Bacillus atrophaeus, and preparation and application of bacillus atrophaeus
A technology of Bacillus atrophaeus and microbial preparations, applied in the field of aquatic application microorganisms, can solve the problems of lagging disease control technology, restricting the healthy and sustainable development of the industry, and economic losses, etc., and achieve the effects of fast reproduction, ensuring food safety, and reducing drug residues
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Embodiment 1
[0042] Screening and isolation process of Aj080319IA-12 strain:
[0043] The Aj080319IA-12 strain was isolated from the intestines of wild sea cucumbers by the Yellow Sea Fisheries Research Institute of the Chinese Academy of Fishery Sciences. In 2008, the Yellow Sea Fisheries Research Institute of the Chinese Academy of Fishery Sciences captured wild sea cucumbers from the sea area of Taipingjiao, Qingdao. Under sterile conditions, the sea cucumbers were live dissected. The intestinal tract was rinsed three times with 15‰ normal saline, and the washed intestinal homogenate was uniformly diluted and spread on TSB medium and 2216E seawater agar medium, and the plates with appropriate dilution were selected for bacterial isolation and passed multiple Bacteria were cultured and purified by streaking, and the morphological structure and quantity of the bacteria were recorded and photographed; the common pathogenic bacteria of A. japonicus - Vibrio splendidus (Vibrio splendidus),...
Embodiment 2
[0045] Morphological characteristics of Aj080319IA-12 strain
[0046] Gram staining of the cells of the strain Aj080319IA-12 was positive, the cells were rod-shaped, spore staining showed spores, and capsule staining showed that the cells had no capsule. On the TSB solid plate medium, the colony is yellow, the surface is wrinkled, silky, opaque, the edges are irregular, and raised. After 48 hours of culture, the colony is black around the colony, and there is a bacterial film and precipitates during liquid culture.
Embodiment 3
[0048] 16S rDNA sequence determination
[0049] Such as figure 1 Shown is the results of the phylogenetic tree obtained by using the PHYLIP software according to the 16S rDNA sequence described in the embodiment of the present invention.
[0050] The universal primers used to amplify bacterial 16S rDNA were 27F: 5’-AGAGTTTGATC(C / A)TGGCTCAG-3’ and reverse primer 1492R: 5’-TACGG(C / T)TACCTTGTTACGACTT-3’. The 16S rDNA was amplified using the genomic DNA of the Aj080319IA-12 strain as a template and transformed and sequenced. See the sequence table for the obtained sequence. The obtained 16S rDNA sequence of the Aj080319IA-12 strain was compared in Genbank for homology, and Clustalw software was used for multiple sequence matching alignment, and the phylogenetic inference software package PHYLIP4.0 was used for statistical analysis and clustering analysis, and the neighbor linking method was used Obtain a phylogenetic tree ( figure 1 ), the phylogenetic tree was evaluated by boo...
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