BIFIDOBACTERIUM ADOLESCENTIS AND USE THEREOF

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the priority of Chinese Patent Application No. 201710963441.5, filed on Oct. 17, 2017, and the disclosures of which are hereby incorporated by reference.

FIELD

The present disclosure relates to the field of microbe technology, specifically to a Bifidobacterium adolescentis and use thereof, especially to a Bifidobacterium adolescentis that is capable of modulating intestinal flora, modulating brain-gut axis and significantly alleviating metabolic syndrome, and use thereof.

BACKGROUND

Recent years, with the developing economy, life-styles of people in many countries have changed obviously. With the amount of physical activity has decreased, obesity rate has increased significantly, and prevalence rates of diabetes and metabolic syndrome have increased by a large margin. An epidemiology survey shows that 20% to 30% of the adults across the globe are suffering from metabolic syndrome. In 2013, a multicenter, multistage stratified, large-scale sampling survey carried out by Chinese Diabetes Society of Chinese Medical Association showed that among people over 20 years old in large and medium-sized cities, towns and countryside of China, prevalence rates of metabolic syndrome in men and women were 16.7% and 11.7%, respectively, and the total prevalence rate was 13.7%. Furthermore, the rate was continuously increasing. Analyses show that age, blood pressure, diabetes family history, obesity, hyperlipidemia, male, low income and little exercise are main related risk factors for metabolic syndrome. The survey also found that the rates of the overweight and the obesity among people have increased by a large margin, and prevalence rate of metabolic syndrome of male is significant higher than that of female at the same age among middle-aged crowd.

Metabolic syndrome is a clinical syndrome, which has simultaneous symptoms of central obesity, fasting blood glucose rising, high blood pressure, decrease of high-density lipoprotein cholesterol and increase of triacylglycerol, in which numerous hazardous factors basing on the abnormal pathological changes of carbohydrate metabolism, lipid metabolism and protein metabolism aggregates, and which promotes development of diabetes (type II) and cardiovascular diseases such as atherosclerosis and so on. Due to metabolic syndrome is a pathological condition in which numerous metabolic abnormalities aggregates, its occurrence is relates to insulin resistance, becoming a hot spot in the research field of cardiovascular diseases and diabetes, and raising many controversies at home and abroad. In addition, metabolic syndrome accompanies with disorder of intestinal microecosystem. Disorder of intestinal microecosystem may further lead to disorder of intestinal functions, nerve center functions and peripheral nerve functions, for example, intestinal inflammation, inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS), abnormality of neurotransmitter 5-hydroxytryptamine level and level of some hormone and so on. Many researches have shown that abundances of intestinal microbes of some genus have intimate connection with intestinal diseases, for example, the abundance of Blautia genus will rise in intestinal flora of IBS patient. At the same time, researches also show that mental diseases such as depression, anxiety and so on have intimate connection with metabolic syndrome, intestinal flora disorder, and low 5-hydroxytryptamine level in human body. Improving 5-hydroxytryptamine level in peripheral blood helps increasing neurotransmitter level of central nervous system, so that relieving symptoms of anxiety, depression and so on.

At present, all the drug treatments of metabolic syndrome aim to decrease all kinds of risk factors, and these drugs include: anti-obesity drugs for weight loss; dimethylbiguanide and thiazolidinediones for reducing insulin resistance; sulfonylurea and rosiglitazone for controlling blood glucose; fibrate and statin for improving disorder of lipid metabolism; captopril, amlodipine and so on for controlling the blood pressure; drugs for treating intestinal diseases such as IBD, IBS and so on, including glucocorticoid, immunosuppressant, psychotropic drugs and so on; drugs for mental diseases such as anxiety, depression and so on, including selective serotonin reuptake inhibitors such as paroxetine, noradrenaline, and specific 5-hydroxytryptamine antidepressant drugs such as mirtazapine and so on. All of the medicines above have certain therapeutic effects, but as the conditions getting worse, the amounts of medicine used increase, the interactions between the medicines as well as the toxic and side effects of medicines become significant, leading to adverse reaction of digestive tract and showing liver and renal toxicity in some degree. In consideration of problems of the medicines, early intervention in metabolic syndrome, intestinal disease and mental disease can effectively decrease onset risk of cardiovascular and cerebrovascular diseases, diabetes, inflammatory bowel diseases, depression and so on.

Probiotics are edible microbes that are beneficial to human health, which have potential functions of alleviating abnormal metabolism of blood glucose and blood lipid, and modulating intestinal flora proportion and brain-gut axis. Thus, there is important social and economy value to research and develop probiotic products that can effectively intervene the occurrence and development of metabolic syndrome, intestinal diseases and mental diseases.

At present, there is no patent about using probiotics to increase 5-hydroxytryptamine level so as to regulate brain-gut axis and relieve anxiety and depression. Although there is related patent application (CN107083339A) that discloses adding Blautia bacteria to protect piglets from diarrhea, there is no patent about modulating the abundance of Blautia genus bacteria in intestinal tract so as to alleviate intestinal disease by the uptake of edible microbes (list of bacterium that can be permitted to be used in food, infant food, health products by National Health Commission of the People's Republic of China, 2014). In addition, there are some patents or patent application relating to compositions and preparation method thereof for preventing and curing metabolic syndrome. For example, CN104906263A discloses a composition consisting of tea polyphenol, procyanidin and POTENTILLAE DISCOLORIS HERBA extract, which is used to treat metabolic syndrome. CN105796674A discloses a traditional Chinese medicine composition comprising PLANTAGINIS SEMEN, COPTIDIS RHIZOMA and so on, which is capable of preventing and curing metabolic syndrome. In addition, a few patents relate to probiotics-containing compositions that are used to improve metabolic syndrome, and the methods for preparing the same. For example, CN105816623A discloses a probiotic-fermented traditional Chinese medicine composition being used to cure and improve metabolic syndrome, which is made from traditional Chinese medicine such as PANACIS QUINQUEFOLII RADIX, DIOSCOREAE RHIZOMA, MOUTAN CORTEX, PORIA by extracting and fermenting the extract with probiotics. All the above patents and patent applications are using traditional Chinese medicine components or a mixture of bacteria and traditional Chinese medicine to alleviate metabolic syndrome, in which the bacteria and the functions of the bacteria are not clear. CN105567586A discloses a Lactobacillus plantarum NCU116 with antidiabetic function, which achieves antidiabetic effects through modulating body blood glucose, blood lipid, hormone level and body metabolism. The Lactobacillus plantarum is screened and selected from bacteria in kimchi instead of human sources. No evidence shows that Lactobacillus plantarum can colonize in human intestinal tract and take effects. So far, there is no a human-sourced individual probiotic (such as Bifidobacterium) that can colonize in human intestinal tract to relieve the metabolic syndrome, or to alleviate symptoms such as hyperglycemia, hyperlipidemia, intestinal flora imbalance, intestinal inflammation and so on, and related mental diseases.

SUMMARY

In view of above, an object of the present disclosure is to solve the problems in the prior art by providing a probiotics. The probiotics can colonize in intestinal tract of human body, improve 5-hydroxytryptamine level in peripheral blood, regulate brain-gut axis and recovering testosterone level in serum back to normal level, normalize abnormal abundances of Blautia genus and Turicibacter genus in intestinal flora, improve metabolic syndrome, relieve hyperglycemia, hyperlipidemia and inflammation of liver and duodenum, liver fibrosis and other symptoms.

The present disclosure provides a strain CCFM8630 of Bifidobacterium adolescentis, which is deposited at China General Microbiological Culture Collection Center (CGMCC, Address: Beijing Institute of Microbiology, Chinese Academy of Sciences, No. 1, Beichen West Road, Chaoyang District, Beijing, China) on Jul. 7, 2017, with an accession number CGMCC 14395.

In one embodiment, the present disclosure studies effect of strain CCFM8630 of Bifidobacterium adolescentis on intestinal flora imbalance caused by high-carbohydrate and high-fat diet. The results show that the uptake of strain CCFM8630 of Bifidobacterium adolescentis significantly recovers relative abundances of Bifidobacterium genus, Turicibacter genus and Blautia genus in disordered intestinal microbes of rat feces, and the intervention effect is obviously better than that of Bifidobacterium animalis BB12.

In one embodiment, the present disclosure studies protection effects of strain CCFM8630 of Bifidobacterium adolescentis on tissue damages of liver, duodenum and so on in rat with metabolic syndrome. The results show that administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage significantly improves symptoms caused by high-fat high-starch diet, such as hepatocyte microvesicular steatosis, interstitial inflammatory cell infiltration, early fibrosis of liver tissue, duodenum villi broadening, interstitial edema, increasing of inflammatory cells and so on in rats, and the intervention effect is obviously better than that of Bifidobacterium animalis BB12.

In one embodiment, the present disclosure studies effect of strain CCFM8630 of Bifidobacterium adolescentis on (fasting) blood glucose level of rat with metabolic syndrome. The results show that administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage obviously decreases the fasting blood glucose level of the model rat close to the blank control group. The ability of strain CCFM8630 of Bifidobacterium adolescentis on decreasing fasting blood glucose level of rat is better than that of rosiglitazone and Bifidobacterium animalis BB12 by intragastric gavage administration.

In one embodiment, the present disclosure studies effect of strain CCFM8630 of Bifidobacterium adolescentis on oral glucose tolerance of rat with metabolic syndrome. The results show that strain CCFM8630 of Bifidobacterium adolescentis significantly improves oral glucose tolerance and the effect is better than that of Bifidobacterium animalis BB12, indicating that strain CCFM8630 of Bifidobacterium adolescentis can further decrease glucose level by improving glucose tolerance.

In one embodiment, the present disclosure studies effects of strain CCFM8630 of Bifidobacterium adolescentis on total cholesterol (TC) and triglyceride (TG) in serum of rat with metabolic syndrome, respectively. The results show that administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage decreases levels of total cholesterol and triglyceride in serum.

Further, in one embodiment, the present disclosure studies effects of strain CCFM8630 of Bifidobacterium adolescentis on 5-HT and testosterone level in serum of rat with metabolic syndrome, respectively. The results show that administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage improves 5-hydroxytryptamine (5-HT) level in rat serum, and reduces testosterone in serum back to normal level.

Therefore, the present disclosure provides use of strain CCFM8630 of Bifidobacterium adolescentis in preparing products that can improve metabolic syndrome, regulate intestinal flora or regulate brain-gut axis.

Therein, the improving metabolic syndrome is to relieve symptoms of hyperglycemia and hyperlipidemia, inflammation of liver and duodenum, and liver tissue fibrosis; the modulating intestinal flora is to normalize abnormal abundances of Blautia genus and Turicibacter genus in intestinal flora; and the modulating brain-gut axis as well as relieving anxiety and depression is to increase 5-hydroxytryptamine level in peripheral blood.

The product of the present disclosure includes but is not limited to health food or pharmaceutical preparation.

Therein, the health food includes but is not limited to microbial agent or fermented food.

Further, the present disclosure provides a microbial agent comprising the strain CCFM8630 of Bifidobacterium adolescentis.

Preferably, the viable count of the strain CCFM8630 of Bifidobacterium adolescentis in the microbial agent is more than 10⁶CFU/g

The microbial agent of the present disclosure can be prepared by routine methods.

In some embodiments, the method for preparing the microbial agent is:

inoculating the strain CCFM8630 of Bifidobacterium adolescentis to a modified MRS medium at an inoculum size of 2 to 4 wt %, culturing for 24 to 48h at a temperature between 35 and 39° C. under anaerobic conditions, collecting bacteria, resuspending the bacteria with a protectant to reach a bacterial density of 10¹⁰CFU/mL, culturing the suspension at 37° C. for 50 to 70 minutes under anaerobic conditions, and drying the resulting culture.

Therein, the modified MRS medium (mMRS) in the present disclosure is a MRS medium containing 0.05% of L-cysteine hydrochloride. The specific method for preparing the medium is: dissolving 10 g of tryptone, 10 g of beef extract, 5 g of yeast powder, 20 g of glucose, 5 g of sodium acetate, 2 g of ammonium citrate dibasic, 2 g of dipotassium hydrogen phosphate, 0.5 g of magnesium sulfate heptahydrate, 1 mL of Tween-80, 0.25 g of manganese sulfate monohydrate and 0.5 g of cysteine hydrochloride in water, diluting the mixture to 1000 mL with water, modulating the pH to 6.5, and autoclaving at 119-123° C. for 15 to 25 minutes.

The protectant in the method of the present disclosure is an aqueous solution that contains 100 g/L to 150 g/L of nonfat milk powder, 100 g/L to 150 g/L of maltodextrin and 140 g/L to 160 g/L of trehalose. That is, the protectant consists of nonfat milk powder, maltodextrin, trehalose and water, wherein the concentration of nonfat milk powder is from 100 g/L to 150 g/, the concentration of maltodextrin is from 100 g/L to 150 g/L, and the concentration of trehalose is from 140 g/L to 160 g/L.

Preferably, in the method of the present disclosure, bacteria collected after culturing in the modified MRS medium are subjected to washing with phosphate buffer solution for 2 to 4 times, and pH of the phosphate buffer solution is from 6.8 to 7.2.

The drying of the method in the present disclosure can be any of the drying procedures for bacteria solution, for example vacuum freeze-drying. In some embodiments, the drying of the present disclosure is vacuum freeze-drying after pre-freezing the bacteria at −15 to −20° C. for 8 to 14h.

The present disclosure also provides a fermented food which is produced by using the strain CCFM8630 of Bifidobacterium adolescentis as a starter culture.

The fermented food is fermented dairy products, fermented bean products or fermented fruit and vegetable products.

The fermented dairy products include but are not limited to yogurt, sour cream and cheese. The fermented bean products include but are not limited to soymilk, fermented beans and bean paste. The fruits and vegetables in the fermented fruit and vegetable products include but are not limited to cucumber, carrot, beet, celery and cabbage.

The present disclosure also provides a pharmaceutical preparation, comprising an effective amount of the strain CCFM8630 of Bifidobacterium adolescentis and a pharmaceutically acceptable adjuvant.

The pharmaceutically acceptable adjuvant is one or more selected from the group consisting of filler, adhesive, wetting agent, disintegrating agent, lubricant, and flavoring agent.

In some embodiments of the present disclosure, the pharmaceutical preparation is a granule, a capsule, a tablet, a pill or an oral liquid.

The beneficial technical effects of the present disclosure are as follows.

The strain CCFM8630 of Bifidobacterium adolescentis of the present disclosure significantly increases neurotransmitter 5-hydroxytryptamine (5-HT) level in peripheral blood of rat, regulates brain-gut axis, relieves mental illnesses related to metabolic syndrome, for example anxiety, depression and so on, recovers the hormone level, for example testosterone and so on in peripheral blood of rat caused by high-fat high-starch diet, recovers abundances of Bifidobacterium genus, Blautia genus and Turicibacter genus in abnormal intestinal flora of rat caused by high-fat high-starch diet. In addition, strain CCFM8630 of Bifidobacterium adolescentis has pretty good tolerance to simulated gastrointestinal fluid, and quickly colonizes in intestinal, significantly alleviates pathology damages of tissues, such as liver, duodenum and so on of rat with metabolic syndrome caused by high-fat high-starch diet; significantly improves oral glucose tolerance of rat with metabolic syndrome and decreases the under curve area of glucose tolerance test; significantly increases triglyceride and total cholesterol levels in serum of rat with metabolic syndrome caused by high-fat high-starch diet. The strain CCFM8630 of Bifidobacterium adolescentis of the present disclosure can be used to prepare health foods or medicines that improve metabolic syndrome, regulates intestinal flora, relieves irritable bowel syndrome, regulates brain-gut axis and alleviates mental illness such as anxiety, depression and so on, which has a pretty wide application prospect.

Description of Microbiological Preservation

CCFM8630, classification name: Bifidobacterium adolescentis, is deposited at China General Microbiological Culture Collection Center (CGMCC, Address: Beijing Institute of Microbiology, Chinese Academy of Sciences, No. 1, Beichen West Road, Chaoyang District, Beijing, China) on Jul. 7, 2017, with an accession number CGMCC 14395.

BRIEF DESCRIPTION OF DRAWINGS

In order to describe the technical solutions in the examples of the present disclosure or the conventional art more clearly, the accompanying drawings used in description of the embodiments or the prior art will be illustrated briefly.

FIG. 1 shows colony morphology of strain CCFM8630 of Bifidobacterium adolescentis.

FIG. 2 shows effects of strain CCFM8630 of Bifidobacterium adolescentis on tissue structure of liver in rat with metabolic syndrome.

FIG. 3 shows effects of strain CCFM8630 of Bifidobacterium adolescentis on tissue structure of duodenum in rat with metabolic syndrome.

FIG. 4 shows effects of strain CCFM8630 of Bifidobacterium adolescentis on abundances of some intestinal microbes in rat with metabolic syndrome; and there are significant differences (P<0.05) between a, b and c groups.

FIG. 5 shows effects of strain CCFM8630 of Bifidobacterium adolescentis on oral glucose tolerance in rat with metabolic syndrome; and there are significant differences (P<0.05) between a, b and c groups.

FIG. 6 is a curve showing changes of blood glucose level in oral glucose tolerance test.

FIG. 7 shows the area under the curve (AUC_glucose) in oral glucose tolerance test.

FIG. 8 shows effects of strain CCFM8630 of Bifidobacterium adolescentis on total cholesterol (TC) in serum of rat with metabolic syndrome; and there are significant differences (P<0.05) between a, b and c groups.

FIG. 9 shows effects of strain CCFM8630 of Bifidobacterium adolescentis on triglyceride (TG) in serum of rat with metabolic syndrome; and there are significant differences (P<0.05) between a, b and c groups.

FIG. 10 shows effects of strain CCFM8630 of Bifidobacterium adolescentis on 5-hydroxytryptamine (5-HT) and testosterone levels of rat with metabolic syndrome; and there are significant differences (P<0.05) between a, b and c groups.

DETAILED DESCRIPTION

The present disclosure discloses a Bifidobacterium adolescentis and use thereof. One of ordinary skill in the art can learn from the contents herein and improve the process parameters appropriately. In particular, it shall be noted that all the similar substitutions and modifications are apparent to one of ordinary skill in the art and are to be considered within the scope of the present invention. The method and product of the present invention have been described with preferred examples. It is apparent that one of the ordinary skill in the art can make change or modify the combination to the method and product of the present invention without departing from the spirit, scope and spirit of the invention, therefore realizing and applying the techniques of the present invention. www

The strain CCFM8630 of Bifidobacterium adolescentis of the present disclosure has the following biology properties.

(1) Bacterium properties: Gram staining positive, without spore, not moving

(2) Colony properties: colonies are formed after 36-hour anaerobic culture, the diameters of the colonies are between 0.5 and 2 mm, the front view is a circle and the side view is a protuberance, and the edge is smooth, the color is milky and non-transparent, the surface is moist and smooth, and no pigment is produce. See FIG. 1.

(3) Growth properties: the bacteria are cultured in mMRS medium under anaerobic condition for about 24-hour at constant temperature of 37° C. to reach log phase.

(4) Good tolerance to simulate gastrointestinal fluid.

(5) Significantly improve pathological tissue damages such as liver, duodenum and so on of rat with from metabolic syndrome.

(6) Significantly improve oral glucose tolerance of rat with metabolic syndrome.

(7) Decrease area under the curve in glucose tolerance test.

(8) Regulate the levels of triglyceride and total cholesterol in serum back to normal level.

(9) Increase 5-hydroxytryptamine (5-HT) level in peripheral blood and regulate testosterone to normal level.

(10) Significantly recover abundances of Bifidobacterium genus, Turicibacter genus, Blautia genus and so on in abnormal intestinal flora caused by high-fat high-starch diet.

The strain CCFM8630 of Bifidobacterium adolescentis of the present disclosure is obtained by the following method.

I. Isolation and Screening of Lactobacillus

(1) 1 g of fresh feces was diluted in gradient, spreaded on solid mMRS medium, and cultured at 37° C. for 72 hours under anaerobic condition. The feces were obtained from a 95-year-old male from Changshou Village, Chaihu Town in Zhongxiang City of Hubei Province, China.

(2) Morphology of the colonies were observed and recorded, and single colony was picked out and purified by streaking.

(3) The bacteria were cultured at 37° C. for 48 hours in mMRS medium, and the colonies obtained were subjected to Gram Staining. The morphologies of the colonies were recorded.

(4) The Gram-negative strains and Gram-positive cocci were discarded, the Gram-positive bacilli were selected.

(5) The bacteria were subjected to catalase analyzing, the catalase-positive strains were discarded and catalase-negative strains were retained.

II. Preliminary Identification of Bifidobacterium: Fructose-6-Phosphoketolase Assay

(1) The Lactobacillus obtained in Step I was cultured in mMRS liquid medium for 24 hours, and then 1 mL of the culture was taken and centrifuged at 8000 rpm for 2 minutes.

(2) The bacterial pellet was washed two times with 0.05M KH₂PO₄solution (pH 6.5) containing 0.05% (mass percentage) of cysteine hydrochloride.

(3) The bacteria was resuspended in 200 μL of the phosphate buffer above with an addition of 0.25% (mass percentage) Triton X-100.

(4) 50 μL mixture solution of 6 mg/mL sodium fluoride and 10 mg/mL sodium iodoacetate, and 50 μL of 80 mg/mL fructose-6-phosphoric acid were added and incubated at 37° C. for 1 hour.

(5) 300 μL of 0.139 g/mL hydroxylamine hydrochloride solution (pH 6.5) was added and placed at room temperature for 10 minutes.

(6) 200 μL of 15% (mass percentage) trichloroacetic acid and 200 μL of 4M HCl were added, respectively.

(7) 200 μL of 0.1M HCl containing 5% (mass percentage) ferric chloride was added. The color of the system turned red quickly, indicating a F6PPK-positive reaction. Therefore, the bacteria were initially identified as Bifidobacterium.

III. Molecular Biological Identification of Bifidobacterium

(1) Genome extraction of single bacterium (according to operation procedures of TIANamp Bacteria DNA kit)

A. The Lactobacillus obtained in Step II was cultured overnight. 1 mL culture was put into a 1.5 mL centrifuge tube and centrifuged at 10,000 rpm (˜11,500×g) for 1 minute. The supernatant was removed as much as possible.

B. 180 μL buffer (20 mg/mL lysozyme solution with 20 mM Tris (pH 8.0), 2 mM Na₂-EDTA, and 1.2% Triton) was added to the bacteria and incubated at 37° C. for more than 30 minutes. (The lysozyme solution should be prepared by dissolving lysozyme dry powder in the buffer, or the lysozyme would be inactive.

C. 20 μL Proteinase K solution was added to the tube and mixed well.

D. 220 μL buffer GB was added, shaken for 15 seconds, and placed at 70° C. for 10 minutes. The solution turned clean. The tube was centrifuged for a few seconds to remove water drops on inner wall of the tube.

E. 220 μL absolute alcohol was added adequately shaken for 15 seconds. Flocculent precipitates maybe appear. The tube was centrifuged for a few seconds to remove water drops on inner wall of the tube.

F. The solution and flocculent precipitate obtained in the last step were put into an adsorption column CB3 (the absorption column was disposed in a collecting tube), and subjected to centrifugation at 12,000 rpm (˜13,400×g) for 30 seconds. The flow-through liquor was discarded, and the adsorption column was put back into the collecting tube.

G. 500 μL of buffer GD (check for absolute alcohol adding before use) was added to the adsorption column CB3. The column was centrifuged at 12,000 rpm (˜13,400×g) for 30 seconds. The flow-through liquor was discarded, and the adsorption column was put back into the collecting tube.

H. 600 μL of washing solution PW (check for absolute alcohol adding before use) was added to the adsorption column CB3. The column was centrifuged at 12,000 rpm (˜13,400×g) for 30 seconds. The flow-through liquor was discarded, and the adsorption column was put back into the collecting tube. This step was repeated once.

I. The adsorption column CB3 was put back into the collecting tube, centrifuged at 12,000 rpm (˜13,400×g) for 2 minutes, and the flow-through liquor was discarded. The adsorption column CB3 was placed at room temperature for a few minutes to let the adsorption column totally dry.

J. The adsorption column CB3 was transferred to a clean centrifugal tube, and 50 to 200 μL of elution buffer TE was dropped to the middle of the adsorption film. The adsorption column was placed at room temperature for 2 to 5 minutes, and then centrifuged at 12,000 rpm (˜13,400×g) for 2 minutes. The eluted solution was collected into a centrifuge tube.

(2) Whole Genome Sequencing

The extracted whole genome was sent to a professional sequencing company and a second-generation sequencer was used to sequence the whole bacterial genome. The sequencing results were subjected to similarity comparison by BLAST software among GeneBank database. The results show that the strain provided by the present disclosure is a Bifidobacterium adolescentis belonging to Bifidobacterium genus, but different from known Bifidobacterium adolescentis, so that it is identified as a new strain. By blastn algorithm, the genome of strain CCFM8630 of Bifidobacterium adolescentis was compared with that of standard Bifidobacterium adolescentis strain ATCC15703 (https://www.ncbi.nlm.nih.gov/genome/?term=683), 259 genes in total show differences, which are shown in Table 1.

TABLE 1

Differential genes between strain CCFM8630 of Bifidobacterium adolescentis

and standard strain ATCC15703 of Bifidobacterium adolescentis

Correlation

Gene ID
Nr Database Article ID
Note of the Result
Similarity
Length

Z25_GM000009
gi|748204304|ref|WP_039774992.1|
hypothetical protein[Bifidobacterium
76.423
123

adolescentis]

Z25_GM000013
gi|822625123|ref|WP_046999178.1|
hypothetical protein [Bifidobacterium
87.5
128

adolescentis]

Z25_GM000014
gi|822625124|ref|WP_046999179.1|
transcriptional regulator
95.062
81

[Bifidobacterium adolescentis]

Z25_GM000019
gi|748204310| ref|WP_039774998.1|
hypothetical protein [Bifidobacterium
100
81

adolescentis]

Z25_GM000024
gi|748204315|ref|WP_039775003.1|
single-stranded DNA-binding
81.921
177

protein [Bifidobacterium

adolescentis]

Z25_GM000026
gi|748204318|ref|WP_039775006.1|
hypothetical protein [Bifidobacterium
88.889
63

adolescentis]

Z25_GM000029
gi|748204322|ref|WP_039775010.11
hypothetical protein [Bifidobacterium
35
80

adolescentis]

Z25_GM000031
gi|673003250|gb|KFI98296.1|
hypothetical protein BSTER_0878
50.909
55

[Bifidobacterium stercoris JCM 15918]

Z25_GM000032
gi|154085053|gb|EDN84098.1|
hypothetical protein BIFADO_01031
54.545
44

[Bifidobacterium adolescentis L2-32]

Z25_GM000034
gi|748204324|ref|WP_039775012.1|
hypothetical protein [Bifidobacterium
89.787
235

adolescentis]

Z25_GM000035
gi|748204325|ref|WP_039775013.1|
HNH endonuclease [Bifidobacterium
94.495
109

adolescentis]

Z25_GM000038
gi|917738554|ref|WP_052252740.1|
hypothetical protein [Bifidobacterium
34.874
476

adolescentis]

Z25_GM000039
gi|917738475|ref|WP_052252661.1|
hypothetical protein [Bifidobacterium
43.805
226

adolescentis]

Z25_GM000047
gi|917738481|ref|WP_052252667.1|
hypothetical protein [Bifidobacterium
100
206

adolescentis]

Z25_GM000048
gi|748204335|ref|WP_039775023.1|
hypothetical protein [Bifidobacterium
97.619
126

adolescentis]

Z25_GM000049
gi|748204337|ref|WP_039775025.1|
hypothetical protein [Bifidobacterium
100
66

adolescentis]

Z25_GM000053
gi|747124343|gb|AJE05765.1|
Phage tail fiber protein
91.367
139

[Bifidobacterium adolescentis]

Z25_GM000053
gi|747124343|gb|AJE05765.1|
Phage tail fiber protein
85.185
54

[Bifidobacterium adolescentis]

Z25_GM000054
gi|917738485|ref|WP_052252671.1|
hypothetical protein [Bifidobacterium
60.938
192

adolescentis]

Z25_GM000058
gi|917738487|ref|WP_052252673.1|
hypothetical protein [Bifidobacterium
94.318
88

adolescentis]

Z25_GM000059
gi|747124349|gb|AJE05771.1|
Integrase [Bifidobacterium
99.251
267

adolescentis]

Z25_GM000079
gi|489906072|ref|WP_003809496.1|
hypothetical protein [Bifidobacterium
100
78

adolescentis]

Z25_GM000121
gi|747124414|gb|AJE05836.1|
excinuclease subunit A [Bifidobacterium
100
768

adolescentis]

Z25_GM000122
gi|489905980|ref|WP_003809404.1|
MULTISPECIES: GNAT family
100
137

N-acetyltransferase [Bifidobacterium]

Z25_GM000123
gi|705407595|ref|WP_033499438.1|
alpha/beta hydrolase [Bifidobacterium
99.7
333

adolescentis]

Z25_GM000155
gi|154083897|gb|EDN82942.1|
hypothetical protein BIFADO_01229
100
125

[Bifidobacterium adolescentis L2-32]

Z25_GM000158
gi|500063247|ref|WP_011743164.1|
hypothetical protein [Bifidobacterium
100
502

adolescentis]

Z25_GM000206
gi|489905824|ref|WP_003809249.1|
transcriptional regulator [Bifidobacterium
100
198

adolescentis]

Z25_GM000208
gi|737015357|ref|WP_035010987.1|
hypothetical protein [Bifidobacterium
100
49

adolescentis]

Z25_GM000214
gi|489905803|ref|WP_003809228.1|
heat-inducible transcriptional
100
381

repressor HrcA [Bifidobacterium

adolescentis]

Z25_GM000236
gi|673003360|gb|KFI98406.1|
hypothetical protein BSTER_6020
100
36

[Bifidobacterium stercoris

JCM 15918]

Z25_GM000237
gi|748204479| ref|WP_039775167.1|
hypothetical protein [Bifidobacterium
100
84

adolescentis]

Z25_GM000262
gi|500063319| ref|WP_011743236.1|
DNA-deoxyinosine glycosylase
99.441
179

[Bifidobacterium adolescentis]

Z25_GM000266
gi|747124562| gb|AJE05984.1|
hypothetical protein BBMN23_1012
100
38

[Bifidobacterium adolescentis]

Z25_GM000267
gi|757771530|ref|WP_042991197.1|
hypothetical protein [Bifidobacterium
100
71

adolescentis]

Z25_GM000273
gi|740659212|ref|WP_038444512.1|
hypothetical protein [Bifidobacterium
100
112

adolescentis]

Z25_GM000279
gi|118765497|dbj|BAF39676.1|
transcriptional regulator [Bifidobacterium
100
92

adolescentis ATCC 15703]

Z25_GM000289
gi|489931370|ref|WP_003834689.1|
MULTISPECIES: cell division
100
294

protein Fic [Bifidobacterium]

Z25_GM000297
gi|489906825|ref|WP_003810247.1|
hypothetical protein [Bifidobacterium
100
910

adolescentis]

Z25_GM000298
gi|489906826|ref|WP_003810248.1|
hypothetical protein [Bifidobacterium
100
329

adolescentis]

Z25_GM000300
gi|154082753|gb|EDN81798.1|
DNA (cytosine-5-)-methyltransferase
100
234

[Bifidobacterium adolescentis L2-32]

Z25_GM000302
gi|489906833|ref|WP_003810255.1|
hypothetical protein [Bifidobacterium
99.852
674

adolescentis]

Z25_GM000303
gi|740659235|ref|WP_038444535.1|
hypothetical protein [Bifidobacterium
99.342
152

adolescentis]

Z25_GM000304
gi|154082758|gb|EDN81803.1|
hypothetical protein BIFADO_01928
100
33

[Bifidobacterium adolescentis L2-32]

Z25_GM000306
gi|154082760|gb|EDN81805.1|
hypothetical protein BIFADO_01930
100
42

[Bifidobacterium adolescentis L2-32]

Z25_GM000308
gi|154082763|gb|EDN81808.1|
hypothetical protein BIFADO_01933
81.395
43

[Bifidobacterium adolescentis L2-32]

Z25_GM000313
gi|705409744|ref|WP_033500282.1|
NAD(+) kinase [Bifidobacterium
98.726
314

adolescentis]

Z25_GM000321
gi|673003127|gb|KFI98173.1|
transporter [Bifidobacterium
100
243

stercoris JCM 15918]

Z25_GM000322
gi|673003126|gb|KFI98172.1|
HD superfamily metal-dependent
99.408
169

phosphohydrolase [Bifidobacterium

stercoris JCM 15918]

Z25_GM000325
gi|489906866|ref|WP_003810288.1|
arginine repressor [Bifidobacterium
100
172

adolescentis]

Z25_GM000331
gi|489906872|ref|WP_003810294.1|
hypothetical protein [Bifidobacterium
97.357
227

adolescentis]

Z25_GM000332
gi|740659259|ref|WP_038444559.1|
phenylalanine-tRNA ligase subunit
99.654
867

beta [Bifidobacterium adolescentis]

Z25_GM000341
gi|489906895|ref|WP_003810317.1|
membrane protein [Bifidobacterium
100
262

adolescentis]

Z25_GM000345
gi|489906905|ref|WP_003810327.1|
cobalt ABC transporter permease
100
292

[Bifidobacterium adolescentis]

Z25_GM000356
gi|747124649|gb|AJE06071.1|
putative glycosyltransferase
100
360

[Bifidobacterium adolescentis]

Z25_GM000361
gi|671342920|gb|IAII76436.1|
helicase [Bifidobacterium
99.603
1261

adolescentis]

Z25_GM000364
gi|154082824|gb|EDN81869.1|
Ion channel [Bifidobacterium
100
241

adolescentis L2-32]

Z25_GM000373
gi|917265320|ref|WP_051872032.1|
hypothetical protein [Bifidobacterium
100
248

adolescentis]

Z25_GM000375
gi|751368899|gb|KIM01425.1|
hypothetical protein LU08_05925
100
61

[Bifidobacterium adolescentis]

Z25_GM000380
gi|118765587|dbj|BAF39766.1|
putative DNA polymerase III
99.517
207

epsilon subunit [Bifidobacterium

adolescentis ATCC 15703]

Z25_GM000381
gi|751368903|gb|KIM01429.1|
homocysteine methyltransferase
100
294

[Bifidobacterium adolescentis]

Z25_GM000383
gi|747124679|gb|AJE06101.1|
amino acid transport protein
100
109

[Bifidobacterium adolescentis]

Z25_GM000390
gi|118765595|dbj|BAF39774.1|
hypothetical protein BAD_0993
98.551
69

[Bifidobacterium adolescentis

ATCC 15703]

Z25_GM000401
gi|118765606|dbj|BAF39785.1|
hypothetical protein BAD_1004
100
103

[Bifidobacterium adolescentis

ATCC 15703]

Z25_GM000415
gi|154082938|gb|EDN81983.1|
hypothetical protein BIFADO_02108
100
43

[Bifidobacterium adolescentis L2-32]

Z25_GM000420
gi|489907169|ref|WP_003810591.1|
recombination regulator RecX
100
231

[Bifidobacterium adolescentis]

Z25_GM000421
gi|489907170|ref|WP_003810592.1|
DNA recombination/repair protein
100
337

RecA [Bifidobacterium adolescentis]

Z25_GM000423
gi|489907175|ref|WP_003810597.1|
transcriptional regulator [Bifidobacterium
100
169

adolescentis]

Z25_GM000432
gi|740659333|ref|WP_038444633.1|
membrane protein [Bifidobacterium
100
324

adolescentis]

Z25_GM000464
gi|547082934|ref|WP_021913854.1|
hypothetical protein [Bifidobacterium
100
72

adolescentis]

Z25_GM000470
gi|747124764|gb|AJE06186.1|
hypothetical protein BBMN23_1214
100
267

[Bifidobacterium adolescentis]

Z25_GM000472
gi|489907274|ref|WP_003810696.1|
site-specific tyrosine recombinase
100
317

XerD [Bifidobacterium

adolescentis]

Z25_GM000474
gi|489907277|ref|WP_003810699.1|
MULTISPECIES: 50S ribosomal
100
64

protein L35 [Bifidobacterium]

Z25_GM000482
gi|489907294|ref|WP_003810716.1|
hypothetical protein [Bifidobacterium
100
79

adolescentis]

Z25_GM000496
gi|118765699|dbj|BAF39878.1|
DEAD/DEAH box helicase-like
95.745
47

[Bifidobacterium adolescentis

ATCC 15703]

Z25_GM000506
gi|705408403|ref|WP_033499726.1|
cell division protein [Bifidobacterium
99.833
600

adolescentis]

Z25_GM000509
gi|547083061|ref|WP_021913877.1|
division/cell wall cluster tran-
100
171

scriptional repressor MraZ

[Bifidobacterium adolescentis]

Z25_GM000510
gi|740659382|ref|WP_038444682.1|
hypothetical protein [Bifidobacterium
98.462
65

adolescentis]

Z25_GM000513
gi|489907353|ref|WP_003810775.1|
transcriptional regulator NrdR
100
150

[Bifidobacterium adolescentis]

Z25_GM000514
gi|751368419|gb|KIM00984.1|
peptidoglycan-binding protein
100
85

[Bifidobacterium adolescentis]

Z25_GM000561
gi|489904214|ref|WP_003807643.1|
MULTISPECIES: nucleotidyltrans-
100
263

ferase [Bifidobacterium]

Z25_GM000577
gi|736988102|ref|WP_034984157.1|
hypothetical protein [Bifidobacterium
95.683
278

adolescentis]

Z25_GM000579
gi|751369378|gb|KIM01887.1|
hypothetical protein LU08_03085
100
59

[Bifidobacterium adolescentis]

Z25_GM000580
gi|657871024|ref|WP_029575615.1|
MULTISPECIES: polyketide
95
60

cyclase [Terrabacteria group]

Z25_GM000581
gi|490750313|ref|WP_004612621.1|
MULTISPECIES: competence
100
74

protein TfoX [Firmicutes]

Z25_GM000582
gi|154084484|gb|EDN83529.1|
hypothetical protein BIFADO_00438
100
262

[Bifidobacterium adolescentis L2-32]

Z25_GM000583
gi|154084485|gb|EDN83530.1|
hypothetical protein BIFADO_00439
100
48

[Bifidobacterium adolescentis L2-32]

Z25_GM000590
gi|489904267|ref|WP_003807696.1|
50S ribosomal protein L32
100
64

[Bifidobacterium adolescentis]

Z25_GM000655
gi|154084571|gb|EDN83616.1|
hypothetical protein BIFADO_00529
97.778
90

[Bifidobacterium adolescentis L2-32]

Z25_GM000657
gi|489904451|ref|WP_003807880.1|
FmdB family transcriptional regulator
98.333
60

[Bifidobacterium adolescentis]

Z25_GM000667
gi|748204088|ref|WP_039774776.1|
hypothetical protein [Bifidobacterium
96.639
119

adolescentis]

Z25_GM000668
gi|740658926|ref|WP_038444227.1|
ABC transporter substrate-binding
30.162
431

protein [Bifidobacterium adolescentis]

Z25_GM000669
gi|736880360|ref|WP_034879947.1|
peptide ABC transporter permease
98.485
330

[Bifidobacterium pseudocatenulatum]

Z25_GM000670
gi|736880694|ref|WP_034880276.1|
ABC transporter permease
100
191

[Bifidobacterium pseudocatenulatum]

Z25_GM000671
gi|705411392|ref|WP_033500823.1|
ABC transporter ATP-binding
77.061
279

protein [Bifidobacterium

kashiwanohense]

Z25_GM000752
gi|740658820|ref|WP_038444121.1|
oleate hydratase [Bifidobacterium
99.681
626

adolescentis]

Z25_GM000764
gi|489906187|ref|WP_003809611.1|
DoxX [Bifidobacterium adolescentis]
95.181
83

Z25_GM000770
gi|705409239|ref|WP_033500070.1|
hypothetical protein [Bifidobacterium
93.443
122

adolescentis]

Z25_GM000771
gi|736508496|ref|WP_034524549.1|
hypothetical protein [Bifidobacterium
91.971
137

adolescentis]

Z25_GM000772
gi|673000940|gb|KFI96007.1|
hypothetical protein BSTER_1719
91.071
56

[Bifidobacterium stercoris

JCM 15918]

Z25_GM000774
gi|489905678|ref|WP_003809103.1|
nitrate reductase [Bifidobacterium
81.865
193

adolescentis]

Z25_GM000776
gi|705409248|ref|WP_033500073.1|
hypothetical protein [Bifidobacterium
68.121
298

adolescentis]

Z25_GM000780
gi|489906219|ref|WP_003809643.1|
hypothetical protein [Bifidobacterium
41.791
201

adolescentis]

Z25_GM000782
gi|705409263|ref|WP_033500080.1|
hypothetical protein [Bifidobacterium
69.663
89

adolescentis]

Z25_GM000783
gi|705409266|ref|WP_033500081.1|
hypothetical protein [Bifidobacterium
58.333
132

adolescentis]

Z25_GM000784
gi|705409268|ref|WP_033500082.1|
hypothetical protein [Bifidobacterium
99.415
171

adolescentis]

Z25_GM000789
gi|705409278|ref|WP_033500087.1|
terminase [Bifidobacterium
100
537

adolescentis]

Z25_GM000791
gi|705409284|ref|WP_033500090.1|
PhnA protein [Bifidobacterium
97.083
240

adolescentis]

Z25_GM000793
gi|705409286|ref|WP_033500091.1|
GTP-binding protein
90.385
52

[Bifidobacterium adolescentis]

Z25_GM000795
gi|154084106|gb|EDN83151.1|
hypothetical protein BIFADO_00046
92.982
57

[Bifidobacterium adolescentis L2-32]

Z25_GM000796
gi|673000966|gb|KFI96033.1|
hypothetical protein BSTER_1745
90.909
44

[Bifidobacterium stercoris

JCM 15918]

Z25_GM000798
gi|705409296|ref|WP_033500096.1|
hypothetical protein [Bifidobacterium
97.619
84

adolescentis]

Z25_GM000799
gi|705409298|ref|WP_033500097.1|
hypothetical protein [Bifidobacterium
91.176
68

adolescentis]

Z25_GM000800
gi|489905582|ref|WP_003809007.1|
methyltransferase [Bifidobacterium
100
161

adolescentis]

Z25_GM000801
gi|705409306|ref|WP_033500101.1|
hypothetical protein [Bifidobacterium
100
125

adolescentis]

Z25_GM000802
gi|673000975|gb|KFI96042.1|
putative ferredoxin [Bifidobacterium
98.148
54

stercoris JCM 15918]

Z25_GM000808
gi|489905562|ref|WP_003808987.1|
hypothetical protein [Bifidobacterium
83.051
59

adolescentis]

Z25_GM000809
gi|705409319|ref|WP_033500107.1|
hypothetical protein [Bifidobacterium
91.429
70

adolescentis]

Z25_GM000810
gi|705409321|ref|WP_033500108.1|
antirepressor [Bifidobacterium
60.902
266

adolescentis]

Z25_GM000811
gi|489903579|ref|WP_003807009.1|
hypothetical protein [Bifidobacterium
77
200

adolescentis]

Z25_GM000815
gi|705409328|ref|WP_033500111.11
hypothetical protein [Bifidobacterium
97.333
75

adolescentis]

Z25_GM000817
gi|489905540|ref|WP_003808965.1|
anhydro-N-acetylmuramyl-tripep-
90.476
42

tide amidase [Bifidobacterium

adolescentis]

Z25_GM000818
gi|489905537|ref|WP_003808962.1|
transcriptional regulator [Bifidobacterium
93.333
120

adolescentis]

Z25_GM000822
gi|917316857|ref|WP_051923569.1|
hypothetical protein [Bifidobacterium
31.288
163

adolescentis]

Z25_GM000834
gi|154085026|gb|EDN84071.1|
hypothetical protein BIFADO_01003
100
28

[Bifidobacterium adolescentis L2-32]

Z25_GM000835
gi|489905505|ref|WP_003808930.1|
general stress protein
100
171

[Bifidobacterium adolescentis]

Z25_GM000856
gi|747124227|gb|AJE05649.1|
Type I restriction-modification
40.845
213

system specificity subunit S

[Bifidobacterium adolescentis]

Z25_GM000856
gi|747124227|gb|AJE05649.1|
Type I restriction-modification
31.313
198

system specificity subunit S

[Bifidobacterium adolescentis]

Z25_GM000858
gi|737014132|ref|WP_035009793.1|
integrase [Bifidobacterium
99.675
308

adolescentis]

Z25_GM000870
gi|154084979|gb|EDN84024.1|
hypothetical protein BIFADO_00956
96.078
102

[Bifidobacterium adolescentis L2-32]

Z25_GM000873
gi|154084979|gb|EDN84024.1|
hypothetical protein BIFADO_00956
76.19
105

[Bifidobacterium adolescentis L2-32]

Z25_GM000874
gi|154084978|gb|EDN84023.1|
hypothetical protein BIFADO_00955
90.476
63

[Bifidobacterium adolescentis L2-32]

Z25_GM000881
gi|489905358|ref|WP_003808783.1|
histidine kinase [Bifidobacterium
98.619
869

adolescentis]

Z25_GM000915
gi|154084913|gb|EDN83958.1|
universal stress family protein
100
335

[Bifidobacterium adolescentis L2-32]

Z25_GM000921
gi|489905270|ref|WP_003808696.1|
type VII secretion protein
100
96

[Bifidobacterium adolescentis]

Z25_GM000926
gi|489905257|ref|WP_003808683.1|
hypothetical protein [Bifidobacterium
100
317

adolescentis]

Z25_GM000927
gi|489905255|ref|WP_003808681.11
hypothetical protein [Bifidobacterium
100
174

adolescentis]

Z25_GM000928
gi|489905253|ref|WP_003808679.1|
VWA domain-containing protein
99.718
355

[Bifidobacterium adolescentis]

Z25_GM000930
gi|489905249|ref|WP_003808675.1|
cell surface protein [Bifidobacterium
99.459
185

adolescentis]

Z25_GM000932
gi|489905244|ref|WP_003808670.1|
hypothetical protein [Bifidobacterium
99.229
519

adolescentis]

Z25_GM000943
gi|500063070|ref|WP_011742987.1|
phosphoserine phosphatase SerB
100
227

[Bifidobacterium adolescentis]

Z25_GM000966
gi|489905157|ref|WP_003808583.1|
hypothetical protein [Bifidobacterium
100
75

adolescentis]

Z25_GM001016
gi|751369644|gb|KIM02148.1|
fimbrial protein [Bifidobacterium
83.333
534

adolescentis]

Z25_GM001017
gi|748205409|ref|WP_039776097.11
hypothetical protein [Bifidobacterium
100
184

adolescentis]

Z25_GM001020
gi|671342004|gb|AII75520.1|
fimbriae protein with LPXTG
98.077
728

motif and von Willebrand factor

typeA domain [Bifidobacterium

adolescentis]

Z25_GM001042
gi|673001223|gb|KFI96287.1|
TetR-type transcriptional regulator
100
225

[Bifidobacterium stercoris

JCM 15918]

Z25_GM001043
gi|705408020|ref|WP_033499573.11
phage infection protein [Bifidobacterium
99.863
731

adolescentis]

Z25_GM001046
gi|1489903990|ref|WP_003807419.1|
NrdH-redoxin [Bifidobacterium
100
78

adolescentis]

Z25_GM001051
gi|154084351|gb|EDN83396.1|
hypothetical protein BIFADO_00303
98.246
57

[Bifidobacterium adolescentis L2-32]

Z25_GM001067
gi|489933463|ref|WP_003836777.1|
MULTISPECIES: ABC transporter
100
268

permease [Bifidobacterium]

Z25_GM001068
gi|748203972|ref|WP_039774660.1|
hypothetical protein [Bifidobacterium
99.205
629

adolescentis]

Z25_GM001069
gi|747123689|gb|AJE05111.1|
Hypothetical protein BBMN23_0139
98.467
1566

[Bifidobacterium adolescentis]

Z25_GM001073
gi|748203974|ref|WP_039774662.1|
hypothetical protein [Bifidobacterium
99.225
258

adolescentis]

Z25_GM001149
gi|154084733|gb|EDN83778.1|
hypothetical protein BIFADO_00702
100
123

[Bifidobacterium adolescentis L2-32]

Z25_GM001157
gi|489904938|ref|WP_003808365.1|
acetylesterase [Bifidobacterium
100
272

adolescentis]

Z25_GM001178
gi|489904982|ref|WP_003808409.1|
peptidase Ml3 [Bifidobacterium
100
696

adolescentis]

Z25_GM001194
gi|747124065|gb|AJE05487.1|
hypothetical protein BBMN23_0515
97.674
43

[Bifidobacterium adolescentis]

Z25_GM001220
gi|747125053|gb|AJE06475.1|
putative transport protein
98.879
535

[Bifidobacterium adolescentis]

Z25_GM001244
gi|489906423|ref|WP_003809846.1|
hypothetical protein [Bifidobacterium
100
332

adolescentis]

Z25_GM001245
gi|705409011|ref|WP_033499974.1|
hypothetical protein [Bifidobacterium
100
363

adolescentis]

Z25_GM001248
gi|651887219|ref|WP_026646861.1|
ATP synthase F0F1 subunit
100
92

epsilon [Bifidobacterium

ruminantium]

Z25_GM001252
gi|489906439|ref|WP_003809862.1|
ATP synthase subunit delta
100
275

[Bifidobacterium adolescentis]

Z25_GM001266
gi|748205152|ref|WP_039775840.1|
hypothetical protein [Bifidobacterium
100
643

adolescentis]

Z25_GM001268
gi|740659818|ref|WP_038445118.1|
AsnC family transcriptional
100
158

regulator [Bifidobacterium

adolescentis]

Z25_GM001270
gi|154083598|gb|EDN82643.1|
hypothetical protein BIFADO_01696
96.774
93

[Bifidobacterium adolescentis L2-32]

Z25_GM001276
gi|500063830|ref|WP_011743747.1|
aspartyl/glutamyl-tRNA(Asn/Gln)
100
98

amidotransferase subunit C

[Bifidobacterium adolescentis]

Z25_GM001282
gi|489906654|ref|WP_003810077.1|
2-hydroxyhepta-2,4-diene-1,7-dioate
99.634
273

isomerase [Bifidobacterium

adolescentis]

Z25_GM001286
gi|547062388|ref|WP_021912974.1|
YggS family pyridoxal
100
272

phosphate enzyme

[Bifidobacterium adolescentis]

Z25_GM001301
gi|917316766|ref|WP_051923478.1|
hypothetical protein [Bifidobacterium
38.806
67

adolescentis]

Z25_GM001321
gi|154084201|gb|EDN83246.1|
16S rRNA methyltransferase GidB
100
247

[Bifidobacterium adolescentis L2-32]

Z25_GM001341
gi|489903773|ref|WP_003807203.1|
hypothetical protein [Bifidobacterium
100
155

adolescentis]

Z25_GM001366
gi|747123591|gb|AJE05013.1|
hypothetical protein BBMN23_0041
100
49

[Bifidobacterium adolescentis]

Z25_GM001377
gi|822624998|ref|WP_046999053.1|
hypothetical protein [Bifidobacterium
99.715
351

adolescentis]

Z25_GM001381
gi|671341961|gb|AII75477.1|
putative membrane protein
98.708
774

[Bifidobacterium adolescentis]

Z25_GM001382
gi|500062698|ref|WP_011742615.1|
ABC transporter [Bifidobacterium
98.893
903

adolescentis]

Z25_GM001387
gi|154083139|gb|EDN82184.1|
hypothetical protein BIFADO_02312
100
42

[Bifidobacterium adolescentis L2-32]

Z25_GM001396
gi|748204836|ref|WP_039775524.1|
phosphoesterase [Bifidobacterium
100
268

adolescentis]

Z25_GM001397
gi|489907517|ref|WP_003810939.1|
ATP-binding protein
100
74

[Bifidobacterium adolescentis]

Z25_GM001403
gi|705408881|ref|WP_033499925.1|
cell division protein DivIVA
98.473
524

[Bifidobacterium adolescentis]

Z25_GM001406
gi|154083118|gb|EDN82163.1|
hypothetical protein BIFADO_02290
100
541

[Bifidobacterium adolescentis L2-32]

Z25_GM001421
gi|154083101|gb|EDN82146.1|
hypothetical protein BIFADO_02273
98.889
90

[Bifidobacterium adolescentis L2-32]

Z25_GM001422
gi|822625273|ref|WP_046999328.1|
phosphate ABC transporter
99.519
208

[Bifidobacterium adolescentis]

Z25_GM001426
gi|748204805|ref|WP_039775493.1|
hypothetical protein [Bifidobacterium
99.667
601

adolescentis]

Z25_GM001427
gi|489907456|ref|WP_003810878.1|
type II secretion system
100
591

protein E [Bifidobacterium

adolescentis]

Z25_GM001428
gi|489907454|ref|WP_003810876.1|
prepilin-type N-terminal
100
264

cleavage/methylation domain-

containing protein [Bifidobacterium

adolescentis]

Z25_GM001429
gi|822582229|gb|KLE27724.1|
hypothetical protein AAX71_05000
100
191

[Bifidobacterium adolescentis]

Z25_GM001432
gi|917738515|ref|WP_052252701.1|
hypothetical protein [Bifidobacterium
100
805

adolescentis]

Z25_GM001436
gi|489907436|ref|WP_003810858.1|
hypothetical protein [Bifidobacterium
100
203

adolescentis]

Z25_GM001441
gi|500063552|ref|WP_011743469.1|
hemolysin [Bifidobacterium
98.63
73

adolescentis]

Z25_GM001446
gi|118765731|dbj|BAF39910.1|
histidinol dehydrogenase
100
27

[Bifidobacterium adolescentis ATCC

15703]

Z25_GM001449
gi|547074020|ref|WP_021913401.1|
imidazoleglycerol-phosphate
100
199

dehydratase [Bifidobacterium

adolescentis]

Z25_GM001456
gi|740659385|ref|WP_038444685.1|
hypothetical protein [Bifidobacterium
98.895
181

adolescentis]

Z25_GM001457
gi|751368642|gb|KIM01183.1|
hypothetical protein LU08_07305
84.651
215

[Bifidobacterium adolescentis]

Z25_GM001458
gi|747124820|gb|AJE06242.1|
hypothetical protein BBMN23_1270
90
90

[Bifidobacterium adolescentis]

Z25_GM001460
gi|671343072|gb|AII76588.1|
hypothetical protein BADO_1174
91.892
185

[Bifidobacterium adolescentis]

Z25_GM001467
gi|489907882|ref|WP_003811303.1|
hypothetical protein [Bifidobacterium
100
83

adolescentis]

Z25_GM001481
gi|489907849|ref|WP_003811270.1|
sugar ABC transporter permease
100
299

[Bifidobacterium adolescentis]

Z25_GM001483
gi|154083302|gb|EDN82347.1|
haloacid dehalogenase-like hydrolase
99.598
249

[Bifidobacterium adolescentis L2-32]

Z25_GM001489
gi|737015814|ref|WP_035011431.1|
adhesin [Bifidobacterium
98.382
309

adolescentis]

Z25_GM001496
gi|705409435|ref|WP_033500154.1|
MerR family transcriptional regulator
100
178

[Bifidobacterium adolescentis]

Z25_GM001497
gi|651887462|ref|WP_026647098.1|
MULTISPECIES: hypothetical
100
55

protein [Bifidobacterium]

Z25_GM001499
gi|651887464|ref|WP_026647100.11
MULTISPECIES: ATPase
99.758
413

[Bifidobacterium]

Z25_GM001508
gi|748205242|ref|WP_039775930.1|
alpha-xylosidase
100
804

[Bifidobacterium adolescentis]

Z25_GM001520
gi|154083261|gb|EDN82306.1|
hypothetical protein BIFADO_02438
97.059
34

[Bifidobacterium adolescentis L2-32]

Z25_GM001532
gi|154083241|gb|EDN82286.1|
hypothetical protein BIFADO_02416
100
69

[Bifidobacterium adolescentis L2-32]

Z25_GM001533
gi|500063643|ref|WP_011743560.1|
alpha-mannosidase
99.579
949

[Bifidobacterium adolescentis]

Z25_GM001536
gi|500063640|ref|WP_011743557.1|
membrane protein
100
149

[Bifidobacterium adolescentis]

Z25_GM001540
gi|500063637|ref|WP_011743554.1|
DNA mismatch repair protein
21.366
454

MutH [Bifidobacterium

adolescentis]

Z25_GM001557
gi|154083218|gb|EDN82263.1|
anaerobic ribonucleoside-triphos-
100
237

phate reductase activating

protein [Bifidobacterium

adolescentis L2-32]

Z25_GM001559
gi|671343544|gb|AII77060.1|
glutamate-cysteine ligase
100
423

[Bifidobacterium adolescentis]

Z25_GM001567
gi|740659868|ref|WP_038445168.1|
AMP-binding protein
29.651
172

[Bifidobacterium adolescentis]

Z25_GM001569
gi|154083208|gb|EDN82253.1|
hypothetical protein BIFADO_02381
100
27

[Bifidobacterium adolescentis L2-32]

Z25_GM001573
gi|489907668|ref|WP_003811090.1|
AcrR family transcriptional regulator
100
211

[Bifidobacterium adolescentis]

Z25_GM001581
gi|705408834|ref|WP_033499905.1|
MarR family transcriptional regulator
99.561
228

[Bifidobacterium adolescentis]

Z25_GM001595
gi|154083714|gb|EDN82759.1|
dCTP deaminase
99.512
205

[Bifidobacterium adolescentis L2-32]

Z25_GM001609
gi|747125322|gb|AJE06744.1|
hypothetical protein BBMN23_1772
97.368
38

[Bifidobacterium adolescentis]

Z25_GM001611
gi|736987863|ref|WP_034983920.1|
hypothetical protein [Bifidobacterium
99.676
309

adolescentis]

Z25_GM001612
gi|751369357|gb|KIM01867.1|
MFS transporter [Bifidobacterium
98.446
193

adolescentis]

Z25_GM001613
gi|489903120|ref|WP_003806552.1|
hypothetical protein [Bifidobacterium
97.214
323

adolescentis]

Z25_GM001647
gi|747124986|gb|AJE06408.1|
hypothetical protein BBMN23_1436
99.432
176

[Bifidobacterium adolescentis]

Z25_GM001648
gi|705409118|ref|WP_033500018.1|
hypothetical protein [Bifidobacterium
98.905
274

adolescentis]

Z25_GM001649
gi|705409855|ref|WP_033500321.1|
glycosyl transferase family 2
99.543
657

[Bifidobacterium adolescentis]

Z25_GM001651
gi|705409864|ref|WP_033500324.1|
sugar ABC transporter
79.621
422

[Bifidobacterium adolescentis]

Z25_GM001652
gi|705409866|ref|WP_033500325.1|
glycosyl transferase family 9
88.129
278

[Bifidobacterium adolescentis]

Z25_GM001653
gi|705409873|ref|WP_033500328.1|
galactofuranosyltransferase
89.338
619

[Bifidobacterium adolescentis]

Z25_GM001655
gi|751368680|gb|KIM01218.1|
deoxyribonuclease [Bifidobacterium
60.656
61

adolescentis]

Z25_GM001658
gi|705409861|ref|WP_033500323.1|
hypothetical protein [Bifidobacterium
34.84
376

adolescentis]

Z25_GM001660
gi|705409873|ref|WP_033500328.1|
galactofuranosyltransferase
31.613
620

[Bifidobacterium adolescentis]

Z25_GM001662
gi|705409861|ref|WP_033500323.1|
hypothetical protein [Bifidobacterium
32.68
153

adolescentis]

Z25_GM001664
gi|751369058|gb|KIM01578.1|
glycosyl transferase [Bifidobacterium
88.818
313

adolescentis]

Z25_GM001665
gi|737015663|ref|WP_035011282.1|
hypothetical protein [Bifidobacterium
28.689
366

adolescentis]

Z25_GM001677
gi|489903795|ref|WP_003807225.1|
hypothetical protein [Bifidobacterium
56.559
709

adolescentis]

Z25_GM001678
gi|747124812|gb|AJE06234.1|
L-lactate dehydrogenase 2
32.646
291

[Bifidobacterium adolescentis]

Z25_GM001686
gi|154084014|gb|EDN83059.1|
ABC 3 transport family protein
100
280

[Bifidobacterium adolescentis L2-32]

Z25_GM001691
gi|500063170|ref|WP_011743087.1|
membrane protein [Bifidobacterium
100
330

adolescentis]

Z25_GM001694
gi|747124275|gb|AJE05697.1|
hypothetical protein BBMN23_0725
100
106

[Bifidobacterium adolescentis]

Z25_GM001698
gi|914800850|ref|WP_050731476.1|
transcriptional regulator
99.794
486

[Bifidobacterium adolescentis]

Z25_GM001702
gi|489906165|ref|WP_003809589.1|
MULTISPECIES: WhiB family
100
71

transcriptional regulator

[Bifidobacterium]

Z25_GM001710
gi|489903317|ref|WP_003806748.1|
ligase [Bifidobacterium
97.692
260

adolescentis]

Z25_GM001716
gi|500063597|ref|WP_011743514.1|
LacI family transcriptional
99.712
347

regulator [Bifidobacterium

adolescentis]

Z25_GM001722
gi|705408847|ref|WP_033499910.1|
deoxyribonuclease HsdR
98.844
1038

[Bifidobacterium adolescentis]

Z25_GM001732
gi|751369233|gb|KIM01747.1|
alpha/beta hydrolase [Bifidobacterium
97.902
286

adolescentis]

Z25_GM001737
gi|748204973|ref|WP_039775661.1|
hypothetical protein [Bifidobacterium
99.643
561

adolescentis]

Z25_GM001738
gi|822581535|gb|KLE27047.1|
exopolysaccharide biosynthesis
100
554

polyprenyl glycosylphosphotransferase

[Bifidobacterium adolescentis]

Z25_GM001741
gi|671343263|gb|AII76779.1|
hypothetical protein BADO_1371
97.692
130

[Bifidobacterium adolescentis]

Z25_GM001742
gi|822625628|ref|WP_046999683.1|
hypothetical protein [Bifidobacterium
100
61

adolescentis]

Z25_GM001744
gi|822625629|ref|WP_046999684.1|
hypothetical protein [Bifidobacterium
100
191

adolescentis]

Z25_GM001745
gi|920095976|ref|WP_052946241.1|
hypothetical protein [Bifidobacterium
99.296
142

adolescentis]

Z25_GM001761
gi|695759186|ref|WP_032682397.1|
nitroreductase [Bifidobacterium
44.326
282

longum]

Z25_GM001762
gi|651887594|ref|WP_026647221.1|
MULTISPECIES: multidrug
98.96
481

transporter [Bifidobacterium]

Z25_GM001763
gi|547053514|ref|WP_021912793.1|
MULTISPECIES: hypothetical
99.257
269

protein [Bifidobacterium]

Z25_GM001765
gi|822625640|ref|WP_046999695.1|
integrase [Bifidobacterium
100
31

adolescentis]

Z25_GM001771
gi|748204999|ref|WP_039775687.1|
hypothetical protein [Bifidobacterium
97.727
264

adolescentis]

Z25_GM001774
gi|748205357|ref|WP_039776045.1|
MFS transporter [Bifidobacterium
100
394

adolescentis]

Z25_GM001776
gi|917738583|ref|WP_052252769.1|
hypothetical protein [Bifidobacterium
100
69

adolescentis]

Z25_GM001782
gi|489906168|ref|WP_003809592.1|
hemolysin III [Bifidobacterium
100
295

adolescentis]

Z25_GM001787
gi|489905617|ref|WP_003809042.1|
hypothetical protein [Bifidobacterium
75.972
283

adolescentis]

Z25_GM001790
gi|673000931|gb|KFI95998.1|
hypothetical protein BSTER_1710
90.566
53

[Bifidobacterium stercoris

JCM 15918]

Z25_GM001794
gi|747123691|gb|AJE05113.1|
Mobile element protein [Bifidobacterium
99.664
298

adolescentis]

Initial
End

Initial
End
Database
Database

Gene ID
Mismatching
Gap
Gene
Gene
Article
Article
E
Score

Z25_GM000009
29
0
37
405
19
141
2.65E−66
196

Z25_GM000013
16
0
1
384
1
128
1.10E−70
206

Z25_GM000014
4
0
1
243
1
81
2.17E−51
154

Z25_GM000019
0
0
1
243
1
81
6.14E−58
170

Z25_GM000024
32
0
1
531
1
177
4.98E−79
231

Z25_GM000026
7
0
1
189
1
63
2.54E−36
114

Z25_GM000029
48
2
49
285
6
82
5.56E−09
47.4

Z25_GM000031
26
1
1
162
1
55
2.22E−11
51.2

Z25_GM000032
20
0
1
132
1
44
1.57E−13
56.6

Z25_GM000034
24
0
154
858
1
235
1.33E−157
437

Z25_GM000035
6
0
1
327
1
109
4.39E−67
196

Z25_GM000038
287
11
97
1464
16
488
7.28E−77
249

Z25_GM000039
116
5
70
738
24
241
5.77E−59
199

Z25_GM000047
0
0
1
618
1
206
2.58E−153
421

Z25_GM000048
3
0
1
378
1
126
1.77E−90
256

Z25_GM000049
0
0
1
198
1
66
4.55E−46
139

Z25_GM000053
9
1
1
408
1
139
4.92E−65
202

Z25_GM000053
8
0
676
837
202
255
1.28E−23
95.1

Z25_GM000054
62
5
1
549
1
188
2.63E−29
105

Z25_GM000058
5
0
25
288
1
88
1.82E−57
170

Z25_GM000059
2
0
154
954
1
267
0
538

Z25_GM000079
0
0
1
234
1
78
1.70E−47
144

Z25_GM000121
0
0
1
2304
60
827
0
1498

Z25_GM000122
0
0
1
411
1
137
2.21E−102
287

Z25_GM000123
1
0
1
999
1
333
0
627

Z25_GM000155
0
0
133
507
1
125
2.93E−88
252

Z25_GM000158
0
0
1
1506
1
502
0
979

Z25_GM000206
0
0
1
594
9
206
3.90E−149
410

Z25_GM000208
0
0
1
147
15
63
2.87E−32
103

Z25_GM000214
0
0
1
1143
1
381
0
713

Z25_GM000236
0
0
158
51
8
43
1.89E−20
74.3

Z25_GM000237
0
0
1
252
1
84
1.96E−57
169

Z25_GM000262
1
0
1
537
1
179
1.51E−132
367

Z25_GM000266
0
0
114
1
1
38
4.15E−21
76.3

Z25_GM000267
0
0
1
213
1
71
9.96E−50
149

Z25_GM000273
0
0
1
336
1
112
8.14E−79
225

Z25_GM000279
0
0
1
276
1
92
7.79E−66
191

Z25_GM000289
0
0
1
882
1
294
0
613

Z25_GM000297
0
0
1
2730
1
910
0
1768

Z25_GM000298
0
0
1
987
1
329
0
677

Z25_GM000300
0
0
1
702
272
505
1.69E−176
493

Z25_GM000302
1
0
1
2022
1
674
0
1389

Z25_GM000303
1
0
1
456
1
152
5.39E−111
310

Z25_GM000304
0
0
31
129
4
36
1.26E−19
70.9

Z25_GM000306
0
0
1
126
1
42
6.35E−25
84.3

Z25_GM000308
8
0
1
129
1
43
2.79E−20
72.8

Z25_GM000313
4
0
1
942
1
314
0
633

Z25_GM000321
0
0
58
786
18
260
1.58E−160
444

Z25_GM000322
1
0
1
507
1
169
1.01E−126
351

Z25_GM000325
0
0
1
516
1
172
2.34E−123
343

Z25_GM000331
6
0
1
681
1
227
9.73E−147
408

Z25_GM000332
3
0
1
2601
1
867
0
1765

Z25_GM000341
0
0
1
786
1
262
1.94E−167
462

Z25_GM000345
0
0
1
876
1
292
0
590

Z25_GM000356
0
0
1
1080
1
360
0
743

Z25_GM000361
5
0
1
3783
1
1261
0
2506

Z25_GM000364
0
0
1
723
12
252
2.05E−161
445

Z25_GM000373
0
0
1
744
1
248
4.34E−167
460

Z25_GM000375
0
0
1
183
25
85
3.63E−41
127

Z25_GM000380
1
0
1
621
1
207
2.78E−153
421

Z25_GM000381
0
0
1
882
23
316
0
615

Z25_GM000383
0
0
1
327
1
109
3.25E−77
221

Z25_GM000390
1
0
1
207
29
97
2.31E−47
144

Z25_GM000401
0
0
1
309
1
103
4.96E−74
213

Z25_GM000415
0
0
1
129
1
43
4.04E−27
90.1

Z25_GM000420
0
0
1
693
1
231
5.65E−154
425

Z25_GM000421
0
0
49
1059
17
353
0
684

Z25_GM000423
0
0
1
507
1
169
1.18E−102
290

Z25_GM000432
0
0
1
972
1
324
0
588

Z25_GM000464
0
0
1
216
14
85
7.51E−49
147

Z25_GM000470
0
0
1
801
3
269
0
521

Z25_GM000472
0
0
1
951
1
317
0
654

Z25_GM000474
0
0
1
192
1
64
1.06E−40
125

Z25_GM000482
0
0
1
237
5
83
7.26E−38
120

Z25_GM000496
2
0
53
193
1
47
9.18E−29
96.7

Z25_GM000506
1
0
1
1800
1
600
0
1159

Z25_GM000509
0
0
1
513
1
171
4.07E−126
350

Z25_GM000510
1
0
1
195
1
65
2.19E−44
135

Z25_GM000513
0
0
1
450
21
170
2.59E−111
311

Z25_GM000514
0
0
1
255
17
101
4.79E−60
176

Z25_GM000561
0
0
1
789
1
263
0
514

Z25_GM000577
12
0
1
834
1
278
0
541

Z25_GM000579
0
0
1
177
17
75
7.99E−39
121

Z25_GM000580
3
0
1
180
72
131
3.69E−38
121

Z25_GM000581
0
0
1
222
26
99
6.30E−50
151

Z25_GM000582
0
0
40
825
14
275
0
546

Z25_GM000583
0
0
1
144
1
48
1.13E−30
99.4

Z25_GM000590
0
0
1
192
1
64
7.89E−44
133

Z25_GM000655
2
0
1
270
1
90
1.83E−62
182

Z25_GM000657
1
0
1
180
1
60
9.01E−41
125

Z25_GM000667
4
0
1
357
56
174
4.09E−82
236

Z25_GM000668
210
24
172
1380
98
465
7.16E−26
110

Z25_GM000669
5
0
1
990
1
330
0
593

Z25_GM000670
0
0
1
573
66
256
4.03E−134
375

Z25_GM000671
64
0
10
846
1
279
2.38E−165
459

Z25_GM000752
2
0
1
1878
1
626
0
1306

Z25_GM000764
4
0
1
249
10
92
1.03E−53
160

Z25_GM000770
8
0
1
366
1
122
3.64E−82
234

Z25_GM000771
10
1
1
408
1
137
1.80E−90
258

Z25_GM000772
5
0
1
168
1
56
2.67E−31
101

Z25_GM000774
34
1
1
579
1
192
3.83E−115
324

Z25_GM000776
92
2
16
903
5
301
9.22E−146
410

Z25_GM000780
110
4
1
588
1
199
3.32E−48
155

Z25_GM000782
27
0
13
279
6
94
7.78E−41
128

Z25_GM000783
53
2
1
390
1
132
1.84E−41
132

Z25_GM000784
1
0
1
513
1
171
5.25E−125
347

Z25_GM000789
0
0
1
1611
1
537
0
1111

Z25_GM000791
7
0
1
720
11
250
7.22E−175
479

Z25_GM000793
5
0
1
156
1
52
9.01E−33
105

Z25_GM000795
4
0
4
174
1
57
3.49E−35
111

Z25_GM000796
4
0
1
132
1
44
1.78E−26
88.6

Z25_GM000798
2
0
1
252
1
84
3.16E−58
171

Z25_GM000799
6
0
1
204
1
68
9.58E−43
131

Z25_GM000800
0
0
490
972
1
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6.20E−122
345

Z25_GM000801
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1
125
1.91E−91
259

Z25_GM000802
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1
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3.24E−34
108

Z25_GM000808
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1
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1.71E−31
102

Z25_GM000809
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3
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1.09E−44
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Z25_GM000810
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1
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1
261
4.83E−108
311

Z25_GM000811
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1
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2.83E−109
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Z25_GM000815
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1
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1.17E−50
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Z25_GM000817
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3.67E−25
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Z25_GM000818
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3.53E−83
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Z25_GM000822
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2.24E−10
54.7

Z25_GM000834
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2.75E−16
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Z25_GM000835
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2.17E−128
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Z25_GM000856
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7.45E−31
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Z25_GM000856
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204
382
1.92E−16
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Z25_GM000858
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639

Z25_GM000870
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1.86E−57
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Z25_GM000873
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28
132
4.52E−57
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Z25_GM000874
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1.81E−18
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Z25_GM000881
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Z25_GM000915
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Z25_GM000921
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4.31E−64
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Z25_GM000926
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Z25_GM000927
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6.61E−106
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Z25_GM000928
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Z25_GM000930
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Z25_GM000932
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Z25_GM000943
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Z25_GM000966
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Z25_GM001016
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Z25_GM001017
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Z25_GM001020
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Z25_GM001042
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Z25_GM001043
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Z25_GM001046
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Z25_GM001051
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Z25_GM001067
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Z25_GM001068
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Z25_GM001069
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Z25_GM001073
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Z25_GM001149
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Z25_GM001157
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Z25_GM001178
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Z25_GM001194
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Z25_GM001220
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Z25_GM001244
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Z25_GM001245
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Z25_GM001248
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1.44E−63
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Z25_GM001252
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565

Z25_GM001266
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Z25_GM001268
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6.53E−115
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Z25_GM001270
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6.71E−55
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Z25_GM001276
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4.48E−67
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Z25_GM001282
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559

Z25_GM001286
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1.57E−178
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Z25_GM001301
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3.90E−10
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Z25_GM001321
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495

Z25_GM001341
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8.77E−100
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Z25_GM001366
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6.56E−31
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Z25_GM001377
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Z25_GM001381
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Z25_GM001382
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Z25_GM001387
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Z25_GM001396
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300
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507

Z25_GM001397
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1
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1.52E−49
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Z25_GM001403
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710

Z25_GM001406
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Z25_GM001421
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1
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2.11E−62
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Z25_GM001422
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1.54E−138
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Z25_GM001426
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Z25_GM001427
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Z25_GM001428
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Z25_GM001429
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Z25_GM001432
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Z25_GM001436
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1.76E−139
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Z25_GM001441
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1
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2.47E−46
142

Z25_GM001446
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1
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7.28E−15
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Z25_GM001449
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1
199
6.41E−147
405

Z25_GM001456
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38
218
4.72E−137
380

Z25_GM001457
33
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4
218
8.47E−135
375

Z25_GM001458
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922
1191
10
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2.90E−55
176

Z25_GM001460
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624
52
236
7.40E−125
351

Z25_GM001467
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1
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4.13E−61
178

Z25_GM001481
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1
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576

Z25_GM001483
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1
249
8.18E−170
467

Z25_GM001489
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109
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49
357
0
543

Z25_GM001496
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25
202
2.64E−98
281

Z25_GM001497
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20
74
4.83E−37
117

Z25_GM001499
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1
413
0
848

Z25_GM001508
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1
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1
804
0
1629

Z25_GM001520
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1
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1
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7.55E−19
68.6

Z25_GM001532
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82
5.66E−46
145

Z25_GM001533
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126
1074
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1842

Z25_GM001536
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1
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1
149
7.52E−93
264

Z25_GM001540
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4
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47
477
4.60E−15
75.5

Z25_GM001557
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1
237
0
495

Z25_GM001559
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1
423
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876

Z25_GM001567
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127
603
379
537
6.65E−07
51.2

Z25_GM001569
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1
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31
7.82E−15
58.9

Z25_GM001573
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1
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1
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1.15E−159
438

Z25_GM001581
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1
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1
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1.18E−154
427

Z25_GM001595
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1
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1.38E−154
424

Z25_GM001609
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1
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1.15E−22
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Z25_GM001611
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1
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0
637

Z25_GM001612
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201
393
6.42E−132
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Z25_GM001613
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1
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0
649

Z25_GM001647
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8.95E−131
362

Z25_GM001648
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6.97E−157
436

Z25_GM001649
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19
675
0
1338

Z25_GM001651
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1
1
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1
418
0
697

Z25_GM001652
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25
858
1
278
5.94E−165
457

Z25_GM001653
66
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1
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4
622
0
1181

Z25_GM001655
24
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159
219
4.31E−20
78.2

Z25_GM001658
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42
389
1.42E−53
189

Z25_GM001660
341
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4
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52
621
1.36E−79
261

Z25_GM001662
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1
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301
6.23E−17
75.5

Z25_GM001664
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1
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2
314
0
552

Z25_GM001665
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157
509
1.95E−28
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Z25_GM001677
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1
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1
705
0
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Z25_GM001678
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30
315
2.44E−44
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Z25_GM001686
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1
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2.97E−164
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Z25_GM001691
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29
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0
628

Z25_GM001694
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1
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1.83E−74
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Z25_GM001698
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2
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0
891

Z25_GM001702
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1
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4.51E−50
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Z25_GM001710
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186
445
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Z25_GM001716
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1
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0
711

Z25_GM001722
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1
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0
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Z25_GM001732
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0
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Z25_GM001737
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35
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Z25_GM001738
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1
554
0
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Z25_GM001741
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1
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1.96E−94
266

Z25_GM001742
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185
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1
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7.52E−28
101

Z25_GM001744
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28
218
1.22E−133
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Z25_GM001745
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18
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3.99E−90
257

Z25_GM001761
157
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1
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52
333
1.64E−86
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Z25_GM001762
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825

Z25_GM001763
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103
371
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554

Z25_GM001765
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28
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3.38E−18
68.2

Z25_GM001771
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Z25_GM001774
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18
411
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Z25_GM001776
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6.78E−48
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Z25_GM001782
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529

Z25_GM001787
68
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3.32E−161
448

Z25_GM001790
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1
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2.69E−32
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Z25_GM001794
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584

The beneficial technical effects of the present disclosure are as follows.

In order to understand the present disclosure further, the technical solutions in the examples of the present disclosure will be described clearly and completely herein in conjunction with the examples of the present disclosure. Apparently, the described examples are only a part of the examples of the present disclosure, rather than all examples. Based on the examples in the present disclosure, all of other examples, made by one of ordinary skill in the art without any creative efforts, fall into the protection scope of the present disclosure.

All of the reagents related to examples of the present disclosure are commercial products without special description, which can be purchased on market. All of the following examples are completed by theory and technology research group of probiotics of Research Center of Food Biotechnology in School of Food Science and Technology, Jiangnan University.

Example 1: Strain CCFM8630 of Bifidobacterium adolescentis has Good Tolerance to Simulated Gastrointestinal Fluid

The cryopreserved strain CCFM8630 of Bifidobacterium adolescentis were inoculated in the mMRS medium (MRS medium containing 0.05% cysteine hydrochloride) and cultured at 37° C. for 48 hours under anaerobic cultivation, followed by 2 to 3 times subculture in mMRS liquid medium. The medium with strain CCFM8630 of Bifidobacterium adolescentis was taken and centrifuged for 5 minutes at a speed of 8000×g, and then resuspended (1:1) in an artificial simulated gastric juice (mMRS medium containing 1% pepsin, pH 2.5), followed by anaerobic cultivation at 37° C. Sampling was carried out at 0 hour, 0.5 hour, 1 hour and 2 hours, and the samples were cultured on mMRS medium agar plate for colony counting. The viability numbers were counted and the survival rates were calculated. The survival rate is the rate of the viable count at the desired time point to the viable count at the 0 hour, which was expressed in %.

The medium with cultured strain CCFM8630 of Bifidobacterium adolescentis was taken and centrifuged at a speed of 8000×g for 5 minutes. The bacteria were collected and resuspened (1:1) in artificial simulated intestinal fluid (mMRS medium containing 0.3% bile salt from ox, 1% trypsin, pH 8.0), followed by anaerobic cultivation at 37° C. Sampling was carried out at 0 hour, 0.5 hour, 1 hour, 2 hours, 3 hours and 4 hours, and the samples were cultured on mMRS medium agar plate for colony counting. The viability numbers were counted and the survival rates were calculated. The survival rate is the rate of the viable count at the desired time point to the viable count at the 0 hour, which was expressed in %.

The experiment results were shown in Table 2 and Table 3. The results showed that strain CCFM8630 of Bifidobacterium adolescentis has a relative good tolerance to simulated gastrointestinal fluid.

TABLE 2

Tolerance of strain CCFM8630 of Bifidobacterium adolescentis to

simulated gastrointestinal fluid

Simulated Gastric Fluid

Treatment Time (h)

0.5
1
2

Survival Rate (%)
56.9
39.5
12.6

TABLE 3

Tolerance of strain CCFM8630 of Bifidobacterium adolescentis to

simulated intestinal fluid

Simulated Intestinal Fluid

Treatment Time (h)

0.5
1
2
3
4

Survival Rate (%)
100
100
49.1
51.6
25.9

Example 2: Strain CCFM8630 of Bifidobacterium adolescentis has No Toxic and Side Effects on SD Rat

The strain CCFM8630 of Bifidobacterium adolescentis bacteria were resuspended in 2% sucrose solution to give a bacterial suspension with a concentration of 3.0×10⁹CFU/mL. 8 healthy male SD rats with a weight between 180 and 200 g were chosen and acclimated for 1 week before experiments. The rats were administered with the above bacteria suspension by intragastric gavage once daily at a dose of 2 mL/day/rat. The death and weight of the rats were observed and recorded for one week. The results were shown in Table 4.

TABLE 4

Death and changes of body weight in rats

Time (day)
1
2
3
4
5
6
7

Weight (g)
230.2 ± 1.2
234.8±1.7
240.9 ± 1.4
246.2 ± 1.1
251.1 ± 0.8
257.2 ± 0.6
263.7 ± 0.9

Death
—
—
—
—
—
—
—

Comment: “—”, no death.

The results showed that administration of strain CCFM8630 of Bifidobacterium adolescentis with a concentration of 3.0×10⁹CFU/mL did not have significant influences on rats, there was no significant change on the body weight and no death. There were no obvious pathological symptoms in the appearance of the rats.

Example 3: Strain CCFM8630 of Bifidobacterium adolescentis has Good Recovery Effect on Tissue Damages of Liver, Duodenum and so on in Rats with Metabolic Syndrome

48 healthy male SD rats with weight from 180 to 200 g were chosen and acclimated for 1 week. The rats were divided into 6 groups randomly: non-specific control group (NC), high-fat high-starch (HFHS) diet model control group, simvastatin control group (SC), rosiglitazone hydrochloride control group (RH), strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), Bifidobacterium animalis BB12 control group (BB12), 8 rats per group. The rats were administered with the bacteria suspension (3.0×10⁹CFU/mL, in 2% sucrose solution) by intragastric gavage. Grouping and treatment method were shown in Table 5.

TABLE 5

Grouping and treatment method of the experiment

Number of
Treatment

Treatment Method: administrated by

Group
rats/Group
Duration
Feed
intragastric gavage daily

NC
8
12 Weeks
Normal feed
2 ml of 2% sucrose solution

HFHS
8
12 Weeks
High-fat high-starch
2 ml of 2% sucrose solution

feed

SC
8
12 Weeks
High-fat high-starch
2 ml of 2% sucrose solution containing

feed
3 mg/kg/BW/d of simvastatin

RH
8
12 Weeks
High-fat high-starch
2 ml of 2% sucrose solution containing

feed
10 mg/kg/BW/d of rosiglitazone hydrochloride

CCFM8630
8
12 Weeks
High-fat high-starch
2 ml of 2% sucrose solution containing

feed
3.0 × 10⁹CFU/mL of CCFM8630

BB12
8
12 Weeks
High-fat high-starch
2 ml of 2% sucrose solution containing

feed
3.0 × 10⁹CFU/mL of BB12 of

At the end of the experiment, the rats were fasted (with access to water) for 12 hours. After administering 10% chloral hydrate by peritoneal injection, the rats were anesthetized, the blood samples were collected from the hearts, and the rats were sacrificed by cervical dislocation. The blood samples were centrifuged at a speed of 3000×g at 4° C. for 10 minutes. The supernatant was collected and frozen at −80° C. for later use. Liver, duodenum and so on were collected and quickly put into ice-cold physiological saline to wash away the blood, followed by fixation in paraformaldehyde. In addition, small intestine was collected and immediately frozen in liquid nitrogen.

Intestine, duodenum and so on were taken and prepared as paraffin sections, followed by HE staining. Morphology of the tissues were observed and imaged under optical microscope for pathological evaluation. The results were shown in FIGS. 2 and 3. The HE staining was performed by the following steps.

(1) Fixation: the tissue samples were washed with physiological saline and immediately put into neutral paraformaldehyde solution (4%) for fixation, and the duration of fixation was generally within 72 hours.

(2) Washing: the tissue samples were washed with running water or immersed in water for a few hours or overnight.

(3) Dehydration: the tissue samples were dehydrated by successively immersing in ethanol solutions of 70%, 80% and 90%, each for 30 minutes, and then immersing in 95% ethanol solution once for 20 minutes, immersing in 100% ethanol solution twice, each time for 10 minutes.

(4) Transparency: the tissue samples were immersed in a mixture of ½ absolute ethanol and ½ xylene for 10 minutes, xylene I for 10 minutes, and xylene II for 10 minutes (until the samples became transparent).

(5) Waxing: the tissue samples were placed in paraffin (at 62° C.) for 2 hours.

(6) Embedding: the largest side of the sample was placed in the bottom so that the sections have the largest tissue surface.

(7) Cutting: the wax blocks were cut by a manually operating microtome into slices with a thickness of 5 μm.

(8) Floatation and adhesion of sections (slice-salvaging): a water bath was used and the water was maintained at 42° C.; sections were placed onto the water surface smoothly.

(9) Drying: slides and slide rack were put into a 55° C. drying oven for about 2 hours until the wax melted.

(10) Hydration: slides were immersed in xylene I and II for 10 minutes respectively for dewaxing, and then immersed in ethanol solutions of 100%, 95%, 90%, 80% and 70% for 5 minutes respectively, and then immersed in distilled water for 3 minutes.

(11) Primary stain: the slides were put into hematoxylin solution and stained for about 20 seconds.

(12) Washing: the slides were washed with tap water for about 15 minutes until the slices became blue. Pay attention to the water flow to avoid the sections detaching from the slide.

(13) Differentiation: the slides were put into ethanol solution with 1% hydrochloric acid for 7 seconds until the slices turned red (the color became light).

(14) Rinsing: the slides were washed with tap water for 15 to 20 minutes until the color recovered blue.

(15) Re-stain: the slides were immersed in eosin solution and immediately taken out for dehydration.

(16) Dehydration: the slides were immersed in 95% ethanol solution I, 95% ethanol solution II and 70% ethanol solution successively, followed by immersing in 80% ethanol solution for 50 seconds and absolute ethanol for 2 minutes.

(17) Transparency: the slides were immersed in ½ of absolute ethanol and ½ of xylene for 1 minute, xylene I for 2 minutes and xylene II for 2 minutes, respectively.

(18) Sealing: after the treatment of xylene, the neutral balsam was used as mounting medium, which could be diluted to appropriate consistency with xylene.

FIG. 2 showed that high-fat high-starch diet caused hepatocyte microvesicular steatosis, and a number of rats have infiltration of inflammatory cell and hyperplasia of fibrous tissue. In high-fat high-starch diet model control group (HFHS), there was significant hyperplasia of fibrous tissue in liver tissue and morphologic features of early fibrosis. In strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage significantly improved the lesions above, and the effects were significantly better than that of group BB12. FIG. 3 showed that under optical microscope, lesions of duodenum were villi broadening, interstitial edema, increase of inflammatory cells and increase of interstitial macrophages in a few cases. In strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage improved the lesions above, and the effects were significantly better than that of group BB12.

Example 4: Strain CCFM8630 of Bifidobacterium adolescentis has Recovery Effect on Intestinal Flora Imbalance Caused by High-Fat High-Starch Diet

Grouping, molding and treatment processes using SD rats were the same as described in Example 3. Before the end of the experiment, fresh feces of the rats were taken and metagenome samples were extracted. A second-generation sequencer was used for sequencing and the microbial community structure was analyzed.

The experiment results were shown in FIG. 4. In feces of high-fat high-starch diet model control group (HFHS), relative abundances of intestinal microbes Bifidobacterium genus and Turicibacter genus significantly decreased. In strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), intake of strain CCFM8630 of Bifidobacterium adolescentis leaded to a significant recovery of the relative abundances of these two genera, while drugs and BB12 did not show recovery effect on the abundances of these two genera intestinal microbes. This also indicated that strain CCFM8630 of Bifidobacterium adolescentis can not only colonize in intestine, but also improve proportions of other species of Bifidobacterium genus in the intestine. In addition, in rat feces of the high-fat high-starch diet model control group, the relative abundance of intestinal microbes of Blautia genus significantly increased. In strain CCFM8630 of Bifidobacterium adolescentis intervention group, intake of strain CCFM8630 of Bifidobacterium adolescentis regulated the abundance of Blautia genus back to normal level, and the effects were better than that of the drugs and BB12.

Example 5: Strain CCFM8630 of Bifidobacterium adolescentis Reduced (Fasting) Blood Glucose Level of Rats with Metabolic Syndrome

Grouping, molding and treatment processes using SD rats were the same as described in Example 3.

At the end of the experiment the rats were fasted (with access to water) for 12 hours and fasting blood glucose level of the rats was tested. The results were shown in FIG. 5.

In high-fat high-starch diet model control group (HFHS), fasting blood glucose level of rats significantly increased. In strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage significantly decreased fasting blood glucose level of model rats, approximately to non-specific control group, and its ability to decrease fasting blood glucose level of rat is better than that of rosiglitazone hydrochloride control group (RH) and Bifidobacterium animalis BB12 control group (BB12).

Example 6: Strain CCFM8630 of Bifidobacterium adolescentis Increased Glucose Tolerance of Rat with Metabolic Syndrome

Grouping, molding and treatment processes using SD rats were the same as described in Example 3. At the end of the experiment, the rats were fasted (with access to water) for 12 hours. Glucose solution (2 g/kg) was injected by intraperitoneal injection and the blood glucose level was measured at 0, 30, 60 and 120 minutes. The experiment results were shown in FIGS. 6 and 7.

As shown in FIG. 6, glucose tolerance of rats in high-fat high-starch diet model control group (HFHS) was poor. After administration of glucose by intragastric gavage, blood glucose level rose significantly and decreased slowly. As shown in FIG. 7, in strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage significantly decreased AUC_glucosearea, and there was no significant difference comparing with that of rosiglitazone hydrochloride control group (RH) and non-specific control group (NC). This indicated that strain CCFM8630 of Bifidobacterium adolescentis significantly improves oral glucose tolerance, and the effect was better than that of Bifidobacterium animalis BB12. These results were consistent with that of the blood glucose indexes, indicating that strain CCFM8630 of Bifidobacterium adolescentis further decreased blood glucose level by increasing glucose tolerance.

Example 7: Strain CCFM8630 of Bifidobacterium adolescentis Decreased Total Cholesterol (TC) Level in Serum of Rat with Metabolic Syndrome

Grouping, molding and treatment processes using SD rats were the same as described in Example 3. At the end of the experiment, the rats were fasted (with access to water) for 12 hours. After administering 10% chloral hydrate by peritoneal injection for anesthetizing, blood sample was collected from the heart, and the rats were sacrificed by cervical dislocation. The blood samples were centrifuged at a speed of 3000×g at 4° C. for 10 minutes, and the supernatant was collected. The total cholesterol (TC) in the blood was measured according to the protocol of the detection kit. The experiment results were shown in FIG. 8.

As shown in FIG. 8, total cholesterol in serum of rats in high-fat high-starch diet model control group (HFHS) significantly increased. In strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage decreased level of total cholesterol in serum.

Example 8: Strain CCFM8630 of Bifidobacterium adolescentis Decreased Triglyceride (TG) Level in Serum of Metabolic Syndrome Rat

Grouping, molding and treatment processes using SD rats were the same as described in Example 3. At the end of the experiment, the rats were fasted (with access to water) for 12 hours. After administering 10% chloral hydrate by peritoneal injection for anesthetizing, blood sample was collected from the heart, and the rats were sacrificed by cervical dislocation. The blood samples were centrifuged at a speed of 3000×g at 4° C. for 10 minutes, and the supernatant was collected. The triglyceride (TG) level in the blood was measured according to the protocol of the detection kit. The experiment results were shown in FIG. 9.

As shown in the experiment results, comparing with non-specific control group (NC), triglyceride level in serum of rats in high-fat high-starch diet model control group significantly increased. In strain CCFM8630 of Bifidobacterium adolescentis intervention group, administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage decreased triglyceride level in serum, and the effect was equivalent to that of rosiglitazone hydrochloride control group. The administration of BB12 by intragastric gavage did not show significant effect.

Example 9: Strain CCFM8630 of Bifidobacterium adolescentis Affected 5-HT and Testosterone Levels in Serum of Rat with Metabolic Syndrome

Grouping, molding and treatment processes using SD rats were the same as described in Example 3. At the end of the experiment, the rats were fasted (with access to water) for 12 hours. After administering 10% chloral hydrate by peritoneal injection for anesthetizing, blood sample was collected from the heart, and the rats were sacrificed by cervical dislocation. The blood samples were centrifuged at a speed of 3000×g at 4° C. for 10 minutes, and the supernatant was collected. The 5-HT and testosterone levels in the blood were measured according to the protocol of the detection kit. The experiment results were shown in FIG. 10.

As shown in the experiment results, strain CCFM8630 of Bifidobacterium adolescentis significantly increased 5-HT level in serum of rats, while BB12 has no significant improvement on 5-HT level. Comparing with non-specific control group (NC), testosterone level in serum of rats in high-fat high-starch diet model control group (HFHS) significantly increased. In strain CCFM8630 of Bifidobacterium adolescentis intervention group (CCFM8630), administration of strain CCFM8630 of Bifidobacterium adolescentis by intragastric gavage reduced the testosterone level in serum back to normal.

BIFIDOBACTERIUM ADOLESCENTIS AND USE THEREOF

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