CN106974262B

CN106974262B - Application of intestinal probiotic bacillus in treating and preventing obesity and related diseases

Info

Publication number: CN106974262B
Application number: CN201610028004.XA
Authority: CN
Inventors: 冯强; 张东亚
Original assignee: BGI Shenzhen Co Ltd
Current assignee: BGI Shenzhen Co Ltd
Priority date: 2016-01-15
Filing date: 2016-01-15
Publication date: 2021-05-25
Anticipated expiration: 2036-01-15
Also published as: CN106974262A

Abstract

The invention discloses application of probiotic bacillus in treating and preventing obesity-related diseases, in particular to application of probiotic bacillus in preparing a composition or a preparation, wherein the composition or the preparation is used for one or more of the following purposes: (a) prevention and/or treatment of obesity; (b) reducing blood fat; (c) preventing or treating cardiovascular diseases; and/or (d) the prevention and/or treatment of diabetes. The intestinal probiotic bacillus of the invention can obviously reduce weight, blood fat and body fat ratio.

Description

Application of intestinal probiotic bacillus in treating and preventing obesity and related diseases

Technical Field

The invention belongs to the field of microbiology, and particularly relates to application of probiotic bacillus in treating and preventing obesity and related diseases, and also relates to a composition containing probiotic bacillus in intestines and application thereof.

Background

There are a large number of commensal microorganisms in humans, most of which reside in the human intestinal tract, in amounts exceeding 1000 trillion (10 trillion)¹⁴Order of magnitude) of more than 10 times the total number of human cells. In a long evolutionary process, intestinal microorganisms and human beings achieve good cooperation and play a crucial role in nutrition, metabolism and immunity of human bodies, and many researchers regard the intestinal microflora of the human bodies as an organ of the human bodies or a second genome of the human bodies, wherein the abundant genetic information contained in the intestinal microflora is closely related to the health of the human bodies. Through research on hundreds of diseases such as diabetes, coronary heart disease, obesity, colon cancer and the like, some specific species show remarkable association with the diseases, and the results provide a completely new direction in clinical assessment and diagnosis of the diseases and later intervention treatment.

Obesity is a chronic disease, and various factors can cause obesity, and the origin of the obesity is not clearly researched. Obesity is also an inducing factor of a series of diseases such as hypertension, diabetes, coronary heart disease, cystic disease, osteoarthritis, sleep apnea, respiratory disorders, uterine tumors, prostate cancer, breast cancer, colon cancer and the like. According to NIH reports, about 9700 million americans are currently overweight and obese, with type II diabetes associated with obesity reaching about 1510 million people, and about 20 million people die of obesity-related diseases each year.

Obesity is usually an excess of body fat due to a change in physiological or biochemical functions. Fats generally include neutral lipids, phospholipids, and cholesterol. The increase in fat is due to energy intake being greater than energy expenditure. Obesity is, in principle, of two types: (a) simple obesity (simple obesity) and (b) secondary obesity (secondary obesity). Simple obesity can be classified into congenital obesity (obesity) and acquired obesity (obesity), and the number of simple obese patients can account for more than 95% of the total obesity. Congenital obesity is caused by a large number of adipocytes and is common in childhood obesity. Acquired obesity is caused by larger size adipocytes and is common in adult obesity. Secondary obesity, also known as symptomatic obesity, is usually caused by endocrine or metabolic disorders.

There are currently five strategies for treating obesity: diet, exercise, behavior therapy, medication and rehabilitation (theripteic operation). Which strategy is taken depends primarily on the patient's health risk factors and the rate and effect of weight loss, and these strategies may be selected or combined to treat obese patients. The rate and effect of weight loss is influenced by a number of factors such as age, height, family history and risk factors. Diet-exercise therapy, i.e., eating low-calorie, low-fat foods and performing aerobic exercise, but this method is generally considered unsuccessful for the general public and requires constant adherence for a long period of time; the operation for removing the fat in the body can achieve the effect of immediate effect, but has a plurality of limitations, such as operation risk, difficulty in lasting fat removal effect, high cost and the like.

Drug therapy is currently the primary clinical treatment for obesity and its obesity-related diseases (e.g., diabetes). The mechanisms of drug therapy include appetite suppression, increased energy expenditure, stimulation of fat movement, reduction of triglyceride synthesis, and inhibition of fat absorption. The main drugs at present are: phenylpropanolamine (PPA), freshman (orlistat, xenolith iii) and nominatine (sibutramine, reductil tm). Hyperglycemia in some diabetic patients is still not adequately controlled by dietary and/or exercise therapy or the use of the above therapeutic compounds. For these patients, exogenous insulin should be used. The use of exogenous insulin is an expensive and painful procedure for the patient and can cause a number of complications for the patient. For example, an incorrect calculation of the insulin dose may result in an insulin response (hypoglycemia) due to no meals or abnormal exercise. In addition, local or systemic allergies or immune resistance to drugs may also occur with drugs.

At present, no effective method and medicament for treating and preventing obesity and related diseases with small side effect exist in the field.

There is therefore a great need in the art to develop new, non-toxic side effects for the treatment and prevention of obesity and its related diseases.

Disclosure of Invention

It is an object of the present invention to provide the use of probiotic bacteria in the treatment and prevention of obesity and its related diseases.

It is another object of the present invention to provide a pharmaceutical product, or an animal feed composition, for the treatment and prevention of obesity and related diseases, which is effective without toxic and side effects.

It is another object of the present invention to provide a method for reducing body weight and/or blood glucose and uses thereof.

The present invention provides in a first aspect the use of a probiotic bacterium of the gut for the manufacture of a composition or formulation for one or more uses selected from the group consisting of: (a) prevention and/or treatment of obesity; (b) reducing blood fat; (c) preventing or treating cardiovascular diseases; and/or (d) preventing and/or treating diabetes, wherein the probiotic bacteria of the intestinal tract are selected from the group consisting of: oxalic acid bacterium formigenes (Oxalobacter formigenes), Rabeyerba hominis (Roseburia hominis), coprinus prausnitzii (Faecalibacterium prausnitzii), Haemophilus parainfluenza (Haemophilus parainfluenza), or combinations thereof.

In another preferred example, the probiotic bacteria comprise oxalic acid producing bacteria (Oxalobacter formigenes).

In another preferred embodiment, the oxalic acid producing bacterium is selected from the group consisting of: oxalobacter formigenes OXCC13, Oxalobacter formigenes DSM 4420, Oxalobacter formigenes ATCC35274, or combinations thereof.

In another preferred example, the probiotic bacillus comprises human Roseburia hominis (Roseburia hominis).

In another preferred embodiment, the human raspberrii is selected from the group consisting of: roseburia hominis A2-183, Roseburia hominis A2-181, or a combination thereof.

In another preferred example, the probiotic bacteria comprise coprinus praussnitius (Faecalibacterium praussnidii).

In another preferred embodiment, the A.prowazekii comprises Faecalibacterium praussnitzii L2-6.

In another preferred embodiment, the probiotic bacteria comprise Haemophilus parainfluenzae (Haemophilus parainfluenzae).

In another preferred embodiment, the Haemophilus parainfluenza comprises Haemophilus parainfluenza strain T3T 1.

In another preferred embodiment, the probiotic bacillus of intestinal tract comprises one or more selected from table 3.

In another preferred embodiment, said probiotic bacteria of the intestinal tract are selected from table 3 and from the same or different genera.

In another preferred embodiment, the composition is selected from the group consisting of: a pharmaceutical composition, a feed composition, or a combination thereof.

In another preferred embodiment, the composition is an oral preparation.

In another preferred embodiment, the composition is a liquid preparation, a solid preparation or a semi-solid preparation.

In another preferred embodiment, the dosage form of the composition is selected from the group consisting of: powders, tablets, dragees, capsules, granules, suspensions, solutions, syrups, drops, and sublingual tablets.

In another preferred embodiment, the liquid formulation is selected from the group consisting of: solution preparations or suspension preparations.

In a second aspect the present invention provides the use of a probiotic bacterium of the gut for the manufacture of a composition or formulation for one or more uses selected from the group consisting of: (i) reducing the level of monocyte chemotactic protein-1 (MCP-1) in a mammal; and/or (ii) improving Leptin resistance and increasing in vivo sensitivity to Leptin, wherein said probiotic bacteria are selected from the group consisting of: oxalic acid bacterium formigenes (Oxalobacter formigenes), Rabeyerba hominis (Roseburia hominis), coprinus prausnitzii (Faecalibacterium prausnitzii), Haemophilus parainfluenza (Haemophilus parainfluenza), or combinations thereof.

In another preferred embodiment, the composition or formulation is also used independently or additionally for one or more uses selected from the group consisting of:

(iii) inhibiting weight gain in a mammal;

(iv) reducing the body-to-fat ratio (fat weight/body weight ratio) of a mammal;

(v) lowering blood lipid levels in a mammal;

(vi) increasing the level of High Density Lipoprotein (HDLC) in a mammal;

(vii) reducing Low Density Lipoprotein (LDLC) levels in a mammal.

In another preferred embodiment, the mammal includes a human, a rodent (e.g., rat, mouse).

In another preferred embodiment, said lowering of blood lipid levels in a mammal comprises lowering Total Cholesterol (TC) levels and/or triglyceride levels.

In a third aspect the present invention provides a composition for use in the treatment and/or prevention of obesity, the composition comprising: (i) a safe and effective amount of probiotic bacteria; and (ii) a pharmaceutically acceptable carrier; wherein the probiotic bacteria are selected from the group consisting of: oxalic acid bacterium formigenes (Oxalobacter formigenes), Rabeyerba hominis (Roseburia hominis), coprinus prausnitzii (Faecalibacterium prausnitzii), Haemophilus parainfluenza (Haemophilus parainfluenza), or combinations thereof.

In another preferred embodiment, the composition comprises 1X 10 to 1X 10²⁰cfu/mL or cfu/g of probiotic bacteria, preferably 1X 10⁴-1×10¹⁵cfu/mL or cfu/g of probiotic bacteria in the intestinal tract, based on the total volume or total weight of the composition.

In another preferred embodiment, said composition comprises 0.0001-99 wt%, preferably 0.1-90 wt% of said probiotic bacteria in intestinal tract, based on the total weight of said composition.

In another preferred embodiment, the composition is in unit dosage form (tablet, capsule or vial), and the mass of the composition in each unit dosage form is 0.05-5g, preferably 0.1-1 g.

In another preferred embodiment, the composition further comprises other intestinal probiotics and/or prebiotics.

In another preferred embodiment, said other probiotic bacteria are selected from the group consisting of: lactic acid bacteria, bifidobacteria, Lactobacillus acidophilus, or combinations thereof.

In another preferred embodiment, the prebiotic is selected from the group consisting of: fructooligosaccharides (FOS), Galactooligosaccharides (GOS), Xylooligosaccharides (XOS), Lactosucrose (LACT), Soy Oligosaccharides (SOS), Inulin (Inulin), or combinations thereof.

In a fourth aspect, the present invention provides a method for preparing a composition according to the third aspect of the present invention, comprising the steps of:

mixing (i) probiotic bacteria of the gut with (ii) a pharmaceutically acceptable carrier, thereby forming a composition according to the third aspect of the invention.

In another preferred embodiment, the composition is an oral preparation.

In a fifth aspect, the present invention provides a method of reducing body weight and/or blood lipids, by administering to said subject (i) a probiotic bacterium of the gut or a composition according to the third aspect of the invention.

In another preferred embodiment, said administering comprises oral administration.

In another preferred embodiment, the dosage is 0.01-5g/50kg body weight/day, preferably 0.1-2g/50kg body weight/day.

In another preferred embodiment, the subject comprises a mammal, such as a human.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Drawings

FIG. 1 shows the weight gain of the mice in each group after intragastric administration of oxalic acid bacterium formiate compared to before intragastric administration.

FIG. 2 shows the weight gain of groups of mice after gavage of human Roseburia bacilli compared to before gavage.

FIG. 3 shows the weight gain of groups of mice after gavage with C.provenii, H.parainfluenza or a combination thereof compared to before gavage.

FIG. 4 shows the weight gain of the groups of mice after the gavage combination compared to before gavage.

FIG. 5 shows the body-fat ratio of the mice in each group 9 weeks after the administration of Oxobacterium formigenes.

FIG. 6 shows the body-to-lipid ratio of groups of mice after 9 weeks of gavage of human Roseburia.

FIG. 7 shows the body-to-lipid ratio of groups of mice 9 weeks after gavage of C.provenii, H.parainfluenza or combinations thereof.

FIG. 8 shows the body-to-fat ratio of the groups of mice after 9 weeks of the gastric lavage of the combination bacteria.

FIG. 9 shows the effect of intragastrically administered oxalic acid bacterium formate on blood lipids.

FIG. 10 shows the effect of periwinia gastricis on blood lipids.

FIG. 11 shows the effect of Pectinatus gasseri, Haemophilus parainfluenza or a combination thereof on blood lipids.

FIG. 12 shows the effect of the Zygosaccharomyces on blood lipid.

FIG. 13 shows the effect of oxalic acid bacterium formigenes on monocyte chemotactic protein-1 (MCP-1) by gavage.

FIG. 14 shows the effect of periwinia gastricis on monocyte chemotactic protein-1 (MCP-1).

FIG. 15 shows the effect of C.gasseri, H.parainfluenza or a combination thereof on monocyte chemotactic protein-1 (MCP-1).

FIG. 16 shows the effect of Comptobacterium gavage on monocyte chemoattractant protein-1 (MCP-1).

FIG. 17 shows the effect of Oxalobacter formigenes on Leptin (LEP) by gavage.

FIG. 18 shows the effect of periwinia gastricis on Leptin (LEP).

FIG. 19 shows the effect of C.gasseri, H.parainfluenza or a combination thereof on Leptin (LEP).

FIG. 20 shows the effect of COMBINATION OF GASTRIC on Leptin (LEP).

Detailed Description

The present inventors have conducted extensive and intensive studies and experiments, and as a result, surprisingly found that oxalic acid bacterium formate (Oxalobacter formigenes), human raspberrii (Roseburia hominis), coprinus prausnitzii (Faecalibacterium prausnitzii), and/or Haemophilus parainfluenzae (Haemophilus parainfluenzae) have/has an effect of preventing and treating obesity and related diseases (e.g., cardiovascular diseases), and found that the composition can inhibit weight gain, reduce body-fat ratio, reduce blood lipid, and effectively reduce cardiovascular and obesity, etc., when the active composition containing the above-mentioned probiotic bacteria is fed to a feeding subject. The present invention has been completed based on this finding.

As used herein, the term "comprising" means that the various ingredients can be applied together in a mixture or composition of the invention. Thus, the terms "consisting essentially of and" consisting of are encompassed by the term "comprising.

As used herein, the "body-to-fat ratio" refers to the ratio of fat weight/body weight.

The invention relates to intestinal probiotic bacillus and application thereof

As used herein, the "probiotic bacterium of the present invention" refers to a mixture of one or more of the four genera of oxalic acid producing bacterium, Rabyella humane, coprinus pustulatus, Haemophilus parainfluenzae, or a mixture of one or more of a plurality of bacteria in each genus.

Wherein, the oxalic acid producing bacterium is an anaerobic bacterium, gram staining is negative, and oxalate can be metabolized in intestinal tract.

The human Roseburia hominis is first separated from human excrement, and is anaerobic, gram staining negative or changeable, rod-shaped and slightly bent, and capable of moving. The cell size varies from 0.5-1.5 μm to 5.0. mu.m.

The fecal bacillus prosperius is a strict anaerobic bacterium, gram staining is positive, can not move and can not produce spores. Rod-shaped with rounded ends.

Parainfluenza is a gram-negative bacterium, unable to move, does not produce spores, and has various forms such as globular rod-shaped and filamentous forms. The bacteria are microaerophilic bacteria, and the optimal growth temperature is 35-37 ℃.

The invention provides application of probiotic bacillus in treating and preventing obesity and related diseases (such as cardiovascular diseases). The subject ingests a high fat food and the probiotic intestinal bacteria have (i) an effect of inhibiting weight gain in the subject; (ii) reducing blood fat; and (iii) the ability to reduce body-to-fat ratio. According to a preferred embodiment of the present invention, C57BL/6J male mice fed with a high-fat diet that causes obesity, treated with a probiotic bacterium of the present invention (e.g., oxalic acid bacterium formate, human rashibacter, coprinus pustulus, haemophilus parainfluenzae, or a combination thereof) have a reduced weight gain and reduced blood lipid levels and decreased various indicators associated with obesity or cardiovascular disease, such as Leptin (LEP) and monocyte chemotactic protein-1 (MCP-1), as compared to untreated controls. Therefore, the probiotic bacillus (such as oxalic acid bacterium formigenes, Rabyella humane, coprinus pustulosa, haemophilus parainfluenza, or the combination thereof) in intestinal tract can be used for preventing and treating obesity and diseases caused by obesity, such as cardiovascular diseases and the like.

Composition and application thereof

The invention also provides a composition, preferably a pharmaceutical composition. The composition comprises an effective amount of a probiotic bacterium of the present invention (e.g., oxalic acid bacterium, human ralstonia fortunata, coprinus pratense, haemophilus parainfluenzae, or a combination thereof), and in a preferred embodiment, the composition further comprises a probiotic bacterium of the intestinal tract selected from the group consisting of: lactic acid bacteria, bifidobacteria, lactobacillus acidophilus, or combinations thereof; and/or a prebiotic selected from the group consisting of: fructooligosaccharides (FOS), Galactooligosaccharides (GOS), Xylooligosaccharides (XOS), Lactosucrose (LACT), Soy Oligosaccharides (SOS), Inulin (Inulin), or combinations thereof.

In a preferred embodiment, the composition is a liquid preparation, a solid preparation or a semisolid preparation.

In a preferred embodiment, the liquid formulation is selected from the group consisting of: solution preparations or suspension preparations.

In a preferred embodiment, the dosage form of the composition is selected from the group consisting of: powders, tablets, dragees, capsules, granules, suspensions, solutions, syrups, drops, and sublingual tablets.

The pharmaceutical composition of the present invention may be administered in any form of pharmaceutical tablets, injections or capsules, which includes excipients, pharmaceutically acceptable vehicles and carriers, which may be selected according to the administration route. The pharmaceutical preparation of the present invention may further comprise auxiliary active ingredients.

Lactose, glucose, sucrose, sorbitol, mannose, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinylpyrrolidone (PVP), cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, or the like can be used as the carrier, excipient, diluent, or the like of the pharmaceutical composition of the present invention.

In addition, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, emulsifiers, suspension stabilizers, preservatives, sweeteners, flavors, and the like. The pharmaceutical compositions of the present invention may be manufactured in enteric-coated formulations by a variety of well-known methods so that the active ingredient of the pharmaceutical composition, i.e., the microorganism, passes through the stomach without being destroyed by stomach acid.

In addition, the microorganism of the present invention can be used in the form of a capsule prepared by a conventional method. For example, standard excipients are mixed with the lyophilized microorganisms of the present invention to form pellets, which are then filled into gelatin capsules. In addition, the microorganisms of the present invention and the pharmaceutically acceptable excipients such as liquid gums, celluloses, silicates or mineral oils are mixed to make a suspension or dispersion, which can be filled into soft gelatin capsules.

The pharmaceutical composition of the present invention can be made into enteric coated tablets for oral administration. The term "enteric coating" in the present application includes all coatings which are approved for use with conventional drugs, which are not degraded by gastric acid, but which are sufficiently decomposed in the small intestine to rapidly release the microorganisms of the present invention. The casings of the invention can be maintained at 36-38 ℃ for more than 2 hours in synthetic gastric acid, e.g. HCl solution at pH 1, and preferably disintegrate within 1.0 hour in synthetic intestinal fluid, e.g. buffer at pH 7.0.

The enteric coating of the invention is coated at about 16-30mg, preferably 16-25mg, more preferably 16-20mg per tablet. The thickness of the casing is 5-100 μm, and the ideal thickness is 20-80 μm. The enteric coating composition is selected from conventional polymers known per se.

Preferred casings for use in the present invention are prepared from cellulose acetate phthalate polymers or trimellitate polymers and copolymers of methacrylic acid (e.g., copolymers containing greater than 40% methacrylic acid and methacrylic acid containing hydroxypropyl methylcellulose phthalate or its ester derivatives).

The cellulose acetate phthalate used in the enteric coating of the present invention has a viscosity of about 45 to 90cp, an acetyl content of 17 to 26%, and a phthalic acid content of 30 to 40%. The cellulose acetate trimellitate used in the enteric coating has a viscosity of about 5-21cp and an phthalide content of 17-26%. Cellulose acetate trimellitate is manufactured by Eastman Kodak company and can be used for the casing material in the present invention.

The hydroxypropyl methyl cellulose phthalate used in the enteric coating of the invention has a molecular weight of generally 20,000-.

Hydroxypropyl methylcellulose phthalate, which is used in the casing of the present invention, was HP50, produced by Shin-Etsu chemidl co.ltd. HP50 contains 6-10% hydroxypropyl, 20-24% methoxy, and 21-27% propyl, and has a molecular weight of 84,000 daltons. Another enteric material is HP55, HP55 contains 5-9% hydroxypropyl methylcellulose phthalate, 18-22% methoxyl, 27-35% phthalic acid, and has a molecular weight of 78,000 daltons.

The sausage casing of the invention is prepared as follows: the enteric coating solution is sprayed onto the core using conventional methods. All solvents in the enteric coating process are alcohols (e.g., ethanol), ketones (e.g., acetone), halogenated hydrocarbon compounds (e.g., dichloromethane), or combinations thereof. Softeners, such as di-n-butyl phthalate and glyceryl triacetate, are added to the enteric coating solution in a ratio of 1 part of the garment to about 0.05 parts or about 0.3 parts softener. The spraying process is preferably carried out continuously, the amount of material sprayed being controlled according to the conditions employed for coating. The spray pressure can be adjusted at will, and in general, the desired results are obtained at an average pressure of 1-1.5 Pa.

The term "pharmaceutically effective amount" as used herein refers to an amount that is functional or active in humans and/or animals and is acceptable to humans and/or animals. For example, in the present invention, a composition containing 1X 10 to 1X 10²⁰cfu/ml or cfu/g (in particular, may contain 1X 10)⁴-1×10¹⁵cfu/ml or cfu/g; more particularly, it may contain 1X 10⁶-1×10¹¹cfu/ml or cfu/g) of a probiotic bacterium of the invention (e.g. oxalic acid producing bacterium, human raspberri, coprinus pratense, haemophilus parainfluenza, or a combination thereof).

When used to prepare a pharmaceutical composition, the effective dose of the probiotic bacteria of the present invention (e.g., oxalic acid bacterium formiate, ralstonia herbecteri, coprinus pratense, haemophilus parainfluenza, or a combination thereof) used may vary with the mode of administration and the severity of the disease to be treated. Dosage forms suitable for oral administration comprising about 1X 10 to 1X 10 in intimate admixture with a solid or liquid pharmaceutically acceptable carrier²⁰cfu/ml or cfu/g (in particular, may contain 1X 10)⁴-1×10¹⁵cfu/ml or cfu/g; more particularly, it may contain 1X 10⁶-1×10¹¹cfu/ml or cfu/g) of active intestinal probiotic bacteria (such as oxalic acid producing bacillus, human raspberri, coprinus pratense, haemophilus parainfluenza, or combinations thereof). The dosage regimen may be adjusted to provide optimal treatmentResponse to therapy. For example, divided doses may be administered several times per day, or the dose may be proportionally reduced, as may be required by the urgency of the condition being treated.

The probiotic bacteria (e.g., oxalic acid bacterium formigenes, Rabyella hominis, C.purposis, Haemophilus parainfluenza, or a combination thereof) can be administered orally, or the like. The solid support comprises: starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, and liquid carriers include: culture medium, polyethylene glycol, nonionic surfactant, and edible oil (such as corn oil, peanut oil, and sesame oil) as appropriate for the characteristics of the probiotic bacterium in the intestinal tract (such as oxalic acid bacterium formiate, Ralstonia hominis, C.provenii, H.parainfluenzae, or combinations thereof) and the particular mode of administration desired. Adjuvants commonly used in the preparation of pharmaceutical compositions may also advantageously be included, for example flavouring agents, colouring agents, preservatives and antioxidants such as vitamin E, vitamin C, BHT and BHA.

Preferred pharmaceutical compositions are solid compositions, especially tablets and solid-filled or liquid-filled capsules, from the standpoint of ease of preparation and administration. Oral administration is preferred.

The composition of the present invention is administered to the subject 1 or more times per day. Dosage units for administration represent dosages which can be divided formally and which are suitable for human beings or all other mammalian subjects. Each unit containing a pharmaceutically acceptable carrier and a therapeutically effective amount of a microorganism of the invention. The amount administered will vary with the weight and severity of the obesity of the patient, the supplemental active ingredients included and the microorganism used. Furthermore, the administration can be divided, if possible, and can be continued, if desired. Therefore, the amount to be administered is not a limitation of the present invention. In a preferred embodiment, the composition comprises: drugs, animal feeds, and the like.

In a preferred embodiment, the formulation of the composition is as follows:

1×10-1×10²⁰cfu/mL of probiotic bacteria in the intestinal tract (e.g., Oxalobacter formigenes, Rabyia hominis, Shigella prosperi, Haemophilus parainfluenza, or combinations thereof); and a pharmaceutically acceptable carrierAnd/or an excipient.

In another preferred embodiment, the formulation of the composition is as follows:

1×10⁶-1×10¹¹cfu/mL of probiotic bacteria in the intestinal tract (e.g., Oxalobacter formigenes, Rabyia hominis, Shigella prosperi, Haemophilus parainfluenza, or combinations thereof); and a pharmaceutically acceptable carrier, and/or excipient.

Method for reducing body weight and/or blood lipids

In another preferred example, the method comprises: ingesting a pharmaceutical composition of the present invention. The subject is a human.

In another preferred example, the method comprises: ingesting a pharmaceutical composition or animal feed of the invention, or a combination thereof. The experimental object is an animal, preferably a mouse or a rabbit.

The main advantages of the invention include:

(a) the intestinal probiotics (such as oxalic acid producing bacillus, human Rabyia, coprinus pusillus, haemophilus parainfluenza or the combination thereof) can obviously reduce the weight, the blood fat and the body-to-fat ratio.

(b) The probiotic bacteria of the present invention (e.g., oxalic acid bacterium formigenes, ralstonia herbecteri, coprinus pratense, haemophilus parainfluenza, or a combination thereof) are capable of significantly reducing indicators (e.g., cholesterol and triglycerides) associated with obesity and its associated diseases (e.g., cardiovascular diseases).

(c) The probiotic bacillus (such as oxalic acid producing bacillus, Rabyia hominis, coprinus pusillus, haemophilus parainfluenza or the combination thereof) in the intestinal tract can obviously reduce the level of total cholesterol, triglyceride and low-density lipoprotein.

(d) The probiotic bacteria of the present invention (e.g., oxalic acid bacterium formigenes, ralstonia herbecteri, coprinus pratense, haemophilus parainfluenza, or combinations thereof) are capable of significantly increasing the level of high density lipoproteins.

(e) The probiotic bacillus (such as oxalic acid producing bacillus, Rabyia hominis, coprinus pusillus, haemophilus parainfluenza or the combination thereof) in the intestinal tract can improve insulin resistance and reduce the risk of atherosclerosis and cardiovascular diseases.

(f) The probiotic bacillus (such as oxalic acid bacterium formigenes, Rabyella humane, coprinus pratense, haemophilus parainfluenza, or the combination thereof) in the intestinal tract can obviously reduce the level of monocyte chemotactic protein-1 (MCP-1).

(g) The intestinal probiotics (such as oxalic acid producing bacillus, human Rabyia, coprinus pratense, haemophilus parainfluenza or the combination thereof) can effectively improve Leptin resistance accompanied by obesity and improve the sensitivity to Leptin in vivo.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, molecular cloning is generally performed according to conventional conditions such as Sambrook et al: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the protocol of microorganisms: the conditions described in the handbook of experiments (James Cappuccino and Natalie Sherman eds., Pearson Edurion Press) or as recommended by the manufacturer.

Example 1 compositions containing probiotic bacteria of the intestinal tract (e.g., oxalic acid bacterium, Rabyia hominis, C.provenii, Haemophilus parainfluenzae, or combinations thereof)

The raw material formulation is shown in table 1.

TABLE 1 composition formula

Raw materials	Mass percent (%)
		Bacterial composition	0.5
Milk	90.0
		White sugar	9.5

The bacterial components in the formulations 1-7 are single bacterial components and respectively contain Oxalobacter formigenes OXCC13, Oxalobacter formigenes DSM 4420, Oxalobacter formigenes ATCC35274, Roseburia hominis A2-183, Roseburia hominis A2-181, Faecalibacterium praerunsi L2-6, Haemophilus parainfluenza T3T 1.

The bacteria component in the formula 8 is a mixture of any two or more (preferably 2, 3 or 4) of the 7 bacteria (in a weight ratio of 1:1 or 1:1:1: 1).

Mixing milk and white sugar according to the above formula ratio, stirring to completely mix, preheating, homogenizing under 20Mpa, sterilizing at 90 deg.C for 5-10 min, cooling to 40-43 deg.C, inoculating 1-100 × 10⁶cfu/g of bacterial component, i.e., made into a composition containing bacterial component (e.g., oxalic acid bacterium formigenes, Rabyia hominis, C.putrescentiae, Hemophilus parainfluenza, or combinations thereof).

Example 2

Pharmaceutical composition containing probiotic bacteria (such as oxalic acid producing bacillus, Rabeyerba human, coprinus pulcheri, haemophilus parainfluenza, or their combination)

The raw material ratio is shown in table 2.

TABLE 2 pharmaceutical composition formulations

Raw materials	Mass percent (%)
		Bacterial composition	1.0％
Lactose	2.0％
		Yeast powder	2.0％
Peptone	1.0％
		Purified water	94.0％

The bacteria component in the formula 8 is a mixture of any two or more (preferably 2 or 3) of the above 7 bacteria (in a weight ratio of 1:1 or 1:1: 1).

Mixing lactose, yeast powder, and peptone with purified water at a certain proportion, preheating to 60-65 deg.C, homogenizing under 20Mpa, sterilizing at 90 deg.C for 20-30 min, cooling to 36-38 deg.C, inoculating bacteria components (1-50 × 10)⁶cfu/mL), fermenting at 36-38 deg.C to pH of 6.0, centrifuging, and freeze drying to water content less than 3% to obtain freeze-dried product containing bacteria components. Weighing 0.5 g of lyophilized product containing bacteria component, mixing with maltodextrin in equal amount, and encapsulating to obtain medicinal composition containing bacteria component (such as oxalic acid bacterium formiate, Ralstonia hominis, coprinus praecox, Haemophilus parainfluenzae, or their combination).

Example 3 therapeutic Effect on obese model mice

Experimental materials:

mice: c57BL/6J male mice (purchased from Guangdong provincial animal center for medical laboratory) were purchased as normal breeding mice, 6 weeks old. The mice were grown in the same environment and fed the same diet.

The inventor obtains 7 strains of intestinal probiotic bacilli from a depository institution and saves the intestinal probiotic bacilli in Shenzhen Hua Dagenen research institute. Meanwhile, Lactobacillus plantarum (Lactobacillus plantarum) is selected from China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.8198 and used as a control group (LP group) to be cultured in MRS culture solution for 24-48h at 37 ℃.

Wherein, the source information of the 7 strains of intestinal probiotics is shown in table 3. All strains were verified by 16S rDNA sequencing and the experiment was started.

The strain information is shown in Table 3.

TABLE 3 information on the strains

Wherein Oxalobacter formigenes OXCC13 (fungus 1) is derived from Burmingham division of university of Alabama, USA (Ellis, M.L., Shaw, K.J., Jackson, S.B., Daniel, S.L., & Knight, J. (2015.) Analysis of Commercial kinetic Stone biological supplement.Urology, 85(3): 517) 521); roseburia hominis A2-181 (strain 5) was from the Rowell institute of Rovint, UK (Duncan SH, Aminov RI, Scott KP et al (2006), Proposal of Roseburia faecis sp.nov., Roseburia hominis sp.nov.and Roseburia inulvorans sp.nov., base on isolates from human beings facial aspects. International Journal of Systematic and evolution Microbiology,56: 2437-2441.); faecalibacterium prausnitzii L2-6 (strain 6) coprinus pratense is also from the Rowett Research Institute (rocean, s.h., Hold, g.l., Harmsen, h.j.m., Stewart, c.s., Flint, H.J (2002), Growth requirements and transfer products of fusarium prausnitzii, and a prosal to rectangle of Faecalibacterium prausnitzii gen. nov., comb. nov. int.j.syol. evolution. Microbiol.52: 2141-; haemophilus parinfluenzae T3T1 (bacterium 7) is derived from Sanger Centre (Sanger Centre) (Juhas, M., Power, P.M., Harding, R.M., Ferguson, D.J., Dimopoulu, I.D., Elamin, A.R., & Smith, A. (2007), Sequence and functional analyses of Haemophilus spp. genetic islands. genome Biol,8(11): R237.).

High fat diet (HF): containing 78.8% basal diet, 1% cholesterol, 10% egg yolk powder, 10% lard and 0.2% bile salts, purchased from Nantong Temilion feed science and technology, Inc.

Ordinary maintenance feed: purchased from the centre of medical laboratory animals in Guangdong province.

The experimental method comprises the following steps:

selecting normally fed C57BL/6J adult male mice, randomly grouping, and respectively using a control group (CK), a microbial inoculum group (a bacterium 1 group, a bacterium 2 group, a bacterium 3 group, a bacterium 4 group, a bacterium 5 group, a bacterium 4+5 group, a bacterium 6 group, a bacterium 7 group, a bacterium 6+ bacterium 7 group, a bacterium 1+ bacterium 4 group, a bacterium 3+ bacterium 6 group, a bacterium 2+ bacterium 5+ bacterium 7 group, a bacterium 3+ bacterium 6+ bacterium 7 group, a bacterium 2+ bacterium 4+ bacterium 6+ bacterium 7 group), a control microbial inoculum group (LP, Lactobacillus plantarum CC No.8198) and an obesity model group (HF), wherein 10 mice in each group freely feed and drink water in an SPF specific pathogen free environment. The LP group, HF group and microbial agent group were fed with high-fat diet, and CK group was fed with ordinary maintenance diet. After feeding for 4 weeks, the bacterium agent group and the LP group begin to perfuse corresponding strain liquid; the HF group and the CK group were gavaged with the same amount of medium for 9 weeks.

The bacteria feeding amount is 0.15mL/10g body weight, and the bacteria concentration is 1 × 10⁷cfu/mL, concentration 1X 10 after concentration⁸cfu/mL, at a frequency of once every other day. The bacterial liquid needs to be cultured in advance, activated every week to ensure freshness, the concentration is respectively measured and adjusted to 1 × 10⁸cfu/mL. For a single microbial inoculum group, taking corresponding bacterial liquid and irrigating the stomach according to the dosage; for the mixed bacteria group, the single bacteria liquid is mixed in equal proportion and then is irrigated into the stomach according to the dosage.

In the experimental period, the weight, state, food intake and other data of the mice are recorded every week. The last week of the experiment each group of mice was subjected to a glucose tolerance (OGTT) test. After the experiment, the mice were sacrificed, the fat weight was recorded, and serum was taken and the blood lipid and protein factor content were measured using an Elisa kit.

The experimental results are as follows:

(1) the effect of oxalic acid bacterium formigenes, human Rascherzilla, fecal prowell, Haemophilus parainfluenza or a combination thereof on the body weight of mice.

TABLE 4 weight gain of mice in each group after intragastric administration of oxalic acid bacterium formiate compared to before intragastric administration (see FIG. 1)

Note: the data in the tables are mean ± standard deviation, and no identical letter after the number indicates significant difference (p <0.05) for any two sets of data in each column, as is the case for tables 5-19.

TABLE 5 weight gain after gavage of human Roseburia Ribes compared to before gavage in groups of mice (see FIG. 2)

TABLE 6 weight gain of groups of mice after gavage with C.provenii, H.parainfluenza or combinations thereof compared to before gavage (see FIG. 3)

TABLE 7 weight gain of mice in each group after the administration of the gastric lavage combination bacteria compared to before the administration of gastric lavage (see FIG. 4)

The results are shown in tables 4-7 and FIGS. 1-4. The results show that the oxalic acid producing bacterium, the human Raosbai Rumeji, the coprinus pustulatus, the haemophilus parainfluenza or the combination bacteria thereof can effectively slow down the weight gain of the obesity model mice (P < 0.05).

(2) The effect of oxalic acid bacterium formigenes, human Rascherzilla, fecal prowell, Haemophilus parainfluenza or a combination thereof on the body-to-lipid ratio.

TABLE 8 body-to-fat ratio of mice in each group 9 weeks after intragastric administration of oxalic acid bacterium formis (see FIG. 5)

Grouping	Fat weight/body weight × 100%
		CK	2.89±0.12d
Bacterium
	1	3.92±0.21c
Bacterium
	2	4.05±0.23c
Bacterium
	3	3.94±0.20c
LP			5.31±0.24b
	HF	7.42±0.41a

TABLE 9 body fat ratio of groups of mice after 9 weeks of gavage human Roseburia

Grouping	Fat weight/body weight × 100%
		CK	2.87±0.12e
Bacterium
	4	3.98±0.25c
Bacterium
	5	3.87±0.18cd
Bacterium
	4+5	3.73±0.19d
LP			5.29±0.16b
	HF	7.54±0.46a

TABLE 10 body-to-fat ratio of groups of mice 9 weeks after gavage of fecal prestonii, haemophilus parainfluenza or combinations thereof (see FIG. 7)

Grouping	Fat weight/body weight × 100%
		CK	2.87±0.16d
Bacterium
	6	3.92±0.19c
Bacterium
	7	3.82±0.19c
Bacterium
	6+7	3.77±0.35c
LP			5.69±0.45b
	HF	7.53±0.58a

TABLE 11 body-fat ratio of mice in each group 9 weeks after gavage of Comptobacterium (see FIG. 8)

Grouping	Fat weight/body weight × 100%
		CK	2.83±0.15e
Bacterium
	1+4	3.97±0.21c
Bacterium
	3+6	3.94±0.24c
Bacterium
	2+5+7	3.85±0.22c
Bacterium
	3+6+7	3.28±0.17d
Bacterium
	2+4+6+7	3.21±0.10d
LP			5.42±0.46b
	HF	7.39±0.44a

The results are shown in tables 8 to 11 and FIGS. 5 to 8. The results show that the oxalic acid producing bacillus, the human Roseburia bacilli, the coprinus pratense, the haemophilus parainfluenza or the combination thereof can obviously reduce the body-to-fat ratio of the obesity model mice (P < 0.05).

(3) The effect of oxalic acid bacterium formigenes, Rascherzbacilus humanus, Excreobacter prowazei, Haemophilus parainfluenza, or a combination thereof on blood lipids.

TABLE 12 blood lipid levels of mice in each group 9 weeks after intragastric administration of oxalic acid producing bacterium formate (see FIG. 9)

Grouping	TC(mmol/L)	TG(mmol/L)	LDLC(mmol/L)	HDLC(mmol/L)
					CK	3.865±0.178d	0.954±0.056d	1.239±0.039d	3.319±0.262a
Bacterium 1	4.850±0.192c	1.083±0.075bc	1.466±0.071c	3.257±0.169a
					Bacterium 2	4.905±0.186c	1.025±0.060c	1.442±0.084c	3.273±0.147a
Bacterium 3	4.884±0.240c	1.095±0.062b	1.420±0.089c	3.323±0.159a
					LP	5.315±0.286b	1.233±0.087a	1.803±0.135b	2.723±0.141b
HF	6.459±0.393a	1.285±0.092a	2.385±0.174a	2.048±0.110c

TABLE 13 lipid levels in groups of mice 9 weeks after gavage of Roseburia hominis (see FIG. 10)

Grouping	TC(mmol/L)	TG(mmol/L)	LDLC(mmol/L)	HDLC(mmol/L)
					CK	3.877±0.166d	0.961±0.056d	1.288±0.055d	3.364±0.190a
Bacterium 4	4.730±0.151c	1.051±0.075c	1.454±0.060c	3.359±0.131a
					Bacterium 5	4.625±0.196c	1.078±0.073c	1.451±0.081c	3.412±0.144a
Bacterium 4+5	4.553±0.181c	1.067±0.056c	1.432±0.064c	3.365±0.182a
					LP	5.189±0.346b	1.148±0.083b	1.826±0.124b	2.706±0.143b
HF	6.297±0.309a	1.263±0.091a	2.438±0.161a	2.189±0.058c

TABLE 14 blood lipid content of mice in each group 9 weeks after gavage of fecal pratense, haemophilus parainfluenza or combinations thereof (see FIG. 11)

Grouping	TC(mmol/L)	TG(mmol/L)	LDLC(mmol/L)	HDLC(mmol/L)
					CK	3.859±0.192d	1.004±0.077c	1.251±0.066c	3.308±0.219a
Bacterium 6	4.615±0.129c	1.017±0.064c	1.310±0.061c	3.376±0.104a
					Bacterium 7	4.601±0.185c	1.017±0.067c	1.303±0.103c	3.429±0.085a
Bacterium 6+7	4.518±0.170c	1.021±0.062c	1.374±0.094c	3.368±0.124a
					LP	5.146±0.256b	1.225±0.080b	1.911±0.113b	2.833±0.162b
HF	6.290±0.334a	1.310±0.089a	2.472±0.154a	2.196±0.095c

TABLE 15 blood lipid content of mice in each group 9 weeks after gavage of the combination bacteria (see FIG. 12)

Grouping	TC(mmol/L)	TG(mmol/L)	LDLC(mmol/L)	HDLC(mmol/L)
					CK	3.910±0.194g	0.923±0.046d	1.237±0.051c	3.229±0.238a
Bacterium 1+4	4.735±0.187cd	1.044±0.058c	1.247±0.059c	3.249±0.192a
					Bacterium 3+6	4.834±0.176c	1.035±0.064c	1.230±0.090c	3.298±0.151a
Bacterium 2+5+7	4.722±0.185de	1.018±0.067c	1.206±0.063c	3.354±0.226a
					Bacterium 3+6+7	4.513±0.195ef	1.037±0.058c	1.165±0.046c	3.431±0.141a
Bacterium 2+4+6+7	4.324±0.131f	0.987±0.030d	1.175±0.077c	3.452±0.131a
					LP	5.249±0.313b	1.213±0.081b	1.722±0.105b	2.609±0.153b
HF	6.398±0.378a	1.307±0.089a	2.357±0.137a	2.133±0.112c

The results are shown in FIGS. 9 to 12 and tables 12 to 15. The main components of blood lipids are cholesterol and triglycerides, and elevated plasma cholesterol and triglyceride levels are associated with the development of atherosclerosis. The results show that the oxalic acid producing bacillus, the human Roseburia bacilli, the coprinus pustulatus, the haemophilus parainfluenza or the combination bacteria thereof can reduce the blood fat and reduce the related indexes of atherosclerosis related diseases (such as cardiovascular diseases). And, the oxalic acid producing bacterium, the human Roseburia bacilli, the fecal strain of Fomitopsis, the Haemophilus parainfluenzae or the combination thereof can remarkably reduce the content of Total Cholesterol (TC), total Triglyceride (TG) and Low Density Lipoprotein (LDLC), and remarkably increase the content of High Density Lipoprotein (HDLC) (P < 0.05).

(4) Effect of oxalic acid bacterium formigenes, human Rascherzbacilus robustus, Exiguobacter przewalskii, Haemophilus parainfluenza or a combination thereof on Leptin (LEP), monocyte chemotactic protein-1 (MCP-1).

TABLE 16 Leptin (LEP) and monocyte chemotactic protein-1 (MCP-1) content of each group of mice 9 weeks after the administration of oxalic acid bacterium formigenes (FIG. 13, FIG. 17)

Grouping	MCP-1(pg/ml)	LEP(pg/ml)
			CK	333.13±37.24c	1183.00±92.43c
Bacterium
	1	343.35±33.65c	1219.52±98.29c
Bacterium
	2	329.80±46.40c	1204.80±128.01c
Bacterium
	3	341.62±34.19c	1198.74±118.59c
LP				361.86±24.06b	1277.72±110.30b
	HF	385.08±32.09a	1380.57±177.93a

TABLE 17 Leptin (LEP), monocyte chemoattractant protein-1 (MCP-1) levels in groups of mice 9 weeks after gavage of human Roseburia

Grouping	MCP-1(pg/ml)	LEP(pg/ml)
			CK	328.29±28.71b	1159.90±94.14c
Bacterium
	4	323.52±29.00b	1160.44±116.75c
Bacterium
	5	320.63±41.82b	1178.92±112.36c
Bacterium
	4+5	320.66±28.16b	1150.63±101.64c
LP				377.84±35.87a	1278.43±78.00b
	HF	380.66±29.85a	1388.41±70.66a

TABLE 18 content of Leptin (LEP), monocyte chemoattractant protein-1 (MCP-1) in groups of mice 9 weeks after gavage of C.provenii, H.parainfluenza or combinations thereof (see FIGS. 15 and 19)

Grouping	MCP-1(pg/ml)	LEP(pg/ml)
			CK	334.91±35.60c	1194.32±109.69c
Bacterium
	6	337.22±25.76c	1189.56±107.81c
Bacterium
	7	336.94±33.58c	1155.88±141.71c
Bacterium
	6+7	337.77±34.50c	1160.91±117.87c
LP				375.21±28.55b	1263.23±122.81b
	HF	386.05±49.66a	1389.64±141.06a

TABLE 19 Leptin (LEP) and monocyte chemotactic protein-1 (MCP-1) content of each group of mice 9 weeks after gavage combined bacteria (see FIG. 16, FIG. 20)

The results are shown in FIGS. 13 to 20 and tables 16 to 19. The results show that the content of Leptin (LEP) and monocyte chemotactic protein-1 (MCP-1) in the serum of the obesity model mouse can be obviously reduced by oxalic acid bacterium formate, human Raosbai-rui bacillus, promiscus putrescentiae, parainfluenza haemophilus or the combination bacteria thereof (P is less than 0.05).

The results show that the oxalic acid producing bacillus, the human Raosbai-rui bacillus, the coprinus pustulatus, the haemophilus parainfluenza or the combination bacteria thereof can obviously improve the leptin resistance and improve the sensitivity to the Leptin (LEP) in vivo; in addition, the serum MCP-1 level is obviously reduced after the treatment of the oxalic acid producing bacillus, the human Roseburia bacilli, the coprinus praecox, the haemophilus parainfluenza or the combination bacteria thereof, which is beneficial to improving the insulin resistance and reducing the risk of atherosclerosis and cardiovascular diseases.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. Use of probiotic bacteria of the intestinal tract for the preparation of a composition or formulation, characterized in that said composition or formulation is used for one or more uses selected from the group consisting of: (a) prevention and/or treatment of obesity; (b) reducing blood fat; (c) preventing and/or treating cardiovascular diseases, wherein the cardiovascular diseases are atherosclerosis; wherein the probiotic bacteria are selected from the group consisting of: oxalic acid bacterium formigenes (Oxalobacter formigenes), or a combination of oxalic acid bacterium formigenes (Oxalobacter formigenes) with one or more bacteria selected from the group consisting of: human Rabeyeri (Roseburia hominis), fecal putrescentiae (Faecalibacterium prausnitzii) and Haemophilus parainfluenza (Haemophilus parainfluenza).

2. The use according to claim 1, wherein the oxalate-producing bacterium is selected from the group consisting of: oxalobacter formigenes OXCC13, Oxalobacter formigenes DSM 4420, Oxalobacter formigenes ATCC35274, or combinations thereof.

3. Use of probiotic bacteria of the intestinal tract for the preparation of a composition or formulation, characterized in that said composition or formulation is used for one or more uses selected from the group consisting of:

(i) inhibiting weight gain in a mammal;

(ii) reducing the body-to-fat ratio, i.e. the ratio of fat weight/body weight, of the mammal;

(iii) reducing blood lipid levels in a mammal, wherein the probiotic bacteria are selected from the group consisting of: oxalic acid bacterium formigenes (Oxalobacter formigenes), or a combination of oxalic acid bacterium formigenes (Oxalobacter formigenes) with one or more bacteria selected from the group consisting of: human Rabeyeri (Roseburia hominis), fecal putrescentiae (Faecalibacterium prausnitzii) and Haemophilus parainfluenza (Haemophilus parainfluenza).

4. The use of claim 3, wherein said lowering the blood lipid level of a mammal comprises:

(iv) increasing the level of High Density Lipoprotein (HDLC) in a mammal;

(v) reducing Low Density Lipoprotein (LDLC) levels in a mammal.

5. A composition for the treatment and/or prevention of obesity, the composition comprising: (i) a safe and effective amount of probiotic bacteria; and (ii) a pharmaceutically acceptable carrier; wherein the probiotic bacteria are selected from the group consisting of: a combination of oxalic acid bacterium formigenes (Oxalobacter formigenes) with one or more bacteria selected from the group consisting of: human Rabeyeri (Roseburia hominis), fecal putrescentiae (Faecalibacterium prausnitzii) and Haemophilus parainfluenza (Haemophilus parainfluenza).

6. The composition of claim 5, wherein said composition comprises 1 x 10 to 1 x 10²⁰cfu/mL or cfu/g of probiotic bacteria in the intestinal tract, based on the total volume or total weight of the composition.

7. The composition of claim 6, wherein said composition comprises 1 x 10⁴-1×10¹⁵cfu/mL or cfu/g of probiotic bacteria in the intestinal tract, based on the total volume or total weight of the composition.

8. The composition of claim 5, wherein said composition comprises 0.0001-99 wt% of said probiotic bacteria, based on the total weight of said composition.

9. The composition of claim 8, wherein said probiotic bacteria are present in an amount of 0.1 to 90 wt.%, based on the total weight of the composition.

10. The composition of claim 5, wherein the composition further comprises other probiotic bacteria and/or prebiotics.

11. A method of making the composition of claim 5, comprising the steps of:

mixing (i) probiotic bacteria of the intestinal tract, with (ii) a pharmaceutically acceptable carrier, thereby forming the composition of claim 5.