Bacillus pumilus shou002 and application thereof
A technology of Bacillus pumilus and aquatic pathogens, applied in the field of microorganisms, can solve problems such as drug residues, environmental hazards, high morbidity and mortality
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Embodiment 1
[0017] The screening of embodiment 1 bacillus pumilus shou002
[0018] Using manganese nutrient agar, thiosulfate citrate bile salt sucrose agar (TCBS), eosin methylene blue agar (EMB), seawater agar (2216E) and freshwater agar (FWA) medium to separate and screen from large yellow croaker intestine Antagonistic bacteria with strong antagonistic effect on Vibrio parahaemolyticus (Vp1.2164) and Vibrio alginolyticus (Va-Y) were produced.
[0019] In vitro antagonistic experiments were carried out on the screened strains by the spot method, and the strains with the broadest antibacterial spectrum and the strongest effect were screened. The strains were identified by bacteria identification instrument and 16Sr RNA gene sequence analysis to determine their taxonomic status.
[0020] Select a strain with strong antagonistic effect and wide antibacterial spectrum, and through the VITEK-32 rapid bacterial identification instrument and 16Sr RNA gene sequence analysis, it is determined ...
Embodiment 2
[0021] Example 2 Bacillus pumilus shou002 extracellular product inhibits pathogenic Vibrio
[0022] (1) Referring to Sugita et al. (Sugita H, Hirose Y, Matsue N, et al. Vibrio sp.strain NM10, isolated from the intestine of a Japanese coastal fish, has an inhibitory effect against Pasteurella piscicida[J].Appli Environ Microbiol, 1998, 64: 231-237), the specific steps are as follows: activate the screened antagonistic bacteria on the slant of nutrient agar and inoculate them in nutrient broth medium, culture at 28°C for a certain period of time with constant temperature shaking, 4°C, 9600rpm / Centrifuge under the condition of 20 min for 20 min, discard the cell pellet, and filter the supernatant through a 0.22 μm microporous membrane, and the obtained liquid is the extracellular product of Bacillus pumilus shou002, which is stored at 4 °C for later use.
[0023] (2) Antibacterial activity was detected by plate diffusion method.
[0024] Vibrio parahaemolyticus (Vibrio parahaem...
Embodiment 3
[0026] The safety of embodiment 3 bacillus pumilus shou002
[0027] The nutrient broth culture medium (including 10g of peptone, 3g of beef extract powder, 5g of sodium chloride, 1000ml of distilled water, heated and dissolved, divided into flasks, and sterilized at 121°C for 15 minutes) was streaked on sheep blood. On the plate, cultured at 28°C for 24 hours, the production of hemolysin was judged according to the formation of a transparent circle around the colony, and Vibrio Vp1.2164 was used as a positive control. The test results showed that Vibrio parahaemolyticus 1.2164 had hemolysis on the blood plate, but Bacillus pumilus shou002 had no hemolysis on the blood plate, indicating that the strain did not produce hemolysin and had no potential pathogenicity.
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