Bifidobacterium bifidum for the treatment of diabetes and related diseases

Abstract

Provided is a Bifidobacterium bifidum strain ibiome00l, which can significantly reduce the body weight and the weight of white fat in high-fat diet-induced obese diabetic mice, and enhance the expression of lipolysis genes ATGL and HSL. In addition, it significantly reduced the fasting blood glucose, the area under the curve of the oral glucose tolerance test, the plasma glycated hemoglobin and the insulin content in the mice. This strain is a potential functional strain for treating metabolic syndromes such as obesity and diabetes, and is more effective than combinations with other Bifidobacterium bifidum.

Description

Technical Field

[0001] The present invention belongs to the field of microbial technology, and specifically relates to Bifidobacterium bifidum for the treatment of diabetes and related diseases.

Background Art

[0002] With the development of society, people's living standards have improved and their diet structures have been adjusted. During this process, diets tend to be enriched, and high-fat diets containing a lot of fish and meat have become an important daily diet style. However, this has led to an increase in the incidence of metabolic diseases. In particular, metabolic syndromes such as obesity and diabetes are the most common, having a profound impact on people's physical and mental health.

[0003] There are about 1.5 kg of bacteria in the intestine, and the huge intestinal flora forms a close structural and functional relationship through long-term coevolution with the host. The intestinal flora contributes to the homeostasis balance of the host through a series of functions such as digestion of nutrients, supply of vitamins and energy to the host, and participation in the construction of normal immunity. The imbalance of the intestinal flora is considered to be related to more than 50 types of diseases. Recent research has found that metabolic diseases, especially obesity and diabetes, are closely related to the intestinal flora, and the intestinal flora can regulate the host's fat accumulation and insulin sensitivity. Clinical FMT research has shown that insulin resistance was significantly improved after obese patients received intestinal FMT from normal individuals for 6 weeks. Another study identified 52,484 intestinal bacterial genes related to type 2 diabetes in the intestinal flora of 171 Chinese adult type 2 diabetes patients and 174 healthy volunteers. Therefore, probiotics have great potential in the treatment of obesity and diabetes.

[0004] GPR120 is a long-chain unsaturated free fatty acid receptor and has various physiological functions such as regulating the secretion of gastrointestinal hormones and the development and differentiation of adipocytes. Animal experiments have shown that a high-fat diet causes more severe obesity, insulin resistance, and hepatic steatosis in GPR120-deficient mice. A series of preclinical studies have shown that GPR120 agonists can regulate glucose and energy homeostasis, including improving chronic inflammation and insulin resistance caused by obesity, regulating thermogenesis in adipocytes, and regulating appetite. Human cohort studies have also shown that the R270H mutation in the GPR120L amino acid sequence is significantly associated with obesity. Therefore, GPR120 is a potentially important target in the treatment of metabolic syndromes such as obesity and diabetes.

[0005] There are two types of triglyceride lipases in adipose tissue, namely adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL). The former has activity without the need for hormone activation, so it is very important in basal lipolysis and is the main lipolytic enzyme. The latter needs to be activated by lipolytic hormones. The triglycerides (TG) hydrolyzed by both account for about 95% of the total hydrolysis.

[0006] HSL was discovered in 1962 and was named because its lipase activity is greatly affected by hormones. According to research, HSL activators phosphorylate HSL via PKA and transfer it to lipid droplets to promote the lipolysis process. Insulin is its most important inhibitor.

[0007] ATGL was discovered in 2004. Since its C-terminus contains a hydrophobic lipid droplet binding region, it is mainly localized on the surface of lipid droplets. ATGL can specifically hydrolyze the first ester bond of TG and is considered to be the rate-limiting enzyme in the TG hydrolysis process. When its expression decreases, a large amount of TG accumulates in adipocytes and other tissues, leading to obesity and other metabolic complications.

Summary of the Invention

Problems to be Solved by the Invention

[0008] An object of the present invention is to provide Bifidobacterium bifidum for use in the treatment of diabetes and related diseases.

Means for Solving the Problems

[0009] The present invention is realized by the following technical means. Bifidobacterium bifidum ibiome001 is preserved in the Guangdong Provincial Microbial Culture Collection Center, with the address being "5th Floor, Building 59, 100 Xianlie Middle Road, Guangzhou, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences", the preservation date being May 17, 2022, and the preservation number being GDMCC No. 62473. The whole genome sequence is shown in SEQ ID NO.2.

[0010] Bifidobacterium bifidum ibiome001 contains at least one specific gene fragment or its complementary fragment among SEQ ID NOs. 2-5.

[0011] The present invention protects the use of the above Bifidobacterium bifidum ibiome001 and its metabolites, or a mixture containing the bacterium and / or its metabolites, in the manufacture of a functional bacterial agent or a pharmaceutical for the prevention or treatment of one or more of the following diseases and symptoms (a) to (j) in mammals: (a) Diabetes (b) Metabolic syndrome (c) Abnormality of glycated hemoglobin (d) Abnormality of insulin sensitivity (e) Abnormality of fasting blood glucose level (f) Abnormality of oral glucose tolerance (g) Abnormality of fasting insulin content (h) Obesity (i) Weight gain (j) Increase in the weight of adipose tissue.

[0012] The present invention protects the use of the above Bifidobacterium bifidum ibiome001 and its metabolites, or a mixture containing the bacterium and / or its metabolites, in the manufacture of a functional bactericide or pharmaceutical that activates GPR120.

[0013] The present invention protects the use of the above Bifidobacterium bifidum ibiome001 and its metabolites, or a mixture containing the bacterium and / or its metabolites, in the manufacture of a functional bactericide or pharmaceutical that enhances the expression of lipolytic genes ATGL and / or HSL.

[0014] Furthermore, the diabetes is type 2 diabetes.

[0015] Furthermore, the mammal is a high-fat diet mammal.

[0016] The present invention protects the use of the above Bifidobacterium bifidum ibiome001 and its metabolites, or a mixture containing the bacterium and / or its metabolites, in the manufacture of a food or dietary supplement.

[0017] The present invention also protects a composition containing the above Bifidobacterium bifidum ibiome001 or its metabolites, and a composition containing a pharmaceutically acceptable carrier.

[0018] Furthermore, the pharmaceutically acceptable carrier includes one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, diluents, and absorption enhancers that are commonly used in medicine.

Advantages of the Invention

[0019] The beneficial effects of the present invention are as follows. Bifidobacterium bifidum ibiome001 of the present invention can significantly reduce the body weight and the weight of white fat in obese and diabetic mice induced by a high-fat diet, and enhance the expression of lipolysis genes ATGL and HSL. In addition, it can significantly reduce the fasting blood glucose of mice, the area under the curve of the oral glucose tolerance test, the glycated hemoglobin and insulin content in plasma. Bifidobacterium bifidum ibiome001 is a potential functional strain in the treatment of metabolic syndromes such as obesity and diabetes, and is more effective than combinations with other Bifidobacterium bifidum.

[0020] Biological preservation For Bifidobacterium bifidum ibiome001, the preservation date is May 17, 2022, the preservation location is the Guangdong Provincial Center for Microbial Culture Collection, the address is the 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, and the preservation number is GDMCC No. 62473.

Brief Description of the Drawings

[0021]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Figure 14

Figure 15

Mode for Carrying Out the Invention

[0022] To better understand the present invention, the present invention will be further described below based on examples and drawings. The following examples are for explaining the present invention and are not intended to limit it. Also, unless otherwise specified, all conditions, procedures, and methods not specifically described are general methods, and the reagents and materials used are commercially available.

[0023] Example 1 Isolation and Identification of Strains 1 Isolation Fecal samples from 10 healthy volunteers were taken and stored in 20% (v / v) glycerol phosphate buffer, and each fecal sample was serially diluted to 10 -5 , 10 -6 , 10 -7 . Each concentration was plated on MRS broth medium (manufactured by Solarbio, product number M8540) and cultured anaerobically at 37°C for 48 hours. Single clones were transferred to MRS broth liquid medium for culture, and PCR amplification was performed using general primers for 16s rRNA (upstream primer 27F: AGAGTTTG ATCCTGGCTCAG, downstream primer 1492R: GGTTA CCTTGTTACGACTT).

[0024] 2 Identification 2.1 16s rRNA Sequencing The amplified product was sent for sequencing, and Bifidobacterium bifidum was selected by comparison of the 16s rRNA gene sequence, and after in vitro screening, it was named Bifidobacterium bifidum ibiome001. Testing Method: Take 100 μL of the bacterial solution, centrifuge at 12,000 rpm for 2 minutes, discard the medium, and resuspend the bacteria in sterile ddH2O for use in PCR. PCR system (20 μL): 2×Taq Master Mix: 10 μL, primer 1 (341F): 1 μL, primer 2 (1492R): 1 μL, ddH2O: 6 μL, bacterial solution: 2 μL. PCR reaction program: 95°C for 3 minutes, 95°C for 15 seconds, 58°C for 15 seconds, 72°C for 30 seconds, steps 2-4 were 35x, and 72°C for 5 minutes. The PCR products of the 16S rRNA gene were sequenced and the results are shown in SEQ ID NO.1.

[0025] 2.2 Microscopic examination of smears Using a microscope, ibiome001 was examined by 40x magnification in a smear, and the microscopic image shown in Figure 1 was obtained. From the image, ibiome001 was Gram-stain positive, exhibiting short rod-like, thin rod-like or spherical shapes, forming pleomorphic forms such as various branches and ramifications, not having spores, and not being motile.

[0026] 2.3 Photographs of single colonies After culturing ibiome001 in MRS medium for 48 hours, photographs were taken, and a photograph of a single colony is shown in Figure 2. The colony is white, round, with regular edges and a moist surface.

[0027] 2.4 Hydrogen peroxide enzyme test and detection of sugar alcohol fermentation biochemical reaction Transfer 300 μL of cryopreserved ibiome001 into 1 mL of MRS medium to revive the strain. Streak the liquid-cultured ibiome001 on MRS solid medium for purification, select a single colony and inoculate it into 1 mL of MRS liquid medium, and culture for 24 hours. Then, serially dilute the liquid-cultured strain, spread it on MRS solid medium, and culture for 72 hours. Next, perform the catalase test and detect the sugar alcohol fermentation biochemical reaction. Catalase test: 2 - 3 drops of catalase reaction reagent (purchased from Qingdao Haibo Biotechnology, product number HB8650) were dropped onto the ibiome001 colony. Since no bubbles were observed in the result, it was judged to be negative. Detection of sugar alcohol fermentation biochemical reaction: Use a sterilized pipette tip to transfer a single colony into commercial bacterial biochemical test ampoules (purchased from Qingdao Haibo Biotechnology, product numbers GB057, GB102 - 1, GB104 - 1, GB176, GB178, GB188, GB189, GB193, GB195, GB196, GB197, GB199, GB200, GB201, GB202, GB203, GB204, GB206, GB207), and incubate at 37°C under anaerobic culture for 48 hours after inoculation. Judge the test results based on the instructions of the reagent kit, as shown in Table 1. [Table 1] JPEG2025519946000002.jpg83170

[0028] 2.5 Whole-genome sequencing Send ibiome001 to a genetic testing company for whole-genome sequencing, and compare the obtained whole-genome sequence with the sequences of strains such as ATCC 29521 (=JCM 1255, GenBank Assembly Accession: GCA_001025135.1), YIT 10347 (GenBank Assembly Accession: GCA_020892075.1), TMC 3115 (GenBank Assembly Accession: GCA_003573895.1), NCTC13001 (GenBank Assembly Accession: GCA_900637095.1), JCM 7004 (GenBank Assembly Accession: GCA_003573955.1), PRL2010 (GenBank Assembly Accession: GCA_000165905.1), HN002 (GenBank Assembly Accession: GCA_016838705.1), BGN4 (GenBank Assembly Accession: GCA_000265095.1), BF3 (GenBank Assembly Accession: GCA_001281345.1), S17 (GenBank Assembly Accession: GCA_000164965.1), S6 (GenBank Assembly Accession: GCA_003390735.1) of Bifidobacterium bifidum to obtain the specific sequence fragments shown in SEQ ID NO.2-5, as shown in Figures 3-7.

[0029] Example 2 Comparison of GPR120 activation by different Bifidobacterium bifidum (1) Culture the HEK293 cell line stably expressing β-arrestin-TEV and tTA-luciferase in DMEM medium containing 10% fetal bovine serum and 1% penicillin / streptomycin. (2) Preparation of the transfection mixture: Mix 200 ng / well of the GPR120-tango plasmid in 20 μL of DMEM with 400 ng of polyethylenimine (dissolved in 20 μL of DMEM) and incubate at room temperature for 20 minutes. After culturing the HEK293 cells in step (1) for 2 days (until about 90% confluence), the transfection mixture in step (2) is added to the HEK293 cells in step (1). (4) 24 hours after transfection, the culture medium is replaced with 180 mL of DMEM medium containing 1% penicillin / streptomycin and 10 mM HEPES and 20 μL of bacterial culture supernatant (Bb-1, Bb-2, Bb-3 or ibiome001, among which Bb-1, Bb-2, Bb-3 are other Bifidobacterium bifidum screened in the laboratory, and the 16s rDNA sequence is identical to SEQ ID NO.1). (5) After stimulating with the bacterial culture supernatant for 24 hours, the supernatant is discarded, and 50 μL of Bright-Glo solution (manufactured by Promega) diluted 20-fold with PBS containing 20 mM HEPES is added per well. (6) After incubation at room temperature for 20 minutes, fluorescence quantification is performed using Spectramax i3. As shown in the experimental results in Figure 8, compared with the culture medium, ibiome001 has the strongest activation effect on GPR120, which is 4 times that of the culture medium control. The activation effects of other Bifidobacterium bifidum strains are all less than 2 times, proving that the activation effect of ibiome001 on GPR120 is significantly superior to that of other Bifidobacterium bifidum strains.

[0030] Example 3 Effects of the strain on mouse body weight, adipose tissue weight, lipolysis gene expression, oral glucose tolerance, fasting blood glucose, glycated hemoglobin, and plasma insulin C57BL / 6J (10-week-old, male) mice of SPF grade were selected and purchased from "Jiangsu Jicui Yakang Biotechnology Co., Ltd.". After the mice were adapted for 1 week, the mice were administered a 60% high-fat diet (purchased from Madison, product number MD12033) and medicated simultaneously. The experiment was divided into 4 groups in total. (1) control group: that is, the control group, and 0.2 mL of PBS solution is administered intragastrically. (2) Bb group: 10 9Administer CFU Bifidobacterium bifidum ibiome001 to the stomach. (3) Bb + BI group: Administer Bifidobacterium bifidum ibiome001 + Bifidobacterium longum (strains are mixed at a ratio of 1:1, and the total colony count is 10^9 CFU) to the stomach. (4) 5mix (Ba + Bf + BI + Bb + Bp) group: Mix five types of Bifidobacterium bifidum, namely Bifidobacterium bifidum ibiome001 (Bb), Bifidobacterium longum (BI), Bifidobacterium adolescentis (Ba), Bifidobacterium faecale (Bf), and Bifidobacterium pseudocatenulatum (Bp) (strains are mixed at a ratio of 1:1:1:1:1, and the total colony count is 10^9 CFU).

[0031] Effect on the body weight of mice Before the start of the experiment, the above mice were grouped by body weight, and after the start of the experiment, the body weight of the mice was recorded weekly for a total of 13 weeks. The body weight change rate and absolute body weight of the mice are shown in Figure 9. As can be seen from Figure 9a, when Bifidobacterium bifidum ibiome001 (Bb) was administered to the stomach, the increase in the body weight of the mice could be significantly suppressed from the 4th week, and this effect continued until the end of the experiment. On the other hand, when combined with other Bifidobacterium bifidum, the effect of suppressing body weight gain was not significant. From Figure 9b, it can be seen that 13 weeks after administering Bifidobacterium bifidum ibiome001 to the stomach, the absolute body weight of the mice decreased by 5.53 g compared with the control group.

[0032] Effect on the fasting blood glucose level and oral glucose tolerance of mice Measure the fasting blood glucose level of the mice at the 10th week of the experiment and conduct an oral glucose tolerance test (OGTT). OGTT experimental method: The animals were fasted for 12 hours, blood was collected from the tip of the mouse's tail, and the fasting blood glucose level (at 0 hour) of the mouse was measured with a blood glucose test strip. At the same time, the mouse was administered a glucose solution at a dosage of 2 g / kg, and then blood was collected from the tip of the tail every 30 minutes, 60 minutes, and 120 minutes after administering glucose to the stomach, respectively, to measure the blood glucose level. A change curve of blood glucose was drawn over time, and the area under the curve was calculated. The results of the fasting blood glucose level are shown in Figure 10: Administering Bifidobacterium bifidum ibiome001 (Bb) to the stomach significantly decreased the fasting blood glucose level of high-fat diet-induced obese diabetic mice, but there was no improvement effect after combination with other Bifidobacterium. The results of the oral glucose tolerance test (OGTT) are shown in Figure 11a: Gastric administration of Bifidobacterium bifidum ibiome001 (Bb) significantly decreased the blood glucose level 30 minutes after glucose loading (P = 0.051) and significantly decreased the area under the curve of the oral glucose tolerance test (Figure 11b), but there was no significant improvement effect after combination with other Bifidobacterium.

[0033] 3 Effect on mouse adipose tissue weight After 13 weeks of administration, the mice were fasted for 12 hours, the mice were sacrificed, plasma and adipose tissues (mesenteric white fat, subcutaneous white fat, epididymal white fat, brown fat) were collected, and the adipose tissues were weighed respectively. The results are shown in Figure 12: Administering Bifidobacterium bifidum ibiome001 (Bb) to the stomach could significantly reduce the weights of subcutaneous white fat and mesenteric white fat in mice. Administering a combination of Bifidobacterium bifidum and Bifidobacterium longum (Bb + BI) to the stomach could only significantly reduce the weight of subcutaneous white fat in mice. Administration of a mixture of five types of Bifidobacterium into the stomach had no effect on the fat in each part of the mice.

[0034] 4 Effect on the expression of lipid decomposition genes in mice RNA was extracted from the above-mentioned white adipose tissue, and the expression levels of lipid decomposition-related genes were measured by qPCR method. The specific method is as follows. Extraction of tissue RNA: RNA in the tissue was extracted using a total RNA extraction kit for animal tissues (purchased from "Tiangen Biochemical Technology (Beijing) Co., Ltd.", product number DP424). The specific operation was referred to the kit instruction manual. Next, the RNA concentration and purity were measured using NanoDrop and agarose gel electrophoresis. Reverse transcription: 2 μg of total RNA in the tissue was added to 2 μL of 5×g DNA Buffer, supplemented to 10 μL with RNase-Free ddH2O, centrifuged briefly, placed at 42°C, incubated for 3 minutes, then placed on ice for 10 minutes. Subsequently, 2 μL of 10×Fast RT Buffer, 1 μL of RT Enzyme Mix, and 2 μL of FQ-RT Primer Mix were added, supplemented to 20 μL with RNase-Free ddH2O, reacted in a 42°C water bath for 15 minutes, then reacted at 95°C for 3 minutes, and stored at -80°C. qPCR: 2 μg of cDNA, 10 μL of SYBR staining solution, and 0.8 μL of primer, and the remaining volume was supplemented with ddH2O (total reaction system: 20 μL). The reaction conditions were denaturation at 95°C for 10 minutes and amplification (15 seconds at 95°C, 1 minute at 60°C, a total of 40 cycles). Two replicates were set for each gene, and relative quantitative analysis was performed using 2 -Δ Δ CT. As shown in the results of qPCR in Figure 13, the expression of lipid metabolism genes ATGL and HSL increased by gastric administration of Bifidobacterium bifidum ibiome001 (Bb), and there was a significant difference in the degree of increase in ATGL (P < 0.05).

[0035] Effects on glycated hemoglobin and plasma insulin in 5 mice The above plasma was collected, and the contents of glycated hemoglobin and insulin in the plasma were measured using reagent kits (purchased from "Huamei Bio", product numbers CSB-E08141m and CSB-E05071m). Glycated hemoglobin level is the gold standard for blood glucose control. As shown in Figure 14, administering Bifidobacterium bifidum ibiome001 (Bb) into the stomach significantly decreased the amount of glycated hemoglobin in the plasma of high-fat diet-induced obese diabetic mice. However, when combined with other Bifidobacterium species, it had no effect on the amount of glycated hemoglobin in the plasma. As shown in Figure 15, administering Bifidobacterium bifidum ibiome001 (Bb) into the stomach significantly decreased the insulin content in the plasma of high-fat diet-induced obese diabetic mice. However, administering a combination of Bifidobacterium bifidum and Bifidobacterium longum (Bb+BI) into the stomach had no effect on the insulin content in the plasma.

[0036] In summary, administering Bifidobacterium bifidum ibiome001 (Bb) into the stomach can significantly reduce the body weight and the weight of white fat of high-fat diet-induced obese diabetic mice, and enhance the expression of lipolysis genes ATGL and HSL. Also, it significantly decreased the fasting blood glucose, the area under the curve of the oral glucose tolerance test, the plasma glycated hemoglobin and the insulin content of the mice. Bifidobacterium bifidum ibiome001 is a potential functional strain for the treatment of metabolic syndromes such as obesity and diabetes, and is more effective than combinations with other Bifidobacterium bifidum species.

[0037] The above embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, all kinds of modifications and improvements made by those skilled in the art to the technical solution of the present invention should fall within the protection scope defined by the claims of the present invention.

Industrial Applicability

[0038] By administering Bifidobacterium bifidum ibiome001 of the present invention to the stomach, the body weight and the weight of white fat of high-fat diet-induced obese diabetic mice can be significantly reduced, and the expression of lipolysis genes ATGL and HSL can be enhanced. In addition, the fasting blood glucose, the area under the curve of the oral glucose tolerance test, the plasma glycated hemoglobin and the insulin content of the mice were significantly reduced. Bifidobacterium bifidum ibiome001 is a potential functional strain for treating metabolic syndromes such as obesity and diabetes, and is more effective than combinations with other Bifidobacterium bifidum, and has obvious industrial applicability.

Claims

1. Bifidobacterium bifidum ibioime001, wherein the strain of Bifidobacterium bifidum is preserved in the "Guangdong Provincial Center for Microbial Culture Collection", with the address being "5th Floor, Building 59, No. 100 Dashanyuan, Xianlie Middle Road, Guangzhou, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences", the preservation date being May 17, 2022, and the preservation number being GDMC No. 62473, and the Bifidobacterium bifidum is characterized by this.

2. The method for detecting Bifidobacterium bifidum ibioime001 according to claim 1, wherein the strain of Bifidobacterium bifidum contains at least one specific gene fragment or its complementary fragment among SEQ ID NOs. 2 - 5.

3. A composition, wherein the composition contains Bifidobacterium bifidum ibioime001 according to claim 1 and a pharmaceutically acceptable carrier, and the composition is characterized by this.

4. The pharmaceutically acceptable carrier according to claim 3 contains one or more of the fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, diluents, and absorption promoters commonly used in medicine, and the composition is characterized by this.

5. Use in the preparation of a functional bacterium agent or drug of Bifidobacterium bifidum ibioime001 according to claim 1, or the composition according to claim 3 or 4, wherein among them, the functional bacterium agent or the drug is used for the prevention or treatment of one or more of the following diseases and symptoms (a) to (j) in mammals, and the use is characterized by this. (a)Diabetes (b)Metabolic syndrome (c)Abnormal glycated hemoglobin (d)Abnormal insulin sensitivity (e)Abnormal fasting blood glucose level (f)Abnormal oral glucose tolerance (g)Abnormal fasting insulin content (h)Obesity (i)Weight gain (j)Increased weight of adipose tissue.

6. Use in the preparation of a functional bacterium agent or drug that activates GPR120 of Bifidobacterium bifidum ibioime001 according to claim 1, or the composition according to claim 3 or 4.

7. Use of the Bifidobacterium bifidum ibime001 according to claim 1, or the composition according to claim 3 or 4, in the manufacture of a functional bactericide or pharmaceutical product that enhances the expression of the lipolytic genes ATGL and / or HSL.

8. The use according to claim 5, wherein the diabetes is type 2 diabetes.

9. The use according to claim 5 or 8, wherein the mammal is a high-fat diet mammal.

10. Use of the Bifidobacterium bifidum ibime001 according to claim 1, or the composition according to claim 3 or 4, in the manufacture of a food or dietary supplement.

Images