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A kind of feed bacillus licheniformis that inhibits the formation of bacterial biofilm and its application

A bacillus licheniformis, the technology of inhibiting bacteria, applied in the field of microbiology, can solve the problems of head skin injury, loss of pig farming, Staphylococcus aureus infection, etc.

Active Publication Date: 2021-06-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] In recent years, with the expansion of the pig breeding scale and the increase of the breeding density in my country, the piglets will be grouped and put into the nursery after weaning. During the process of biting each other, it is easy to cause the skin on the head. Injuries, etc., and then Staphylococcus aureus infection occurred, which caused huge losses to my country's pig industry

Method used

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  • A kind of feed bacillus licheniformis that inhibits the formation of bacterial biofilm and its application
  • A kind of feed bacillus licheniformis that inhibits the formation of bacterial biofilm and its application
  • A kind of feed bacillus licheniformis that inhibits the formation of bacterial biofilm and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Isolation and Identification of Bacillus licheniformis RM17

[0032] 1. Isolation and cultivation of strains

[0033]Take 10ml of dairy cow rumen juice sample, filter, take 1ml of the filtrate and put it into a test tube with 9ml of normal saline, vortex and shake to mix it well, that is a 1:10 dilution, then take the dilution for 10-fold incremental dilution, and then choose 3 1ml of appropriate gradient dilutions were spread on LB medium. Cultivate at 37°C for 48-72 hours, observe and record the colony shape, pick a single colony with good growth, and separate and purify by streaking.

[0034] 2. UV mutagenesis and screening of strains

[0035] Pour the sterilized LB medium into a petri dish, and after solidification, take the bacterial suspension obtained in step 1 and spread it on a plate. Control the number of colonies in each petri dish to about 50. 20cm, mutagenesis 30s.

[0036] Pick the mutagenized strains, inoculate them in LB liquid medium, and ...

Embodiment 2

[0049] Stress resistance detection of embodiment 2 bacillus licheniformis RM17

[0050] 1. Heat resistance test

[0051] The Bacillus licheniformis RM17 (CGMCC No.16730) bacterial solution was treated in a water bath at 60°C, 80°C, and 100°C for 20 minutes, and each treatment was repeated three times. After the treatment, the number of viable bacteria was determined by pouring method.

[0052] The survival rate of Bacillus licheniformis RM17 CGMCC No.16730 can reach 80% after being treated at 60°C for 20 minutes, and the survival rate is still higher than 60% after being treated at 80°C for 20 minutes. The number of live bacteria is >10 6 CFU / ml.

[0053] 2. Acid resistance test

[0054] Bacillus licheniformis RM17 was inoculated into LB medium with pH values ​​of 2.0, 3.0, and 4.0, respectively, and the number of viable bacteria was determined by plate pouring method at 0h, 1h, 2h, 3h, and 4h, respectively.

[0055] Bacillus licheniformis RM17 CGMCC No.16730 can grow norm...

Embodiment 3

[0058] The growth curve determination of embodiment 3 bacillus licheniformis RM17

[0059] The growth curve represents the dynamic changes in the whole process of bacteria growing and multiplying in a new suitable environment until aging and death. Bacillus licheniformis RM17 CGMCC No.16730 was inoculated into LB medium at an inoculation amount of 1% (v / v), and cultivated at 37°C for 24 hours. The LB medium without bacterial solution was used as a blank control, and every 2 Hourly determination of OD600 value. The experiment was repeated three times, the results were averaged, the data were recorded and the growth curve was drawn. Such as Figure 5 As shown, in 2-14 hours, Bacillus licheniformis RM17 was in the logarithmic growth phase, and the reproduction rate was higher. The growth of Bacillus licheniformis RM17 entered the stationary phase at 14-24 hours, and the number of viable bacteria tended to be stable.

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Abstract

The invention discloses a feed bacillus licheniformis for inhibiting bacterial biofilm formation and application thereof. The bacillus licheniformis of the invention is RM17, and its preservation number is CGMCC NO.16730. The Bacillus licheniformis RM17 of the present invention is a Gram-positive bacterium, which has the ability to inhibit the formation of bacterial biofilms, is resistant to high temperatures of 80°C, can grow in an acidic environment with a pH value of 2.0 or above, and is resistant to bile salts. The ability is strong, and the bacterial agent of Bacillus licheniformis RM17 is used to feed animals, which is safe and reliable, promotes the digestion and absorption of nutrients, reduces the rate of diarrhea, improves feed conversion efficiency, and has positive effects in promoting growth.

Description

technical field [0001] The invention belongs to the field of microbiology, and in particular relates to a feeding bacillus licheniformis capable of inhibiting bacterial biofilm formation, high temperature resistance, acid resistance and bile salt resistance and application thereof. Background technique [0002] In recent years, with the expansion of the pig breeding scale and the increase of the breeding density in my country, the piglets will be grouped and put into the nursery after weaning. During the process of biting each other, it is easy to cause the skin on the head. Injuries, etc., and then Staphylococcus aureus infection occurs, which has caused huge losses to my country's pig industry. As an opportunistic pathogenic bacteria, Staphylococcus aureus exists widely in nature, but one of the related reasons that causes the reproduction of animal-derived Staphylococcus aureus can cause great harm is the formation of bacterial biofilm. [0003] Bacterial biofilm refers t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18A23K50/30A61K35/742A61P31/04C12R1/10
CPCA61K35/742A23K10/18A23K50/30A61P31/04C12N1/205C12R2001/10
Inventor 马曦孙美鸽姬琳堡李雪
Owner CHINA AGRI UNIV