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A strain of Bacillus coagulans hew-b379 with probiotic effect and its application

A technology of HEW-B379 and Bacillus coagulans, applied in the field of microbiology, can solve the problems of less research and application

Active Publication Date: 2019-05-24
BEIJING HESWOF BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research and application of Bacillus coagulans as a probiotic preparation or feed additive is less, especially in the aspects of replacing antibiotics in feeding, promoting the absorption of nutrients by animals, and improving animal production performance.

Method used

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  • A strain of Bacillus coagulans hew-b379 with probiotic effect and its application
  • A strain of Bacillus coagulans hew-b379 with probiotic effect and its application
  • A strain of Bacillus coagulans hew-b379 with probiotic effect and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Isolation and Identification of Bacillus coagulans HEW-B379

[0047] 1. Isolation and purification of Bacillus: Collect 1g of the intestinal contents of healthy pigs under aseptic conditions, put them in a centrifuge tube filled with 4.5mL sterilized physiological saline, shake and mix well, and bathe in a water bath at 85°C for 15min , and then draw 0.5mL of the mixture into a centrifuge tube filled with 4.5mL of sterilized saline, the dilution is 10 -2 , repeat the above process to do 10-fold serial dilution, to 10 -6 Dilution concentration, choose 10 -4 ~10 -6 For three dilutions, draw 0.1mL and spread it on the nutrient agar plate. After the plate is coated, culture it upside down at 40°C for 24 hours. Use an inoculation loop to pick up the colonies of suspected Bacillus bacteria and place them on the nutrient agar plate for streak separation and culture. After 24 hours of culture Finally, the colony with better separation effect was picked, transferred...

Embodiment 2

[0062] The preparation of embodiment 2 Bacillus coagulans HEW-B379 fermented liquid

[0063] Get Bacillus coagulans HEW-B379 (preservation number is CGMCC No.12553) seed solution (viable bacteria concentration is 10 9 CFU / mL) 3mL, inoculated in 300mL medium for shake flask fermentation culture, the fermentation temperature was 40°C, pH 6.8, 200r / min, and the fermentation time was 10h.

[0064] Among them, the shake flask fermentation medium is composed of the following components: 1.2% soluble starch, 1.8% sucrose, 1.0% yeast extract powder, 1.5% peptone, 0.08% dipotassium hydrogen phosphate, 0.2% magnesium sulfate, and 0.5% sodium chloride , calcium carbonate 0.2%, the balance is water, pH6.8±0.2.

[0065] After the shake flask fermentation, carry out the first-level seed culture in the fermenter, take 0.6L shake flask fermentation seed liquid and inoculate it into a 50L fermenter, the liquid volume is 35L medium, the inoculum size is 600mL, the fermentation temperature is 4...

Embodiment 3

[0069] Example 3 Probiotic verification of Bacillus coagulans HEW-B379

[0070] On the aseptic operating table, the concentration of pathogenic bacteria (enteropathogenic E. 10 9 The Bacillus coagulans HEW-B379 bacterium suspension 0.1mL that the embodiment 2 of CFU / mL makes is evenly spread on the nutrient agar plate, is prepared into several pathogenic bacteria plates, and each pathogenic bacterium nutrient agar Use sterile tweezers to clamp two sterilized Oxford cups on the flat plate symmetrically and place them on the flat plate so that they are in contact with the culture medium without gaps. After a few minutes, add 200 μL of the fermentation liquid prepared by the present invention dropwise to each small tube respectively. Make it overflow, incubate at 25°C-45°C for 20-36h, and then measure the diameter of the inhibition zone. Each experiment was repeated three times and the average value was taken. The results are shown in Table 2.

[0071] Table 2 Bacteriostatic ...

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Abstract

The invention provides a strain of Bacillus coagulans with probiotic effect, the strain is named HEW-B379, and the preservation number is CGMCC No.12553. Bacillus coagulans HEW‑B379 of the present invention has significant probiotic properties, and can effectively inhibit intestinal pathogenic Escherichia coli, Staphylococcus aureus, Salmonella typhi, Salmonella, Shigella, Proteus, Shewanella putrefaction, Growth and reproduction of Pseudomonas aeruginosa. Bacillus coagulans HEW-B379 of the present invention has strong stress resistance, can withstand high temperature, simulated gastric juice, simulated bile salt environment, and can maintain a survival rate of 99%-100%, and the bacteria can effectively regulate animal intestinal microorganisms Balance, inhibit the growth of harmful microorganisms, promote the absorption of nutrients by animals, improve the feed conversion rate, and improve animal production performance.

Description

technical field [0001] The invention belongs to the field of microbiology, and in particular relates to a bacillus coagulans HEW-B379 with probiotic effect and application thereof. Background technique [0002] As people's awareness of animal product safety and environmental protection continues to strengthen, as a substitute for antibiotics, probiotics are increasingly used in animal nutrition and feed. Probiotics are a class of microbial additives that can improve the microecological balance of the gastrointestinal tract of animals and are beneficial to animal health and production performance. Its main effect is reflected in improving animal metabolism, improving the absorption and utilization of nutrients, improving immunity, and playing an important role in reducing environmental pollution. [0003] Since Bacillus is highly resistant to adverse environments such as dryness, high temperature, high pressure, and oxidation, this stability increases its potential as a prob...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18A61K35/742A61P1/12A61P31/04C12R1/07
CPCA61K35/742C12N1/205C12R2001/07Y02A50/30
Inventor 李雪平
Owner BEIJING HESWOF BIOTECH CO LTD
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