[0054] III. Verification test on the effect of the complex composition in regulating intestinal flora

[0055] 1. Test materials and animals

[0056] Test sample: the complex powder prepared in this embodiment. Test animals: 40 SPF-grade male BALB / c mice, 6-8 weeks old, weighing 18-22 g, adaptively fed for 3 days. The breeding environment was maintained at a temperature of 22±2 °C, a humidity of 50±5%, and a 12 h light / dark cycle, with free access to food and water.

[0057] 2. Grouping and treatment

[0058] The mice were randomly divided into 4 groups, with 10 mice in each group. The grouping and treatment methods were as follows:

[0059] Blank control group: intragastric administration of normal saline, 0.2 mL / mouse / day

[0060] Low-dose group: intragastric administration of a suspension of the complex powder at a dose of 0.5 g / kg bw / day

[0061] Medium-dose group: intragastric administration of a suspension of the complex powder at a dose of 1.0 g / kg bw / day

[0062] High-dose group: intragastric administration of a suspension of the complex powder at a dose of 2.0 g / kg bw / day DESCRIPTION

[0063] The intragastric administration was continued for 28 days. During this period, the body weight, feed intake, and defecation status of the mice were recorded daily, and no mouse died or showed abnormal adverse reactions.

[0064] 3. Detection indicators and specific effect parameters

[0065] After completion of the intragastric administration, fresh feces and cecal contents of the mice were aseptically collected, and the intestinal flora counts, short-chain fatty acid contents, and cecal pH were determined respectively. The specific effect parameters were as follows:

[0066] (1) Effect on the core intestinal flora in mice. In the blank control group, the counts of Bifidobacterium spp., Lactobacillus spp., Escherichia coli, and Clostridium perfringens in the feces of the mice were 7.21±0.35 IgCFU / g wet feces, 6.84±0.42 IgCFU / g wet feces, 8.32±0.38 IgCFU / g wet feces, and 5.76±0.29 IgCFU / g wet feces, respectively. After intervention, the low-dose group showed an increase in Bifidobacterium spp. to 8.87±0.31 IgCFU / g wet feces, representing a 23.02% increase over the blank control group; an increase in Lactobacillus spp. to 8.12±0.28 IgCFU / g wet feces, representing an 18.71% increase over the blank control group; a decrease in Escherichia coli to 7.25±0.30 IgCFU / g wet feces, representing a 12.86% decrease over the blank control group; and a decrease in Clostridium perfringens to 4.87±0.23 IgCFU / g wet feces, representing a 15.45% decrease over the blank control group. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in Bifidobacterium spp. to 9.82±0.28 IgCFU / g wet feces, representing a 36.20% increase; an increase in Lactobacillus spp. to 9.05±0.25 IgCFU / g wet feces, representing a 32.31% increase; a decrease in Escherichia coli to 6.58±0.26 IgCFU / g wet feces, representing a 20.91% decrease; and a decrease in Clostridium perfringens to 4.21 ±0.20 IgCFU / g wet feces, representing a 26.91% decrease. All indices showed extremely significant differences as compared with the blank control group (PO.01). After DESCRIPTION intervention, the high-dose group showed an increase in Bifidobacterium spp. to 10.58±0.24 IgCFU / g wet feces, representing a 46.74% increase; an increase in Lactobacillus spp. to 9.81±0.30 IgCFU / g wet feces, representing a 43.42% increase; a decrease in Escherichia coli to 5.94±0.28 IgCFU / g wet feces, representing a 28.61% decrease; and a decrease in Clostridium perfringens to 3.62±0.21 IgCFU / g wet feces, representing a 37.15% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the overall efficacy exhibited a clear dose-dependent trend.

[0067] (2) Effect on intestinal metabolism and environment in mice. In the blank control group, the total short-chain fatty acid content in the cecal contents of the mice was 32.15±2.47 pmol / g, including 18.62±1.53 pmol / g acetic acid, 7.54±0.82 pmol / g propionic acid, and 5.99±0.64 pmol / g butyric acid, and the pH of the cecal contents was 7.12±0.15. After intervention, the low-dose group showed an increase in total short-chain fatty acids to 48.76±2.23 pmol / g, representing a 51.66% increase over the blank control group, including 27.35±1.41 pmol / g acetic acid, 11.72±0.73 pmol / g propionic acid, and 9.69±0.56 pmol / g butyric acid, representing increases of 46.88%, 55.44%, and 61.77%, respectively; the pH of the cecal contents decreased to 6.47±0.11, representing a 9.13% decrease over the blank control group. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in total short-chain fatty acids to 63.52±2.18 pmol / g, representing a 97.57% increase, including 35.41±1.36 pmol / g acetic acid, 15.38±0.76 pmol / g propionic acid, and 12.73 ±0.59 pmol / g butyric acid, representing increases of 90.17%, 104.00%, and 112.52%, respectively; the pH of the cecal contents decreased to 6.02±0.09, representing a 15.45% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in total DESCRIPTION short-chain fatty acids to 78.39±2.12 pmol / g, representing a 143.83% increase, including 43.26±1.32 pmol / g acetic acid, 19.24±0.74 pmol / g propionic acid, and 15.89±0.57 pmol / g butyric acid, representing increases of 132.33%, 155.17%, and 165.28%, respectively; the pH of the cecal contents decreased to 5.72±0.10, representing a 19.66% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). Among them, the increase in butyric acid was significantly greater than that in acetic acid and propionic acid, reflecting the specific enhancement effect of Bifidobacterium longum on intestinal butyrate-production capacity.

[0068] (3) Effect on intestinal physiological functions in mice. In the blank control group, the average daily number of fecal pellets was 12.35±1.22, and the fecal moisture content was 42.36±2.15%. After intervention, the low-dose group showed an average daily fecal pellet number of 15.24±1.13, representing a 23.40% increase over the blank control group, and a fecal moisture content of 50.17±1.92%, representing an 18.44% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an average daily fecal pellet number of 17.36±1.06, representing a 40.57% increase, and a fecal moisture content of 56.38±1.83%, representing a 33.09% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an average daily fecal pellet number of 19.08±1.09, representing a 54.49% increase, and a fecal moisture content of 61.25±1.88%, representing a 44.59% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01), indicating that the composition can effectively improve intestinal peristaltic rhythm and enhance intestinal excretory function.

[0069] (4) Food-use safety evaluation. During the 28-day continuous intragastric administration period, no mice in any dose group showed death, lethargy, io DESCRIPTION abnormal food or water intake, diarrhea, abdominal distension, or other adverse reactions. There were no significant differences (P>0.05) between the dose groups and the blank control group in final body weight or organ coefficients of the heart, liver, kidneys, spleen, and lungs. No acute or subacute toxic side effect was observed, indicating good food-use safety.

[0070] Embodiment 2

[0071] I. Formulation composition

[0072] Active probiotic component

[0073] Bifidobacterium longum lyophilized powder: 20 parts, viable count in powder >1. Ox 10A12 CFU / g

[0074] Bifidobacterium adolescentis lyophilized powder: 10 parts, viable count in powder >1.0xl0A12 CFU / g

[0075] Lactobacillus plantarum lyophilized powder: 10 parts, viable count in powder >1. Ox 10A12 CFU / g

[0076] The viable-count ratio of the above three strains is 2:1 :1. Per gram of the final complex composition, the viable count of Bifidobacterium longum is 2.0xl0A10 CFU, and the viable counts of Bifidobacterium adolescentis and Lactobacillus plantarum are both 1.0xl0A10 CFU.

[0077] Prebiotic component

[0078] Fructooligosaccharide: 180 parts

[0079] Stachyose: 120 parts

[0080] Filler

[0081] Maltodextrin: 640 parts

[0082] Glidant

[0083] Silicon dioxide: 20 parts

[0084] II. Preparation method

[0085] 1. Preparation of single-strain lyophilized powders

[0086] (1) Strain activation: glycerol-preserved tubes of the corresponding strains were taken and streaked onto MRS solid medium. Bifidobacterium DESCRIPTION longum and Bifidobacterium adolescentis were cultured anaerobically at 37 °C for 48 h, and Lactobacillus plantarum was cultured at 37 °C for 24 h. Single colonies with full morphology were picked and inoculated into MRS liquid medium, and then subcultured for two generations under the same culture conditions to obtain seed cultures with stable activity. (2) Fermentation culture: the seed cultures were inoculated into optimized MRS fermentation medium at an inoculation amount of 3% (v / v). Bifidobacterium longum and Bifidobacterium adolescentis were anaerobically fermented at 37 °C for 24 h, and Lactobacillus plantarum was fermented at 37 °C for 18 h. After fermentation, the cultures were centrifuged at 8000 r / min for 15 min using a disc centrifuge, the supernatant was discarded, and highly active bacterial sludge was collected. (3) Vacuum freeze-drying: a sterile freeze-drying protectant was added to the bacterial sludge, and the mass ratio of bacterial sludge to freeze-drying protectant was 1:3. After thorough mixing, a uniform bacterial suspension was obtained. The suspension was pre-frozen in a -40 °C low-temperature freezer for 8 h and then transferred to a vacuum freeze-dryer for drying for 24 h under conditions of a cold-trap temperature of -55 °C and a vacuum degree of 10 Pa, thereby obtaining the corresponding single-strain lyophilized powder, which was stored at 4 °C in a sealed light-proof state for later use.

[0087] 2. Preparation of the powder form of the complex composition

[0088] (1) Sieving pretreatment: the three single-strain lyophilized powders prepared above, together with fructooligosaccharide, stachyose, maltodextrin, and silicon dioxide, were separately passed through an 80-mesh sieve to remove agglomerates for later use. (2) Equal-increment mixing: according to the formulation in parts by weight, the three strain powders and an equal mass of maltodextrin were first pre-mixed in a three-dimensional mixer for 10 min. The remaining maltodextrin, fructooligosaccharide, and stachyose were then added successively by the equal-increment method and mixed for 20 min, DESCRIPTION after which silicon dioxide was added and mixing was continued for another 10 min. The final material -mixing uniformity had an RSD of <5%. (3) Packaging and storage: the uniformly mixed material was aseptically packed into aluminum foil bags in a Class 10,000 clean environment at 1 g per bag, sealed, and stored in a cool and dry place, thereby obtaining the Bifidobacterium-Lactobacillus plantarum complex powder for regulating intestinal flora.

[0089] III. Verification test on the effect of the complex composition in regulating intestinal flora

[0090] 1. Test materials and animals

[0091] Test sample: the complex powder prepared in this embodiment. Test animals: 40 SPF-grade male BALB / c mice, 6-8 weeks old, weighing 18-22 g, adaptively fed for 3 days. The breeding environment was maintained at a temperature of 22±2 °C, a humidity of 50±5%, and a 12 h light / dark cycle, with free access to food and water.

[0092] 2. Grouping and treatment

[0093] The mice were randomly divided into 4 groups, with 10 mice in each group. The grouping and treatment methods were as follows:

[0094] Blank control group: intragastric administration of normal saline, 0.2 mL / mouse / day

[0095] Low-dose group: intragastric administration of a suspension of the complex powder at a dose of 0.5 g / kg bw / day

[0096] Medium-dose group: intragastric administration of a suspension of the complex powder at a dose of 1.0 g / kg bw / day

[0097] High-dose group: intragastric administration of a suspension of the complex powder at a dose of 2.0 g / kg bw / day

[0098] The intragastric administration was continued for 28 days. During this period, the body weight, feed intake, and defecation status of the mice were recorded daily, and no mouse died or showed abnormal adverse reactions. DESCRIPTION

[0099] 3. Detection indicators and specific effect parameters

[0100] (1) Effect on the core intestinal flora in mice. In the blank control group, the counts of Bifidobacterium spp., Lactobacillus spp., Escherichia coli, and Clostridium perfringens in mouse feces were 7.21 ±0.35 IgCFU / g wet feces, 6.84±0.42 IgCFU / g wet feces, 8.32±0.38 IgCFU / g wet feces, and 5.76±0.29 IgCFU / g wet feces, respectively. After intervention, the low-dose group showed an increase in Bifidobacterium spp. to 9.12±0.29 IgCFU / g wet feces, representing a 26.49% increase over the blank control group; an increase in Lactobacillus spp. to 8.31±0.26 IgCFU / g wet feces, representing a 21.49% increase; a decrease in Escherichia coli to 7.12±0.28 IgCFU / g wet feces, representing a 14.42% decrease; and a decrease in Clostridium perfringens to 4.73±0.22 IgCFU / g wet feces, representing a 17.88% decrease. All indices showed extremely significant differences as compared with the blank control group (PO.Ol). After intervention, the medium-dose group showed an increase in Bifidobacterium spp. to 10.15=1=0.26 IgCFU / g wet feces, representing a 40.78% increase; an increase in Lactobacillus spp. to 9.24±0.24 IgCFU / g wet feces, representing a 35.09% increase; a decrease in Escherichia coli to 6.42±0.25 IgCFU / g wet feces, representing a 22.84% decrease; and a decrease in Clostridium perfringens to 4.05±0.19 IgCFU / g wet feces, representing a 29.69% decrease. All indices showed extremely significant differences as compared with the blank control group (PO.Ol). After intervention, the high-dose group showed an increase in Bifidobacterium spp. to 10.97±0.23 IgCFU / g wet feces, representing a 52.15% increase; an increase in Lactobacillus spp. to 10.03±0.28 IgCFU / g wet feces, representing a 46.64% increase; a decrease in Escherichia coli to 5.78±0.26 IgCFU / g wet feces, representing a 30.53% decrease; and a decrease in Clostridium perfringens to 3.44±0.20 IgCFU / g wet feces, representing a 40.28% decrease. All indices showed extremely significant differences as compared with the blank control group (PO.Ol), and the optimization effect on the intestinal flora structure DESCRIPTION continuously increased with dose.

[0101] (2) Effect on intestinal metabolism and environment in mice. In the blank control group, the total short-chain fatty acid content in the cecal contents of the mice was 32.15±2.47 pmol / g, including 18.62±1.53 pmol / g acetic acid, 7.54±0.82 pmol / g propionic acid, and 5.99±0.64 pmol / g butyric acid, and the pH of the cecal contents was 7.12±0.15. After intervention, the low-dose group showed an increase in total short-chain fatty acids to 51.93±2.19 pmol / g, representing a 61.52% increase over the blank control group, including 28.92±1.38 pmol / g acetic acid, 12.54±0.71 pmol / g propionic acid, and 10.47±0.55 pmol / g butyric acid, representing increases of 55.32%, 66.31%, and 74.79%, respectively; the pH of the cecal contents decreased to 6.39±0.10, representing a 10.25% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in total short-chain fatty acids to 68.27±2.15 pmol / g, representing a 112.35% increase, including 37.68±1.34 pmol / g acetic acid, 16.52±0.74 pmol / g propionic acid, and 14.07±0.58 pmol / g butyric acid, representing increases of 102.36%, 119.10%, and 134.89%, respectively; the pH of the cecal contents decreased to 5.94±0.09, representing a 16.57% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in total short-chain fatty acids to 84.61±2.10 pmol / g, representing a 163.17% increase, including 46.35±1.30 pmol / g acetic acid, 20.87±0.72 pmol / g propionic acid, and 17.39±0.56 pmol / g butyric acid, representing increases of 148.93%, 176.79%, and 190.32%, respectively; the pH of the cecal contents decreased to 5.61±0.09, representing a 21.21% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the intestinal acid-production capacity and microecological environment-optimization effect were significantly superior to those of DESCRIPTION complex compositions with a lower proportion of Bifidobacterium longum.

[0102] (3) Effect on intestinal physiological functions in mice. In the blank control group, the average daily number of fecal pellets was 12.35±1.22, and the fecal moisture content was 42.36±2.15%. After intervention, the low-dose group showed an average daily fecal pellet number of 15.71±1.11, representing a 27.21% increase over the blank control group, and a fecal moisture content of 51.42±1.90%, representing a 21.39% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an average daily fecal pellet number of 17.95±1.05, representing a 45.34% increase, and a fecal moisture content of 57.93 ±1.81%, representing a 36.76% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an average daily fecal pellet number of 19.87±1.08, representing a 60.89% increase, and a fecal moisture content of 63.17 1.86%, representing a 49.13% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01), and the improvements in intestinal peristalsis and excretory function were more pronounced.

[0103] (4) Food-use safety evaluation. During the 28-day continuous intragastric administration period, no mice in any dose group showed death, abnormal behavior, feeding disorder, or adverse intestinal reactions. There were no significant differences (P>0.05) between the groups and the blank control group in body-weight gain trend or major organ coefficients, indicating excellent food-use safety and no potential toxic side effect.

[0104] Embodiment 3

[0105] I. Formulation composition

[0106] Active probiotic component

[0107] Bifidobacterium longum lyophilized powder: 30 parts, viable count in powder >1. Ox 10A12 CFU / g DESCRIPTION

[0108] Bifidobacterium adolescentis lyophilized powder: 10 parts, viable count in powder >1.0><10A12 CFU / g

[0109] Lactobacillus plantarum lyophilized powder: 10 parts, viable count in powder >1.0 10A12 CFU / g

[0110] The viable-count ratio of the above three strains is 3:1 :1. Per gram of the final complex composition, the viable count of Bifidobacterium longum is 3.0 10A10 CFU, and the viable counts of Bifidobacterium adolescentis and Lactobacillus plantarum are both 1.0 10A10 CFU.

[0111] Prebiotic component

[0112] Fructooligosaccharide: 180 parts

[0113] Stachyose: 120 parts

[0114] Filler

[0115] Maltodextrin: 630 parts

[0116] Glidant

[0117] Silicon dioxide: 20 parts

[0118] II. Preparation method

[0119] 1. Preparation of single-strain lyophilized powders

[0120] (1) Strain activation: glycerol-preserved tubes of the corresponding strains were taken and streaked onto MRS solid medium. Bifidobacterium longum and Bifidobacterium adolescentis were cultured anaerobically at 37 °C for 48 h, and Lactobacillus plantarum was cultured at 37 °C for 24 h. Single colonies with full morphology were picked and inoculated into MRS liquid medium, and then subcultured for two generations under the same culture conditions to obtain seed cultures with stable activity. (2) Fermentation culture: the seed cultures were inoculated into optimized MRS fermentation medium at an inoculation amount of 3% (v / v). Bifidobacterium longum and Bifidobacterium adolescentis were anaerobically fermented at 37 °C for 24 h, and Lactobacillus plantarum was fermented at 37 °C for 18 h. After fermentation, the cultures were centrifuged at 8000 r / min for 15 min DESCRIPTION using a disc centrifuge, the supernatant was discarded, and highly active bacterial sludge was collected. (3) Vacuum freeze-drying: a sterile freeze-drying protectant was added to the bacterial sludge, and the mass ratio of bacterial sludge to freeze-drying protectant was 1:3. After thorough mixing, a uniform bacterial suspension was obtained. The suspension was pre-frozen in a -40 °C low-temperature freezer for 8 h and then transferred to a vacuum freeze-dryer for drying for 24 h under conditions of a cold-trap temperature of -55 °C and a vacuum degree of 10 Pa, thereby obtaining the corresponding single-strain lyophilized powder, which was stored at 4 °C in a sealed light-proof state for later use.

[0121] 2. Preparation of the powder form of the complex composition

[0122] (1) Sieving pretreatment: the three single-strain lyophilized powders prepared above, together with fructooligosaccharide, stachyose, maltodextrin, and silicon dioxide, were separately passed through an 80-mesh sieve to remove agglomerates for later use. (2) Equal-increment mixing: according to the formulation in parts by weight, the three strain powders and an equal mass of maltodextrin were first pre-mixed in a three-dimensional mixer for 10 min. The remaining maltodextrin, fructooligosaccharide, and stachyose were then added successively by the equal-increment method and mixed for 20 min, after which silicon dioxide was added and mixing was continued for another 10 min. The final material -mixing uniformity had an RSD of <5%. (3) Packaging and storage: the uniformly mixed material was aseptically packed into aluminum foil bags in a Class 10,000 clean environment at 1 g per bag, sealed, and stored in a cool and dry place, thereby obtaining the Bifidobacterium-Lactobacillus plantarum complex powder for regulating intestinal flora.

[0123] III. Verification test on the effect of the complex composition in regulating intestinal flora

[0124] 1. Test materials and animals DESCRIPTION

[0125] Test sample: the complex powder prepared in this embodiment. Test animals: 40 SPF-grade male BALB / c mice, 6-8 weeks old, weighing 18-22 g, adaptively fed for 3 days. The breeding environment was maintained at a temperature of 22±2 °C, a humidity of 50±5%, and a 12 h light / dark cycle, with free access to food and water.

[0126] 2. Grouping and treatment

[0127] The mice were randomly divided into 4 groups, with 10 mice in each group. The grouping and treatment methods were as follows:

[0128] Blank control group: intragastric administration of normal saline, 0.2 mL / mouse / day

[0129] Low-dose group: intragastric administration of a suspension of the complex powder at a dose of 0.5 g / kg bw / day

[0130] Medium-dose group: intragastric administration of a suspension of the complex powder at a dose of 1.0 g / kg bw / day

[0131] High-dose group: intragastric administration of a suspension of the complex powder at a dose of 2.0 g / kg bw / day

[0132] The intragastric administration was continued for 28 days. During this period, the body weight, feed intake, and defecation status of the mice were recorded daily, and no mouse died or showed abnormal adverse reactions.

[0133] 3. Detection indicators and specific effect parameters

[0134] (1) Effect on the core intestinal flora in mice. In the blank control group, the counts of Bifidobacterium spp., Lactobacillus spp., Escherichia coli, and Clostridium perfringens in mouse feces were 7.21 ±0.35 IgCFU / g wet feces, 6.84±0.42 IgCFU / g wet feces, 8.32±0.38 IgCFU / g wet feces, and 5.76±0.29 IgCFU / g wet feces, respectively. After intervention, the low-dose group showed an increase in Bifidobacterium spp. to 9.54±0.27 IgCFU / g wet feces, representing a 32.32% increase over the blank control group; an increase in Lactobacillus spp. to 8.57±0.25 IgCFU / g wet feces, representing a 25.29% increase; a decrease in Escherichia coli to 6.94±0.27 IgCFU / g wet feces, DESCRIPTION representing a 16.59% decrease; and a decrease in Clostridium perfringens to 4.52±0.21 IgCFU / g wet feces, representing a 21.53% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in Bifidobacterium spp. to 10.63±0.24 IgCFU / g wet feces, representing a 47.43% increase; an increase in Lactobacillus spp. to 9.51±0.23 IgCFU / g wet feces, representing a 39.03% increase; a decrease in Escherichia coli to 6.19±0.24 IgCFU / g wet feces, representing a 25.60% decrease; and a decrease in Clostridium perfringens to 3.82±0.18 IgCFU / g wet feces, representing a 33.68% decrease. All indices showed extremely significant differences as compared with the blank control group (PO.Ol). After intervention, the high-dose group showed an increase in Bifidobacterium spp. to 11.52±0.22 IgCFU / g wet feces, representing a 59.78% increase; an increase in Lactobacillus spp. to 10.38±0.27 IgCFU / g wet feces, representing a 51.75% increase; a decrease in Escherichia coli to 5.52±0.25 IgCFU / g wet feces, representing a 33.65% decrease; and a decrease in Clostridium perfringens to 3.21±0.19 IgCFU / g wet feces, representing a 44.27% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), reaching the optimal level in promoting the proliferation of beneficial bacteria and inhibiting harmful bacteria.

[0135] (2) Effect on intestinal metabolism and environment in mice. In the blank control group, the total short-chain fatty acid content in the cecal contents of the mice was 32.15±2.47 pmol / g, including 18.62±1.53 pmol / g acetic acid, 7.54±0.82 pmol / g propionic acid, and 5.99±0.64 pmol / g butyric acid, and the pH of the cecal contents was 7.12±0.15. After intervention, the low-dose group showed an increase in total short-chain fatty acids to 56.84±2.16 pmol / g, representing a 76.80% increase over the blank control group, including 31.47±1.36 pmol / g acetic acid, 13.79±0.70 pmol / g propionic acid, and 11.58±0.54 pmol / g butyric acid, representing increases of 69.01%, DESCRIPTION

[0136] 82.89%, and 93.32%, respectively; the pH of the cecal contents decreased to 6.28±0.10, representing an 11.80% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in total short-chain fatty acids to 74.69±2.12 pmol / g, representing a 132.32% increase, including 40.83±1.32 pmol / g acetic acid, 18.16±0.72 pmol / g propionic acid, and 15.70±0.57 pmol / g butyric acid, representing increases of 119.28%, 140.85%, and 162.10%, respectively; the pH of the cecal contents decreased to 5.82±0.08, representing an 18.26% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in total short-chain fatty acids to 93.27±2.08 pmol / g, representing a 190.11% increase, including 50.72±1.28 pmol / g acetic acid, 23.04±0.70 pmol / g propionic acid, and 19.51±0.55 pmol / g butyric acid, representing increases of 172.40%, 205.57%, and 225.71%, respectively; the pH of the cecal contents decreased to 5.48±0.08, representing a 23.03% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the optimization effect on the intestinal microecological metabolic environment was the most pronounced, thereby establishing an acidic intestinal environment more favorable for colonization of beneficial bacteria.

[0137] (3) Effect on intestinal physiological functions in mice. In the blank control group, the average daily number of fecal pellets was 12.35±1.22, and the fecal moisture content was 42.36±2.15%. After intervention, the low-dose group showed an average daily fecal pellet number of 16.28±1.10, representing a 31.82% increase over the blank control group, and a fecal moisture content of 52.89±1.88%, representing a 24.86% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an average daily fecal pellet number of 18.64±1.04, representing a 50.93% DESCRIPTION increase, and a fecal moisture content of 59.76±1.80%, representing a 41.08% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an average daily fecal pellet number of 20.79±1.07, representing a 68.34% increase, and a fecal moisture content of 65.42±1.85%, representing a 54.44% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01), and the improvement in intestinal motility function and constipation was the most significant.

[0138] (4) Food-use safety evaluation. During the 28-day continuous intragastric administration period, no mice in any dose group showed death, abnormal behavior, abnormal food or water intake, diarrhea, abdominal distension, or other intestinal adverse reactions. The mice showed normal body -weight gain, and there were no significant differences (P>0.05) in major organ coefficients as compared with the blank control group. No acute or subacute toxicity or potential health risk was observed, meeting the safety application standards for foods and pharmaceuticals.

[0139] Embodiments 4-6 differ only in the amount of Bifidobacterium adolescentis.

[0140] I. Formulation composition

[0141] Active probiotic component

[0142] Bifidobacterium longum lyophilized powder: 10 parts, viable count in powder >1. Ox 10A12 CFU / g

[0143] Bifidobacterium adolescentis lyophilized powder: 15 parts, viable count in powder >1.0xl0A12 CFU / g

[0144] Lactobacillus plantarum lyophilized powder: 10 parts, viable count in powder >1. Ox 10A12 CFU / g

[0145] The viable-count ratio of the above three strains is 1: 1.5:1. Per gram of the final complex composition, the viable counts of Bifidobacterium longum and Lactobacillus plantarum are both 1.0xl0A10 CFU, and the viable count of DESCRIPTION

[0146] Bifidobacterium adolescentis is 1.5><10A10 CFU.

[0147] Prebiotic component

[0148] Fructooligosaccharide: 180 parts

[0149] Stachyose: 120 parts

[0150] Filler

[0151] Maltodextrin: 645 parts

[0152] Glidant

[0153] Silicon dioxide: 20 parts

[0154] II. Preparation method

[0155] 1. Preparation of single-strain lyophilized powders

[0156] (1) Strain activation: glycerol-preserved strains of the corresponding microorganisms were taken and inoculated onto MRS solid-medium plates by the three-zone streaking method. Bifidobacterium longum and

[0157] Bifidobacterium adolescentis were cultured in an anaerobic constant-temperature incubator at 37 °C for 48 h, and Lactobacillus plantarum was cultured in a constant-temperature incubator at 37 °C for 24 h. After culture, single colonies with full morphology and neat edges were picked and inoculated into MRS liquid medium, and were continuously subcultured for two generations under the corresponding culture conditions to obtain seed cultures with stable activity and qualified concentration. (2) Fermentation culture: the prepared seed cultures were inoculated into optimized MRS fermentation medium at an inoculation amount of 3% (v / v). Bifidobacterium longum and Bifidobacterium adolescentis were fermented for 24 h at 37 °C under anaerobic conditions, and Lactobacillus plantarum was fermented for 18 h at 37 °C under constant-temperature conditions. After the end of fermentation, a disc centrifuge was used to centrifuge at 8000 r / min for 15 min, the fermentation supernatant was discarded, and the precipitate was collected to obtain highly active bacterial sludge. (3) Vacuum freeze-drying: a freeze-drying protectant sterilized by high-pressure steam was added to the DESCRIPTION bacterial sludge, and the mass ratio of bacterial sludge to freeze-drying protectant was 1:3. After full stirring and mixing, a uniform and stable bacterial suspension was prepared. The suspension was first pre-frozen in a -40 °C ultra-low-temperature freezer for 8 h and then transferred to a vacuum freeze-dryer, where vacuum drying was carried out for 24 h at a cold-trap temperature of -55 °C and a vacuum degree of 10 Pa, thereby obtaining the corresponding single-strain lyophilized powder. The product was stored at 4 °C in a sealed light-proof state for later use.

[0158] 2. Preparation of the powder form of the complex composition

[0159] (1) Sieving pretreatment: the three single-strain lyophilized powders prepared above, together with fructooligosaccharide, stachyose, maltodextrin, and silicon dioxide, were separately sieved through a standard 80-mesh sieve to remove caking, ensure uniform fineness of the materials, and the sieved materials were collected for later use. (2) Equal-increment mixing: according to the formulation in parts by weight, the three strain powders and an equal mass of maltodextrin were first added to a three-dimensional mixer and pre-mixed for 10 min. The remaining maltodextrin, fructooligosaccharide, and stachyose were then successively added by the equal-increment method and mixed continuously for 20 min, after which silicon dioxide was added and mixing was continued for another 10 min. After completion, the material-mixing uniformity was tested, and the final mixing uniformity had an RSD of <5%. (3) Packaging and storage: the uniformly mixed composite material was aseptically packed into aluminum foil bags at 1 g per bag in a Class 10,000 clean workshop, sealed, and stored in a cool and dry place, thereby obtaining the Bifidobacterium-Lactobacillus plantarum complex powder for regulating intestinal flora.

[0160] III. Verification test on the effect of the complex composition in regulating intestinal flora

[0161] 1. Test materials and animals DESCRIPTION

[0162] Test sample: the complex powder prepared in this embodiment. Test animals: 40 SPF-grade male BALB / c mice, 6-8 weeks old, weighing 18-22 g, adaptively fed for 3 days. The breeding environment was controlled at a temperature of 22±2 °C and a relative humidity of 50±5%, with a 12 h light / dark alternating cycle. During the test period, the mice had free access to food and water.

[0163] 2. Grouping and treatment

[0164] The mice were randomly divided into 4 groups, with 10 mice in each group. The grouping and intragastric administration methods were as follows:

[0165] Blank control group: intragastric administration of normal saline at an administration volume of 0.2 mL / mouse / day

[0166] Low-dose group: intragastric administration of a normal-saline suspension of the complex powder at a dose of 0.5 g / kg bw / day

[0167] Medium-dose group: intragastric administration of a normal-saline suspension of the complex powder at a dose of 1.0 g / kg bw / day

[0168] High-dose group: intragastric administration of a normal-saline suspension of the complex powder at a dose of 2.0 g / kg bw / day

[0169] The intragastric administration intervention was continued for 28 days. During the test period, the body weight, feed intake, mental status, and defecation of the mice were recorded daily. No mouse died and no abnormal adverse reaction occurred throughout the process.

[0170] 3. Detection indicators and specific effect parameters

[0171] After the intragastric administration intervention, fresh feces and cecal-content samples of the mice were aseptically collected, and the intestinal flora counts, short-chain fatty acid contents, and cecal pH were determined, while defecation-related indices were also recorded. The specific effect parameters were as follows: (1) Effect on the core intestinal flora in mice. In the blank control group, the counts of Bifidobacterium spp., Lactobacillus spp., Escherichia coli, and Clostridium perfringens in mouse DESCRIPTION feces were 7.21±0.35 IgCFU / g wet feces, 6.84±0.42 IgCFU / g wet feces, 8.32±0.38 IgCFU / g wet feces, and 5.76±0.29 IgCFU / g wet feces, respectively. After intervention, the low-dose group showed an increase in Bifidobacterium spp. to 8.72±0.30 IgCFU / g wet feces, representing a 20.94% increase over the blank control group; an increase in Lactobacillus spp. to 8.05±0.29 IgCFU / g wet feces, representing a 17.69% increase; a decrease in Escherichia coli to 7.32±0.31 IgCFU / g wet feces, representing a 12.02% decrease; and a decrease in Clostridium perfringens to 4.95±0.24 IgCFU / g wet feces, representing a 14.06% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in Bifidobacterium spp. to 9.64±0.29 IgCFU / g wet feces, representing a 33.70% increase; an increase in Lactobacillus spp. to 8.94±0.26 IgCFU / g wet feces, representing a 30.70% increase; a decrease in Escherichia coli to 6.68±0.27 IgCFU / g wet feces, representing a 19.71% decrease; and a decrease in Clostridium perfringens to 4.29±0.21 IgCFU / g wet feces, representing a 25.52% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in Bifidobacterium spp. to 10.47±0.25 IgCFU / g wet feces, representing a 45.22% increase; an increase in Lactobacillus spp. to 9.72±0.31 IgCFU / g wet feces, representing a 42.11% increase; a decrease in Escherichia coli to 6.00±0.29 IgCFU / g wet feces, representing a 27.88% decrease; and a decrease in Clostridium perfringens to 3.71 ±0.22 IgCFU / g wet feces, representing a 35.59% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the overall effect showed a clear dose-dependent trend.

[0172] (2) Effect on intestinal metabolism and environment in mice. In the blank control group, the total short-chain fatty acid content in the cecal contents was 32.15±2.47 pmol / g, including 18.62±1.53 pmol / g acetic acid, 7.54±0.82 DESCRIPTION pmol / g propionic acid, and 5.99±0.64 pmol / g butyric acid, and the pH of the cecal contents was 7.12±0.15. After intervention, the low-dose group showed an increase in total short-chain fatty acids to 47.35±2.21 pmol / g, representing a 47.28% increase over the blank control group, including 26.54±1.39 pmol / g acetic acid, 11.38±0.74 pmol / g propionic acid, and 9.43±0.57 pmol / g butyric acid, representing increases of 42.53%, 50.93%, and 57.43%, respectively; the pH of the cecal contents decreased to 6.51±0.12, representing an 8.57% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in total short-chain fatty acids to 61.49±2.19 pmol / g, representing a 91.26% increase, including 34.27±1.37 pmol / g acetic acid, 14.95±0.77 pmol / g propionic acid, and 12.27±0.60 pmol / g butyric acid, representing increases of 84.05%, 98.28%, and 104.84%, respectively; the pH of the cecal contents decreased to 6.08±0.10, representing a 14.61% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in total short-chain fatty acids to 75.82±2.13 pmol / g, representing a 135.83% increase, including 41.83±1.33 pmol / g acetic acid, 18.64±0.75 pmol / g propionic acid, and 15.35±0.58 pmol / g butyric acid, representing increases of 124.65%, 147.21%, and 156.26%, respectively; the pH of the cecal contents decreased to 5.79±0.10, representing an 18.68% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the increase in butyrate, a core functional intestinal metabolite, was particularly prominent.

[0173] (3) Effect on intestinal physiological functions in mice. In the blank control group, the average daily number of fecal pellets was 12.35±1.22, and the fecal moisture content was 42.36±2.15%. After intervention, the low-dose group showed an average daily fecal pellet number of 15.03±1.14, representing a 21.70% increase over the blank control group, and a fecal DESCRIPTION moisture content of 49.52±1.94%, representing a 16.90% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an average daily fecal pellet number of 17.12±1.07, representing a 38.62% increase, and a fecal moisture content of 55.64±1.85%, representing a 31.35% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an average daily fecal pellet number of 18.69±1.10, representing a 51.34% increase, and a fecal moisture content of 59.83±1.89%, representing a 41.24% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01), indicating that the composition can effectively improve intestinal peristaltic capacity, normalize intestinal excretory rhythm, and relieve constipation.

[0174] (4) Food-use safety evaluation. During the 28-day continuous intragastric administration period, no mice in the low-, medium-, or high-dose groups showed death, lethargy, abnormal food or water intake, diarrhea, abdominal distension, or other intestinal adverse reactions. There were no significant differences (P>0.05) between the groups and the blank control group in the body -weight gain trend or the major organ coefficients of the heart, liver, spleen, lungs, and kidneys. No acute or subacute toxic side effect was observed, indicating good food-use safety.

[0175] Embodiment 5

[0176] I. Formulation composition Active probiotic component Bifidobacterium longum lyophilized powder: 10 parts, viable count in powder >1. Ox 10A12 CFU / g

[0177] Bifidobacterium adolescentis lyophilized powder: 20 parts, viable count in powder >1.0xl0A12 CFU / g

[0178] Lactobacillus plantarum lyophilized powder: 10 parts, viable count in DESCRIPTION powder >1. Ox 10A12 CFU / g

[0179] The viable-count ratio of the above three strains is 1:2:1. Per gram of the final complex composition, the viable counts of Bifidobacterium longum and Lactobacillus plantarum are both 1.0x l0A10 CFU, and the viable count of Bifidobacterium adolescentis is 2.0xl0A10 CFU.

[0180] Prebiotic component

[0181] Fructooligosaccharide: 180 parts

[0182] Stachyose: 120 parts

[0183] Filler

[0184] Maltodextrin: 640 parts

[0185] Glidant

[0186] Silicon dioxide: 20 parts

[0187] II. Preparation method

[0188] 1. Preparation of single-strain lyophilized powders

[0189] (1) Strain activation: glycerol-preserved tubes of the corresponding strains were taken and inoculated onto MRS solid medium by the streak-inoculation method. Bifidobacterium longum and Bifidobacterium adolescentis were cultured anaerobically at 37 °C for 48 h, and Lactobacillus plantarum was cultured at 37 °C for 24 h. After culture, single colonies showing good growth and uniform morphology were picked and transferred into MRS liquid medium, and were continuously passaged twice under the corresponding culture conditions to obtain seed cultures with stable activity. (2) Fermentation culture: the seed cultures were inoculated into optimized MRS fermentation medium at an inoculation amount of 3% (v / v). Bifidobacterium longum and Bifidobacterium adolescentis were fermented for 24 h at 37 °C under anaerobic conditions, and Lactobacillus plantarum was fermented for 18 h at 37 °C under constant-temperature conditions. After fermentation, a disc centrifuge was used to centrifuge at 8000 r / min for 15 min, the supernatant was discarded, and the precipitate was collected to obtain DESCRIPTION highly active bacterial sludge. (3) Vacuum freeze-drying: a sterile freeze-drying protectant was added to the bacterial sludge, and the mass ratio of bacterial sludge to freeze-drying protectant was 1:3. After full stirring and mixing, a uniform bacterial suspension was prepared. The suspension was pre-frozen in a -40 °C ultra-low-temperature freezer for 8 h and then transferred to a vacuum freeze-dryer, where drying was carried out for 24 h at a cold-trap temperature of -55 °C and a vacuum degree of 10 Pa, thereby obtaining the corresponding single-strain lyophilized powder, which was stored at 4 °C in a sealed light-proof state for later use.

[0190] 2. Preparation of the powder form of the complex composition

[0191] (1) Sieving pretreatment: the three single-strain lyophilized powders, fructooligosaccharide, stachyose, maltodextrin, and silicon dioxide were separately sieved through an 80-mesh sieve to remove agglomerated materials, ensure consistent fineness of all raw materials, and the sieved materials were collected for later use. (2) Equal-increment mixing: according to the formulation in parts by weight, the three strain powders and an equal mass of maltodextrin were first added to a three-dimensional mixer and pre-mixed for 10 min. The remaining maltodextrin, fructooligosaccharide, and stachyose were then successively added by the equal-increment method and mixed for 20 min, after which silicon dioxide was added and mixing was continued for another 10 min. The final material -mixing uniformity had an RSD of <5%. (3) Packaging and storage: the uniformly mixed material was aseptically packed into aluminum foil bags in a Class 10,000 clean environment at 1 g per bag, sealed, and stored in a cool and dry place, thereby obtaining the Bifidobacterium-Lactobacillus plantarum complex powder for regulating intestinal flora.

[0192] III. Verification test on the effect of the complex composition in regulating intestinal flora

[0193] 1. Test materials and animals DESCRIPTION

[0194] Test sample: the complex powder prepared in this embodiment. Test animals: 40 SPF-grade male BALB / c mice, 6-8 weeks old, weighing 18-22 g, adaptively fed for 3 days. The breeding environment was maintained at a temperature of 22±2 °C, a humidity of 50±5%, and a 12 h light / dark cycle, with free access to food and water.

[0195] 2. Grouping and treatment

[0196] The mice were randomly divided into 4 groups, with 10 mice in each group. The grouping and treatment methods were as follows:

[0197] Blank control group: intragastric administration of normal saline, 0.2 mL / mouse / day

[0198] Low-dose group: intragastric administration of a suspension of the complex powder at a dose of 0.5 g / kg bw / day

[0199] Medium-dose group: intragastric administration of a suspension of the complex powder at a dose of 1.0 g / kg bw / day

[0200] High-dose group: intragastric administration of a suspension of the complex powder at a dose of 2.0 g / kg bw / day

[0201] The intragastric administration was continued for 28 days. During this period, the body weight, feed intake, mental status, and defecation of the mice were recorded daily, and no mouse died or showed abnormal adverse reactions.

[0202] 3. Detection indicators and specific effect parameters

[0203] After the intragastric administration, fresh feces and cecal contents of the mice were aseptically collected, and the intestinal flora counts, short-chain fatty acid contents, cecal pH, and defecation-related indices were determined. The specific effect parameters were as follows: (1) Effect on the core intestinal flora in mice. In the blank control group, the counts of Bifidobacterium spp., Lactobacillus spp., Escherichia coli, and Clostridium perfringens in mouse feces were 7.21±0.35 IgCFU / g wet feces, 6.84±0.42 IgCFU / g wet feces, 8.32±0.38 IgCFU / g wet feces, and 5.76±0.29 IgCFU / g wet DESCRIPTION feces, respectively. After intervention, the low-dose group showed an increase in Bifidobacterium spp. to 8.98±0.28 IgCFU / g wet feces, representing a 24.55% increase over the blank control group; an increase in Lactobacillus spp. to 8.26±0.27 IgCFU / g wet feces, representing a 20.76% increase; a decrease in Escherichia coli to 7.18±0.29 IgCFU / g wet feces, representing a 13.70% decrease; and a decrease in Clostridium perfringens to 4.81±0.23 IgCFU / g wet feces, representing a 16.49% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in Bifidobacterium spp. to 9.93±0.27 IgCFU / g wet feces, representing a 37.73% increase; an increase in Lactobacillus spp. to 9.16±0.25 IgCFU / g wet feces, representing a 33.92% increase; a decrease in Escherichia coli to 6.53±0.26 IgCFU / g wet feces, representing a 21.51% decrease; and a decrease in Clostridium perfringens to 4.12±0.20 IgCFU / g wet feces, representing a 28.47% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in Bifidobacterium spp. to 10.86±0.24 IgCFU / g wet feces, representing a 50.62% increase; an increase in Lactobacillus spp. to 9.98±0.30 IgCFU / g wet feces, representing a 45.91% increase; a decrease in Escherichia coli to 5.81±0.28 IgCFU / g wet feces, representing a 30.17% decrease; and a decrease in Clostridium perfringens to 3.53±0.21 IgCFU / g wet feces, representing a 38.72% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the efficacy increased steadily with the administered dose.

[0204] (2) Effect on intestinal metabolism and environment in mice. In the blank control group, the total short-chain fatty acid content in the cecal contents was 32.15±2.47 pmol / g, including 18.62±1.53 pmol / g acetic acid, 7.54±0.82 pmol / g propionic acid, and 5.99±0.64 pmol / g butyric acid, and the pH of the DESCRIPTION cecal contents was 7.12±0.15. After intervention, the low-dose group showed an increase in total short-chain fatty acids to 50.27±2.18 pmol / g, representing a 56.36% increase over the blank control group, including 28.06±1.37 pmol / g acetic acid, 12.17±0.72 pmol / g propionic acid, and 10.04±0.56 pmol / g butyric acid, representing increases of 50.70%, 61.41%, and 67.61%, respectively; the pH of the cecal contents decreased to 6.42±0.11, representing a 9.83% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in total short-chain fatty acids to 65.84±2.16 pmol / g, representing a 104.79% increase, including 36.42±1.35 pmol / g acetic acid, 15.84±0.75 pmol / g propionic acid, and 13.58±0.59 pmol / g butyric acid, representing increases of 95.60%, 110.08%, and 126.71%, respectively; the pH of the cecal contents decreased to 5.99±0.09, representing a 15.87% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in total short-chain fatty acids to 81.16±2.11 pmol / g, representing a 152.44% increase, including 44.69±1.31 pmol / g acetic acid, 19.97±0.73 pmol / g propionic acid, and 16.50±0.57 pmol / g butyric acid, representing increases of 140.01%, 164.85%, and 175.46%, respectively; the pH of the cecal contents decreased to 5.67±0.09, representing a 20.37% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the optimization effect on the intestinal microecological metabolic environment was significantly enhanced.

[0205] (3) Effect on intestinal physiological functions in mice. In the blank control group, the average daily number of fecal pellets was 12.35±1.22, and the fecal moisture content was 42.36±2.15%. After intervention, the low-dose group showed an average daily fecal pellet number of 15.52±1.12, representing a 25.67% increase over the blank control group, and a fecal moisture content of 50.86±1.91%, representing a 20.07% increase. Both DESCRIPTION indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an average daily fecal pellet number of 17.69±1.06, representing a 43.24% increase, and a fecal moisture content of 57.12±1.82%, representing a 34.84% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an average daily fecal pellet number of 19.43±1.09, representing a 57.33% increase, and a fecal moisture content of 61.91±1.87%, representing a 46.15% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01), and the improvements in intestinal motility and excretory function were more pronounced.

[0206] (4) Food-use safety evaluation. During the 28-day continuous intragastric administration period, no mice in any dose group showed death, abnormal behavior, feeding disorder, or adverse intestinal reactions. There were no significant differences (P>0.05) between the dose groups and the blank control group in body -weight gain trend or major organ coefficients, indicating excellent food-use safety and no acute or subacute toxic side effect.

[0207] Embodiment 6

[0208] I. Formulation composition

[0209] Active probiotic component (all being single-strain powders prepared by vacuum freeze-drying)

[0210] Bifidobacterium longum lyophilized powder: 10 parts, viable count in powder >1. Ox 10A12 CFU / g

[0211] Bifidobacterium adolescentis lyophilized powder: 30 parts, viable count in powder >1.0xl0A12 CFU / g

[0212] Lactobacillus plantarum lyophilized powder: 10 parts, viable count in powder >1. Ox 10A12 CFU / g

[0213] The viable-count ratio of the above three strains is 1:3:1. Per gram of the final complex composition, the viable counts of Bifidobacterium longum and DESCRIPTION

[0214] Lactobacillus plantarum are both 1.0>< 10A10 CFU, and the viable count of Bifidobacterium adolescentis is 3.0 10A10 CFU.

[0215] Prebiotic component

[0216] Fructooligosaccharide: 180 parts

[0217] Stachyose: 120 parts

[0218] Filler

[0219] Maltodextrin: 630 parts

[0220] Glidant

[0221] Silicon dioxide: 20 parts

[0222] II. Preparation method

[0223] 1. Preparation of single-strain lyophilized powders

[0224] (1) Strain activation: glycerol-preserved strains of the corresponding microorganisms were taken and streaked onto MRS solid medium. Bifidobacterium longum and Bifidobacterium adolescentis were cultured anaerobically at 37 °C for 48 h, and Lactobacillus plantarum was cultured at 37 °C for 24 h. Single colonies with full morphology and good activity were picked and inoculated into MRS liquid medium, and were subcultured for two generations under the corresponding culture conditions to obtain seed cultures with stable activity. (2) Fermentation culture: the seed cultures were inoculated into optimized MRS fermentation medium at an inoculation amount of 3% (v / v). Bifidobacterium longum and Bifidobacterium adolescentis were anaerobically fermented at 37 °C for 24 h, and Lactobacillus plantarum was fermented at 37 °C for 18 h. After fermentation, the culture supernatant was discarded after centrifugation at 8000 r / min for 15 min using a disc centrifuge, and highly active bacterial sludge was collected. (3) Vacuum freeze-drying: a sterile freeze-drying protectant was added to the bacterial sludge, and the mass ratio of bacterial sludge to freeze-drying protectant was 1:3. After full mixing, a uniform bacterial suspension was prepared. The suspension was pre-frozen in a -40 °C low-temperature freezer DESCRIPTION for 8 h and then transferred to a vacuum freeze-dryer for drying for 24 h under conditions of a cold-trap temperature of -55 °C and a vacuum degree of 10 Pa, thereby obtaining the corresponding single-strain lyophilized powder, which was stored at 4 °C in a sealed light-proof state for later use.

[0225] 2. Preparation of the powder form of the complex composition

[0226] (1) Sieving pretreatment: the three single-strain lyophilized powders prepared above, together with fructooligosaccharide, stachyose, maltodextrin, and silicon dioxide, were separately passed through an 80-mesh sieve to remove agglomerates and ensure uniform material fineness for later use. (2) Equal-increment mixing: according to the formulation in parts by weight, the three strain powders and an equal mass of maltodextrin were first pre-mixed in a three-dimensional mixer for 10 min. The remaining maltodextrin, fructooligosaccharide, and stachyose were then successively added by the equal-increment method and mixed for 20 min, after which silicon dioxide was added and mixing was continued for another 10 min. The final material-mixing uniformity had an RSD of <5%. (3) Packaging and storage: the uniformly mixed material was aseptically packed into aluminum foil bags in a Class 10,000 clean environment at 1 g per bag, sealed, and stored in a cool and dry place, thereby obtaining the Bifidobacterium-Lactobacillus plantarum complex powder for regulating intestinal flora.

[0227] III. Verification test on the effect of the complex composition in regulating intestinal flora

[0228] 1. Test materials and animals

[0229] Test sample: the complex powder prepared in this embodiment. Test animals: 40 SPF-grade male BALB / c mice, 6-8 weeks old, weighing 18-22 g, adaptively fed for 3 days. The breeding environment was maintained at a temperature of 22±2 °C, a humidity of 50±5%, and a 12 h light / dark cycle, with free access to food and water.

[0230] 2. Grouping and treatment DESCRIPTION

[0231] The mice were randomly divided into 4 groups, with 10 mice in each group. The grouping and treatment methods were as follows:

[0232] Blank control group: intragastric administration of normal saline, 0.2 mL / mouse / day

[0233] Low-dose group: intragastric administration of a suspension of the complex powder at a dose of 0.5 g / kg bw / day

[0234] Medium-dose group: intragastric administration of a suspension of the complex powder at a dose of 1.0 g / kg bw / day

[0235] High-dose group: intragastric administration of a suspension of the complex powder at a dose of 2.0 g / kg bw / day

[0236] The intragastric administration was continued for 28 days. During this period, the body weight, feed intake, and defecation status of the mice were recorded daily, and no mouse died or showed abnormal adverse reactions.

[0237] 3. Detection indicators and specific effect parameters

[0238] After the intragastric administration, fresh feces and cecal contents of the mice were aseptically collected, and the intestinal flora counts, short-chain fatty acid contents, cecal pH, and defecation-related indices were determined. The specific effect parameters were as follows: (1) Effect on the core intestinal flora in mice. In the blank control group, the counts of Bifidobacterium spp., Lactobacillus spp., Escherichia coli, and Clostridium perfringens in mouse feces were 7.21±0.35 IgCFU / g wet feces, 6.84±0.42 IgCFU / g wet feces, 8.32±0.38 IgCFU / g wet feces, and 5.76±0.29 IgCFU / g wet feces, respectively. After intervention, the low-dose group showed an increase in Bifidobacterium spp. to 9.36±0.27 IgCFU / g wet feces, representing a 29.82% increase over the blank control group; an increase in Lactobacillus spp. to 8.49±0.25 IgCFU / g wet feces, representing a 24.12% increase; a decrease in Escherichia coli to 7.01±0.27 IgCFU / g wet feces, representing a 15.75% decrease; and a decrease in Clostridium perfringens to 4.62±0.22 IgCFU / g wet feces, representing a 19.79% decrease. All indices showed DESCRIPTION extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in Bifidobacterium spp. to 10.41±0.25 IgCFU / g wet feces, representing a 44.38% increase; an increase in Lactobacillus spp. to 9.39±0.23 IgCFU / g wet feces, representing a 37.28% increase; a decrease in Escherichia coli to 6.32±0.24 IgCFU / g wet feces, representing a 24.04% decrease; and a decrease in Clostridium perfringens to 3.94±0.19 IgCFU / g wet feces, representing a 31.60% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in Bifidobacterium spp. to 11.38±0.22 IgCFU / g wet feces, representing a 57.84% increase; an increase in Lactobacillus spp. to 10.28±0.27 IgCFU / g wet feces, representing a 50.29% increase; a decrease in Escherichia coli to 5.59±0.25 IgCFU / g wet feces, representing a 32.81% decrease; and a decrease in Clostridium perfringens to 3.31±0.20 IgCFU / g wet feces, representing a 42.53% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), reaching the optimal level in promoting the proliferation of beneficial bacteria and inhibiting harmful bacteria.

[0239] (2) Effect on intestinal metabolism and environment in mice. In the blank control group, the total short-chain fatty acid content in the cecal contents was 32.15±2.47 pmol / g, including 18.62±1.53 pmol / g acetic acid, 7.54±0.82 pmol / g propionic acid, and 5.99±0.64 pmol / g butyric acid, and the pH of the cecal contents was 7.12±0.15. After intervention, the low-dose group showed an increase in total short-chain fatty acids to 54.62±2.15 pmol / g, representing a 69.89% increase over the blank control group, including 30.29±1.35 pmol / g acetic acid, 13.26±0.71 pmol / g propionic acid, and 11.07±0.55 pmol / g butyric acid, representing increases of 62.67%, 75.86%, and 84.81%, respectively; the pH of the cecal contents decreased to 6.31±0.10, representing an 11.38% decrease. All indices showed extremely significant differences as compared DESCRIPTION with the blank control group (P<0.01). After intervention, the medium-dose group showed an increase in total short-chain fatty acids to 71.95 ±2.13 pmol / g, representing a 123.79% increase, including 39.37±1.33 pmol / g acetic acid, 17.49±0.73 pmol / g propionic acid, and 15.09±0.57 pmol / g butyric acid, representing increases of 111.44%, 131.96%, and 151.92%, respectively; the pH of the cecal contents decreased to 5.88±0.08, representing a 17.42% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an increase in total short-chain fatty acids to 89.48±2.09 pmol / g, representing a 178.32% increase, including 48.54±1.29 pmol / g acetic acid, 22.13 ±0.71 pmol / g propionic acid, and 18.81 ±0.55 pmol / g butyric acid, representing increases of 160.69%, 193.50%, and 214.02%, respectively; the pH of the cecal contents decreased to 5.52±0.08, representing a 22.47% decrease. All indices showed extremely significant differences as compared with the blank control group (P<0.01), and the optimization effect on the intestinal microecological metabolic environment was the most pronounced.

[0240] (3) Effect on intestinal physiological functions in mice. In the blank control group, the average daily number of fecal pellets was 12.35±1.22, and the fecal moisture content was 42.36±2.15%. After intervention, the low-dose group showed an average daily fecal pellet number of 16.04±1.10, representing a 29.88% increase over the blank control group, and a fecal moisture content of 52.35±1.89%, representing a 23.58% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the medium-dose group showed an average daily fecal pellet number of 18.37±1.04, representing a 48.74% increase, and a fecal moisture content of 58.94±1.80%, representing a 39.14% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01). After intervention, the high-dose group showed an average daily fecal pellet number of 20.33±1.07, representing a DESCRIPTION

[0241] 64.62% increase, and a fecal moisture content of 64.12±1.85%, representing a 51.37% increase. Both indices showed extremely significant differences as compared with the blank control group (P<0.01), and the improvement in intestinal motility function and constipation was the most significant.

[0242] (4) Food-use safety evaluation. During the 28-day continuous intragastric administration period, no mice in any dose group showed death, abnormal behavior, abnormal food or water intake, diarrhea, abdominal distension, or other intestinal adverse reactions. The mice showed normal body -weight gain, and there were no significant differences (P>0.05) in major organ coefficients as compared with the blank control group. No acute or subacute toxicity or potential health risk was observed, meeting the safety application standards for foods and pharmaceuticals.

[0243] The specific contents are shown in the following table:

[0244] In the six embodiments, one set only gradually adjusted the amount of Bifidobacterium longum, with viable-count ratios of 1.5:1 :1, 2:1: 1, and 3:1 :1, DESCRIPTION while the other set only gradually adjusted the amount of Bifidobacterium adolescentis, with ratios of 1:1.5: 1, 1:2: 1, and 1:3:1. All of these fall within the ratio range of claim 2, and the viable counts per gram of the finished products comply with the standard of claim 4. The tests show that, with increasing amounts of Bifidobacterium longum or Bifidobacterium adolescentis, the intestinal flora-regulating efficacy of the complex composition shows a dose-dependent enhancement in core flora regulation, optimization of the intestinal metabolic environment, and improvement of intestinal physiological function, while all embodiments also exhibit excellent food-use safety and no toxic side effect after 28 days of continuous intervention. This demonstrates that these two strains are core functional strains and that increasing their addition amounts can strengthen efficacy, thereby providing multi-gradient formulations for product development and verifying the rationality and inventiveness of the scope of protection of the claims.

[0245] The above descriptions are merely preferred embodiments of the present invention and do not limit the present invention in any form. Although the present invention has been disclosed above with reference to preferred embodiments, the present invention is not limited thereto. Any person skilled in the art may make some changes or modifications to equivalent embodiments by using the technical contents disclosed above without departing from the scope of the technical solutions of the present invention. Any simple modification, equivalent variation, or modification made to the above embodiments according to the technical essence of the present invention without departing from the contents of the technical solutions of the present invention shall still fall within the scope of the technical solutions of the present invention.

Claims

CLAIMS1. A Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora, characterized in that it is prepared from the following raw materials in parts by weight:The active probiotic component of the complex composition consists of Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum.

2. The Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 1, characterized in that the viable-count ratio of Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum is (1 -3): (1 -3): (1 -2).

3. The Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 2, characterized in that the viable-count ratio of Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum is 1:1 :1.

4. The Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 1, characterized in that, per gram of the complex composition, each of Bifidobacterium longum, Bifidobacterium adolescentis, and Lactobacillus plantarum has a viable count of 1.0xl0A9-5.0xl0A10 CFU.

5. The Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 1, characterized in that the active probiotic components are all single-strain powders prepared by vacuum freeze-drying.

6. The Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 1, characterized in that the complex composition further comprises a prebiotic, and the prebiotic is one or more selected from fructooligosaccharide, galactooligosaccharide, inulin, isomaltooligosaccharide, and stachyose.

7. The Bifidobacterium-Lactobacillus plantarum complex composition forCLAIMS regulating intestinal flora according to claim 1, characterized in that the complex composition further comprises a freeze-drying protectant, and the freeze-drying protectant is one or more selected from skimmed milk powder, trehalose, mannitol, and glycerol.

8. The Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 1, characterized in that the complex composition further comprises a food-grade or pharmaceutically acceptable filler, and the filler is one or more selected from maltodextrin, corn starch, and microcrystalline cellulose.

9. The Bifidobacterium-Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 1, characterized in that the complex composition further comprises a food-grade or pharmaceutically acceptable glidant, and the glidant is one or more selected from magnesium stearate, silicon dioxide, and micronized silica gel.

10. The Bifidobacterium -Lactobacillus plantarum complex composition for regulating intestinal flora according to claim 1, characterized in that the dosage form of the complex composition is any one of powder, capsule, tablet, granule, and oral liquid.

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