Application of adopting Mojave Bacillus KJS-3 as feed additive
A technology of Bacillus virulina and Bacillus, applied in application, animal feed, animal feed, etc., can solve the problems of complex fermentation production process and difficult separation of active substances, and achieve the effects of short fermentation cycle, low production cost and high concentration
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Embodiment 1
[0016] Example 1. Attachment rate experiment of Bacillus mojave KJS-3
[0017] experiment method:
[0018] 1. First set the concentration to 1X10 5 Cells / 4ml Caco-2 cells are cultured in tissue culture dishes.
[0019] 2. Prepare the strain and the control strain (L.rhamnosus GG) respectively into 4.1X10 5 cfu / ml, 4.9X10 5 The solution with a concentration of cfu / ml is added to the pre-prepared Caco-2 cell culture medium.
[0020] 3. At 37℃, 5% CO 2 In the incubator, culture the two groups of petri dishes for 2 hours.
[0021] 4. After the culture is completed, treat it with sterile phosphate buffer (pH=7.4) for 3 times, then recover the attached cells with trypsin, wash with PBS, and then cover with TSA medium.
[0022] Experimental results: The control strain (L.rhamnosus GG) is recognized as the strain with the highest attachment rate in the world, and its attachment rate reaches 83.6±6.8%, but the commonly used strain with high attachment rate is Lactobacillus caseiTMC0409, and its a...
Embodiment 2
[0023] Example 2, Antibacterial Experiment of Bacillus Mojave KJS-3
[0024] In order to test the antibacterial characteristics of bacillus mojavensis KJS-3, bacillus mojavensis KJS-3 was tested against two subtypes of super virus resistant vancomycin-resistant enterococcus (VRE) Experiment with 4 subtypes of drug-resistant Staphylococcus aureus (MRSA):
[0025] Test method—take an appropriate amount of strains from a petri dish containing Bacillus mojave KJS-3 and place them in a test tube containing TSB culture medium to make 4 ml of bacterial culture medium. After overnight culture, inoculate it in 100 ml on the next day Continue to culture in TSB culture broth for more than 3 hours. After the culture is completed, put 100 ml of bacterial broth into the fermentor to prepare 4 liters of fermentation broth. After another 2 days, centrifuge, take the supernatant and add 3 moles of hydrochloric acid for pH. After adjusting the pH to 2, put it in a 4°C freezer for storage, and then ...
Embodiment 3
[0030] Example 3 Experiment of primary metabolite of Bacillus mojave KJS-3
[0031] The primary metabolite of bacillus mojavensis KJS-3 produces various enzymes, vitamins and other nutrients. The present invention carries out API50CHB enzyme activity test and vitamin test experiment.
[0032] API50CHB enzyme activity test: This test is a common typical experiment to identify bacterial enzyme activity. It is a simple medium composed of API50CH test strips of 49 fermentable carbohydrates. First inoculate the strain of Bacillus mojave KJS-3 into 100 ml of TSB culture medium and cultivate for more than 8 hours, centrifuge to obtain the supernatant, and inoculate an appropriate amount of the suspension of this strain into each tube of the test strip. , Due to the acid production of fermented carbohydrates and the decrease of the pH value, the indicator changes color. The darker the color, the stronger the enzyme activity, so as to identify the type and strength of enzymes produced by B...
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