Bacillus velezensis and application thereof in prevention and treatment of porcine diarrhea
A technology of Bacillus Velez and Bacillus, applied in the field of microbiology, can solve the problem that no one isolates the strain of Bacillus Velez, and achieve the effects of not being easy to die, reducing the rate of diarrhea, and reducing diarrhea
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Embodiment 1
[0037] Example 1: Isolation and identification of Bacillus sp. strain LL2018013
[0038] Step 1) Screening of Bacillus strains
[0039] Take the contents of the small intestine, cecum and large intestine of Diqing Tibetan pigs in sterilized saline (w / v=1%), shake fully, bathe in water at 80°C for 10 minutes, and after gradient dilution, transfer to LB solid medium for culture. After culturing at 35°C for 12 hours, a single colony with typical characteristics of Bacillus was picked, purified and cultured to obtain Bacillus.
[0040] Step 2) screening of antagonistic strains
[0041] Use the bacillus obtained in step 1) to prepare the bacteria solution to be tested, use Escherichia coli K88, Salmonella typhimurium and Staphylococcus aureus as indicator bacteria, prepare a suspension with 0.9% saline and adjust the concentration, and add it to the agar In the culture medium, prepare about 10 indicator bacteria 5 CFU / mL agar plate, after cooling, use the gel punching method to ...
Embodiment 2
[0058] Embodiment 2 weaned piglet feeding experiment
[0059] Preparation of A1 fresh bacterial solution: Inoculate the above-mentioned Bacillus veleus strain LL2018013 into LB liquid medium at a volume percentage of 0.1%, and culture at 30-35° C. for 20-24 hours to obtain A1 fresh bacterial solution.
[0060] Feeding experiment: 40 piglets weaned on 7 days of Diqing Tibetan pig were selected and randomly divided into two groups, half male and half female. 8 CFU / L), the control group was fed milk replacer, and the test period was 21 days, including 7 days in the pre-test and 14 days in the main test to observe the morbidity and mortality.
[0061] Diarrhea rate = number of diarrhea episodes / (total number of test piglets × positive test days) × 100%
[0062] Table 3 Diarrhea rate and survival rate of weaned piglets
[0063]
Embodiment 3
[0064] Example 3 Breeding pig intestinal flora abundance test
[0065] 1) Prepare A1 dry bacteria powder:
[0066] Inoculate the Bacillus veleus strain LL2018013 described in Example 1 into LB liquid medium according to 0.1% by volume, cultivate at 35°C for 20h, and then centrifuge at 10000rpm to obtain the thallus; add an appropriate amount of purified water to the above thalline and mix to obtain Mud, mixed with protective agent, then add appropriate amount of purified water, spray dry with embedding material to obtain dry bacterial powder;
[0067] Among them, the protective agent components and proportions are: 15% skimmed milk powder, 5% glycerin, 8% R3 resistant starch, 2% vitamin E, 3% maltodextrin, 3% agar powder, and the rest is potato resistant Starch; the embedding material is potato resistant starch, pectin, sodium hydroxymethylcellulose, and carrageenan, and the embedding material is prepared into a 5% aqueous solution according to the ratio of 1:1:1:1 for later ...
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