CN111543640A - Application of bifidobacterium animalis subsp lactis i797 for improving infantile diarrhea and dyspepsia - Google Patents
Application of bifidobacterium animalis subsp lactis i797 for improving infantile diarrhea and dyspepsia Download PDFInfo
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- CN111543640A CN111543640A CN202010382762.8A CN202010382762A CN111543640A CN 111543640 A CN111543640 A CN 111543640A CN 202010382762 A CN202010382762 A CN 202010382762A CN 111543640 A CN111543640 A CN 111543640A
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- Prior art keywords
- bifidobacterium animalis
- animalis subsp
- lactis
- oligosaccharide
- subsp lactis
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Abstract
The invention discloses an application of bifidobacterium animalis subsp lactis i797 in preparing food, health-care products, medicaments and pharmaceutical compositions, wherein the bifidobacterium animalis subsp lactis i797 has the strain preservation number of CGMCC NO.18403, and the effect of improving infantile diarrhea and/or dyspepsia is achieved. The invention also discloses a formula milk powder with the effect of improving infantile diarrhea and/or dyspepsia, which takes the bacterial liquid or bacterial powder of bifidobacterium animalis subsp lactis i797 as an active component. The bifidobacterium animalis subsp lactis i797 has the effect of improving infantile diarrhea and dyspepsia, and is widely applied to the field of food.
Description
Technical Field
The invention belongs to the technical field of food engineering, and particularly relates to application of bifidobacterium animalis subsp lactis i797 for improving infantile diarrhea and dyspepsia.
Background
The bifidobacteria are the main flora in intestinal tracts of human beings and other animals, account for more than 90 percent of the total bacteria in the bodies of breast-fed infants, are often added into food, and have the characteristics of high GC content, high nutritional value, strict anaerobism, cell terminal bifurcation and the like. The bifidobacterium plays an important role in regulating intestinal flora, improving intestinal inflammation, resisting aging, resisting tumor, reducing blood fat, resisting obesity and the like. The bifidobacteria which are found comprise 7 genera and 32 species and are widely present in human intestinal tracts, milk and other environments. At present, the isolated culture medium of bifidobacterium is a selective culture medium, and the bifidobacterium can produce acid on the culture medium added with bromocresol purple to generate an obvious color change ring around a bacterial colony, so that the bifidobacterium is screened. As the bifidobacteria is extremely sensitive to oxygen, 0.5 per mill of L-cysteine is added on the basis of MRS, and the bifidobacteria of different strains are screened from the excrement of the infants.
The bifidobacteria are probiotics which do not produce internal and external toxins, pathogenic substances and harmful gases, and have important significance for maintaining the health of organisms. The bifidobacterium plays an important role in maintaining the micro-ecological balance of the intestinal tract of the organism, inhibiting the invasion and colonization of pathogenic bacteria, regulating the immunity of the organism, reducing the cholesterol content and the like, thereby having wide application prospect. In addition, vitamins and amino acid nutrients generated by the bifidobacteria in the growth process can improve the nutritional value of the milk, so the bifidobacteria is widely applied to the production of the milk and dairy products.
The infantile diarrhea and dyspepsia are common diseases in pediatrics, and the sick infants have increased stool frequency, are fluid and have certain seasonal and periodic characteristics. Because the children patients are younger and have low immunity, the children patients often have symptoms of vomiting, inappetence and the like after the diseases are attacked, and serious children can cause dehydration and even threaten life. The long-term diarrhea easily affects the growth and development of children patients, and particularly the children patients have weak ability to drug metabolism, so that the clinical treatment needs to keep a cautious attitude and avoid using chemical drugs used for adult treatment.
Disclosure of Invention
The inventor selects a strain from the feces of the breast-fed infants, and the strain is identified as Bifidobacterium animalis subsp.lactis i797(Bifidobacterium animalis subsp.lactis i797), and the preservation number of the strain is CGMCC NO. 18403. In the process of researching the functions of the bacterial strain, the bacterial strain has good effect of regulating the intestinal tract, can regulate the balance of intestinal flora and improve the characteristics of excrement.
Therefore, the first object of the invention is to provide the application of bifidobacterium animalis lactis i797 in preparing foods, health-care products, medicaments and pharmaceutical compositions, wherein the bifidobacterium animalis lactis i797 has the effect of improving infantile diarrhea and/or dyspepsia. The second purpose of the invention is to provide a formula milk powder, wherein the formula milk powder contains the bifidobacterium animalis subsp lactis i 797. The third purpose of the invention is to provide a preparation method of the formula milk powder.
In order to achieve the purpose, the invention provides the following technical scheme:
as a first aspect of the invention, the application of bifidobacterium animalis subsp lactis i797 with the effect of improving infantile diarrhea and/or dyspepsia in preparing food, health-care products, medicaments and pharmaceutical compositions, wherein the strain preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
According to the invention, the food comprises formula milk powder, compound probiotics, dairy products, drinks and the like.
According to the invention, the dosage form of the medicament can be capsules, tablets, pills, powder and the like.
As a second aspect of the invention, the biological product has the effect of improving infantile diarrhea and/or dyspepsia, the biological product takes bacterial liquid or bacterial powder of bifidobacterium animalis subspecies lactis i797 as an active component, and the preservation number of the bifidobacterium animalis subspecies lactis i797 is CGMCC NO. 18403.
According to the invention, the biological product also comprises galacto-oligosaccharide and fructo-oligosaccharide, the mass ratio of the bifidobacterium animalis subsp lactis i797 to the galacto-oligosaccharide to the fructo-oligosaccharide is 3:9:8, and the addition amount of the bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g。
According to the invention, the biological products comprise food, health products, medicines and pharmaceutical compositions.
According to the invention, the 16SrRNA sequence of Bifidobacterium animalis subsp lactis i797 is shown as SEQ ID NO: 1 is shown.
According to the invention, the tuf gene sequence of bifidobacterium animalis subsp lactis i797 is shown in SEQ ID NO: 2, respectively.
As a third aspect of the invention, the intestinal microecological preparation takes a bacterial liquid or bacterial powder of bifidobacterium animalis subsp lactis i797 as an active component, and the bacterial preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
According to the invention, the intestinal micro-ecological preparation further comprises galacto-oligosaccharide and fructo-oligosaccharide, the bifidobacterium animalis subsp lactis i797, galacto-oligosaccharideThe mass ratio of sugar and fructo-oligosaccharide is 3:9:8, and the addition amount of Bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g。
As a fourth aspect of the invention, the formula milk powder with the effect of improving infantile diarrhea and/or dyspepsia takes the bacterial liquid or the bacterial powder of bifidobacterium animalis subspecies lactis i797 as an active component, and the strain preservation number of the bifidobacterium animalis subspecies lactis i797 is CGMCC NO. 18403.
According to the invention, the formula milk powder also comprises galacto-oligosaccharide and fructo-oligosaccharide, the mass ratio of the bifidobacterium animalis subsp lactis i797 to the galacto-oligosaccharide to the fructo-oligosaccharide is 3:9:8, and the addition amount of the bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g。
According to the invention, the formula milk powder with the effect of improving infantile diarrhea and/or dyspepsia comprises the following components in parts by weight:
the preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
According to the invention, the addition amount of bifidobacterium animalis subsp lactis i797 is more than or equal to 1 × 107cfu/g。
Preferably, the formula milk powder with the effect of improving infantile diarrhea and/or dyspepsia comprises the following components in parts by weight:
the preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
According to the invention, the addition amount of bifidobacterium animalis subsp lactis i797 is more than or equal to 1 × 107cfu/g。
Preferably, the formula milk powder with the effect of improving infantile diarrhea and/or dyspepsia comprises the following components in parts by weight:
the preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
According to the invention, the addition amount of bifidobacterium animalis subsp lactis i797 is more than or equal to 1 × 108cfu/g。
As a fifth invention of the present invention, a method for preparing a formula milk powder for improving infantile diarrhea and/or dyspepsia comprises the following steps:
the method comprises the following steps: weighing raw milk, desalted whey powder, skim milk powder, soybean oil, concentrated whey protein powder, galacto-oligosaccharide, fructo-oligosaccharide and phospholipid according to the weight parts for later use;
step two, respectively weighing arachidonic acid grease, compound mineral substances, compound vitamins and bifidobacterium animalis subsp lactis i797 in parts by weight for later use;
step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: and weighing, bagging or canning and sealing the composition A to obtain the formula milk powder.
The invention has the beneficial effects that: the bifidobacterium animalis subsp lactis i797 has high oxygen resistance activity, can improve infantile diarrhea and dyspepsia, has strong intestinal juice tolerance capability and better digestive juice survival rate, can be used for preparing foods, health-care products, medicaments, pharmaceutical compositions and the like, and has wide market application value.
Detailed Description
The present invention will be further described with reference to the following examples. It should be understood that the following examples are illustrative only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, which are not specified under specific conditions, are generally carried out under conventional conditions.
1. Bifidobacterium animalis subsp.lactis i797, also known as Bifidobacterium lactis i797, or alternatively: bifidobacterium animalis subsp.lactis JMCC0025 is selected from intestinal flora of infant, and the preservation number of the strain is CGMCC NO. 18403. The strain is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation address is No. 3 Xilu No.1 of Beijing, Chaoyang, and the preservation date is 2019, 8 months and 20 days.
Through molecular biological identification, the 16SrRNA sequence of bifidobacterium animalis subsp lactis i797 is as follows:
ACGGCTCCCCCACAAGGGTCGGGCCACCGGCTTCGGGTGCTACCCACTTTCATGACTTGACGGGCGGTGTGTACAAGGCCCGGGAACGCATTCACCGCGGCGTTGCTGATCCGCGATTACTAGCGACTCCGCCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGACCGGTTTTCAGCGATCCGCCCCACGTCACCGTGTCGCACCGCGTTGTACCGGCCATTGTAGCATGCGTGAAGCCCTGGACGTAAGGGGCATGATGATCTGACGTCATCCCCACCTTCCTCCGAGTTGACCCCGGCGGTCCCACATGAGTTCCCGGCATCACCCGCTGGCAACATGCGGCGAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACGACCATGCACCACCTGTGAACCGGCCCCGAAGGGAAACCGTGTCTCCACGGCGATCCGGCACATGTCAAGCCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAATCCGCATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTTCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGATGCTTAACGCGTTGGCTCCGACACGGGACCCGTGGAAAGGGCCCCACATCCAGCATCCACCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTGACGGCCCAGAGACCTGCCTTCGCCATTGGTGTTCTTCCCGATATCTACACATTCCACCGTTACACCGGGAATTCCAGTCTCCCCTACCGCACTCCAGCCCGCCCGTACCCGGCGCAGATCCACCGTTAGGCGATGGACTTTCACACCGGACGCGACGAACCGCCTACGAGCCCTTTACGCCCAATAAATCCGGATAACGCTCGCACCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGTGCTTATTCGAACAATCCACTCAACACGGCCGAAACCGTGCCTTGCCCTTGAACAAAAGCGGTTTACAACCCGAAGGCCTCCATCCCGCACGCGGCGTCGCTGCATCAGGCTTGCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTATCTCAGTCCCAATGTGGCCGGTCACCCTCTCAGGCCGGCTACCCGTCAACGCCTTGGTGGGCCATCACCCCGCCAACAAGCTGATAGGACGCGACCCCATCCCATGCCGCAAAAGCATTTCCCACCCCACCATGCGATGGAGCGGAGCATCCGGTATTACCACCCGTTTCCAGGAGCTATTCCGGTGCACAGGGCAGGTTGGTCACGCATTACTCACCCGTTCGCCACTCTCACCCGACAGCAAGCTGCCAGGGATCCCGTTCGACTGCATGTGTAAG
SEQ ID NO:1
the tuf gene sequence of bifidobacterium animalis subsp lactis i797 is as follows:
GGATCTCGATGAGAGCAGCGTGGTATCACCATCAACATTGCCCACATCGAGTACCAGACGGCCAAGCGTCACTACGCCCACGTCGACTGCCCGGGCCACGCCGACTTCGTGAAGAACATGATCACCGGCGCTGCCCAGATGGATGGCGCCATCCTCGTTGTGGCCGCCACCGACGGCCCGATGGCCCAGACCCGCGAGCACGTGCTGCTCGCCCGTCAGGTCGGCGTCCCGAAGATCCTCGTCGCTCTGAACAAGTGCGATATGGTCGATGACGAAGAGCTCATCGAGCTCGTCGAAGAAGAGGTCCGCGACCTCCTCGACGAGAACGGCTTCGACCGCGACTGCCCGGTCGTGCACACCTCCGCTTACGGCGCTCTGCATGACGACGCTCCCGGATCACGACAAGTGGGTTGCCACCATCAAGGAGCTCATGGACGACGTCGACGAGTACATCCCGACCCCGGTCCACGACCTCGACAAGCCGTTCCTGATGCCGATCGAGGACGTCTTCACCATCTCCGGCCGTGGCACCGTCGTCACCGGTCGTGTCGAGCGCGGCAAGCTGCCGATCAACACGAACGTCGAGATCGTCGGCATCCGCCCGACCCAGACCACCACCGTCACCTCCATCGAGACCTTCCACAAGCAGATGGATGAGTGCGAGGCCGGCGACAACACCGGTCTGCTGCTCCGCGGCATCAACCGCACCGACGTCGAGCGTGGCCAGGTCGTGGCTGCTCCGGGTTCGGTCACCCCGCACACCAAGTTCGAAGGCGAAGTCTACGTCCTTACCAAGGATGAGGGCGGCCGTCACTCGCCGTTCTTCTCGAACTACCGTCCGCAGTTCTACTTCCGCACCACCGACGTCACCGGCGTCATCACGCTGCCGGAAGGCGTCGAGATGGTTCAGCCTGGCGATCACGCGACCTTCACGGTTGAGCTGATCCAGCCGATCGCTATGGAAGAGGGCTTCACCTTCCCAGTGCTTGAAGGC
SEQ ID NO:2
example 1 method for isolation and purification of Bifidobacterium animalis subsp
1. Collecting samples
Taking 1g of intestinal excrement of infants or young children, adding the intestinal excrement into 9mL of physiological saline, and fully and uniformly mixing to obtain a sample A;
2. enrichment of samples
Get V1Sample a 2mL, added to V2Modified MRS liquid Medium 100mL in T1Anaerobic culture at 37 ℃1Obtaining a culture solution B after 72 hours;
3. separating and screening strains
Taking 1ml of culture solution B1, diluting with sterile physiological saline with concentration of 0.9% by gradient multiplication by 10 times, sequentially diluting with gradient 10-1、10-2、10-3、10-4、10-5Doubling to obtain bacterial suspension C1～C5；
Taking the improved MRS solid culture medium, melting, pouring into the first to fifth culture dishes respectively, and obtaining a culture medium D after cooling and complete solidification1～D5Respectively sucking the bacterial suspension C1～C50.1mL each and applied to the medium D in one-to-one correspondence1～D5On, then invert the plate and put it at T2Anaerobic culture at 37 deg.C2Observing the growth of the colonies for 72 h;
after the plate has the typical bacteria, selecting a corresponding single bacterial colony E according to the bacterial colony characteristics of the standard bifidobacterium and the reference of related literature pictures;
4. strain purification
Selecting single colony E, streaking the single colony E onto improved MRS solid culture medium, and inoculating3Culturing under anaerobic condition at 37 deg.C3Obtaining a single colony F after 72 hours;
the single colony F is continuously streaked and inoculated on an improved MRS solid culture medium, T4Culturing under anaerobic condition at 37 deg.C4Obtaining a single colony G after 72 hours;
then, continuously streaking and inoculating the single colony G on an improved MRS solid culture medium, T5Culturing under anaerobic condition at 37 deg.C5Obtaining a pure culture H which is the strain of the bifidobacterium animalis subsp lactis JMCC0025 after 72H;
fifthly, storing
Mixing pure culture H and sterile 50% glycerol at a ratio of 1:1, placing in strain storage tube, mixing, and placing in T6Storing at-70 deg.c while inoculating the test tube slant of modified MRS solid culture medium for temporary storage.
In this embodiment, the raw materials of the improved MRS liquid medium include: casein peptone, beef extract, yeast extract, glucose, sodium acetate, diamine citrate, Tween-80, and K2HPO4、MgSO4·7H2O、MnSO4·7H2O, cysteine and distilled water; wherein casein peptone, beef extract, yeast extract, glucose, sodium acetate, citric acid diamine, and Tween-80、K2HPO4、MgSO4·7H2O、MnSO4·7H2The proportion of the O, the cysteine and the distilled water is 10 g: 10 g: 5 g: 20 g: 5 g: 2 g: 1 g: 2 g: 0.2 g: 0.05 g: 0.5 g: 1000 mL; the modified MRS solid culture medium is prepared by adding agar with the mass part of 15% to each 1000mL of modified MRS liquid culture medium.
Example 2-5A method for separating and purifying Bifidobacterium animalis subsp
Examples 2 to 5 are the separation and purification method of example 1, and are substantially the same as the method of example 2, except that the technical parameters of the separation and purification process are different, and the specific parameters are shown in table 1.
TABLE 1 isolation and purification procedure and parameters for Bifidobacterium animalis strain JMCC0025 from examples 2 to 5
Example 6 bacteriological basic characteristics of Bifidobacterium animalis strain JMCC0025
This example shows the basic bacteriological characteristics of bifidobacterium animalis strain JMCC0025, which are shown in table 2.
TABLE 6 basic characteristics of Bifidobacterium animalis Lactobacillus subsp.lactis JMCC0025
| Experimental project | Results | Experimental project | Results |
| Gram stain | Positive for | Cell morphology | Polymorphic rod shape |
| Oxidase enzyme | - | Contact enzyme | - |
Example 7 sugar fermentation characteristics of Bifidobacterium animalis strain JMCC0025
This example shows the sugar fermentation characteristics of Bifidobacterium animalis strain JMCC 0025. The experimental method for the sugar fermentation characteristics comprises the following steps: a single colony of the strain of Bifidobacterium animalis Lactobacillus subsp JMCC0025 obtained by the separation and purification method in example 2 is selected and inoculated into a sterilized improved MRS liquid culture medium, cultured for 24h at 37 ℃, the bacterial suspension is inoculated into a sugar fermentation tube, and anaerobically cultured for 48h at 37 ℃ to observe the color change. The results of the identification of the sugar fermentation characteristics are shown in Table 3:
TABLE 3 identification of Bifidobacterium animalis lactococcus JMCC0025 sugar fermentation characteristics
Note: "+" indicates fermentation utilization; "-" indicates no fermentative utilization.
The modified MRS liquid medium used in this example has the same composition as the modified MRS liquid medium of example 2.
Example 8 molecular biological identification of species of Bifidobacterium animalis, Lactobacillus subsp
The strain of bifidobacterium animalis subsp. lactis JMCC0025 obtained by the separation and purification method in example 4 is identified in molecular biology, and is finally determined as the bifidobacterium animalis subsp. lactis by DNA extraction, PCR amplification, 16SrRNA sequencing and NCBI website blast.
Example 9 study of intestinal Condition Regulation of Bifidobacterium animalis Lactobacillus subsp lactococcus JMCC0025 Strain
The drink containing the bifidobacterium animalis strain JMCC0025 is drunk by testers, and the drinking results of 10 testers are counted. The statistical results are shown in table 4.
TABLE 4 analysis of differences in various indicators after trial drinking of Bifidobacterium lactis strain JMCC0025
Remarking: significant changes (statistical significance P <0.05) are differences.
In the statistical process:
(1) the difference analysis finds that the defecation times, the defecation difficulty degree, the stool hardness and the stool shape have significant differences;
(2) through the conversation of drinkers, the frequency of the excrement after drinking is increased, the color of the excrement turns yellow, and the index change of the excrement softening is obvious.
From table 4, it can be found that, in all the indexes, four indexes (defecation frequency, defecation difficulty degree, stool hardness and stool shape) have significant changes (with statistical significance P <0.05), which indicates that bifidobacterium animalis lactobacillus subsp JMCC0025 has a regulating effect on intestinal tracts.
Example 10 post-acidification profiling of species of Bifidobacterium animalis subsp
Mixing 97-98 parts of fresh milk and 2-3 parts of white granulated sugar, uniformly blending, homogenizing at 65 ℃ and 15MPa, sterilizing at 95 ℃ for 300s, cooling to 37 ℃ to obtain sterilized milk, inoculating the bifidobacterium animalis subsp lactis JMCC0025 to the sterilized milk, wherein the inoculation amount is 3% of the mass of the sterilized milk, fermenting at 36 ℃, and detecting the pH change condition.
And performing a control experiment by taking a standard strain bifidobacterium BB-12 as a control strain, wherein the experimental parameters are the same as those of the strain of the bifidobacterium animalis subsp.
The results of the acid production analysis are shown in Table 5.
TABLE 5 acidity Change in Bifidobacterium animalis Lactobacillus subsp.lactis JMCC0025
| Time of fermentation | BB-12 | HMCC0025 |
| 0h | 6.4 | 6.32 |
| 18h | 6.13 | 6.06 |
| 24h | 5.55 | 4.66 |
| 42h | 3.97 | 3.92 |
| 49h | 3.56 | 3.87 |
| 66h | 3.82 | 3.87 |
| 90h | 3.82 | 3.88 |
As can be seen from Table 5, Bifidobacterium animalis strain JMCC0025 provides good control of post-acidification at higher storage conditions suitable for growth of lactic acid bacteria at 37 ℃.
EXAMPLE 11 preparation of formula
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 0.7g of galacto-oligosaccharide, 0.97g of fructo-oligosaccharide and 1g of phospholipid for later use;
step two, respectively weighing 0.1g of arachidonic acid grease and 0.03g of bifidobacterium lactis for later use, wherein the addition amount of bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g；
Step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
EXAMPLE 12 preparation of formula milk powder
Respectively weighing 79g of raw milk, 10g of desalted whey powder, 6g of skim milk powder, 1.1g of soybean oil, 0.9g of concentrated whey protein powder, 0.96g of galacto-oligosaccharide, 0.7g of fructo-oligosaccharide and 0.9g of phospholipid for later use;
step two, respectively weighing 0.1g of arachidonic acid grease and 0.04g of bifidobacterium lactis for later use, wherein the addition amount of bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g；
Step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
EXAMPLE 13 preparation of formula
Weighing 85g of raw milk, 6g of desalted whey powder, 4g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 0.8g of galacto-oligosaccharide, 0.77g of fructo-oligosaccharide and 1.1g of phospholipid respectively;
step two, respectively weighing 0.1g of arachidonic acid grease and 0.03g of bifidobacterium lactisBacillus, wherein the content of Bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g；
Step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
EXAMPLE 14 preparation of formula
Respectively weighing 75g of raw milk, 11.7g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 0.9g of galacto-oligosaccharide, 0.8g of fructo-oligosaccharide and 1g of phospholipid;
step two, respectively weighing 0.1g of arachidonic acid grease and 0.3g of bifidobacterium lactis, wherein the addition amount of bifidobacterium animalis subsp lactis i797 is not less than 1 × 108cfu/g；
Step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
EXAMPLE 15 preparation of formula
Adding 0.032g Bifidobacterium animalis subspecies lactis i797 into 100g calcium ferrum zinc concocted milk powder, wherein the addition amount of Bifidobacterium animalis subspecies lactis i797 is not less than 1 × 107cfu/g. The le Ca Fe Zn modified milk powder is purchased from Shijiazhuang Jun le Bao milk industry Co.
Comparative example 1 preparation of formula milk powder
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 0.9g of galacto-oligosaccharide, 0.8g of fructo-oligosaccharide and 1g of phospholipid;
step two, respectively weighing 0.1g of arachidonic acid oil and 0.032 of bifidobacterium lactis JMCC0778 (the source is animal bifidobacterium in the strain resource library JMCC of Shijiazhuang Junlebao Dairy Co., Ltd.), wherein the addition amount of the bifidobacterium lactis JMCC0778 is not less than 1 × 107cfu/g；
Step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
Comparative example 2 preparation of formula milk powder
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 0.9g of galacto-oligosaccharide, 0.8g of fructo-oligosaccharide and 1g of phospholipid for later use;
step two, weighing 0.1g of arachidonic acid grease for later use;
step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
Comparative example 3 preparation of formula milk powder
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder and 1g of phospholipid for later use;
step two, respectively weighing 0.1g of arachidonic acid grease and 0.04g of bifidobacterium lactis for later use, wherein the addition amount of bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g；
Step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
Comparative example 4 preparation of formula milk powder
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 1.8g of glucose oligosaccharide and 1g of phospholipid for later use;
step two, respectively weighing 0.1g of arachidonic acid grease and 0.04g of bifidobacterium lactis for later use, wherein the addition amount of bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g；
Step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
Comparative example 5 preparation of formula milk powder
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 0.45g of galacto-oligosaccharide, 0.4g of fructo-oligosaccharide and 1g of phospholipid for later use;
step two, weighing 0.1g of arachidonic acid grease and 0.04g of bifidobacterium lactis for later use;
step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
Comparative example 6 preparation of formula milk powder
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 1.5g of galacto-oligosaccharide and 1g of phospholipid for later use;
step two, weighing 0.1g of arachidonic acid grease and 0.04g of bifidobacterium lactis for later use;
step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
Comparative example 7 preparation of formula milk powder
Respectively weighing 75g of raw milk, 12g of desalted whey powder, 8g of skim milk powder, 1g of soybean oil, 1g of concentrated whey protein powder, 1.5g of fructo-oligosaccharide and 1g of phospholipid for later use;
step two, weighing 0.1g of arachidonic acid grease and 0.04g of bifidobacterium lactis for later use;
step three, mixing materials: uniformly mixing the materials weighed in the first step and the second step to obtain a composition A;
step four, packaging: weighing the composition A, bagging or canning, and sealing.
It should be noted that the formula milk powder of the above embodiments may further include complex minerals, complex vitamins, etc. during actual production and processing to meet the product requirements.
The formulations of examples 1 to 5 and comparative examples 1 to 3 are shown in Table 6.
TABLE 6 formulations of examples 1-5 and comparative examples 1-3
EXAMPLE 16 study of high antioxidant Activity of Bifidobacterium lactis in formula milk powder
Viable cell count was measured on the formulas prepared in examples 11 to 12, examples 14 to 15, comparative example 1 and comparative example 3. The results of viable cell count measurement are shown in Table 7.
TABLE 7 study of high oxygen activity
The results in Table 7 show that the viable count of the milk powder formulas of examples 11-12 and 14-15 does not change obviously after being stored for one year at normal temperature, and the results show that the Bifidobacterium animalis subsp lactis i797 has good high oxygen-resistant survival characteristics and is suitable for being used in the milk powder formulas.
EXAMPLE 17 test of improving infantile diarrhea and dyspepsia with formula milk powder containing Bifidobacterium lactis
This example is an experiment relating to the improvement of gastrointestinal conditions such as infantile diarrhea and dyspepsia with bifidobacterium animalis subsp.
88 infants who had increased stool recruitment times, had symptoms of fluid state, abdominal distention, anorexia, crying, etc., and had no red blood cells or bloody purulent stool in routine stool examination. Meanwhile, children suffering from other systemic serious diseases and children who are treated by using antibacterial drugs are excluded from being combined. The change of each index was observed by feeding the milk powders of comparative examples 1 to 7 and examples 11 to 12 and 13 to 14 for 21 days. The analysis of each index after feeding is shown in Table 8.
TABLE 8 analysis of differences in the respective indices after feeding the formula
Remarking: indicates significant changes (statistical significance P <0.05) as differences.
And (3) displaying the data in the table:
(1) the indexes of abdominal distension, diarrhea, crying, anorexia, sleep and the like are found to have significant differences through difference analysis;
(2) through revisitation and conversation of the family members of the drinkers, the indexes of crying and screaming, frequency of excrement and anorexia of the fed infants are found to be obviously changed.
As can be seen from table 8, comparative example 1 and comparative example 2 showed significant changes in only the index of emesis among all indexes;
comparative example 3 only the "vomiting" and "abdominal distension" indices were significantly changed in all indices;
comparative examples 4 to 7 showed no significant change in all indices.
Example 11, of all the indices, 3 indices (abdominal distension, vomiting, anorexia) showed significant changes (statistically significant P < 0.05);
example 12, of all the indexes, 3 indexes (abdominal distension, diarrhea, crying) showed significant changes (statistical significance P < 0.05);
example 14, of all the indices, 5 indices (abdominal distension, diarrhea, vomiting, crying, anorexia) showed significant changes (statistically significant P < 0.05);
example 15, of all the indices, 4 indices (bloating, diarrhea, crying, anorexia) showed significant changes (statistically significant P < 0.05).
The result shows that the bifidobacterium animalis subsp lactis i797 has a synergistic effect after being compounded with galacto-oligosaccharide and fructo-oligosaccharide, and has a regulating effect on gastrointestinal conditions such as infantile diarrhea, dyspepsia and the like. Particularly, when the mass ratio of bifidobacterium animalis subsp lactis i797 to galacto-oligosaccharides to fructo-oligosaccharides is 3:9:8, the composition has good effects of improving abdominal distension, diarrhea, vomiting, crying and anorexia.
Example 18 tolerance properties of simulated gastrointestinal fluids of Bifidobacterium animalis subsp.
Activating 3 generations of bifidobacterium lactis i797 strain to be tested, then taking 1mL of the strain, respectively placing the strain in artificial gastric juice containing 9mL of filter-sterilized bacteria and having a pH value of 3.0, uniformly shaking the strain, carrying out anaerobic culture at 37 ℃, respectively sampling when the culture and the culture are started for 2 hours, and determining the viable count. Then 1mL of culture solution digested in artificial gastric juice with pH value of 3.0 for 2h was inoculated into 9mL of filter-sterilized artificial intestinal juice with pH value of 8.0, and cultured at 37 deg.C, and viable count was measured at 0h, 4h, and 6h, respectively.
The control experiment is carried out by taking bifidobacterium BB-101 as a standard strain, and the experimental parameters are the same as those of the bifidobacterium animalis subsp.
Survival (%) (cfu N1/cfu N0) × 100%
In the formula, N1 represents the viable count of 6 hours after being treated by the artificial digestive juice, and N0 represents the viable count of 0 hour after being treated by the artificial digestive juice.
b. Intestinal juice: bile Salts (Difco)0.9g/100mL, adjusted to pH 8.0, and filter sterilized for use.
The results are shown in Table 9.
TABLE 9 simulated gastric fluid intestinal tolerance results for Bifidobacterium animalis subsp
The result shows that BB-101 has stronger gastric acid tolerance and poorer intestinal fluid tolerance, while the bifidobacterium animalis lactis I797 has poorer gastric acid tolerance; the intestinal juice tolerance is strong; has better simulated digestive juice survival rate reaching 7.4 percent.
In conclusion, the bifidobacterium animalis subsp lactis i797 strain has good high oxygen-resistant survival characteristic, has good effect of regulating the intestinal tract, and can regulate the balance of intestinal flora and improve the characteristics of excrement. Therefore, it is obvious to those skilled in the art that the bifidobacterium animalis subsp lactis i797 strain can be further prepared into drinks, foods, medicines, or the like by methods known in the art. For example: bifidobacterium animalis subsp lactis i797 strain can be used for preparing cereal food and its derivatives, probiotic preparation, and formula milk powder; can also be used for preparing beverage and fermented yogurt for improving gastrointestinal function. When used to prepare a probiotic formulation, may comprise only the probiotic of bifidobacterium animalis subsp lactis i 797; or the compound probiotics is prepared by mixing bifidobacterium animalis subsp lactis i797, lactobacillus paracasei N1115, lactobacillus plantarum N3117, lactobacillus rhamnosus and the like, and the dosage form of the compound probiotics can be compound probiotic microcapsule powder.
The foregoing is merely an example of the embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> Shijiazhuang Junle Baoru Co Ltd
<120> application of bifidobacterium animalis subsp lactis i797 for improving infantile diarrhea and dyspepsia
<141>2020-05-08
<160>2
<170>SIPOSequenceListing 1.0
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<211>1411
<212>DNA
<213> Bifidobacterium animalis subsp
<400>1
acggctcccc cacaagggtc gggccaccgg cttcgggtgc tacccacttt catgacttga 60
cgggcggtgt gtacaaggcc cgggaacgca ttcaccgcgg cgttgctgat ccgcgattac 120
tagcgactcc gccttcacgc agtcgagttg cagactgcga tccgaactga gaccggtttt 180
cagcgatccg ccccacgtca ccgtgtcgca ccgcgttgta ccggccattg tagcatgcgt 240
gaagccctgg acgtaagggg catgatgatc tgacgtcatc cccaccttcc tccgagttga 300
ccccggcggt cccacatgag ttcccggcat cacccgctgg caacatgcgg cgagggttgc 360
gctcgttgcg ggacttaacc caacatctca cgacacgagc tgacgacgac catgcaccac 420
ctgtgaaccg gccccgaagg gaaaccgtgt ctccacggcg atccggcaca tgtcaagccc 480
aggtaaggtt cttcgcgttg catcgaatta atccgcatgc tccgccgctt gtgcgggccc 540
ccgtcaattt ctttgagttt tagccttgcg gccgtactcc ccaggcggga tgcttaacgc 600
gttggctccg acacgggacc cgtggaaagg gccccacatc cagcatccac cgtttacggc 660
gtggactacc agggtatcta atcctgttcg ctccccacgc tttcgctcct cagcgtcagt 720
gacggcccag agacctgcct tcgccattgg tgttcttccc gatatctaca cattccaccg 780
ttacaccggg aattccagtc tcccctaccg cactccagcc cgcccgtacc cggcgcagat 840
ccaccgttag gcgatggact ttcacaccgg acgcgacgaa ccgcctacga gccctttacg 900
cccaataaat ccggataacg ctcgcaccct acgtattacc gcggctgctg gcacgtagtt 960
agccggtgct tattcgaaca atccactcaa cacggccgaa accgtgcctt gcccttgaac 1020
aaaagcggtt tacaacccga aggcctccat cccgcacgcg gcgtcgctgc atcaggcttg 1080
cgcccattgt gcaatattcc ccactgctgc ctcccgtagg agtctgggcc gtatctcagt 1140
cccaatgtgg ccggtcaccc tctcaggccg gctacccgtc aacgccttgg tgggccatca 1200
ccccgccaac aagctgatag gacgcgaccc catcccatgc cgcaaaagca tttcccaccc 1260
caccatgcga tggagcggag catccggtat taccacccgt ttccaggagc tattccggtg 1320
cacagggcag gttggtcacg cattactcac ccgttcgcca ctctcacccg acagcaagct 1380
gccagggatc ccgttcgact gcatgtgtaa g 1411
<210>2
<211>994
<212>DNA
<213> Bifidobacterium animalis subsp
<400>2
ggatctcgat gagagcagcg tggtatcacc atcaacattg cccacatcga gtaccagacg 60
gccaagcgtc actacgccca cgtcgactgc ccgggccacg ccgacttcgt gaagaacatg 120
atcaccggcg ctgcccagat ggatggcgcc atcctcgttg tggccgccac cgacggcccg 180
atggcccaga cccgcgagca cgtgctgctc gcccgtcagg tcggcgtccc gaagatcctc 240
gtcgctctga acaagtgcga tatggtcgat gacgaagagc tcatcgagct cgtcgaagaa 300
gaggtccgcg acctcctcga cgagaacggc ttcgaccgcg actgcccggt cgtgcacacc 360
tccgcttacg gcgctctgca tgacgacgct cccggatcac gacaagtggg ttgccaccat 420
caaggagctc atggacgacg tcgacgagta catcccgacc ccggtccacg acctcgacaa 480
gccgttcctg atgccgatcg aggacgtctt caccatctcc ggccgtggca ccgtcgtcac 540
cggtcgtgtc gagcgcggca agctgccgat caacacgaac gtcgagatcg tcggcatccg 600
cccgacccag accaccaccg tcacctccat cgagaccttc cacaagcaga tggatgagtg 660
cgaggccggc gacaacaccg gtctgctgct ccgcggcatc aaccgcaccg acgtcgagcg 720
tggccaggtc gtggctgctc cgggttcggt caccccgcac accaagttcg aaggcgaagt 780
ctacgtcctt accaaggatg agggcggccg tcactcgccg ttcttctcga actaccgtcc 840
gcagttctac ttccgcacca ccgacgtcac cggcgtcatc acgctgccgg aaggcgtcga 900
gatggttcag cctggcgatc acgcgacctt cacggttgag ctgatccagc cgatcgctat 960
ggaagagggc ttcaccttcc cagtgcttga aggc 994
Claims (10)
1. An application of bifidobacterium animalis subsp lactis i797 in preparing food, health-care products, medicines and medicinal compositions for improving infantile diarrhea and/or dyspepsia is disclosed, wherein the strain preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
2. The use of claim 1, wherein the food product comprises a formula, a complex probiotic, a dairy product, a beverage.
3. A biological product with the effect of improving infantile diarrhea and/or dyspepsia is characterized in that the biological product takes bacterial liquid or bacterial powder of bifidobacterium animalis subsp lactis i797 as an active component, and the preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
4. The biological product with the effect of improving infantile diarrhea and/or dyspepsia of claim 3, further comprising galacto-oligosaccharide and fructo-oligosaccharide, wherein the mass ratio of the bifidobacterium animalis subsp lactis i797 to the galacto-oligosaccharide to the fructo-oligosaccharide is 3:9:8, and the addition amount of the bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g。
5. The biological product for improving infantile diarrhea and/or dyspepsia of claim 3, wherein the biological product comprises food, health product, medicine and pharmaceutical composition.
6. An intestinal microecological preparation is characterized in that a bacterial liquid or bacterial powder of bifidobacterium animalis subsp lactis i797 is used as an active component, and the preservation number of the strain of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
7. The intestinal microecological preparation of claim 6, further comprising galacto-oligosaccharide and fructo-oligosaccharide, wherein the mass ratio of the bifidobacterium animalis subsp lactis i797 to the galacto-oligosaccharide to the fructo-oligosaccharide is 3:9:8, and the addition amount of the bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g。
8. The formula milk powder with the effect of improving infantile diarrhea and/or dyspepsia is characterized in that a bacterial liquid or bacterial powder of bifidobacterium animalis subsp lactis i797 is used as an active component, and the preservation number of the bifidobacterium animalis subsp lactis i797 is CGMCC NO. 18403.
9. The formula milk powder with effect of improving infantile diarrhea and/or dyspepsia of claim 8, further comprising galacto-oligosaccharide and fructo-oligosaccharide, wherein the mass ratio of bifidobacterium animalis subsp lactis i797 to the galacto-oligosaccharide to the fructo-oligosaccharide is 3:9:8, and the addition amount of the bifidobacterium animalis subsp lactis i797 is not less than 1 × 107cfu/g。
10. The formula for improving infantile diarrhea and/or dyspepsia according to claim 9, wherein the formula comprises the following components in parts by weight:
the preservation number of the bifidobacterium animalis subspecies lactis i797 is CGMCC NO.18403, and the addition amount of the bifidobacterium animalis subspecies lactis i797 is more than or equal to 1 × 107cfu/g。
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Address after: 050221 No. 68 stone Copper Road, Hebei, Shijiazhuang Applicant after: JUNLEBAO Dairy Group Co.,Ltd. Address before: 050221 No. 68 stone Copper Road, Hebei, Shijiazhuang Applicant before: THE SHIJIAZHUANG JUNLEBAO DAIRY Co.,Ltd. |
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Application publication date: 20200818 |