Rainbow trout-derived bacillus subtilis strain as well as screening method and application thereof
A technology derived from Bacillus subtilis and rainbow trout, applied in the field of microorganisms, can solve problems such as inability to obtain better effects and promotion, and achieve the improvement of immunity and digestion-related enzyme activities in rainbow trout-related tissues, improvement of anti-infection ability, and promotion of body length. Effect
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Embodiment 1
[0042] Example 1 Screening and identification of Bacillus subtilis strain RT-BS07
[0043] 1. Isolation and purification of RT-BS07 strain
[0044]Representative rainbow trout farms in Qinghai, Liaoning and other cold-water fish main breeding areas were selected as sampling sites, and the probiotics in the intestinal tract of rainbow trout were initially screened and isolated. Under aseptic conditions, the body surface of rainbow trout was disinfected, the intestinal tissue of rainbow trout was dissected, washed three times with sterile phosphate buffered saline (PBS), the intestinal tissue was broken with a tissue homogenizer to obtain a tissue homogenate, and the tissue was homogenized. The slurry was collected in 1 mL of sterile saline, and after serial dilution, 100 μL was spread on the surface of the selective medium, and placed in a constant temperature incubator at 28°C for upside-down culture. Pick a single colony on the plate and perform pure culture by streak cultur...
Embodiment 2
[0063] Example 2 Analysis of biological characteristics of Bacillus subtilis strain RT-BS07
[0064] 1. Analysis of antibacterial properties of RT-BS07 strain in vitro
[0065] Polymyxin B was used as positive control, PBS was used as negative control, and common disease microorganisms in the breeding process of rainbow trout were selected, and the effects of RT-BS07 strain on Aeromonas salmonicida (Aeromonas salmonicida), Pseudomonas fluorescens (Pseudomonas fluorescens), A.sobria, A.veronii, Yersinia ruckeri, and A.hydrophila (For specific information on the strains used, see: Zhang Xiao, Li Shaowu, Wang Di, Cao Yongsheng, Lu Tongyan. Establishment and application of qPCR rapid detection method for Yersinia ruckerii [J]. Chinese Fisheries Science, 2017, 24(6) :1254-1260.) Antagonistic antibacterial effect.
[0066] The specific measurement method is: add the selected representative pathogenic microorganisms to the solid medium and pour the plate, punch holes in the Oxford ...
Embodiment 3
[0099] Example 3 Evaluation of the safety and application effect of RT-BS07 strain on rainbow trout
[0100] 1. Preparation of bait containing probiotic RT-BS07 strain and immune feeding of experimental fish
[0101] Take out the RT-BS07 strain frozen at -80°C, and inoculate 5 μL of the frozen bacteria into 5 mL of LB liquid medium (yeast extract 5 g; tryptone 10 g; sodium chloride 10 g; adjust the pH to 7.0 with NaOH, Add water to make up to 1L, keep at 121°C for 15min (autoclave), shake the bacteria at 28°C at 120 rpm for 12h to obtain a culture solution, then take 2mL of the culture solution and add it to 2L liquid LB medium, continue for 120 rpm Shake the bacteria per minute for 18 hours to prepare the bacterial suspension.
[0102] Aile silver fish feed was selected as the basic feed, and a feed with a diameter of 2 mm was selected according to the size of the experimental fish. The active ingredients of the feed included: 45% crude protein, 20% crude fat, 17.9% nitrogen...
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