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Application of Bacillus subtilis ANSB060

A technology of Bacillus subtilis and Escherichia coli, applied in application, animal feed, additional food elements, etc., to achieve strong stress resistance, high spore survival rate, and the effect of promoting animal growth

Active Publication Date: 2012-11-28
计成 +6
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no report on the probiotics and stress resistance of Bacillus subtilis ANSB060, as well as its application in drinking water and feed for farmed animals

Method used

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  • Application of Bacillus subtilis ANSB060
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  • Application of Bacillus subtilis ANSB060

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The preparation of embodiment 1 Bacillus subtilis ANSB060 fermented liquid

[0047] Get Bacillus subtilis ANSB060 CGMCC No.3440 5ml (viable bacteria concentration is 10 9 CFU / ml), inoculated in 50ml medium for shake flask fermentation culture, the fermentation temperature is 37°C, the pH value is 7.0, the rotation speed is 200r / min, and the fermentation time is 24h.

[0048] Among them, the shake flask fermentation medium is composed of the following components: peptone 10g, glucose 5g, dipotassium hydrogen phosphate 0.1g, potassium dihydrogen phosphate 1.0g, manganese sulfate monohydrate 0.5g, magnesium sulfate heptahydrate 1.0g, distilled water 1000mL ; pH value is 7.0.

[0049] After the shake flask fermentation, carry out the fermenter pilot test, take 50ml shake flask fermentation seed liquid and inoculate it into a 10L fermenter, the liquid volume is 5L, the fermentation temperature is 37°C, the pH value is 7.0, the stirring speed is 300r / min, the fermentation ti...

Embodiment 2

[0052] Example 2 Probiotic verification of Bacillus subtilis ANSB060

[0053] Take 3 nutrient agar plates and culture them at 37°C for 24 hours, and remove part of the water to facilitate the diffusion of antibacterial substances in the medium. Take a small amount of Bacillus subtilis ANSB060 with an inoculation stick, and plant them in the center of the poured nutrient agar plate, culture at 37°C for 24 hours, then put the plate upside down on a petri dish containing a thin layer of chloroform and cover it for fumigation for about 30 minutes to kill the live bacteria. cells and fix the colonies to the surface of the plate. Remove the lid of the Petri dish and allow the residual chloroform in the plate to evaporate completely for 20 minutes. Spread a layer of freshly cultured Escherichia coli, Salmonella, and Staphylococcus aureus culture solution on the surface of the three plates, pour out the remaining bacteria solution after covering evenly, dry the plate, and place the p...

Embodiment 3

[0055] Example 3 Verification of Stress Resistance of Bacillus subtilis ANSB060

[0056] 1. Tolerance test of simulated gastric juice environment: Dissolve pepsin in 0.5% normal saline to make the final concentration 3g / L, adjust the pH value to 2.0 with concentrated hydrochloric acid or 10% NaOH. Count the live bacteria liquid of Bacillus subtilis ANSB060 before the test. During the test, take 0.5mL live bacteria solution and add it to 4.5mL simulated gastric juice, and quickly mix it fully on a shaker, then place it at 37°C for 2 hours to obtain a culture solution, and count the number of viable bacteria remaining in the spores.

[0057] The results showed that the spore survival rate reached 98.8%.

[0058] 2. Tolerance test of simulated bile salt environment: use pancreatin to make a 1g / L solution, add 0.3% pig bile salt to the solution, adjust the pH to 8.0 with 10% NaOH, and prepare a bile salt solution Sterilize by filtration with a 0.45 μm microfiltration membrane. ...

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Abstract

The present invention relates to application of Bacillus subtilis ANSB060 (Preservation code number of CGMCC NO. 3440), especially to application of the bacterial strain in drinking water and / or feed of breeding animals. The Bacillus subtilis ANSB060 can be obtained by such techniques as repeated separation, purification and rejuvenation and the like; the Bacillus subtilis ANSB060 has the advantages of strong bioactivity, excellent health benefit property, and good stress resistance and the like. Fermentation liquor of the Bacillus subtilis ANSB060 obtained by the production technique like deep fermentation and the like is added to the drinking water or the feed for the animals; after entering into the animal intestinal canals, the bacterial strain can quickly activate, reproduce and form preponderant profitable strain such that the application of the Bacillus subtilis ANSB060 can reduce the harmful strain, adjust the intestinal microecological balance, substitute drugs like antibiotics, and improve the animal weight increment and the feed utilization ratio and the like.

Description

technical field [0001] The invention relates to the application of bacillus subtilis ANSB060, in particular to the application of the bacterial strain as a feed additive for breeding animals. Background technique [0002] Modern intensive animal husbandry restricts animals from contacting the soil and the surrounding natural environment. This feeding condition is not conducive to the establishment of normal flora in the gastrointestinal tract of animals. When the external environmental conditions change, it is easy to cause the imbalance of micro-ecological flora in the gastrointestinal tract of animals, thereby reducing the resistance to pathogenic bacteria. At present, this loss is mainly reduced by improving feeding management and adding antibiotics to the diet. However, with the widespread use of antibiotics, the problems of drug resistance and residues are becoming more and more prominent. High attention of ordinary consumers. Prebiotics have become the first choice f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A23K1/17A23K20/195
Inventor 计成马秋刚高欣董军王强张艳黎敏
Owner 计成
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