LU503459B1 - Application of Bifidobacteriumlactis in improving intestinal flora disorder - Google Patents
Application of Bifidobacteriumlactis in improving intestinal flora disorder Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1238—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using specific L. bulgaricus or S. thermophilus microorganisms; using entrapped or encapsulated yoghurt bacteria; Physical or chemical treatment of L. bulgaricus or S. thermophilus cultures; Fermentation only with L. bulgaricus or only with S. thermophilus
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/531—Lactis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
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Abstract
The present invention belongs to the technical field of microorganism applications, and specifically relates to an application of Bifidobacteriumlactis in improving intestinal flora disorder. Bifidobacteriumlactis is assigned with the accession number of CGMCC No.21255. The Bifidobacteriumlactis is applied to improvement of intestinal flora disorder, and may restore Alpha diversity and Beta diversity of intestinal flora on an abnormal level to a normal level. That is to say, the diversity of disordered intestinal flora may be effectively restored, such that the diversity of the intestinal flora may be maintained at a balanced state. In particular, when being applied to preparation of yogurt, the Bifidobacteriumlactis may also improve the stability of the yogurt and reduce curding time.
Description
Application of Bifidobacteriumlactis in improving intestinal flora disorder LU503459
The present invention belongs to the technical field of microorganism applications, and specifically relates to an application of Bifidobacteriumlactis in improving intestinal flora disorder.
Intestinal microorganisms (flora) are closely related to health and diseases. The intestinal flora is complicated and numerous, and the main flora are Firmicutes,
Bacteroidetes and Proteobacteria. The intestinal flora and metabolites thereof play an important role in human energy metabolism, nutrient absorption, innate immunity, acquired immunity and gastrointestinal functions, for example, providing energy and nutrients by means of metabolic activities, for example, preventing the invasion of foreign microbes, for example, regulating immune factors, repairing an intestinal barrier function, and maintaining a steady state of an intestinal environment. In normal cases, the intestinal environment is relatively stable, but when being subjected to strong exogenous stimuli (for example, alcohol), the intestinal environment may change drastically to go beyond a regulation range of the intestinal flora, leading to the development of various diseases. For example, high short-term alcohol intake or long-term alcohol abuse may lead to a variety of health problems. Alcohol is in direct contact with the digestive tract during drinking, such that microorganisms in the mouth, esophagus and intestines are affected, and the steady state of the environment therein is destroyed. As a result, an imbalance in the internal environment may lead to damage to the digestive tract, and trigger diseases such as damage of esophageal and intestinal mucosa or gastric ulcers.
Probiotics are a class of bacteria that may regulate the total disorder of host intestinal flora and maintain the balance of human and animal intestinal flora by directly or indirectly adjusting the composition of host intestinal microorganisms and activating the activity of host endogenous microbiota or an immune system. The probiotics may effectively improve the structure of the intestinal flora, correct intestinal flora disorder of abdominal infections and reduce bacteria translocation, so as to protect the intestinal mucosa, and also have the functions of inhibiting pathogenic bacteria, eliminating carcinogenic factors and improving the immunity of the body, such that more and more studies use the probiotics to improve the intestinal flora disorder. However, there are numerous existing strains, and not all of the strains have the effect of regulating the intestinal flora disorder. For example, LU503459
Bifidobacteriumlactis WBBROS or Bifidobacterium longum ZCC14 cannot regulate the intestinal flora disorder. It may be seen that only some strains may adjust the balance of the intestinal flora by restoring the community diversity and composition of intestinal microorganisms, thereby improving the intestinal flora disorder and promoting human health.
Therefore, a strain that may improve intestinal flora disorder or discovery of new applications of existing strains in improving intestinal flora disorder is necessary.
In view of the above problems, the present invention is intended to provide a new application of Bifidobacteriumlactis (which is also called Bifidobacteriumlactis TY-SO1) with the accession number CGMCC No.21255 in improving intestinal flora disorder. The
Bifidobacteriumlactis may restore Alpha diversity and Beta diversity of intestinal flora on an abnormal level to a normal level. That is to say, the diversity of disordered intestinal flora may be effectively restored, such that the diversity of the intestinal flora may be maintained at a balanced state.
In order to achieve the above purpose, the present invention may use the following technical solutions.
The present invention provides an application of Bifidobacteriumlactis in improving intestinal flora disorder. Bifidobacteriumlactis is assigned with the accession number of
CGMCC No.21255.
Further, intestinal flora disorder may be alcoholic intestinal flora disorder.
Further, the application may include an application of Bifidobacteriumlactis in preparation of food, health care products or drugs.
Further, the food may be yogurt.
Further, a method for preparing the yogurt includes: inoculating the
Bifidobacteriumlactis and a starter culture into a yogurt base for fermentation.
Further, the addition of the Bifidobacteriumlactis is 1.0x10°CFU/mL-1.0x108CFU/mL.
Further, yeast power may be selected as the starter culture, of which addition is 0.01 g/L-0.03 g/L.
Further, fermentation may include: performing fermentation at 41°C-43°C until pH is 4446.
The present invention has the following beneficial effects. The Bifidobacteriumlactis LU503459 with the accession number of CGMCC No.21255 is applied to improvement of intestinal flora disorder. The Bifidobacteriumlactis may restore Alpha diversity and Beta diversity of intestinal flora on the abnormal level to the normal level. That is to say, the flora diversity of disordered intestinal flora may be effectively restored, such that the diversity of the intestinal flora can be maintained at the balanced state.
Fig. 1 shows a Shannon index of intestinal flora in mice.
Fig. 2 shows a Simpson index of intestinal flora in mice.
Fig. 3 shows a Chao index of intestinal flora in mice.
Fig. 4 shows Principal Coordinate Analysis (PCoA) of intestinal flora in mice.
Fig. 5 shows taxonomic distribution of phylum-level bacterial communities in mice.
Fig. 6 shows the relative abundance of Firmicutes in mice.
Fig. 7 shows a ratio of Firmicutes to Bacteroidetes in mice.
Fig. 8 shows the relative abundance of Proteobacteria in mice.
Fig. 9 shows difference analysis of genus-level bacterial communities in mice.
Fig. 10 shows taxonomic distribution of genus-level bacterial communities in mice.
Fig. 11 shows the curding time of yogurt in each group.
Fig. 12 shows a curding state of yogurt in each group.
Fig. 13 shows Water Holding Capacity (WHC) of yogurt in each group.
Fig. 14 shows the viscosity of yogurt in each group.
Fig. 15 shows the hardness of yogurt in each group.
Fig. 16 shows the pH of yogurt in each group.
Fig. 17 shows the acidity of yogurt in each group.
Fig. 18 shows the impact of Bifidobacteriumlactis TY-SO1 on the sense of yogurt.
In the figures, "*" represents that there is a statistical difference (P < 0.05) between two groups.
The embodiments are given to better describe the present invention, but the content of the present invention is not limited only to the given embodiments. Therefore, non-essential improvements and adjustments to the embodiments made by a person skilled in the art in accordance with the content of the above present invention still fall within the scope of protection of the present invention. LU503459
The terms used herein are only intended to describe specific embodiments and are not intended to limit the present invention. Expressions in the singular form include those in the plural form unless the expressions have a distinctly different meaning in the context. As used herein, it is to be understood that terms such as "include", "have", "contain", and the like are intended to indicate the presence of features, figures, operations, components, parts, elements, materials, or combinations. The terms of the present invention are disclosed in the specification and are not intended to exclude the possibility that one or more other features, figures, operations, components, parts, elements, materials, or combinations thereof may exist or may be added. As used here, "/" may be interpreted as "and" or "or", as appropriate.
The term “CFU/mL” in the present invention refers to the number of colonies in 1 ml of liquid. For example, when the addition of the Bifidobacteriumlactis in milk is 1.0x10°CFU/mL, it indicates that 1.0x10°CFU of the Bifidobacteriumlactis is added to 1 ml of milk.
An embodiment of the present invention provides an application of
Bifidobacteriumlactis in improving intestinal flora disorder. The Bifidobacteriumlactis is assigned with the accession number of CGMCC No.21255. It is to be noted that, the
Bifidobacteriumlactis is a strain which has been deposited in the workplace of the applicant, which was deposited with the China General Microbiological Culture Collection
Center (CGMCC) at No. 3, Yard 1, BeiChen West Road, Chaoyang District, Beijing on
November 27, 2020 and was assigned with the accession number of CGMCC No.21255, with the classification name being Bifidobacteriumlactis TY-SO1. In addition, in the present invention, it is discovered that an application of Bifidobacteriumlactis in improving intestinal flora disorder belongs to a new application of the Bifidobacteriumlactis. The
Bifidobacteriumlactis is applied to improvement of intestinal flora disorder. The
Bifidobacteriumlactis may restore Alpha diversity and Beta diversity of intestinal flora on the abnormal level to the normal level. That is to say, the flora diversity of disordered intestinal flora may be effectively restored, such that the diversity of the intestinal flora may be maintained at the balanced state.
In some specific embodiments, in the application of the Bifidobacteriumlactis in improving the intestinal flora disorder, the Bifidobacteriumlactis has been shown to improve the intestinal flora disorder in any condition known in the art, especially in alcoholic intestinal flora disorder. The Bifidobacteriumlactis has more significant effect on improving the alcoholic intestinal flora disorder than other disorder conditions, such as LU503459 intestinal flora disorder caused by antibiotics, intestinal flora disorder caused by diet, or intestinal flora disorder caused by abnormal intestinal motility. Specifically, alcohol addiction is not only a brain problem, but also related to the intestinal flora disorder. 5 Alcohol disrupts the balance of intestinal flora, and the intestinal flora in turn affects the brain, making people crave the alcohol even more, thereby creating a vicious cycle.
Therefore, the Bifidobacterium may be used to regulate the intestinal flora to a normal level, so as to affect the brain, thereby abstaining from the alcohol.
In some specific embodiments, in the application of the Bifidobacteriumlactis in improving the intestinal flora disorder, the application may be specifically embodied in preparation of food, health care products or drugs. It is to be noted that, the
Bifidobacteriumlactis may be added to the food, the health care products or the drugs in the form of viable bacteria, starter culture or auxiliary starter culture, such that, in addition to effects carried by the food, the health care products or the drugs, the food, the health care products or the drugs also have the effect of the Bifidobacteriumlactis in improving the intestinal flora disorder, especially the alcoholic intestinal flora disorder. That is to say, the
Bifidobacteriumlactis may enrich the effect of the food, the health care products or the drugs. It is to further be noted that, since the performance of the Bifidobacteriumlactis affects the stability and flavor and odor of the food, the health care products or the drugs, producing unpleasant odor and affecting overall taste, not any Bifidobacteriumlactis may be applied to the food, the health care products or the drugs. For example,
Bifidobacteriumlactis B04, Bifidobacteriumlactis WLABO9 or Bifidobacterium bifidum
B96 is difficultly applied to the food, the health care products or the drugs due to the impact of flavor and odor. The Bifidobacteriumlactis in the present invention does not affect the stability and flavor and odor of the food, the health care products or the drugs, and also makes the food, the health care products or the drugs have the effect of improving the intestinal flora disorder.
In some specific embodiments, in the application of the Bifidobacteriumlactis in improving the intestinal flora disorder, the Bifidobacteriumlactis may be applied to the preparation of yogurt. Specifically, in recent years, probiotics increasingly appear in the field of food. The yogurt is an ideal carrier delivering the probiotics to the human gastrointestinal tract, which is an important application form of the probiotics in the field of functional food. The probiotics may not only improve the functionality of the yogurt, but also improve the quality of the yogurt and give the yogurt a unique flavor. The yogurt prepared by the Bifidobacteriumlactis in the present invention is high in safety, and may LU503459 also prevent and improve the alcoholic intestinal flora disorder, which has a very important meaning and a wide market prospect.
In some specific embodiments, in the application of the Bifidobacteriumlactis in improving the intestinal flora disorder, the method that the Bifidobacteriumlactis may be applied to the preparation of the yogurt may include: inoculating the Bifidobacteriumlactis and a starter culture into a yogurt base for fermentation. It is to be noted that, by applying the Bifidobacteriumlactis to the preparation of the yogurt, in addition to high safety and effects of improving the intestinal flora disorder, the WHC, hardness, viscosity and curding time of the yogurt may also be obviously improved, so as to cause the yogurt to have good stability. Specifically, compared with the yogurt prepared by using the
Bifidobacteriumlactis as the auxiliary starter culture, the WHC, hardness and viscosity of the yogurt prepared with the Bifidobacteriumlactis are obviously improved, the curding time is obviously reduced, and the yogurt shows obvious dose-dependence. In addition, changes in the pH and acidity of the yogurt prepared with the Bifidobacteriumlactis during storage at 4°C are relatively small, indicating that the post-acidification of the yogurt may not be caused. Furthermore, the prepared yogurt is excellent in color, taste and flavor. It is to further be noted that, the starter culture is fermentative bacteria for the yogurt known in the art, such as Lactobacillus bulgaricus or Streptococcus thermophilus. The yogurt base at least includes raw milk or mixed milk, or may include additives such as sweetening agents and stabilizing agents.
In some specific embodiments, in the application of the Bifidobacteriumlactis in improving the intestinal flora disorder, when the Bifidobacteriumlactis and the starter culture are inoculated into the yogurt base for fermentation, the addition of the
Bifidobacteriumlactis is 1.0x10°CFU/mL-1.0x10*CFU/mL, for example, 1.0x10’CFU/mL.
It is to be noted that, if the addition of the Bifidobacteriumlactis is too little, and when the addition is lower than 1.0x10°CFU/mL, the effect of improving the intestinal flora disorder may not be effectively achieved, the reduction of the curding time is not significant, the increase of WHC, viscosity and hardness is not significant, and the improvement in the nature of yogurt is not obvious, thereby affecting the stability of the yogurt. If the addition of the Bifidobacteriumlactis is too much, and the addition is higher than 1.0x10*CFU/mL, the flavor and odor of the Bifidobacteriumlactis in the yogurt are too strong, and overall acceptance is significantly reduced. The addition is preferably 1.0x108CFU/mL, which may make the curding time of the prepared yogurt shortest and the WHC, hardness and viscosity excellent, and the overall acceptance is also high (about 8.5 points). It is to be LU503459 understood that, 1.0x10°CFU/mL-1.0x10°CFU/mL is the amount of bacterial fluid added.
In some other specific embodiments of the Bifidobacteriumlactis in other forms (for example, bacterial powder), conversion may be performed according to the amount of this embodiment.
In some specific embodiments, in the application of the Bifidobacteriumlactis in improving the intestinal flora disorder, the starter culture may select yeast power or bacterial fluid. When the yeast power is selected, the addition may be 0.01g/L-0.03g/L, for example, 0.02g/L.
In some specific embodiments, in the application of the Bifidobacteriumlactis in improving the intestinal flora disorder, fermentation includes: performing fermentation at 41°C-43°C until pH is 4.4-4.6. It is to be noted that, 41°C-43°C is the optimal temperature for fermentation, facilitating the fermentation of the yogurt; and pH is 4.4-4.6, which is an isoelectric point of the yogurt, facilitating the curding of the yogurt.
In order to better understand the present invention, the content of the present invention is further described below with reference to specific embodiments, but is not only limited to the following examples.
In the following embodiments, the used Bifidobacteriumlactis TY-SO1 is isolated from the intestinal tract of the longevous in Bama, Guangxi, is deposited with the CGMCC, and is assigned with the accession number of CGMCC No.21255.
In the following embodiments, the used milk is taken from Chongqing Tianyou Dairy
Co., Ltd..
In the following embodiments, a method for measuring the pH and acidity of the yogurt includes: using an iCinac dairy fermentation monitor to continuously monitor the pH of a yogurt sample; using phenolphthalein as an indicator; mixing 10 g of the yogurt sample and 20 ml of pure water; and performing titration with 0.1mol/L NaOH until the mixture turns red and does not fade within 30s, which is the end point of the titration.
In the following embodiments, a method for measuring the WHC of the yogurt includes: recording the weight of a 50ml centrifuge tube as Wo; recording the weight of a centrifuge tube holding 30 g of the yogurt sample as Wi; performing centrifugation on the yogurt sample for 25 min at 4000r/min at 4°C; and discarding supernatant, and recording the weight of the centrifuge tube as W2. À calculation formula is shown as follows.
Wa -Wo LU503459
WHC (%) = wow * 100
In the following embodiments, a method for measuring the viscosity of the yogurt includes: using a DV-II+Pro viscometer to measure the viscosity of the yogurt sample.
In the following embodiments, a method for measuring the hardness of the yogurt includes: using a TAXTexpressC texture analyzer to measure the hardness of the yogurt sample. A cylinder probe is selected; a testing distance is 15 mm; a trigger point is 5 g; and the speed before, during and after measurement is 1 mm/s.
In the following embodiments, a method for evaluating the sense of the yogurt includes: performing sense evaluation by a group of 10 trained sense evaluators. The color, odor, taste, tissue status and overall acceptability of the yogurt sample are evaluated according to the RHB103-2004 Detailed Rules for Sensory Quality Evaluation of Sour
Milk. Scoring is performed by using a nine-point scoring method, where 1 point represents very dislike, 5 points represent average, and 9 points represent like the above item very much.
Embodiment 1: An improvement effect of Bifidobacteriumlactis TY-SO1 on alcoholic intestinal flora disorder (1) Culture of strains
The strains deposited at -80°C is inoculated into 5 ml of an MRS liquid medium added with 0.05% cysteine at an inoculation amount of 2%, for anaerobic culture at 37°C for 48-72h. Then, second generation culture is continuously performed for 48-72h under an anaerobic condition of 37°C to a logarithmic phase. Centrifugation is performed on the bacterial suspension for 10 min at 4°C with a speed of 6000 r/min, and then bacteria are collected by discarding supernate. The bacteria are washed with normal saline, and then centrifugation is performed and repeated twice, so as to obtain bacterial sludge. Gradient dilution is performed on the bacterial sludge with the normal saline before use, and the number of viable bacteria is counted by means of a pour-plate method to calculate a
Colony Forming Unit (CFU). (2) Establishment of a model of alcohol intestinal flora disorder in mice
Before a formal experiment, mice are adaptively fed for a week. After an adaptive phase ends, the mice are randomly grouped into 3 groups (n=8) according to weight, that is a Group Control, a Group EtOH, and a Group TY-SO1. The mice in the Group TY-SO1 are continuously intragastrically administered with 1.0x10°CFU/kg of TY-SO1 bacterial fluid for 28 days; and the mice in the Group Control and the Group EtOH are intragastrically LU503459 administered with equal amount of normal saline. At Day 29, the mice in each group are fasted for 12 hours before model establishment; and then the mice in the Group EtOH and the Group TY-SO1 are intragastrically administered with an ethanol solution (12ml/kg) with volume fraction of 50% at one time, and the mice in the Group Control is given equal amount of normal saline. The mice are sacrificed after 10 hours of intragastric administration with the ethanol solution, and then the small intestines of all mice are collected for subsequent experiments. (3) Diversity analysis of intestinal microorganisms in mice
The total genomic DNA of microbial communities is extracted from cecal content samples of mice; the quality of the extracted genomic DNA is detected by means of 1% agarose gel electrophoresis; and the concentration and purity of the DNA are measured by means of NanoDrop2000. Illumina-MiSeq/NovaSeq high throughput sequencing is performed by Shanghai Majorbio Technology Co., Ltd.. Data analysis is performed on an online platform of a Majorbio cloud platform (www.Majorbio.com). (a) Impact of Bifidobacteriumlactis TY-SO1 on the diversity of intestinal microbial communities in mice
A 16SrDNA sequencing technology is used to determine the impact of the TY-SO1 on the intestinal microbiota in mice; and results are shown in Fig. 1, Fig. 2, Fig. 3 and Fig. 4.
A Shannon index and a Chao index of the Group EtOH are significantly reduced, and a Simpson index is significantly increased (P<0.05) (the Shannon index, the Simpson index and the Chao index belong to Alpha diversity indexes and may be used to evaluate the Alpha diversity of intestinal microbial communities). However, after the intervention of the TY-SO1, the Shannon index, the Chao index and the Simpson index are restored to a normal level (P<0.05) (Referring to Fig. 1, Fig. 2 and Fig. 3). Then, Beta diversity among different groups is evaluated by means of Principal Coordinate Analysis (PCoA). The clustering of the Group TY-SO1 is restored and similar to that of the Group Control (Referring to Fig. 4).
The above results show that alcohol reduces the diversity of intestinal flora, and
TY-SO1 has a significant regulation effect on restoring the diversity of flora, such that the diversity of intestinal microbial communities in alcohol-exposed mice may be maintained in a balanced state. (b) Impact of Bifidobacteriumlactis TY-SO1 on the composition of intestinal microbial communities in mice
The impact of the TY-SO1 on the composition of intestinal microbial communities in LU503459 mice is tested on phylum and genus levels; and results are shown in Fig. 5, Fig. 6, Fig. 7,
Fig. 8, Fig. 9 and Fig. 10.
On the phylum level, Firmicutes (67.30%) and Bacteroidetes (10.92%) in the Group
Control are dominant bacterial phyla (Fig. 5). After alcohol intervention, compared with the Group Control, the abundance of the Firmicutes is obviously reduced (P<0.05, referring to Fig. 6), and a ratio of the Firmicutes to the Bacteroidetes is significantly reduced (P<0.05, referring to Fig. 7). In addition, the abundance of Proteobacteria in the
Group EtOH is significantly increased (P<0.05), and in the Group TY-S01, the normal abundance of these bacteria is maintained (referring to Fig. 8). It is to be noted that, the
Firmicutes can inhibit the invasion of opportunistic pathogens; the ratio of the Firmicutes to the Bacteroidetes is an indicator of the health of intestinal microorganisms; and the
Proteobacteria is considered to be a potential pro-inflammatory phylum, including many pathogenic bacteria.
On the genus level, Lactobacillus, Bacillus and unclassified c Bacilli in the Group
EtOH are significantly lower than those in the Group Control and the Group TY-SO01 (referring to Fig. 9). The growth of the Lactobacillus beneficial to intestinal tract not only reduces the ratio of harmful bacteria, but also facilitates the resistance of the body to the invasion of pathogenic microorganisms. The Bacillus may maintain the balance of intestinal microbiota by resisting pathogens (referring to Fig. 10), and protect the health of the intestinal tract.
The above results indicate that the TY-SO1 can maintain the composition of the intestinal microbial communities in the alcohol-exposed mice and effectively improve intestinal flora disorder.
Embodiment 2: An application of Bifidobacteriumlactis TY-SO1 in functional yogurt (1) Preparation of yogurt 6% of white granulated sugar (6%w/w) is added to 1 L of milk, pasteurization is performed for 10 min at 90°C after homogenization, the temperature is rapidly cooled to 42°C, and then 0.02g/L of a CHR HANSEN Mild 1.0 starter culture is inoculated. At the same time, different doses of the TY-SO1 are added to the milk; an experiment is divided into 4 groups (n=3), that is, a blank group (not including the TY-S01), a low-dose group (including 1.0x10°CFU/mL of the TY-S01), a medium-dose group (including 1.0x10’CFU/m of the LTY-S01) and a high-dose group (including 1.0x108CFU/m of the
LTY-S01). A yogurt sample is fermented at 42°C until the pH is about 4.5, curding time is LYS03459 recorded, and then the yogurt sample is refrigerated at 4°C. (2) Performance measurement of the yogurt (a) Impact of Bifidobacteriumlactis TY-SO1 on the curding time, WHC, viscosity and hardness of the yogurt
The yogurt is an ideal carrier delivering the probiotics to the human gastrointestinal tract, and is an important application form of the probiotics in the field of functional food.
In the embodiments of the present invention, according to the method for measuring the
WHC, viscosity and hardness of the yogurt, the yogurt prepared in (1) is measured, and the curding time and appearance of the yogurt are simultaneously recorded. Results are shown in Fig. 11, Fig. 12, Fig. 13, Fig. 14 and Fig. 15.
By adding the TY-SO1 in the yogurt, the curding time may be significantly shortened (P<0.05) (Fig. 11). In addition, the yogurt in different groups shows a smooth surface with no cracks or air bubbles without whey precipitation (Fig. 12). In addition, the WHC, viscosity and hardness of the yogurt inoculated with the TY-SO1 are all higher than those in the blank group (Fig. 13, Fig. 14 and Fig. 15), and show obvious dose-dependence (P<0.05). The curding time in the medium-dose group and the high-dose group reduced by about 19% compared with the blank group, such that in industrial production, production cycles may be significantly reduced. The WHC of the high-dose group is increased by about 5% compared with the blank group, the hardness in the high-dose group is increased by about 31% compared with the blank group, and the viscosity in the high-dose group is increased by about 60% compared with the blank group, thereby greatly improving the stability of the yogurt. Therefore, the WHC of the yogurt is related to the tissue status. If the WHC is strong, the tissue status of the yogurt is good, such that whey precipitation may be prevented; and the viscosity and the hardness are also important indicators for reflecting the quality of the yogurt.
The above results show that, by means of adding the TY-SOI in the yogurt, the stability of the yogurt may be enhanced, and the quality of the yogurt may be improved.
The high-dose group shows the best effects, where the curding time is reduced by about 19% compared with the blank group; the WHC is increased by about 5% compared with the blank group; the hardness is increased by about 31% compared with the blank group; the viscosity is increased by about 60% compared with the blank group. That is to say, the overall stability of the prepared yogurt in the high-dose group is optimal.
(b) Impact of Bifidobacteriumlactis TY-SO1 on pH and acidity of the yogurt LU503459
The pH and acidity affect the viability of probiotics during storage and the quality of the yogurt. In the embodiments of the present invention, the pH and acidity of the yogurt prepared in (1) are measured according to the above method, and results are shown in Fig. 16 and Fig. 17. Compared with the blank group, changes in the pH and acidity of the yogurt inoculated with the TY-SO1 during storage at 4°C are relatively small, such that the yogurt may be stored for nearly 20 days, the reduction of the largest change in pH is only 0.3, and the acidity is increased by only 4°T, hardly affecting the taste of the yogurt. The results show that the addition of the TY-SO1 to the yogurt does not lead to post-acidification of the yogurt, and the TY-SO1 has an application potential as an auxiliary fermentation strain in the yogurt. (c) Impact of the Bifidobacteriumlactis TY-SO1 on the sense of the yogurt
In the embodiments of the present invention, the sense evaluation of the yogurt added with the TY-SO1 is performed in five aspects, namely color, odor, taste, tissue status and overall acceptability, according to the sense evaluation method of the yogurt described above. Results are shown in Fig. 18. The addition of the TY-SO1 may significantly improve the tissue status of the yogurt (P<0.05) and has dose-dependence. Compared with the blank group, the addition of low and medium doses of TY-SO1 may make the yogurt with a delicate and lubricious taste and a unique aroma of fermented milk, such that a overall acceptability score is relatively high (P<0.05), which may reach 8.5 points. The results show that the addition of appropriate amount of TY-SO1 to the yogurt may improve the tissue status, flavor, taste and overall acceptability of the yogurt.
In conclusion, the Bifidobacteriumlactis TY-SO1 in the present invention can significantly restore the diversity of intestinal microbial communities in alcohol-exposed mice and the composition of intestinal microbial communities. By means of adding an appropriate amount of TY-SO1, the stability of the yogurt may be enhanced, the tissue status may be improved, and the flavor and taste of the yogurt may be improved. Therefore, the TY-SO1 may be used to improve alcohol intestinal flora disorder and may be applied to yogurt fermentation as the auxiliary starter culture, without affecting the stability of the yogurt. In addition, the TY-SO1 may also make the prepared yogurt have the flavor of the yogurt and have the effect of improving the intestinal flora disorder (especially the alcohol intestinal flora disorder).
It is finally to be noted that, the above embodiments are merely for describing and not intended to limit the technical solutions of the present invention. Although the present invention is described in detail with reference to the preferred embodiments, those of LU503459 ordinary skill in the art should understand that the technical solutions of the present invention may be modified or equivalently replaced without departing from the purpose and scope of the technical solutions of the present invention, and shall all fall within the scope defined by the claims of the present invention.
Claims (8)
1. An application of Bifidobacteriumlactis in improving intestinal flora disorder, wherein Bifidobacteriumlactis is assigned with the accession number of CGMCC No.21255.
2. The application as claimed in claim 1, wherein intestinal flora disorder is alcoholic intestinal flora disorder.
3. The application as claimed in claim 1 or 2, comprising an application of Bifidobacteriumlactis in preparation of food, health care products or drugs.
4. The application as claimed in claim 3, wherein the food is yogurt.
5. The application as claimed in claim 4, wherein a method for preparing the yogurt comprises: inoculating the Bifidobacteriumlactis and a starter culture into a yogurt base for fermentation.
6. The application as claimed in claim 5, wherein the addition of the Bifidobacteriumlactis is 1.0x10°CFU/mL-1.0x103CFU/mL.
7. The application as claimed in claim 6, wherein yeast power is selected as the starter culture, of which addition is 0.01 g/L-0.03 g/L.
8. The application as claimed in claim 5, 6 or 7, wherein fermentation comprises: performing fermentation at 41°C-43°C until pH is 4.4-4.6.
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