Feeding Bacillus licheniformis for inhibiting formation of bacterial biofilm, and application thereof
A Bacillus licheniformis, anti-bacterial technology, applied in the field of microbiology, can solve the problems of pig industry loss, head skin damage, Staphylococcus aureus infection, etc., to increase daily weight gain, reduce feed-to-meat ratio, and inhibit intestinal Effects of Pathogen Biofilm Formation
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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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