A microcystin-degrading bacterium and its application
A technology of microcystin and microbacteria, which is applied in the field of microbial application and environmental engineering, can solve the problems of little research on bacterial algicides, achieve a virtuous circle of ecological security, improve economic benefits, and facilitate colonization
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Embodiment 1
[0044] Example 1: Preparation of Microcystis Crude Extract and Detection of Microcystin Content in Microcystis Crude Extract
[0045] 1. Experimental materials: Microcystis crude extract, microcystin MC-LR chromatographically pure (Wuhan Institute of Hydrobiology), and experimental chemicals are analytical reagent grade;
[0046] 2. Sample preparation:
[0047] (1) Preparation of Microcystis crude extract: Add Microcystis aeruginosa FACHB 915 (Wuhan Institute of Hydrobiology) into a 10L glass bottle containing 8L Microcystis medium BG11 (Stanier et al., 1971), Light and ventilation culture, culture temperature is 25°C, light intensity is 20μE / (m2 s), light-dark cycle is 12h:12h, culture time is 14 days, the culture is centrifuged at 8000r / min, the precipitate is taken, and the precipitate is divided into equal parts 20 mL of 5% v / v acetic acid solution was added to 0.2 g of frozen-thawed algae cells, stirred and extracted for 20 min, centrifuged at 8000 r / min for 5 min to obt...
Embodiment 2
[0053] Example 2: Enrichment culture, separation and purification of algal toxin-degrading bacteria
[0054] Water sample collection: Breed eutrophic fish ponds (shrimp ponds), take one liter of surface water, store and transport them in plastic bottles, and operate aseptically.
[0055] For each water sample, two different experiments were designed, each experiment was repeated once, and each flask was statically cultured with a 50mL Erlenmeyer flask, and each Erlenmeyer flask was equipped with a 30mL water sample culture (water sample: EBB liquid medium=1:10 , v / v), wherein 30 mg of Microcystis crude extract was added to one of the conical flasks, so that the final concentration of MC-LR in the Microcystis crude extract was 100 μg / mL. Another conical flask was added with 30 μg MC-LR (purchased from Wuhan Institute of Hydrology), so that the final concentration of MC-LR in this conical flask was 1 μg / mL. 26-28°C, enrichment culture. Sampling 0.5mL every 3-4 days and storing...
Embodiment 3
[0060] Example 3: Identification of algal toxin-degrading bacteria
[0061] Morphological, physiological, biochemical, and molecular biological identifications were carried out on the target strain Exiguobacterium BIDE11 screened above, and the results showed that:
[0062] The bacterium is aerobic and grows well at 37°C. The plate colonies are translucent, irregular, raised, drop-shaped, positive for peroxidase, and can utilize a variety of oligosaccharides and polysaccharides. Gram staining is positive in microscopic examination, and a large amount of extracellular secretion material, not very viscous.
[0063] The above screened target strains were purely cultured, and their 16SrDNA partial sequence (shown in SEQ ID NO.1) was amplified by PCR using 16SrDNA universal primers, and registered in the gene bank, and compared and analyzed.
[0064] After comparison and analysis, the identification showed that the target strain (Exiguobacterium sp.) BIDE11 was of the genus Exiguo...
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