Bifidobacterium longum BL-113 for improving and promoting growth and development of adolescents, product and application

Abstract

The application provides long bifidobacterium BL-113 for improving and promoting growth and development of teenagers. The long bifidobacterium can improve or promote height growth of teenagers, improve or promote absorption and utilization of trace elements such as calcium and zinc by teenagers, improve intestinal flora, resist pathogenic bacteria, help intelligence development of teenagers, promote perfection of the immune system of teenagers, and has great application prospect in the fields of food and medicine.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of microorganisms, and particularly relates to Bifidobacterium longum BL-113 for improving and promoting growth and development of teenagers, products and application. BACKGROUND

[0002] Teenagers aged 6-18 years old are in the peak period of height growth, and if the opportunity window can be seized, the maximum height growth and growth and development can be achieved. Researches show that the height growth and growth and development of teenagers can be promoted by the following three ways: improving the levels of hormones and metabolic factors related to growth and development, promoting the absorption of nutritional elements and calcium, and regular exercise.

[0003] The intestinal microbiome can dynamically regulate the host physiology and development, and the changes in the abundance and composition thereof can affect the health, growth and development of the host and even cause diseases. A large number of studies have shown that the intestinal flora can regulate the immune system, host metabolism and endocrine environment, and maintain bone health and growth. Different metabolic products produced by the intestinal flora continuously regulate the levels of various metabolic factors, metabolic pathways and the absorption of nutritional elements, and thus affect bone growth.

[0004] Short-chain fatty acids (SCFA), including acetate, propionate, and butyrate, are the main products of indigestible carbohydrates broken down by intestinal bacteria. Many recent studies have confirmed that intestinal flora can play a role in the regulation of SCFA on bone homeostasis through various pathways. First, SCFA produced by some intestinal flora can accelerate the dissolution of minerals and the absorption of calcium ions by reducing the pH of the intestine, ultimately achieving the purpose of regulating bone density. Second, SCFA can induce the increase of insulin-like growth factor-1 (IGF-1), a key mediator that affects bone growth, which can promote the longitudinal growth of the femur. IGF-1 also has the effect of protecting brain neurons and promoting the growth of new motor neurons, which can improve the speed of learning new skills and motor ability. It can also improve the body's ability to absorb amino acids, glucose, and fatty acids to promote growth and development. In addition, the activity of histone deacetylase (HDAC) is inhibited by butyrate and propionate, which in turn affects the differentiation ability of Treg cells, ultimately inhibits osteoclast differentiation, prevents bone resorption, and promotes bone formation. The Wnt signaling pathway plays an important role in the development and metabolism of bones in animals and humans, and activation of this pathway can promote bone mass increase. SCFA can induce T cells to produce Wnt10b protein, thereby activating the Wnt signaling pathway, reducing osteoblast apoptosis and promoting osteoblast production, and increasing bone mass. On the other hand, intestinal microorganisms can polarize colonic macrophages to the M1 state, produce endogenous inflammatory cytokines, cause microbial-induced bystander effect (MIBE), and activate the Wnt / β-catenin signaling pathway with the help of TNF-α. This typical pathway regulates osteoblasts throughout human infancy to adulthood.

[0005] The increase of glucagon-like peptide-1 (GLP-1) can inhibit bone resorption and stimulate the proliferation and differentiation of osteoblasts. Secondary bile acids, which can activate G protein-coupled bile acid receptors (TGR5) as activators, can activate TRG5 to promote the production of GLP-1. Intestinal flora plays an important role in the conversion of primary bile acids to secondary bile acids. Lipopolysaccharide (LPS) derived from intestinal flora can up-regulate the expression of Sox-9 and inhibit the hypertrophy of growth plate chondrocytes. Intestinal flora can also affect the level of sex hormones in the host, thereby regulating bone homeostasis and promoting the growth and development of adolescents and children.

[0006] Probiotics are live symbiotic microorganisms that can regulate the intestinal microbial composition by colonizing the intestinal tract, increase bone mass in children, and promote their growth. On the other hand, probiotics can reduce intestinal permeability, inhibit intestinal inflammation, thereby protecting children's gastrointestinal health, enhancing resistance, and reducing intestinal flora imbalance. Bifidobacterium is a very common probiotic in the human body, which can improve the body's immunity and anti-infection ability, inhibit intestinal putrefactive pathogenic microorganisms, and regulate cell differentiation and apoptosis. With age, the number of bifidobacterium in the human intestine gradually decreases, and the dominant species changes from infant bifidobacterium and short bifidobacterium to adolescent bifidobacterium and long bifidobacterium. Long bifidobacterium is a probiotic widely used in functional foods. Increasing the intake of long bifidobacterium can change the intestinal microbial composition and maintain intestinal barrier homeostasis.

[0007] Therefore, it is of great significance for the healthy growth of children and adolescents to develop a probiotic that can promote the growth and development of adolescents, maintain intestinal homeostasis, improve sports ability, and enhance immunity. SUMMARY

[0008] The present application provides a long bifidobacterium and its application in promoting the growth and development of adolescents, increasing height, and enhancing immunity. The long bifidobacterium can maintain intestinal homeostasis, promote the absorption and utilization of nutrients, produce a variety of metabolites beneficial to the human body and involved in human metabolism, thereby increasing the bone mass and osteoblast number of adolescents, and promoting their height growth.

[0009] The first aspect of the present application provides a long bifidobacterium BL-113 for improving and promoting the growth and development of adolescents, characterized in that the long bifidobacterium BL-113 has been deposited with the China General Microbiological Culture Collection Center on November 2, 2023, at the address of No. 1, Beichen West Road, Hua-yuan District, Beijing, at the Institute of Microbiology of the Chinese Academy of Sciences, with the deposit number of CGMCC NO. 28848, and the classification name of Bifidobacterium longum.

[0010] The second aspect of the present application provides a preparation method of the long bifidobacterium BL-113 of claim 1, comprising:

[0011] (1) Screening and isolation of target strain

[0012] The fecal sample from a healthy infant not taking antibiotics is serially diluted in sterile normal saline, and the dilutions are selected for plate coating, the medium is selected as MRS medium, culture, observation and recording of bacterial morphology; single colonies with bifidobacterium colony morphology are subjected to gram staining and catalase experiment, and are subjected to microscopic examination; strains with positive gram staining results and negative catalase experiment (no bubbles are produced) are selected as the preliminary target strains;

[0013] (2) Strain preliminary identification

[0014] The above-mentioned target strains are added to the API 20A medium to prepare a bacterial suspension, and then are inoculated into the API 20A experimental strip, anaerobic culture, addition of additional reagents BCP, XYL, HER, H2O2 for result interpretation, and API 20A V3.0 is used for result identification, and strains identified as bifidobacterium are further purified;

[0015] (3) Strain identification

[0016] L-arabinose, maltose, salicin, sucrose, xylose and starch are added to the liquid basic medium as carbon sources, and the above-mentioned strains preliminarily identified as bifidobacterium are inoculated into the liquid medium with different carbon sources for culture, and BTB-MR reagent is used to check the acid production of different strains;

[0017] Strains capable of hydrolyzing L-arabinose, sucrose and maltose, partially hydrolyzing xylose, and not hydrolyzing starch and salicin are further cultured, and are sent to Shengong Bioengineering (Shanghai) Co., Ltd. for sequencing identification, the sequencing results are subjected to nucleic acid sequence comparison in NCBI, and finally the target bifidobacterium BL-113 is obtained.

[0018] In the step (1) of the above preparation method:

[0019] The concentration of the dilution is: 1x10 -6 ~ 1x10 -4 mg / ml;

[0020] The culture condition of the MRS medium is 36±1℃ for 48±2~72±2h.

[0021] In the step (2) of the above preparation method:

[0022] The anaerobic culture condition is 36±1℃ for 48±2~72±2h.

[0023] In the step (3) of the above preparation method:

[0024] The formula of the liquid base medium is casein peptone 0.5 g, tryptone 0.5 g, yeast extract 1.0 g, and salt mixed solution 4 ml (CaCl2 0.2 g, magnesium sulfate heptahydrate 0.48 g, potassium phosphate dibasic 1.0 g, potassium phosphate monobasic 1.0 g, sodium bicarbonate 10.0 g, and NaCl 2.0 g are dissolved in water and then diluted to 1000 ml).

[0025] The culture condition of the liquid medium is 36±1℃ for 48±2-72±2h.

[0026] The formula of the BTB-MR reagent is bromothymol blue (BTB) 0.2 g, methyl red (MR) 0.1 g, 95% ethanol 300 ml, and distilled water 200 ml.

[0027] The culture condition of the strain which does not hydrolyze starch and salicin is 36±1℃ for 48±2-72±2h.

[0028] The third aspect of the present application provides a Bifidobacterium longum BL-113 powder, which is prepared by the above preparation method.

[0029] The fourth aspect of the present application provides a preparation method of the above Bifidobacterium longum BL-113 powder, which comprises the following steps: inoculating the Bifidobacterium longum BL-113 obtained in step (3) of the above preparation method into MRS liquid medium, collecting the bacterial liquid and centrifuging, rinsing the bacterial slurry obtained by centrifugation, adding a protective agent and freeze-drying to obtain the Bifidobacterium longum BL-113 powder. The culture condition of the MRS liquid medium is 36±1℃ for 48±2-72±2h; the protective agent is a composite protective agent composed of skimmed milk powder 10%-15%, trehalose 20%-25%, and Vc-Na2% 4%; the mass ratio of the bacterial slurry and the protective agent is 1:(0.6-1.5); the freeze-drying conditions are as follows: the first stage: -40℃, 4h of pre-freezing, then vacuumizing; the second stage: 5℃, 10h; the third stage: 15℃, 10h, the vacuum degree is 10Pa, and the thickness is about 0.5cm.

[0030] The fifth aspect of the present application provides a Bifidobacterium longum preparation, which comprises the above Bifidobacterium longum BL-113 or the above Bifidobacterium longum BL-113 powder. The preparation can be a solid bacterial powder or a liquid drink.

[0031] The sixth aspect of the present application provides the application of the above-mentioned B. longum BL-113 or the above-mentioned B. longum BL-113 powder or the above-mentioned B. longum preparation in a food composition or a pharmaceutical composition for promoting the growth and development of young children, promoting the height growth of young children, anti-obesity of young children, improving the intestinal metabolic disorder of young children, promoting the normalization of intestinal microorganism composition, improving immunity, improving the sports ability and intelligence development of young children, and improving the absorption of trace elements of young children.

[0032] The B. longum BL-113 provided by the present application can effectively maintain the intestinal homeostasis of young children, maintain the intestinal microorganism composition at a better level, improve immunity, and promote the dual growth and development of intelligence and height of young children. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 is the survival rate detection result of the B. longum BL-113 in Example 1 in artificial gastric juice (pH = 3) and artificial intestinal juice (pH = 8).

[0034] Figure 2 is the change of intestinal flora diversity of Caenorhabditis elegans after being treated by B. longum BL-113 and BL-21 respectively.

[0035] Figure 3 is the analysis of the abundance of phylum level of intestinal flora of Caenorhabditis elegans after being treated by B. longum BL-113 and BL-21 respectively.

[0036] Figure 4 is the change of human height from 0 to 12 weeks before and after taking B. longum BL-113 or placebo. DETAILED DESCRIPTION

[0037] The specific embodiments of the present disclosure will be clearly and completely described below with reference to the accompanying drawings. It should be understood that the embodiments described herein are only a part of the embodiments of the present disclosure, which are used to illustrate and explain the present disclosure, and do not serve as any limitation on the present disclosure and its application or use.

[0038] Example 1

[0039] Screening, strain identification and preservation of B. longum

[0040] 1. Screening and purification

[0041] (1) Strain screening: 1 g of fecal sample from a healthy infant who has not taken antibiotics was continuously diluted in sterile physiological saline until the dilution reached 10 -6, select the appropriate concentration of diluent for plate coating, medium selection MRS medium, 36±1℃ culture 48±2h, observation and record the bacterial morphology. The single colony with bifidobacterium colony morphology was subjected to gram staining and catalase experiment, and was subjected to microscopic examination. The strains with positive gram staining results and negative catalase experiment (no bubble) were selected as the target strains.

[0042] (2) Strain purification: on MRS medium, the above strains were subjected to multiple streaking purification until single colonies with consistent morphology were obtained, and single colonies with colony morphology still consistent with bifidobacterium colony morphology characteristics were selected for culture as strains for subsequent physiological and biochemical identification.

[0043] The colony morphology of the above strain was observed to be smooth and soft, with a milky white color, complete and raised edges.

[0044] 2, identification

[0045] Total DNA was extracted from the isolated strain and sequenced by 16S rDNA (Shanghai Genechem Co., Ltd.), and the sequencing results were subjected to nucleic acid sequence alignment in NCBI, and finally the target Bifidobacterium longum BL-113 was obtained.

[0046] 3, preservation

[0047] The purified Bifidobacterium longum BL-113 was inoculated into MRS liquid medium and cultured at 36±1℃ for 48±2h. The bacterial liquid was collected and centrifuged at 8000g / min for 10min. The bacterial slurry obtained by centrifugation was rinsed twice, and then frozen and dried after adding a protective agent at a mass ratio of 1:1 to the bacterial slurry, to obtain Bifidobacterium longum BL-113 powder.

[0048] Example 2

[0049] This example describes the performance characteristics of Bifidobacterium longum BL-113.

[0050] 1, Bifidobacterium longum BL-113 basic situation

[0051] API identification was used to analyze the physiological and biochemical characteristics of the above isolated strains, such as carbohydrate utilization ability and enzyme activity. A sufficient amount of the above strain was added to the API 20A medium to prepare a bacterial suspension, then inoculated into the API 20A test strip, 36℃ anaerobic culture for 48h, and additional reagents BCP, XYL, HER, H2O2 were added for result interpretation. API 20A V3.0 was used for result identification, and the analysis results are shown in Table 1.

[0052] Table 1

[0053]

[0054] + means able to metabolize; - means unable to metabolize.

[0055] The above Table 1 results show that B. longum BL-113 is non-spore forming, gram-positive, catalase-negative, able to utilize lactose, raffinose, arabinose, sucrose, glucose, maltose, mannitol and salicin, and has hydrolytic action on gelatin and aesculin.

[0056] 2. Verification of the gastric juice and intestinal juice environmental tolerance and colonization ability of B. longum BL-113

[0057] Bifidobacterium is a kind of physiological beneficial bacteria, but its acid tolerance is generally not strong. In this embodiment, the tolerance of B. longum BL-113 to artificial gastric juice and intestinal juice is tested, and another B. longum BL-21 (purchased from Health Probiotics Wecare-probiotics Company) with better acid tolerance and capable of surviving through the gastrointestinal tract is used for comparison. The strain belongs to B. longum longum subsp. and can be used in the fields of food, dairy products and health products. Similar to the BL-113 strain, it also has the effects of improving intestinal flora composition (reducing the ratio of Firmicutes / Bacteroidetes), increasing intestinal peristalsis and reducing fat accumulation, and anti-obesity.

[0058] (1) Gastric juice and intestinal juice environmental tolerance detection

[0059] The initial concentration (i.e. the concentration at 0h) and survival rate of BL-113 strain and BL-21 strain in artificial gastric juice and artificial intestinal juice are shown in Table 3. The results show that after 1h treatment with artificial gastric juice (pH=3), the survival rate of BL-113 strain is 85.1%, which is higher than that of the control group BL-21 strain of 55.2%; after 3h treatment with artificial gastric juice (pH=3), the survival rate of BL-113 strain is 38.7%, which is higher than that of the control group BL-21 strain of 30.2%. After 1h treatment with intestinal juice (pH=8), the survival rate of BL-113 strain is 69.2%, which is higher than that of the control group BL-21 strain of 67.3%; after 3h treatment with intestinal juice (pH=8), the survival rate of BL-113 strain is 47.5%, which is higher than that of the control group BL-21 strain of 35.7%.

[0060] Table 2

[0061]

[0062] (2) Adhesion ability detection

[0063] Take 200ul of normal activity HT-29 cell solution to configure the initial number of cells to be 1×10 5HT-29 cell crawl pieces of 100 cfu / well were prepared in advance. The Bifidobacterium longum BL-113 and BL21 to be determined were respectively resuspended in DMEM medium and added to the 24-well plate containing cell crawl pieces, and the culture conditions were: 37°C, 5% CO2, incubated for 2h. The monolayer cells were washed 4 times with sterile PBS buffer, fixed with methanol for 10min and subjected to Gram staining. The adhesion was observed and recorded by microscope and the number of 50 cell-adherent probiotics was recorded.

[0064] Table 3

[0065] Test time 0.5h 1h BL-113 62.3 per 50 cells 57.4 per 50 cells BL-21 59.7 per 50 cells 55.2 per 50 cells

[0066] The results show that the number of HT-29 cells adhering to bacteria can still reach 57.4 / 50 cells as time increases, indicating that BL-113 has good adhesion ability and is better than the control group in terms of intestinal adhesion.

[0067] The above results show that the BL-113 strain can still survive well after being digested by artificial gastric juice and intestinal juice Figure 2 . Compared with the BL-21 strain, the BL-113 Bifidobacterium longum provided by the present application has stronger digestion liquid resistance and adhesion ability. Therefore, the BL-113 strain can smoothly pass through the digestive tract, enter the intestine and survive and colonize.

[0068] 3. Toxicology test and safety detection of Bifidobacterium longum BL-113

[0069] (1) Antibiotic resistance test

[0070] In order to verify the safety of the Bifidobacterium longum BL-113 provided by the present disclosure, the antibiotic resistance of the Bifidobacterium longum BL-113 was analyzed. The method for evaluating the antibiotic sensitivity of the Bifidobacterium longum BL-113 was selected as SN / T 1944-2007 "Determination of bacterial resistance in animals and their products". In this test, a total of 10 most common antibiotics were used, including gentamicin (GEN), erythromycin (ERM), streptomycin (STM), chloramphenicol (CP), tetracycline (TET), kanamycin (KAN), vancomycin (VAN), clindamycin (CLM), ampicillin (AMP) and quinupristin / dalfopristin (Q / D). The results show that the Bifidobacterium longum BL-113 strain provided by the present application is sensitive to the above antibiotics, indicating that the strain provided by the present application does not contain exogenous antibiotic resistance genes, has good safety and meets the requirements of the edible bacterial resistance evaluation specification.

[0071] (2) Toxicology experiment

[0072] Animal experiments were conducted to test the safety of the Bifidobacterium longum BL-113 provided by the present application.

[0073] Experimental group: 6-8 weeks old healthy male and female mice were given 1.0 x 10 11 The B. longum BL-113 culture stock was continuously administered to 6-8 weeks old healthy male and female mice in an amount of 1.0 x 10

[0074] Table 4

[0075]

[0076] Note: The numbers in Table 4 represent the number of dead animals.

[0077] The results showed that no toxic reactions or deaths were observed in the test mice in the experimental and control groups, and there was no statistical difference between the two groups (p>0.05).

[0078] Example 3

[0079] This example illustrates the effects of B. longum BL-113 on maintaining intestinal homeostasis, improving exercise capacity, and anti-obesity function.

[0080] Caenorhabditis elegans in vitro model test

[0081] Caenorhabditis elegans has a high degree of homology between genes and a wide range of similarities in metabolism, and is mainly colonized by intestinal bacteria. Therefore, Caenorhabditis elegans is an ideal model for studying the effects of human intestinal microorganisms, and the use of this interspecies model helps to understand the physiological relevance of human intestinal microbiota.

[0082] Caenorhabditis elegans fed with 1 ml of E. coli OP50 as a bacterial feed were used as a blank control, and Caenorhabditis elegans fed with 1 ml of E. coli OP50 mixed with 45 mg / L of B. longum BL-113 and B. longum BL-21, respectively, were used as bacterial feed. The growth and development of worms fed with different bacterial feeds, as well as the composition and abundance of intestinal microorganisms, were observed.

[0083] (1) Caenorhabditis elegans movement performance test: 15 worms were randomly selected from different groups and placed in M9 buffer solution. After 30 seconds of free swimming, their movement was observed under a microscope and recorded. The judgment standard was that one continuous rapid sinusoidal movement of the worm was counted as one movement. The results are shown in Table 6.

[0084] (2) Fat accumulation in Caenorhabditis elegans: different groups were stained using oil red O fat staining method, and the fat accumulation in different groups of worms was observed and recorded. The results are shown in Table 6.

[0085] (3) Determination of intestinal flora diversity: Several worms fed with different bacterial diets were selected and sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing to obtain the composition and abundance of intestinal flora of worms under different conditions.

[0086] (4) To test whether the growth and development of Caenorhabditis elegans fed with probiotics was affected by dietary restriction: Using a mature genetic model of dietary restriction, eat-2 (ad116), the worm has a mutation in the nicotinic acetylcholine channel that reduces food intake. The results showed that the growth and development of nematodes fed with probiotics was not affected by dietary restriction. Therefore, no additional dietary control is required when consuming products with added probiotics.

[0087] Table 5

[0088]

[0089] The results showed that *C. elegans* fed with *Bifidobacterium longum* BL-113 mixed with OP50 *Escherichia coli* exhibited better growth and development than other groups. At the phylum level, the Firmicutes / Bacteroidetes ratio decreased, while the Actinobacteria level increased in the *E. coli* OP50+BL-113 group (see...). Figure 3 At the genus level, the levels of *Femtobacter*, *Trichophyton*, and *Sartreus* decreased, while the levels of *Bifidobacterium*, *Lactobacillus*, and *Oscillatoria* increased. These increases in genus levels confirm that *Bifidobacterium longum* BL-113 can indeed increase the level of SCFAs in the gut, reduce intestinal tissue damage, and protect the intestinal barrier.

[0090] Example 4

[0091] This embodiment illustrates how Bifidobacterium longum BL-113 promotes growth and development, increases height, and enhances immunity.

[0092] Various metabolites produced by gut microbiota can promote bone growth and development. In particular, SCFAs can promote osteoblast production, bone mass increase, and bone formation through multiple pathways, thereby promoting bone development in adolescents and children.

[0093] 1. Validation of the effects on bone growth and height increase in a mouse model

[0094] Preparation: 30 mice of 2 months old with similar body size were selected and their leg bone length was measured, and then they were randomly divided into probiotic group, positive control group and blank control group, 10 mice in each group, and they were raised in SPF conditions. During the experiment, the experimental animals drank deionized water and freely ate. From the second week, the probiotic group was given 10 mL / kg.bw of mice by gavage every day, the positive control group was given 40 mg / kg.bw of calcium juice (Beijing Langdi Pharmaceutical Co., Ltd.) to mice by gavage every day, and the blank control group was given normal saline to mice by gavage every day. The body weight of mice was measured every week and recorded.

[0095] SCFA level determination: After continuous gavage feeding for 4 weeks, the mice were anesthetized and sacrificed. The mouse chest was opened, a syringe was inserted into the right ventricle to extract blood samples, and serum was prepared using a serum separation tube. Then the mouse short-chain fatty acid (SCFA) ELISA detection kit (Shanghai Yanqi Biological Technology Co., Ltd.) was used to detect the SCFA level in the mouse serum.

[0096] Bone state determination: After measuring the leg bone length of the mice, the bilateral femurs of the test mice were taken out and dried to constant weight in a 105℃ oven, and the femur density was measured. The femur bone calcium content was measured by atomic absorption spectrophotometry, and the results are shown in Table 6.

[0097] Table 6

[0098]

[0099] The results show that the mice in the probiotic group and the positive control group both show an increase in leg bone length, femur weight and bone calcium content. However, the probiotic group changes more significantly and the SCFA level in the serum increases significantly. Especially butyrate, which has the largest increase in concentration in the serum, this metabolite plays a very important role in bone development. The blank control group has no special performance.

[0100] 2. Clinical verification of growth and development promoting effect

[0101] 70 subjects aged 6-18 years old were recruited and randomly divided into probiotic group and placebo group for a 12-week controlled trial. The experiment followed the principles of randomization and double blinding, and all selected subjects did not take any form of probiotics or prebiotics within four weeks. The parents or guardians of the subjects signed the informed consent form. The probiotic group received Lactobacillus longus BL-113 strips daily, and the placebo group received placebo (maltodextrin) strips daily. The height and weight of the students were measured according to the Technical Specifications for Student Health Examination (GB26343-2010). Combined with body mass index (BMI) = weight (kg) / height (m) 2Height and weight measurements were taken and BMI values were calculated at the start of the test (i.e. week 0) and at week 12, respectively, and the results are shown in Table 7.

[0102] Table 7

[0103]

[0104] The results show that after 12 weeks, the subjects in the probiotic group and the placebo group showed some increase in weight and height, but the subjects in the probiotic group showed a more significant increase in height, a smaller change in weight, and a BMI index that remained within the normal range. Therefore, B. longum BL-113 does have the effect of promoting the growth and development of adolescent children.

[0105] 3. Experimental verification of immune health effects

[0106] An appropriate amount of B. longum BL-113 culture stock solution was added to the RPIM-1640 culture medium (containing 10% FBS) and cultured with Caco-2 cells, while the control group did not add probiotic culture. The culture conditions were 37°C and 5% CO2. The expression level of IL-6 in Caco-2 cells after adding probiotics was detected.

[0107] The experimental results showed that B. longum BL-113 increased the expression of IL-6 from 21 ng / ml to 97 ng / ml. Therefore, B. longum BL-113 does have the function of enhancing human immunity.

Claims

1. A Bifidobacterium longum BL-113 that improves and promotes the growth and development of adolescents, characterized in that, The aforementioned Bifidobacterium longum BL-113 was deposited on November 2, 2023, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC NO.28848 and classified as Bifidobacterium longum.

2. A Bifidobacterium longum BL-113 bacterial powder, characterized in that, Includes Bifidobacterium longum BL-113 as described in claim 1.

3. A method for preparing the Bifidobacterium longum BL-113 bacterial powder according to claim 2, characterized in that, The process includes the following steps: inoculating the Bifidobacterium longum BL-113 of claim 1 into MRS liquid culture medium, collecting the bacterial solution and centrifuging it, rinsing the centrifuged bacterial sludge, adding a protectant and freeze-drying it to obtain Bifidobacterium longum BL-113 bacterial powder.

4. The preparation method according to claim 3, characterized in that, The culture conditions for MRS liquid medium were 36±1℃ for 48±2~72±2h; the cryoprotectant was a composite cryoprotectant consisting of 10%~15% skim milk powder, 20%~25% trehalose, and 2%~4% Vc-Na; the mass ratio of bacterial sludge to cryoprotectant was 1:(0.6~1.5); the freeze-drying conditions were: first stage: -40℃, 4h pre-freezing followed by vacuuming; second stage: 5℃, 10h; third stage: 15℃, 10h, vacuum degree of 10Pa, thickness of approximately 0.5cm.

5. A Bifidobacterium longum preparation, characterized in that, Includes the Bifidobacterium longum BL-113 as described in claim 1 or the Bifidobacterium longum BL-113 powder as described in claim 2.

6. The use of the Bifidobacterium longum BL-113 of claim 1, or the Bifidobacterium longum BL-113 powder of claim 2, or the Bifidobacterium longum preparation of claim 5 in a pharmaceutical composition for promoting growth and development of adolescents and children, promoting height growth of adolescents and children, anti-obesity of adolescents and children, improving intestinal metabolic disorders of adolescents and children, promoting normalization of intestinal microbial composition, improving immunity, improving the motor ability and intellectual development of adolescents and children, and improving the absorption of trace elements in adolescents and children.

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