A simple bacillus and its cultivation method and application
A simple bacillus and culture medium technology, applied in the field of microorganisms, can solve the problems of less research and application of simple bacillus, achieve good market application prospects, meet the needs of industrialization, and have the effects of high fermentation bacteria
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Embodiment 1
[0019] Embodiment 1, the screening of simple bacillus
[0020] 1. Isolation and screening of Bacillus simplex f5
[0021] In July 2014, water samples and sediments from shrimp farming areas were collected from many places along the coast of Jiaonan to screen for spores. The supernatant was taken as a sample by conventional doubling dilution of the sediment; the water sample could be directly used as a sample; after the sample solution was incubated at 75°C for 20 minutes, it was spread on a nutrient agar plate to isolate and purify the strains, and 4 strains of Bacillus were obtained.
[0022] The purified single strain was tested for ammonia nitrogen degradation. Inoculate 4 single strains in LB seed medium, and culture them at 180rpm at 37°C for about 20h until the cell concentration is 10 8 CFU / mL. Transfer to ammonia nitrogen medium (glucose 5g, MgSO 4 ·7H 2 O 0.25g, K 2 HPO 4 0.5g, (NH 4 ) 2 SO 4 0.1g, water 1L, pH 7.2~7.4, cultured at 20°C 150rpm), regularly ...
Embodiment 2
[0027] Embodiment 2, the cultivation of simple bacillus
[0028] Inoculate a single colony of f5 in LB seed medium, and culture at 180 rpm at 37°C for about 20 hours until the cell concentration is 10 8 CFU / mL; transfer to fermentation medium according to 1% inoculum size, the formulation of the fermentation medium is: corn steep liquor 1g, glucose 3g, yeast extract 0.5g, KH 2 PO 4 0.1g, K 2 HPO 4 0.15g, (NH 4 ) 2 SO 4 0.5g, NaCl 0.1g, MgSO 4 ·7H 2 O 0.05g, H 2 O100mL, pH 7.0-7.2.
[0029] Cultured at 37°C and 180rpm for 24 hours, the spore maturity rate of microscopic examination was >90%; counted on nutrient agar plates, the bacterial count in the fermentation broth was >150×10 8 CFU / mL.
Embodiment 3
[0030] Embodiment 3, bacillus simplex f5 are to the degradation measurement of organic matter
[0031] The test was carried out in aquariums, each of which was added with 10 L of culture water (20 g / L of pulverized shrimp feed). The experiment was divided into control group and test group, with 3 parallels in each group. The test group added 1 mL of Bacillus simplex inoculum (the concentration of Bacillus simplex in the inoculum was 10 8 CFU / mL), the control group did not add bacterial agents. During the test, the water body was stirred regularly, observed continuously for 7 days at room temperature, and samples were taken regularly to measure the COD and ammonia nitrogen concentrations of the water body.
[0032] The result is as figure 2 and 3 As shown, strain f5 has obvious removal effect on COD and ammonia nitrogen in aquaculture water. During the test period, the COD and ammonia nitrogen in the water showed a decreasing trend, and the degradation rates were 53.3% an...
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