Bacillus subtilis and application thereof to livestock breeding
A technology of Bacillus subtilis and livestock and poultry feed, which is applied in the fields of application, bacteria, and animal husbandry. It can solve the problems of large differences in probiotic performance and fewer reports on the overall performance of strains, so as to promote the development of immune organs and improve production performance. , the effect of promoting reproduction
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Embodiment 1
[0013] Example 1 Isolation and Identification of Bacillus subtilis WL-04
[0014] 1. Isolation and screening of Bacillus
[0015] Add the sludge collected from the Pingdu Broiler Farm in Qingdao City into sterile water at a ratio of 1:10, shake and mix to prepare a suspension, and treat it in a water bath at 80°C for 10 minutes to kill most of the non-heat-resistant Bacillus with good heat resistance was preliminarily obtained.
[0016] 2. Screening of Bacillus with ability to inhibit pathogenic bacteria and high enzyme production
[0017] 1) Preparation of bacillus bacteria liquid: inoculate the isolated bacillus into nutrient broth respectively, shake and cultivate for 24 hours at 37°C and 200rpm;
[0018] 2) In vitro antibacterial test - micro broth method: Centrifuge the bacillus liquid cultured for 24 hours, collect the supernatant, take a sterile 96-well culture plate, add the bacillus supernatant and pathogenic bacteria liquid (used in the experiment) at a ratio of 1:...
Embodiment 2
[0029] Embodiment 2: Bacillus subtilis WL-04 inhibits the analysis of zoonotic pathogens
[0030] 1. Cultivation of indicator pathogenic bacteria: Randomly select pathogenic bacteria isolated from the intestinal tract of sick chickens and pigs, including Escherichia coli, Salmonella, and Clostridium perfringens, a total of 5 strains (gifted by the Preventive Veterinary Laboratory of Qingdao Agricultural University), of which Escherichia coli, Salmonella was inoculated in nutrient broth, 37°C, 200rpm, cultured for 15h; Clostridium perfringens was inoculated in blister medium, 37°C, cultured for 15h;
[0031] 2. Preparation of Bacillus subtilis WL-04 bacterial liquid: pick a single colony, inoculate it in nutrient broth, culture at 37°C, 200rpm, for 24 hours;
[0032] 3. Antibacterial test in vitro - trace broth method: centrifuge the bacillus liquid cultured for 24 hours, collect the supernatant, take a sterile 96-well culture plate, add the bacillus supernatant and pathogenic ...
Embodiment 3
[0041] Embodiment 3: Bacillus subtilis WL-04 produces enzyme performance analysis
[0042] 1. Prepare Bacillus subtilis WL-04 bacterial liquid: pick a single colony, inoculate it in nutrient broth, culture at 37°C, 200rpm, for 24 hours;
[0043] 2. Obtain the fermentation supernatant: centrifuge the bacillus liquid cultured for 24 hours at 10,000 rpm for 10 minutes, and collect the supernatant;
[0044] 3. Enzyme production: α-amylase activity detection (GB / T24401-2009) and neutral protease activity detection (GB / T28715-2012), a total of 3 batches of determination were carried out, the results are shown in Table 3;
[0045] Table 3: Analysis of enzyme production by Bacillus subtilis WL-04
[0046]
[0047]
[0048] After three batches of repeated experiments and the analysis of enzyme activity, the enzyme production of Bacillus subtilis WL-04 was relatively stable, and the average activity of α-amylase was 22U / mL; the average activity of neutral protease was 170U / mL. M...
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