Bacillus licheniformis and application thereof to aquiculture
A technology of Bacillus licheniformis and Bacillus subtilis, which is applied in application, Lactobacillus, Bifidobacterium and other directions, can solve problems such as limited effect, achieve the effect of improving immunity and disease resistance, and has wide application prospects
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Embodiment 1
[0018] Isolation, screening and identification of embodiment 1 bacterial strain
[0019] 1. Sample
[0020] The intestines of crucian carp were collected from the freshwater aquaculture experimental field in Jiaonan City, Qingdao.
[0021] 2. Medium
[0022] 1) LB medium: 10g of peptone, 10g of sodium chloride, 1000ml of distilled water, pH7.0-7.2, prepare a solid medium, add 20g of agar, and sterilize at 121°C for 20min.
[0023] 2) Starch screening medium: 20g soluble starch, 1g potassium nitrate, 0.5g sodium chloride, 0.5g dipotassium hydrogen phosphate, 0.5g magnesium sulfate, 0.01g iron sulfate, 1000ml distilled water, pH7.0-7.2, prepare solid culture Add 20g of agar to the base, and sterilize at 121°C for 20min.
[0024] 3) Starch utilization medium: 10 g of soluble starch, 5 g of peptone, 10 g of sodium chloride, 1000 ml of distilled water, pH 7.0-7.2.
[0025] 4) Starch-bromocresol purple medium: 10 g of soluble starch, 5 g of peptone, 10 g of sodium chloride, 4 ml...
Embodiment 2
[0037] Embodiment 2 Bacillus licheniformis DN01 produces amylase ability determination
[0038] Inoculate activated Bacillus licheniformis DN01, take 5ml of the activated bacterial solution, inoculate it in a liquid fermentation medium, culture at 37°C, take samples every 8 hours, and measure the amylase enzyme in the fermentation supernatant according to the method of GB / T 24401-2009 live, expressed in U / ml. Amylase activity is defined as one enzyme activity unit (U) when 1 g of soluble starch is liquefied in 1 hour at pH 7.0 and 37°C. For the amylase production curve of Bacillus licheniformis DN01 see figure 1 .
[0039] From figure 1 It can be seen that the Bacillus licheniformis DNO1 provided by the present invention has a strong ability to produce amylase under liquid culture conditions. After 8 hours of cultivation, the activity of amylase produced by fermentation of the strain increased sharply. At 40 hours, the amylase activity reached 2417.5U / ml. Within the range ...
Embodiment 3
[0040] Embodiment 3 bacillus licheniformis DN01 degrades starch and produces short-chain fatty acid situation
[0041] The short-chain fatty acids produced by the decomposition of probiotics can affect the physiological function and growth of the body's intestinal tract. These short-chain fatty acids can provide energy for intestinal epithelial cells, maintain the water and electrolyte balance in the intestinal tract, and also maintain mucosal immune cells. , reduce the production of pro-inflammatory factors, which is beneficial to the repair of mucosal inflammation.
[0042]Bacillus licheniformis DN01 was inoculated in the starch utilization medium, cultured for 24 hours, the bacterial liquid sample was pre-cooled, acidified with concentrated sulfuric acid, extracted with ether, and the supernatant was used for sample detection. Gas chromatography test on the computer, the inlet temperature is 180°C, the column temperature is 80°C, the initial test time is 2min, then rise to ...
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