CN112273657A

CN112273657A - Probiotic composition for preventing or improving allergic diseases, preparation method and application

Info

Publication number: CN112273657A
Application number: CN201910662213.3A
Authority: CN
Inventors: 陈智仙; 俞学锋; 李知洪; 张彦; 朱娅敏
Original assignee: Angel Yeast Co Ltd
Current assignee: Angel Nutt Co.,Ltd.
Priority date: 2019-07-22
Filing date: 2019-07-22
Publication date: 2021-01-29

Abstract

The invention relates to the technical field of food or health food, in particular to a probiotic composition for preventing or improving allergic diseases, a preparation method and application thereof. The invention provides a probiotic composition for preventing or improving allergic diseases, which comprises 0.1-9.9 parts of lactobacillus acidophilus, 0.1-9.9 parts of lactobacillus rhamnosus, 0.1-40 parts of yeast probiotics or yeast extract, 0.1-50 parts of prebiotics and optional probiotics in parts by weight. The probiotic composition has a good treatment effect on allergic diseases of various crowds, can remarkably shorten the duration time of allergic symptoms, relieves the allergic symptoms, and reduces side effects brought by the use of medicines.

Description

Probiotic composition for preventing or improving allergic diseases, preparation method and application

Technical Field

The invention relates to the technical field of food or health food, in particular to a probiotic composition for preventing or improving allergic diseases, a preparation method and application thereof.

Background

Allergic disease has become a common public health problem, and it is estimated that 30% to 40% of the world population is suffering from allergic disease, and the incidence of allergic disease is increasing year by year and has become a global health problem of concern. Allergic diseases are a specific immune response mainly caused by physiological dysfunction or tissue cell damage of a body after the body responds to certain antigens for the first time and is stimulated by the same antigens again, and the specific immune response is also called as hypersensitivity or allergy. The allergic diseases are classified into type I hypersensitivity (immediate type), type II hypersensitivity (cytotoxic type), type III hypersensitivity (immune complex type) and type IV hypersensitivity (delayed type) according to the factors of cells, active substances, a mechanism for damaging tissues and organs, time required for reaction and the like involved in the allergic diseases.

The current treatments for allergic diseases mainly comprise several types, and related allergen vaccines are also used for desensitization treatment, while for allergic symptoms of vaccines which have not been developed, the most important treatment modes are drug therapy for control, including the use of anti-allergic agents, histamine receptor antagonists (antihistamines), steroid agents, and the like. However, the current drug therapy cannot change the progress of allergic diseases, so that the curative effect cannot be achieved fundamentally, and the drug therapy easily causes side effects such as gastrorrhagia, obesity, edema and the like. To this end we propose a probiotic composition for the prevention or treatment of allergic diseases. The anaphylactic reaction in the research of probiotic bacteria for treating anaphylactic diseases belongs to type I hypersensitivity reaction, and is mainly IgE mediated diseases, such as atopic dermatitis, allergic rhinitis, allergic asthma, urticaria, pollinosis and the like.

With the development of biotechnology and intensive research on biopharmaceuticals and the large-scale marketing of probiotic-related products, consumers increasingly agree on the role of probiotics in health conditioning, and taking probiotic foods is becoming an increasingly important choice for preventing and treating allergic diseases.

Probiotics are a class of living microorganisms that, at appropriate intake, bring benefits to the host organism, including lactobacillus, bifidobacterium, saccharomyces, and the like. The probiotic bacteria of the lactobacillus and the bifidobacterium are dominant strains in normal intestinal flora, play a role in benefiting a host by adhering to intestinal mucosa, regulate the balance of the intestinal flora, resist the damage of pathogenic microorganisms, and regulate the immune system of an organism by secreting different cell factors, inhibiting the inflammatory reaction of the intestinal immune system and the like. The oral administration of the probiotics increases the microorganism species lacking in the intestinal flora of people easily suffering from allergic diseases, so that the oral tolerance is obtained, thereby being beneficial to the treatment of the allergic diseases. When foreign antigens enter the intestinal tract again, the probability of allergic reaction of the human body to the foreign antigens can be reduced, and the risk of allergic diseases is reduced.

CN 104093412 a relates to a pharmaceutical composition for treating or preventing allergic (allergy) diseases, in particular to a pharmaceutical composition for treating or preventing allergic diseases, which contains the dead bacteria of lactobacillus acidophilus LB as an active ingredient, and is used for treating or preventing atopic dermatitis, allergic rhinitis, allergic conjunctivitis, urticaria and pollinosis, and belongs to the field of medicines.

CN 108065400A relates to food or health food technical field, specifically relates to an allergy immune regulation and control therapy, adopts lactobacillus paracasei, lactobacillus rhamnosus, lactobacillus reuteri, lactobacillus acidophilus and bifidobacterium longum, five bacteria super-strong combination can effectively improve allergic constitution, but only limited to bacteria probiotics, formed is probiotic powder, and the product is formed only singly.

CN 107114794A belongs to the technical field of food science and relates to a probiotic composition for enhancing anti-allergic ability. The probiotic composition comprises lactobacillus dry powder, bifidobacterium longum dry powder, isomaltooligosaccharide, maltodextrin, glucose and vitamin C, belongs to the technical field of food science, and is limited to the field of bacteria probiotics.

CN 107669706 a relates to a composite probiotic for treating and preventing allergic diseases, a preparation method thereof, and a formulation of food composition and pharmaceutical composition. The probiotics contained in the invention comprise Lactobacillus crispus, Lactobacillus salivarius, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus reuteri and Bifidobacterium animalis, and animal experiments prove that the probiotic composition has the effects of preventing and treating allergic diseases, but the intestinal digestion of animals is greatly different from that of human beings, and meanwhile, different people are also greatly different.

CN 108029923A relates to a production formula of a probiotic solid beverage with an anti-allergic effect, which comprises the following components; the lactobacillus paracasei, the lactobacillus reuteri, the lactobacillus casei, the lactobacillus rhamnosus, the bifidobacterium and the yeast beta-glucan have single product form, and are not suitable for popularization and application of all people.

Disclosure of Invention

Therefore, the technical problem solved by the invention is as follows: the probiotic preparation in the prior art has a single product form, and is not suitable for popularization and application of all people.

The probiotic composition for preventing or improving allergic diseases has a good treatment effect on allergic diseases of various people, and can remarkably shorten the duration of allergic symptoms, relieve the allergic symptoms, reduce the allergic symptoms and reduce side reactions brought by the use of medicaments.

The invention provides a probiotic composition for preventing or improving allergic diseases, which comprises 0.1-9.9 parts of lactobacillus acidophilus, 0.1-9.9 parts of lactobacillus rhamnosus, 0.1-40 parts of yeast probiotics or yeast extract, 0.1-50 parts of prebiotics and optional probiotics in parts by weight.

The invention provides a preparation method of the probiotic composition, which is characterized by mixing the lactobacillus acidophilus, the lactobacillus rhamnosus, the yeast probiotics or the yeast extract, the prebiotics and the optional probiotics to obtain the probiotic composition.

The invention provides an application of the probiotic composition in preventing or improving allergic diseases.

The invention provides a health food composition for preventing or improving allergic diseases, which comprises the probiotic composition.

The yeast extract in the probiotic composition can provide essential nutrients such as amino acid, nucleotide and the like, promote the maturation of intestinal epithelial cells and supply nutrition, and the yeast beta-glucan promotes the generation of in-vivo immunoglobulin M antibodies, provides the immunocompetence of body fluid and can also regulate the colony structure of intestinal tracts.

Specifically, the present invention proposes the following technical solutions.

Preferably, the probiotic composition comprises 0.2-7 parts of lactobacillus acidophilus, 0.2-7 parts of lactobacillus rhamnosus, 2-35 parts of yeast probiotics or yeast extract, 0.3-35 parts of prebiotics and 0.1-50 parts of optional probiotics; preferably, the lactobacillus acidophilus milk powder comprises 0.5 to 5 parts of lactobacillus acidophilus, 1 to 6 parts of lactobacillus rhamnosus, 5 to 30 parts of yeast probiotics or yeast extract, 1 to 30 parts of prebiotics and 1 to 40 parts of optional probiotics; preferably, the lactobacillus acidophilus milk beverage comprises 1-3 parts of lactobacillus acidophilus, 2-5 parts of lactobacillus rhamnosus, 10-20 parts of yeast probiotics or yeast extract, 2-10 parts of prebiotics and 3-20 parts of optional probiotics.

Preferably, the probiotic composition is one or more of brewers' yeast, debaryomyces, candida, pichia, torulopsis, blakeslea and saccharomyces cerevisiae.

Preferably, the probiotic composition described above, wherein the optional probiotic bacteria are selected from one or more of lactobacillus paracasei, lactobacillus reuteri, lactobacillus gasseri, lactobacillus salivarius, lactobacillus johnsonii, lactobacillus fermentum, bifidobacterium longum, bifidobacterium breve, bifidobacterium lactis, and bifidobacterium animalis; preferably, the optional probiotics are selected from one or two of bifidobacterium lactis and bifidobacterium animalis, and more preferably, the probiotics are 1.5-15 parts of bifidobacterium lactis and 1.5-15 parts of bifidobacterium animalis.

Preferably, the probiotic composition is one or more selected from the group consisting of fructooligosaccharide, stachyose, galactooligosaccharide, isomaltooligosaccharide and yeast β -glucan, and preferably is yeast β -glucan.

Preferably, the probiotic composition further comprises an auxiliary material, and preferably, the auxiliary material is one or more selected from anhydrous glucose, maltodextrin, maltitol, xylitol and fruit powder; preferably, the fruit powder is selected from one of hawthorn powder, apple powder, banana powder, cherry powder or sweet orange powder; preferably, the auxiliary material is selected from one or more than two of 2-80 parts of anhydrous glucose, 0.1-15 parts of maltodextrin and 0-5 parts of hawthorn powder, and more preferably, the auxiliary material is selected from 10-60 parts of anhydrous glucose and/or 5-10 parts of maltodextrin.

Preferably, the probiotic composition is contained in an amount of 10% by dry weight²-10¹²Colony forming units, preferably 10⁷-10¹¹A colony forming unit.

Preferably, the probiotic composition is in the form of a powder, a tablet, a solid beverage, a capsule, a soup or a liquid suspension.

Preferably, in the preparation method, the prebiotics and the auxiliary materials are mixed, cooled to room temperature, and then added with the yeast probiotic or yeast extract, lactobacillus acidophilus, lactobacillus rhamnosus and optional probiotic to be mixed to obtain the probiotic mixture.

Preferably, the above preparation method further comprises a drying step after mixing and before cooling to room temperature; preferably, the drying step is drying to a water activity of less than 0.2.

The invention provides an application of the probiotic composition or the probiotic composition prepared by the preparation method in preventing or improving allergic diseases.

The invention provides a health care product composition for preventing or improving allergic diseases, which comprises the probiotic composition or the probiotic composition prepared by the preparation method.

The beneficial effects obtained by the invention are as follows:

the probiotic composition has a good treatment effect on allergic diseases of various crowds, can remarkably shorten the duration time of allergic symptoms, relieves the allergic symptoms, and reduces side effects brought by the use of medicines.

Information on the preservation of the strains

The strain used by the invention, namely the Saccharomyces cerevisiae FX-2(Saccharomyces cerevisiae FX-2), is preserved in China Center for Type Culture Collection (CCTCC) in 2016, 08 and 01, with the preservation number of CCTCC NO: M2016418, and the preservation address is as follows: china, wuhan university, zip code: 430072; telephone: (027) 68754052, which was first disclosed in Chinese patent documents CN107788332A and CN 107788238A.

The strain Saccharomyces cerevisiae Z2.1(Saccharomyces cerevisiae Hansen Z2.1) used by the invention is preserved in China Center for Type Culture Collection (CCTCC) at 25/10/2005, the preservation number is CCTCC NO: M205127, the preservation address is as follows: china, wuhan university, zip code: 430072; telephone: (027) 68754052, which was first disclosed in Chinese patent document CN 101575579A.

The strain beer yeast A3.12(Saccharomyces cerevisiae A3.12) used by the invention is preserved in China Center for Type Culture Collection (CCTCC) in 2017, 12 and 11 months, the preservation number is CCTCC NO: M2017781, the preservation address is as follows: china, wuhan university, zip code: 430072; telephone: (027) -68754052.

The saccharomyces cerevisiae strain is obtained by separating from the soil of the vineyard, and is observed by an optical microscope, appears ellipsoidal and is propagated asexually in a budding mode; after being cultured on a solid medium plate at 28 ℃ for 72 hours, the medium is milk white, opaque and round colony.

The strain Saccharomyces boulardii Angel 1.27(Saccharomyces boulardii Angel 1.27) used in the invention is preserved in China Center for Type Culture Collection (CCTCC) at 16/04/2012, with the preservation number of CCTCC NO: M2012116, and the preservation address: china, wuhan university, zip code: 430072; telephone: (027) -68754052.

The strain is separated from the litchi rind of Lingnan, and the Saccharomyces boulardii is ellipsoid and propagated asexually in a budding mode through observation by an optical microscope; after being cultured on a solid medium plate at 28 ℃ for 72 hours, the solid medium plate is a light yellow, smooth and opaque round colony, and the edge of the colony is complete.

Detailed Description

As described above, the present invention provides a probiotic composition for preventing or improving allergic diseases, which comprises, in parts by weight, 0.1 to 9.9 parts of lactobacillus acidophilus, 0.1 to 9.9 parts of lactobacillus rhamnosus, 0.1 to 40 parts of yeast probiotic or yeast extract, 0.1 to 50 parts of prebiotics, and optionally, probiotics.

The lactobacillus acidophilus L-92 strain is a lactobacillus which is carefully selected and found in long-term lactobacillus research and has an allergy relieving effect, and can relieve allergy symptoms by regulating balance of immune cells. The invention has obtained the invention patents of China, Japan, United states and so on, and has obtained the authentication of Self-Affirmed GRAS in the United states, honor the top prize in eighteenth Nutraward functional raw material field of the American exhibition Engredea. Under a normal state, the immune function protects our body from foreign body injury through the balanced action of three immune cells, namely Th1 cells, Th2 cells and Treg cells. However, in the allergic state, the immune balance is broken and the Th2 cells are excessively affected. Lactobacillus acidophilus L-92 acts to regulate the disrupted immune balance by increasing Treg cells for immune regulation and decreasing excess Th2 cells. Through the above effects, Lactobacillus acidophilus L-92 regulates the balance of Th1 cells and Th2 cells, and relieves anaphylaxis. Can reduce IgE content in blood of allergic patients, and has good relieving effect on pollinosis, perennial allergic rhinitis, and infantile and adult allergic dermatitis.

The lactobacillus rhamnosus LGG was isolated from stool samples of healthy adults in 1985 and met the ideal criteria for probiotic strains: the composition has the advantages of gastric acid and bile resistance, capability of adhering to human intestinal epithelial cells, capability of being planted in human small intestine to maintain undetermined intestinal flora, antibacterial substance production and definite health benefit. LGG is one of 9 probiotics approved by China and used for infant food, and can adsorb glycoprotein and ileum glycoprotein of intestinal mucosa, colonize in intestinal tract, improve damaged intestinal mucosa, balance bacterial phase in intestinal tract, inhibit growth of pathogenic bacteria and inhibit E. Studies have shown that oral administration of lactobacillus rhamnosus (LGG) can reduce Th2 immune responses such as high reactivity scores and whole milk protein (CMP) -specific IgG1 antibodies in serum, while promoting the production of Th1 immune responses including increased interferon- γ (IFN- γ) and CMP-specific IgG2a antibodies. Therefore, the lactobacillus rhamnosus LGG has good prevention and treatment effects on food allergy of infants, particularly milk protein allergy. LGG can increase the proportion of Th1 cells by stimulating the secretion of Th1 cell-associated cytokine IFN-gamma, and inhibit the secretion of Th2 cell-associated cytokine IL-4, thereby reducing the incidence of allergic diseases related to imbalance of the ratio of Th1 cells to Th2 cells. Has good therapeutic effect on food allergy, airway allergy, skin allergy, etc.

The lactobacillus rhamnosus can inhibit many pathogens, including multiple strains of escherichia coli, streptococcus, pseudomonas, salmonella, bacteroides fragilis, and clostridium.

In a preferred embodiment of the present invention, the composition comprises 0.2-7 parts of lactobacillus acidophilus, 0.2-7 parts of lactobacillus rhamnosus, 2-35 parts of yeast probiotic or yeast extract, 0.3-35 parts of prebiotics and 0.1-50 parts of optional probiotic bacteria; preferably, the lactobacillus acidophilus milk powder comprises 0.5 to 5 parts of lactobacillus acidophilus, 1 to 6 parts of lactobacillus rhamnosus, 5 to 30 parts of yeast probiotics or yeast extract, 1 to 30 parts of prebiotics and 1 to 40 parts of optional probiotics; preferably, the lactobacillus acidophilus milk beverage comprises 1-3 parts of lactobacillus acidophilus, 2-5 parts of lactobacillus rhamnosus, 10-20 parts of yeast probiotics or yeast extract, 2-10 parts of prebiotics and 3-20 parts of optional probiotics.

The yeast extract is a product obtained by taking food yeast as a main raw material and carrying out enzymolysis and autolysis (separation and extraction can be carried out again) under the combined action of enzyme of the yeast or additional food-grade enzyme, and is rich in soluble components in yeast cells such as nucleotide, amino acid, peptide, polypeptide and the like.

Under the condition that food allergy occurs to infants, nutrient components are inevitably lost. Among them, protein is one of the most important material bases of life, and therefore, the supplement of nutrients such as amino acids and peptides can make up for the deficiency or lack of nutrients caused by food allergy. The nucleotide is also an essential nutrient substance in the early life of human beings, and although the children can produce the nucleotide in vivo, if the amount of the nucleotide in the diet is proper, the growth and development, the gastrointestinal development and the immune function of the infants can be better. In addition, numerous studies have shown that the addition of nucleotides to the infant's diet contributes to the improvement of the intestinal microflora and the promotion of the growth of bifidobacteria in the intestine. The nucleotide can also inhibit pathogenic bacteria by reducing pH value of feces, thereby regulating intestinal microecological balance. That is, the nucleotides in the yeast extract can also act as prebiotics to some extent.

Preferably, the yeast probiotics are selected from one or more of beer yeast, Debaryomyces, Candida, Pichia, Torulopsis, Saccharomyces boulardii and Saccharomyces cerevisiae.

In a preferred embodiment of the present invention, wherein the optional probiotic bacteria are selected from one or more of lactobacillus paracasei, lactobacillus reuteri, lactobacillus gasseri, lactobacillus salivarius, lactobacillus johnsonii, lactobacillus fermentum, bifidobacterium longum, bifidobacterium breve, bifidobacterium lactis and bifidobacterium animalis; preferably, the probiotics are selected from one or two of bifidobacterium lactis and bifidobacterium animalis, and more preferably, the probiotics are 1.5-15 parts of bifidobacterium lactis and 1.5-15 parts of bifidobacterium animalis.

The lactobacillus paracasei can reduce the content of IgE in blood of allergic patients, increase the concentration of TNF-gamma of the allergic patients and effectively inhibit allergic reactions.

The lactobacillus reuteri can bear the action of gastric acid and bile to reach the upper part of the small intestine and adhere to the wall of the small intestine, has stronger adhesive capacity to the mucous membrane and epithelial cells of the host mucosa, and has special effect of generating broad-spectrum antibacterial substances.

In a preferred embodiment of the present invention, the prebiotic is one or more selected from the group consisting of fructooligosaccharide, stachyose, galactooligosaccharide, isomaltooligosaccharide and yeast β -glucan, preferably yeast β -glucan.

The yeast beta-glucan is a polysaccharide and is derived from yeast cell walls, the active structure of the yeast beta-glucan is polysaccharide consisting of glucose units, the yeast beta-glucan can activate macrophages, neutrophilic leukocytes and the like, so that the content of leukocidins, cytokinins and special antibodies is improved, the immune system of an organism is comprehensively stimulated, and a large number of experiments show that the beta-glucan can promote the generation of in-vivo immunoglobulin M antibodies so as to improve the immunity of body fluid, and can regulate the intestinal flora structure as a prebiotic.

The fructo-oligosaccharide is also called as fructo-oligosaccharide, and is a mixture of kestose, nystose and the like generated by combining 1-3 fructosyl groups with fructosyl groups in sucrose through beta (2-1) glycosidic bonds.

The galacto-oligosaccharide is a functional oligosaccharide with natural properties, and the molecular structure of the galacto-oligosaccharide is generally that 1-7 galactosyl groups are connected to galactose or glucose molecules, namely Gal- (Gal) n-Glc/Gal (n is 0-6).

The isomaltooligosaccharides are also called isomaltooligosaccharides, isomaltooligosaccharides and branched oligosaccharides, and the main components of the isomaltooligosaccharides, panose, isomaltotriose and oligosaccharides with more than tetrasaccharides are formed by combining glucose molecules by alpha-1, 6 glycosidic bonds.

In a preferred embodiment of the present invention, the composition further comprises an adjuvant, wherein the adjuvant is selected from one or more of anhydrous glucose, maltodextrin, maltitol, xylitol and fruit powder; preferably, the fruit powder is selected from one of hawthorn powder, apple powder, banana powder, cherry powder or sweet orange powder; preferably, the auxiliary materials are selected from one or more than two of 2-80 parts of anhydrous glucose, 0.1-15 parts of maltodextrin and 0-5 parts of hawthorn powder, and more preferably, the auxiliary materials are selected from 10-60 parts of anhydrous glucose and/or 5-10 parts of maltodextrin; preferably, the fruit powder is intended to provide a different mouthfeel.

In a preferred embodiment of the present invention, wherein the probiotic composition is present in an amount of 10% by dry weight²-10¹²Colony forming units, preferably 10⁷-10¹¹A colony forming unit.

In a preferred embodiment of the invention, wherein the probiotic composition is in the form of a powder, a tablet, a solid beverage, a capsule, a soup or a liquid suspension, preferably the probiotic composition may be in the form of an infant formula, a follow-up formula, a growing-up milk, an infant cereal, an infant food, a yoghurt, a cereal bar, a breakfast cereal, a dessert, a frozen food, a soup, a liquid suspension, a powder, a tablet, a solid beverage, a capsule, a candy, a nutritional composition or a nutritional supplement.

In a preferred embodiment of the present invention, the preparation method comprises mixing the prebiotics and the adjuvant, cooling to room temperature, and adding the yeast probiotic or yeast extract, lactobacillus acidophilus, lactobacillus rhamnosus, and optionally probiotic bacteria to mix to obtain the probiotic mixture.

In a preferred embodiment of the present invention, the method further comprises a drying step after the mixing and before the cooling to room temperature; preferably, the drying step is drying to a water activity of less than 0.2.

The manufacturers of the raw materials and equipment used in the present example, and the equipment and analysis method used in the product analysis are described below, wherein the chemical substances are not indicated as being chemically pure grades of conventional reagents. Information on the raw materials used in the examples and the experimental equipment are shown in tables 1 and 2

Table 1 information on the raw materials used in the examples

Raw material information	Purity of	Manufacturer of the product
			Yeast extract	100％(KA02)	Angel Yeast Co.,Ltd.
Fructo-oligosaccharide	97.5％	Shandong bowling biological shares Ltd
			Stachyose	99％	XI'AN APP-CHEM BIO(TECH) Co.,Ltd.
Galacto-oligosaccharides	98％	Quantum high tech (China) biology Co.,Ltd.
			Isomaltooligosaccharide	98％	Shandong bowling biological shares Ltd
Yeast beta-glucans	85％	Angel Yeast Co.,Ltd.
			Anhydrous glucose	99％	Wuhan Kafu technologies Co Ltd
Maltodextrin	99.5％	SHANDONG BAILONG CHUANGYUAN BIO-TECH Co.,Ltd.
			Hawthorn powder	99％	Tuofang-Zeshengrong-fruit and vegetable products Limited
Sweet orange powder	99％	Tianjin Xiugu Biotech development Co Ltd

TABLE 2 information of the experimental equipment used in the examples

Device name	Model number	Manufacturer of the product
			Boiling granulator (drying equipment)	FL-300C	CHONGQING JIAMA MECHANICAL MANUFACTURING Co.,Ltd.
Strip-shaped packaging machine	6 rows of	PHARMAPACK TECHNOLOGIES Corp.
			Full-automatic hard capsule filling machine	NJP-2000B	BEIJING HANLIN HANGYU TECHNOLOGY DEVELOPMENT Inc.
Powder packaging machine	PW-129CO	Wuhan Miwei mechanical Co Ltd

The lactobacillus casei Zhang used in the examples was purchased from tokyo koho biotechnology limited;

lactobacillus acidophilus L-92 was purchased from shanghai cheng-food materials ltd;

lactobacillus rhamnosus gg (lgg) purchased from beijing muji health technologies ltd;

lactobacillus reuteri RC14 was purchased from Beijing Muji health science and technology Ltd

Bifidobacterium lactis HN019 was purchased from Kangpini New Material Co., Ltd, Guangzhou City

Lactobacillus rhamnosus HN001 available from Kangpini New Material Co., Ltd

Bifidobacterium animalis Bb-12 was purchased from Beijing Muji health science and technology Ltd

Lactobacillus plantarum P-8 from Beijing Ke Tuhengtong Biotechnology GmbH

Bifidobacterium lactis V9 was purchased from Beijing Ke Tuo Hengtong Biotech GmbH

Example one

Uniformly mixing 39.8g of anhydrous glucose, 0.2g of maltodextrin and 0.1g of fructo-oligosaccharide, drying until the water activity is 0.1, cooling to room temperature, adding and uniformly mixing 0.1g of lactobacillus acidophilus L-92, 0.1g of lactobacillus rhamnosus GG, 9.9g of lactobacillus casei Zhang, 9.9g of lactobacillus reuteri RC14, 9.9g of bifidobacterium lactis HN019 and 30g of saccharomyces cerevisiae Z2.1, and packaging the mixed material into 2g of probiotic solid beverage in one bag by using a strip packaging machine.

The probiotic content was determined as described in GB4789.35-2016, and it was determined that each bag contained Lactobacillus acidophilus L-922 x 10⁹CFU, Lactobacillus rhamnosus GG6 x 10⁸CFU, Lactobacillus casei Zhang2 x 10¹¹CFU, Lactobacillus reuteri RC 142 x 10¹¹CFU, Bifidobacterium lactis HN 0192 x 10¹¹CFU, Saccharomyces cerevisiae Z2.11.2 x 10¹¹CFU。

The lactobacillus is determined by the following steps:

1 sample preparation

1.1 the entire preparation of the samples should follow the sterile procedure.

Weighing 25g of sample by aseptic operation, placing the sample into an aseptic homogenizing cup filled with 225mL of physiological saline, and homogenizing for 1min to 2min at 8000r/min to 10000r/min to prepare 1:10 sample homogenizing solution; or placing the mixture into a sterile homogenizing bag of 225mL physiological saline, and beating the mixture for 1min to 2min by a beating type homogenizer to prepare a 1:10 sample homogenizing solution.

1.2 step

1.2.1 sucking 1:10 sample homogeneous solution 1mL with 1mL sterile pipette or micropipette, slowly injecting into a sterile test tube containing 9mL physiological saline along the tube wall (taking care that the tip of the pipette does not touch the diluent), shaking the test tube or repeatedly blowing and beating with 1 sterile pipette to mix uniformly, and preparing the sample homogeneous solution of 1: 100.

1.2.2 taking another 1mL sterile pipette or micropipette suction head, homogenizing the sample by 10 times according to the operation sequence, and changing the 1mL sterile pipette or suction head for 1 time after diluting once.

1.2.3 lactic acid bacteria count

1.2.3.1 Total number of lactic acid bacteria

According to the estimation of the total number of viable bacteria of a sample to be detected, 2-3 continuous proper dilutions are selected, 0.1mL of sample uniform solution is absorbed by each dilution and is respectively placed on 2 MRS agar plates, and an L-shaped rod is used for surface coating. Counting all colonies on the plate after anaerobic culture for 48h +/-2 h at 36 +/-1 ℃. Dilution from sample to plate coating requires completion within 15 min.

1.2.3.2 Bifidobacterium count

According to the estimation of the content of bifidobacteria in the sample to be detected, 2-3 consecutive suitable dilutions were selected, each dilution sucking 0.1mL of sample homogenate on Mupirocin lithium salt (Li-Mupirocin) modified MRS agar plates, surface coating was performed using sterilized L-bars, and each dilution was made into two plates. Counting all colonies on the plate after anaerobic culture for 48h +/-2 h at 36 +/-1 ℃. Dilution from sample to plate coating requires completion within 15 min.

1.2.3.4 Lactobacillus count

Subtracting the total number of lactobacillus items from the total number of lactobacillus items 1.2.3.1 to obtain the count of lactobacillus.

1.3 colony counting

The dilution factor and the corresponding number of colonies can be recorded visually, if necessary with a magnifying glass or a colony counter. Colony counts are expressed in colony-forming units (CFU).

1.3.1 the total number of colonies was counted on a plate with a colony count of 30CFU to 300CFU and no spread colony growth. Plates below 30CFU record specific colony counts, and plates above 300CFU record as many as not. The number of colonies per dilution should be taken as the average of two plates.

1.3.2 when one of the plates has large flaky colony growth, the plate without the flaky colony growth is not suitable for use, and the plate without the flaky colony growth is used as the colony number of the dilution; if the plate-shaped colonies are less than half of the plate and the colonies in the other half are uniformly distributed, the number of the colonies on one plate can be represented by multiplying half of the plate by 2.

1.3.3 when chain growth occurs on the plate with no distinct boundaries between colonies, each single strand is counted as a colony.

1.4 presentation of results

1.4.1 if the number of colonies on only one dilution plate is within the appropriate count range, the average of the numbers of colonies on both plates is calculated and the average is multiplied by the corresponding dilution factor as a result of the total number of colonies per g (mL).

1.4.2 if the number of plate colonies with two serial dilutions is within the appropriate count range, then N ∑ C/(N) is calculated according to equation (1)₁+0.1n₂)d……………………………(1)

In the formula:

n-number of colonies in the sample;

sigma C-the sum of the number of colonies on a plate (a plate containing the appropriate range of colonies);

n 1-number of plates at first dilution (low dilution factor);

n 2-number of plates at second dilution (high dilution factor);

d-dilution factor (first dilution).

1.4.3 if the number of colonies on all the diluted plates is greater than 300CFU, the plate with the highest dilution is counted, and the other plates can be counted as the number of colonies which is not counted, and the result is calculated by multiplying the average number of colonies by the highest dilution factor.

1.4.4 if the number of plate colonies at all dilutions is less than 30CFU, it should be calculated as the average number of colonies at the lowest dilution times the dilution factor.

1.4.5 if all dilutions (including the liquid sample stock) plates were grown aseptically, then calculated as less than 1 times the lowest dilution factor.

1.4.6 if the colony count of the plate at all dilutions is not between 30CFU and 300CFU, and some of them are less than 30CFU or more than 300CFU, then the average colony count closest to 30CFU or 300CFU is multiplied by the dilution factor.

1.5 colony counts

1.5.1 colonies less than 100CFU, the "round off" rule was modified and reported as an integer.

1.5.2 when the colony number is more than or equal to 100CFU, after the 3 rd digit is reduced by adopting the principle of 'rounding off', the first 2 digits are taken, and the digits are replaced by 0; it can also be expressed in the form of an index of 10, reduced by the "rounding" principle, followed by two significant digits.

1.5.3 weight samples are reported in CFU/g and volume samples are reported in CFU/mL.

2 results and reports

Reports are made based on colony counts, reported in CFU/g (mL).

Example two

Mixing 30.3g anhydrous glucose, 9g maltodextrin and 30g isomaltose hypgather uniformly, drying until the water activity is 0.12, cooling to room temperature, adding and mixing 9.9g lactobacillus acidophilus L-92, 9.9g lactobacillus rhamnosus HN001, 9.9g bifidobacterium animalis Bb-12, 0.2g Saccharomyces boulardii Angel 1.27 and 0.8g yeast extract uniformly, and filling the mixed material into a capsule with the net content of 0.3g by a capsule filling machine.

The content of the probiotic bacteria in each capsule was determined according to the method of example one, and each capsule was determined to contain lactobacillus acidophilus L-923 x 10¹⁰CFU, Lactobacillus rhamnosus HN 0018.9 × 10⁹CFU, Bifidobacterium animalis Bb-128.9 x 10⁹CFU, Saccharomyces boulardii Angel 1.271.2 x 10⁸CFU。

EXAMPLE III

Uniformly mixing 40g of anhydrous glucose, 4.9g of hawthorn powder, 7 parts of maltodextrin and 0.1g of yeast beta-glucan, drying until the water activity is 0.08, cooling to room temperature, adding 8g of lactobacillus acidophilus L-92, 5g of lactobacillus rhamnosus GG, 5g of lactobacillus plantarum P-8, 5g of lactobacillus casei Zhang, 5g of bifidobacterium lactis V9, 5g of bifidobacterium animalis Bb12 and 15g of beer yeast A3.12, uniformly mixing, and packaging the mixed material into 1g of probiotic solid beverage in one bag by using a bar type packaging machine.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain Lactobacillus acidophilus L-928 x 10¹⁰CFU, Lactobacillus rhamnosus GG 1.5 x 10¹⁰CFU, Lactobacillus plantarum P-85 x 10¹⁰CFU, Lactobacillus casei Zhang 5 x 10¹⁰CFU, Bifidobacterium lactis V95 x 10¹⁰CFU, Bifidobacterium animalis Bb121.5 x 10¹⁰CFU and cerevisiae Fermentum A3.123 × 10¹⁰CFU。

Example four

Mixing anhydrous glucose 15g, maltodextrin 15g and yeast beta-glucan 48g, drying until the water activity is 0.15, cooling to room temperature, adding lactobacillus acidophilus L-92 g, lactobacillus rhamnosus HN001 g and bifidobacterium animalis Bb-12 8g, mixing uniformly, and filling the mixed material into capsules with net content of 0.3g by a capsule filling machine.

The probiotic content of each capsule was determined as described in example one, and each capsule was determined to contain lactobacillus acidophilus L-922.4 x 10¹⁰CFU, Lactobacillus rhamnosus HN 0014.5 × 10⁹CFU, Bifidobacterium animalis Bb-126.3 x 10⁹CFU。

EXAMPLE five

Uniformly mixing 25.2g of anhydrous glucose and 5g of galacto-oligosaccharide, drying until the water activity is 0.1, cooling to room temperature, adding and uniformly mixing 9.9g of lactobacillus acidophilus L-92, 9.9g of lactobacillus rhamnosus GG, 5g of lactobacillus plantarum P-8, 5g of lactobacillus reuteri RC14 and 40g of yeast extract, and packaging the mixed material into 1g of probiotic solid beverage in one bag by using a strip packaging machine.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain lactobacillus acidophilus L-929.9 x 10¹⁰CFU, Lactobacillus rhamnosus GG 3 x 10¹⁰CFU, Lactobacillus reuteri P-85 10¹⁰CFU, Lactobacillus plantarum RC 145 x 10¹⁰CFU。

EXAMPLE six

0.1g of sweet orange powder, 1.9g of maltodextrin, 58g of anhydrous glucose and 20g of stachyose are uniformly mixed, dried until the water activity is 0.12, 9.9g of lactobacillus acidophilus L-92, 0.1g of lactobacillus rhamnosus HN001, 5g of lactobacillus casei Zhang and 5g of saccharomyces cerevisiae FX-2 are added and uniformly mixed after being cooled to the room temperature, and the mixed material is filled into 1g of pure-content infant complementary food by a packaging machine.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain lactobacillus acidophilus L-929.9 x 10¹⁰CFU, Lactobacillus rhamnosus HN 0013 x 10⁸CFU, Lactobacillus casei Zhang 5 x 10¹⁰CFU, Saccharomyces cerevisiae FX-21 x 10¹⁰CFU。

EXAMPLE seven

77g of anhydrous glucose and 2g of yeast beta-glucan are uniformly mixed, dried until the water activity is 0.18, 1g of lactobacillus acidophilus L-92, 5g of lactobacillus rhamnosus GG, 3g of bifidobacterium lactis HN019, 2g of bifidobacterium animalis Bb-12 and 10g of yeast extract are added and uniformly mixed after being cooled to the room temperature, and the mixed material is filled into 1g of infant food with net content by a packaging machine.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain Lactobacillus acidophilus L-921 x 10¹⁰CFU, Lactobacillus rhamnosus GG 1.5 x 10¹⁰CFU, Bifidobacterium lactis HN 0193 x 10¹⁰CFU, Bifidobacterium animalis Bb-126 x 10⁹CFU。

Example eight

Mixing anhydrous glucose 55g, maltodextrin 3.5g and yeast beta-glucan 10g, drying until the water activity is 0.06, cooling to room temperature, adding lactobacillus acidophilus L-92 g, lactobacillus rhamnosus HN001 g, bifidobacterium lactis HN019 3g, bifidobacterium animalis Bb-12 g, beer yeast A3.12 g and yeast extract 10g, mixing, and filling the mixed material into a packaging machine to obtain 1g pure content infant complementary food.

The probiotic content of each bag was determined as described in example one, and each bag contained lactobacillus acidophilus L-923 x 10¹⁰CFU, Lactobacillus rhamnosus HN 0016 x 10⁹CFU, Bifidobacterium lactis HN 0196 x 10¹⁰CFU, Bifidobacterium animalis Bb-126 x 10⁹CFU, Saccharomyces cerevisiae A3.122 x 10¹⁰CFU。

Example nine

70g of anhydrous glucose, 7.5g of maltodextrin and 1g of galacto-oligosaccharide are uniformly mixed, dried until the water activity is 0.15, cooled to room temperature, 0.5g of lactobacillus acidophilus L-92, 6g of lactobacillus rhamnosus GG, 0.1g of bifidobacterium lactis V9, 9.9g of bifidobacterium animalis Bb-12 and 5g of beer yeast A3.12 are added and uniformly mixed, and the mixed material is filled into a packaging machine to be 1g of infant food supplement with net content.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain Lactobacillus acidophilus L-925 x 10⁹CFU, Lactobacillus rhamnosus GG 1.8 x 10¹⁰CFU, Bifidobacterium lactis V91 x 10⁹CFU, Bifidobacterium animalis Bb-123 x 10¹⁰CFU, Saccharomyces cerevisiae A3.121 x 10¹⁰CFU。

Example ten

Uniformly mixing 10g of fructo-oligosaccharide, 20g of yeast beta-glucan, 10g of maltodextrin, 4g of banana powder and 10g of anhydrous glucose, drying until the water activity is 0.11, cooling to room temperature, adding and uniformly mixing 5g of lactobacillus acidophilus L-92, 1g of lactobacillus rhamnosus HN001, 9.9g of bifidobacterium lactis HN019, 0.1g of bifidobacterium animalis Bb-12, 20g of yeast extract and 10g of saccharomyces cerevisiae FX-2, and filling the mixed material into a capsule with the net content of 0.3g by using a capsule filling machine.

The probiotic content of each capsule was determined as described in example one, and each capsule was determined to contain lactobacillus acidophilus L-921.5 x 10¹⁰CFU, Lactobacillus rhamnosus HN 0019 x 10⁸CFU, Bifidobacterium lactis HN 0193 x 10¹⁰CFU, Bifidobacterium animalis Bb-129 x 10⁷CFU, Saccharomyces cerevisiae FX-26 x 10⁹CFU。

EXAMPLE eleven

Uniformly mixing 80g of anhydrous glucose, 4.4g of maltodextrin, 5g of hawthorn powder and 0.3g of stachyose, drying until the water activity is 0.1, cooling to room temperature, adding and uniformly mixing 7g of lactobacillus acidophilus L-92, 0.2g of lactobacillus rhamnosus GG, 3g of lactobacillus casei Zhang and 0.1g of Saccharomyces boulardii Angel 1.27, and filling the mixed material into a packaging machine to obtain 1g of pure-content infant complementary food.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain lactobacillus acidophilus L-927 x 10¹⁰FU, Lactobacillus rhamnosus GG6 x 10⁸CFU, Lactobacillus casei Zhang 3 x 10¹⁰CFU, Saccharomyces boulardii Angel 1.272 x 10⁸CFU。

Example twelve

Evenly mixing 35g of galacto-oligosaccharide, 2g of anhydrous glucose and 0.8g of apple powder, drying until the water activity is 0.1, cooling to room temperature, adding and evenly mixing 0.2g of lactobacillus acidophilus L-92, 7g of lactobacillus rhamnosus HN001, 5g of lactobacillus reuteri RC14, 10g of lactobacillus casei Zhang, 5g of lactobacillus plantarum P-8, 30g of saccharomyces cerevisiae Z2.1 and 5g of yeast extract, and filling the mixed material into a packaging machine to obtain 1g of pure-content infant complementary food.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain Lactobacillus acidophilus L-922 x 10⁹FU, Lactobacillus rhamnosus HN 0012 x 10¹⁰CFU, Lactobacillus casei Zhang 1 x 10¹¹CFU, Roy's milk rodBacterium RC 145 x 10¹⁰CFU, Saccharomyces cerevisiae Z2.16 x 10¹⁰CFU, Lactobacillus plantarum P-85 x 10¹⁰CFU。

EXAMPLE thirteen

5g of maltodextrin, 39g of anhydrous glucose and 10g of yeast beta-glucan are uniformly mixed, dried until the water activity is 0.15, 3g of lactobacillus acidophilus L-92, 2g of lactobacillus rhamnosus HN001, 1g of bifidobacterium animalis Bb-12 and 40g of yeast powder are added and uniformly mixed after being cooled to the room temperature, and the mixed material is filled into 1g of infant complementary food with net content by a packaging machine.

The probiotic content of each bag was determined as described in example one, and each bag contained lactobacillus acidophilus L-923 x 10¹⁰FU, Lactobacillus rhamnosus HN 0016 x 10⁹CFU, Bifidobacterium animalis Bb-123 x 10⁹CFU。

Example fourteen

5g of galacto-oligosaccharide and 48g of anhydrous glucose are uniformly mixed, dried until the water activity is 0.18, cooled to room temperature, 5g of lactobacillus acidophilus L-92, 1g of lactobacillus rhamnosus GG, 20g of lactobacillus reuteri RC14, 20g of lactobacillus plantarum P-8 and 1g of yeast extract are added and uniformly mixed, and the mixed material is filled into 1g of infant complementary food with net content by a packaging machine.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain Lactobacillus acidophilus L-925 x 10¹⁰FU, Lactobacillus rhamnosus GG 3 x 10⁹CFU, Lactobacillus reuteri RC 142 x 10¹¹CFU, Lactobacillus plantarum P-82 x 10¹¹CFU。

Example fifteen

2g of yeast beta-glucan, 60g of anhydrous glucose and 8.2g of maltodextrin are uniformly mixed, the mixture is dried until the water activity is 0.09, 9.9g of lactobacillus acidophilus L-92, 9.9g of lactobacillus rhamnosus GG and 10g of yeast extract are added and uniformly mixed after being cooled to the room temperature, and the mixed material is filled into 0.5g of infant complementary food with net content by a packaging machine.

The probiotic content of each bag was determined as described in example one, and each bag was determined to contain lactobacillus acidophilusL-92 5*10¹⁰FU, Lactobacillus rhamnosus GG 1.5 x 10¹⁰CFU。

Comparative example 1

69.8g of anhydrous glucose, 0.2g of maltodextrin and 0.1g of fructo-oligosaccharide are uniformly mixed, dried until the water activity is 0.1, cooled to room temperature, 0.1g of lactobacillus acidophilus L-92, 0.1g of lactobacillus rhamnosus GG, 9.9g of lactobacillus casei Zhang, 9.9g of lactobacillus reuteri RC14 and 9.9g of bifidobacterium lactis HN019 are added and uniformly mixed, and the mixed material is divided into 0.5g of probiotics solid beverage in bags by a strip type packaging machine.

The probiotic content was determined as described in example one, and each bag contained Lactobacillus acidophilus L-921 x 10⁹CFU, Lactobacillus rhamnosus GG 3 x 10⁸CFU, Lactobacillus casei Zhang 9.9 x 10¹⁰CFU, Lactobacillus reuteri RC 149.9X 10¹⁰CFU, Bifidobacterium lactis HN 0199.9 x 10¹⁰CFU。

Comparative example No. two

Uniformly mixing 24.8g of anhydrous glucose, 0.2g of maltodextrin and 0.1g of fructo-oligosaccharide, drying until the water activity is 0.1, cooling to room temperature, adding and uniformly mixing 0.1g of lactobacillus acidophilus L-92, 0.1g of lactobacillus rhamnosus HN001, 9.9g of lactobacillus casei Zhang, 9.9g of lactobacillus reuteri RC14, 9.9g of bifidobacterium lactis V9 and 45g of saccharomyces cerevisiae FX-2, and packaging the mixed material into 1g of probiotic solid beverage in one bag by using a strip packaging machine.

The probiotic content was determined as described in example one, and each bag contained Lactobacillus acidophilus L-921 x 10⁹CFU, Lactobacillus rhamnosus HN 0013 x 10⁸CFU, Lactobacillus casei Zhang 9.9 x 10¹⁰CFU, Lactobacillus reuteri RC 149.9X 10¹⁰CFU, Bifidobacterium lactis V99.9 x 10¹⁰CFU, Saccharomyces cerevisiae FX-29 x 10¹⁰CFU。

Comparative example No. three

Uniformly mixing 39.9g of anhydrous glucose and 0.2g of maltodextrin, drying until the water activity is 0.15, cooling to room temperature, adding and uniformly mixing 0.1g of lactobacillus acidophilus L-92, 0.1g of lactobacillus rhamnosus GG, 9.9g of lactobacillus casei Zhang, 9.9g of lactobacillus reuteri RC14, 9.9g of bifidobacterium lactis HN019 and 30g of saccharomyces cerevisiae Z2.1, and packaging the mixed material into 1g of probiotic bar type solid beverage by a packaging machine.

The probiotic content was determined as described in example one, and each bag contained Lactobacillus acidophilus L-921 x 10⁹CFU, Lactobacillus rhamnosus GG 3 x 10⁸CFU, Lactobacillus casei Zhang 9.9 x 10¹⁰CFU, Lactobacillus reuteri RC 149.9X 10¹⁰CFU, Bifidobacterium lactis HN 0199.9 x 10¹⁰CFU, Saccharomyces cerevisiae Z2.16 x 10¹⁰CFU。

Comparative example No. four

39.8g of anhydrous glucose, 0.2g of maltodextrin and 0.1g of fructo-oligosaccharide are uniformly mixed, 0.1g of lactobacillus acidophilus L-92, 0.1g of lactobacillus rhamnosus GG, 9.9g of lactobacillus casei Zhang, 9.9g of lactobacillus reuteri RC14, 9.9g of bifidobacterium lactis HN019 and 30g of saccharomyces cerevisiae Z2.1 are uniformly mixed, the mixture is dried until the water activity is 0.1, and the mixed material is divided into 2g of probiotic solid beverage bags by a strip type packaging machine.

The probiotic content was determined as described in example one, and each bag contained Lactobacillus acidophilus L-921 x 10⁸CFU, Lactobacillus rhamnosus GG 2.5 x 10⁷CFU, Lactobacillus casei Zhang 5 x 10⁸CFU, Lactobacillus reuteri RC 142.5 x 10¹⁰CFU, Bifidobacterium lactis HN 0191.2 x 10¹⁰CFU, Saccharomyces cerevisiae Z2.12 x 10¹⁰CFU。

The difference between the fourth comparative example and the first example is that the operation steps are different, the first example is that auxiliary materials and prebiotics are added firstly for drying, and then probiotics are added, the fourth comparative example is obtained by mixing all the components and then drying, and from the content of the probiotics obtained in each bag, the content of the probiotics obtained in the fourth comparative example is greatly reduced because the probiotics are not resistant to high temperature, and part of the probiotics can die due to drying after being mixed with the auxiliary materials and the prebiotics.

Comparative example five

Uniformly mixing 39.8g of anhydrous glucose, 0.2g of maltodextrin, 0.1g of fructo-oligosaccharide, 0.1g of lactobacillus acidophilus L-92, 0.1g of lactobacillus rhamnosus GG, 9.9g of lactobacillus casei Zhang, 9.9g of lactobacillus reuteri RC14, 9.9g of bifidobacterium lactis HN019 and 30g of saccharomyces cerevisiae Z2.1, drying until the water activity is 0.295, and packaging the mixed material into 2g of probiotic solid beverage bags by using a strip packaging machine.

The probiotic content was determined as described in example one, and each bag contained Lactobacillus acidophilus L-921.5 x 10⁹CFU, rhamnosus GG 5 x 10⁸CFU, Lactobacillus casei Zhang 1.6 x 10¹¹CFU, Lactobacillus reuteri RC 141.8 x 10¹¹CFU, Bifidobacterium lactis HN 0192 x 10¹¹CFU, Saccharomyces cerevisiae Z2.11.2 x 10¹¹CFU。

Comparing the first example with the fifth comparative example, the difference is that the water activities are different, the water activity of the first example is 0.1, and the water activity of the fifth comparative example is 0.295, and the content of the probiotics contained in each capsule is higher than that of the second example, and lower than that of the fifth comparative example.

Each bag of probiotics obtained in example one and comparative example five was tested after 1 week, and each bag in example one contained Lactobacillus acidophilus L-922 x 10⁹CFU, Lactobacillus rhamnosus GG6 x 10⁸CFU, Lactobacillus casei Zhang2 x 10¹¹CFU, Lactobacillus reuteri RC 142 x 10¹¹CFU, Bifidobacterium lactis HN 0192 x 10¹¹CFU, Saccharomyces cerevisiae Z2.11.2 x 10¹¹A CFU; while each bag of comparative example five contained lactobacillus acidophilus L-921.5 x 10⁹CFU, Lactobacillus rhamnosus GG 4 x 10⁸CFU, Lactobacillus casei Zhang 1.2 x 10¹⁰CFU, Lactobacillus reuteri RC 145 x 10¹⁰CFU, Bifidobacterium lactis HN 0199.2 x 10¹⁰CFU, Saccharomyces cerevisiae Z2.13.5 x 10¹⁰CFU。

From the comparison results, it can be seen that the number of the probiotics in the first example is relatively stable, while the probiotics in the fifth example has a certain loss, which indicates that the probiotics have a higher requirement on water activity, the water activity is generally stable below 0.2, and the higher the water activity, the faster the bacteria decay.

Effects of the embodiment

1. The treatment subjects were: 54 cases of allergic rhinitis patients (age 18-60 years) clinically collected in first people hospital in Yichang city in 2017, 1 month-2017, 5 months are selected by the inventor, and the symptoms of the patients are as follows: rhinitis symptoms such as rhinorrhea, nasal obstruction, rhinocnesmus and sneeze, and rhinitis symptoms such as itchy eyes, lacrimation, red and swollen eyes, itchy ears and cough.

2. The treatment method comprises the following steps: 2g of the probiotic composition prepared in the first embodiment of the invention is taken every time, 2 times a day, 2 granules of the probiotic composition prepared in the second embodiment is taken every time, 2 times a day, and 2 weeks continuously; comparative example 1g of the probiotic composition prepared in the first example, 2 times a day; the probiotic composition prepared in the second comparative example is 2g each time and 2 times a day; the probiotic composition prepared in the third comparative example is 2g each time and 2 times a day; 9 cases of each group. The clinical manifestations and treatment responses of the patients were observed on the day of treatment and 2 weeks, respectively, and the treatment results are shown in table 3 below, wherein the treatment criteria are:

and (3) healing: the rhinitis symptom disappears, and the external symptoms of the rhinitis disappear;

the method has the following advantages: relieving rhinitis symptoms, and eliminating external symptoms of rhinitis;

and (4) invalidation: the symptoms are not alleviated.

TABLE 3 therapeutic results

Group of	Recovery/recovery	Effective/example	Invalid/example	Effective rate%
					Example one	5	2	2	78％
Example two	4	3	2	78％
					Comparative example 1	0	5	4	55％
Comparative example No. two	0	4	5	44％
					Comparative example No. three	0	4	5	44％

Wherein the effective rate is the total proportion of the recovery and the effective cases

And (4) conclusion: as can be seen from the above table, the probiotic compositions obtained in the first and second examples of the present invention have an effective rate of about 78% for allergic rhinitis, and can effectively alleviate the symptoms of allergic rhinitis, while the first to third comparative examples have an effect of less than 60%, which is significantly worse than that of the first and second examples.

Effect of embodiment two

1. The treatment subjects were: the inventor selects 42 clinically accepted atopic dermatitis patients (age 12-45 years) in the Wufeng county hospital of Yichang city in 2017, 1 month-2018, 1 month and 178 clinically accepted allergic children (age 1-14 years) in the child hospital of Wuhan city in 2017, 7 months-2018, 5 months. Symptoms are: itching, erythema, papules, profuse infiltration and lichenification.

2. The treatment method comprises the following steps: 2g of the probiotic composition prepared in the first embodiment of the invention is taken every time, and 1 time is taken every day; example two the resulting probiotic composition was prepared 2 capsules at a time, 1 time a day; 1g of the probiotic composition prepared in the third embodiment of the invention is taken every time, and 1 time is taken every day; taking the probiotic composition prepared in the fourth example 2 capsules each time, 1 time a day; taking the probiotic composition prepared in the fifth step 1g each time and 1 time a day; taking the probiotic composition prepared in the sixth example 1g each time, 1 time per day; taking 1g of the probiotic composition prepared in the seventh embodiment each time, 1 time per day; 1g of the probiotic composition prepared in example eight is taken 1 time a day; taking the probiotic composition prepared in the ninth embodiment 1g each time and 1 time per day; taking 3 granules of the probiotic composition prepared in the example ten, 1 time a day; taking 1g of the probiotic composition prepared in the eleventh example 1 time a day; taking 1g of the probiotic composition prepared in the twelfth example 1 time a day; taking 1g of the probiotic composition prepared in example thirteen every time and 1 time every day; administering 1g of the probiotic composition prepared according to the fourteenth embodiment 1 time a day; taking 1g of the probiotic composition prepared in the example fifteen per time and 1 time per day; comparative example 1g of the probiotic composition prepared in the first example, 2 times a day; the probiotic composition prepared in the second comparative example is 2g each time and 2 times a day; the probiotic composition prepared in the third comparative example is 2g each time and 2 times a day; the probiotic composition prepared in the fourth comparative example is 1 bag each time and 1 time a day; the probiotic composition prepared in the fifth comparative example is 1 bag each time and 1 time a day; each group had 11 doses, and all groups were taken continuously for 2 weeks. The clinical manifestations and treatment responses of patients were observed on the day of treatment, 2 weeks, and 4 weeks, respectively, and the treatment results are shown in table 4, wherein the therapeutic criteria are:

and (3) healing: the clinical symptoms disappear, the primary rash subsides, and no new eruption exists;

the effect is shown: the symptom is obviously relieved, and the obvious effect is that the symptom is relieved and is more than or equal to 70 percent;

improvement: the symptoms are obviously relieved, and the fading rate of the rash is more than or equal to 30 percent;

and (4) invalidation: thus, there was no remission and the rash resolved < 30%.

TABLE 4 therapeutic results

Wherein the effective rate is the proportion of cure, obvious effect and improvement in the total cases of the group.

Comparing the first example with the first comparative example, the difference is that the first example contains yeast probiotics, while the first comparative example does not contain yeast probiotics or yeast extract, and the probiotic composition prepared by the same preparation method, as can be seen from the data in the table, the effective rate in the first example is 73%, and the effective rate in the first comparative example is 36%, which indicates that the probiotic composition containing yeast or yeast extract has a certain effect on improving allergic diseases.

Comparing the first example with the second comparative example, the difference is that the first example contains a certain amount of yeast and the second comparative example contains 45g of yeast, and thus the corresponding probiotic composition is obtained. As can be seen from the data in the table, the effective rate in example one is 73%, and the effective rate in comparative example one is 45%, indicating that the probiotic composition containing specific content of yeast or yeast extract has a certain effect on the improvement of allergic diseases.

Comparing the first example with the third comparative example, the difference is that the first example contains prebiotics, while the third comparative example does not contain prebiotics, and thus the corresponding probiotic composition is obtained. As can be seen from the data in the table, the effective rate in example one is 73%, and the effective rate in comparative example one is 27%, indicating that the probiotic composition containing prebiotics has a certain effect on the improvement of allergic diseases.

In conclusion, the prepared probiotic composition has certain improvement on allergic rhinitis patients and atopic dermatitis patients, and as can be seen from table 3, the probiotic composition has certain improvement effect on allergic rhinitis, the effective rate of the probiotic composition reaches 78%, and the symptoms of the allergic rhinitis patients can be effectively relieved; as can be seen from table 4, the probiotic composition of the present invention has a certain therapeutic effect on atopic dermatitis patients, and the effective rate thereof is above 64% and up to 91%, which indicates that the probiotic composition of the present invention can effectively alleviate the symptoms of atopic dermatitis patients, thereby indicating that the probiotic composition of the present invention has a certain prevention or improvement effect on allergic diseases.

The foregoing is considered as illustrative and not restrictive in character, and that various modifications, equivalents, and improvements made within the spirit and principles of the invention are intended to be included within the scope of the invention.

Claims

1. A probiotic composition for preventing or improving allergic diseases comprises, by weight, 0.1-9.9 parts of Lactobacillus acidophilus, 0.1-9.9 parts of Lactobacillus rhamnosus, 0.1-40 parts of yeast probiotic or yeast extract, 0.1-50 parts of prebiotics and optionally probiotics.

2. The probiotic composition of claim 1 wherein said composition comprises 0.2-7 parts of lactobacillus acidophilus, 0.2-7 parts of lactobacillus rhamnosus, 2-35 parts of yeast probiotic or yeast extract, 0.3-35 parts of prebiotics and 0.1-50 parts of optional probiotic; preferably, the lactobacillus acidophilus milk powder comprises 0.5 to 5 parts of lactobacillus acidophilus, 1 to 6 parts of lactobacillus rhamnosus, 5 to 30 parts of yeast probiotics or yeast extract, 1 to 30 parts of prebiotics and 1 to 40 parts of optional probiotics; preferably, the lactobacillus acidophilus milk beverage comprises 1-3 parts of lactobacillus acidophilus, 2-5 parts of lactobacillus rhamnosus, 10-20 parts of yeast probiotics or yeast extract, 2-10 parts of prebiotics and 3-20 parts of optional probiotics.

3. The probiotic composition according to claim 1 or 2, wherein the yeast probiotic is selected from one or more of brewers yeast, debaryomyces, candida, pichia, torulopsis, blakeslea and saccharomyces cerevisiae.

4. A probiotic composition according to any one of claims 1 to 3, wherein said optional probiotic bacteria are selected from one or more of lactobacillus paracasei, lactobacillus reuteri, lactobacillus gasseri, lactobacillus salivarius, lactobacillus johnsonii, lactobacillus fermentum, bifidobacterium longum, bifidobacterium breve, bifidobacterium lactis and bifidobacterium animalis; preferably, the optional probiotics are selected from one or two of bifidobacterium lactis and bifidobacterium animalis, and more preferably, the probiotics are 1.5-15 parts of bifidobacterium lactis and 1.5-15 parts of bifidobacterium animalis.

5. A probiotic composition according to any one of claims 1 to 4, wherein said prebiotic is selected from one or more of fructooligosaccharides, stachyose, galactooligosaccharides, isomaltooligosaccharides and yeast β -glucans, preferably yeast β -glucans.

6. A probiotic composition according to any one of claims 1 to 5, wherein said composition further comprises an adjuvant, preferably said adjuvant is selected from one or more of anhydrous glucose, maltodextrin, maltitol, xylitol and fruit powder; preferably, the fruit powder is selected from one of hawthorn powder, apple powder, banana powder, cherry powder or sweet orange powder; preferably, the auxiliary material is selected from one or more than two of 2-80 parts of anhydrous glucose, 0.1-15 parts of maltodextrin and 0-5 parts of hawthorn powder, and more preferably, the auxiliary material is selected from 10-60 parts of anhydrous glucose and/or 5-10 parts of maltodextrin.

7. A probiotic composition according to any of claims 1 to 6, wherein the probiotic composition is present in an amount of 10 on a dry weight basis²-10¹²Colony forming units, preferably 10⁷-10¹¹A colony forming unit.

8. A probiotic composition according to any one of claims 1 to 7, wherein said probiotic composition is present in the form of a powder, tablet, solid beverage, capsule, soup or liquid suspension.

9. Method for the preparation of a probiotic composition according to any of claims 1 to 8, by mixing a Lactobacillus acidophilus, a Lactobacillus rhamnosus, a probiotic of yeasts or a yeast extract according to any of claims 1 to 8, a prebiotic and optionally a probiotic to obtain said probiotic composition.

10. The method of claim 9, wherein the prebiotics and the adjuvant are mixed and then cooled to room temperature, and then the probiotic mixture is obtained by adding the yeast probiotic or yeast extract, lactobacillus acidophilus, lactobacillus rhamnosus and optionally the probiotic.

11. The method of claim 10, further comprising a drying step after mixing and before cooling to room temperature; preferably, the drying step is drying to a water activity of less than 0.2.

12. Use of a probiotic composition according to any one of claims 1 to 8 or prepared by the preparation method according to any one of claims 9 to 11 for preventing or ameliorating allergic diseases.

13. A health food composition for preventing or improving allergic diseases, which comprises the probiotic composition of any one of claims 1 to 8 or the probiotic composition prepared by the preparation method of any one of claims 9 to 11.