Composite microecological preparation for multifunctional sea cucumber culture
A compound micro-ecological and multi-functional technology, which is applied in the field of multi-functional compound micro-ecological preparations for sea cucumber cultivation, to achieve the effects of ensuring food safety, improving the quality of sea cucumber products, and reducing drug residues
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Embodiment 1
[0030] Screening and safety experiments of potential probiotics
[0031] Take wild and healthy sea cucumbers, perform vivisection on sea cucumbers under aseptic conditions, take their intestines, squeeze out the contents of the intestines, wash the intestines with sterilized 15‰ normal saline three times, and homogenize the washed intestines , after equal proportion dilution, smear on TSB medium and 2216E seawater agar medium, select appropriate dilution plates to isolate bacteria, and obtain 240 strains of bacteria in total. 240 strains of purely cultured bacteria were cultured with casein hydrolysis medium and starch hydrolysis medium to measure their enzyme production activity. and Aeromonas hydrophlia (Aeromonas hydrophlia) as indicator bacteria, using spot method, cross method and Oxford cup method to detect the antibacterial effect of isolated bacteria on sea cucumber pathogenic bacteria, and screened out 3 strains with good antibacterial effect and Enzyme-producing bac...
Embodiment 2
[0036] Preparation of three active probiotic preparations
[0037] The preparation method of Bacillus atrophaeus CCTCC M 2010316 bacteria powder, Bacillus subtilis CCTCC M 2010314 bacteria powder, and Bacillus cereus CCTCC M 2010315 bacteria powder comprises the following steps:
[0038] A. Plate culture rejuvenation: Inoculate Bacillus atrophaeus, Bacillus subtilis, and Bacillus cereus stored at -80°C on TSB plate medium respectively, and cultivate them at 28°C for 24 hours to rejuvenate the strains and form a single colony;
[0039] B. Preparation of seed liquid: the bacterial classification cultivated in step A is made into 1×10 with 15‰ sterilized physiological saline 5 Inoculate the bacterial suspension of CFU / mL into a 250mL Erlenmeyer flask filled with 100mL of sterile TSB liquid medium at an inoculum size of 5%, and culture at 28°C and 170rpm for 18-20h with shaking;
[0040] C. Fermentation culture: Inoculate the seed solution in step B with 5% inoculum in a 5L Erlen...
Embodiment 3
[0044] Preparation of multi-functional compound micro-ecological preparation for sea cucumber cultivation: Compatibility with Bacillus atrophaeus CCTCC M2010316 bacteria powder, Bacillus subtilis CCTCC M 2010314 bacteria powder, and Bacillus cereus CCTCCM 2010315 bacteria powder according to the weight ratio of 1:1:1 , and mix well to prepare a compound microecological preparation.
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