Degrading bacteria that can efficiently degrade malachite green drugs and their applications
A technology of malachite green and degrading bacteria, which is applied in the direction of microorganism-based methods, methods using microorganisms, bacteria, etc., can solve problems such as unfavorable quality of aquaculture ecosystems and aquatic products, and no public reports of microbial degrading bacteria, etc., to achieve reduction The effects of malachite green drug residue, protection of fishery ecological environment, and easy operation
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Embodiment 1
[0046] Embodiment 1, the screening of malachite green degrading bacterial strain
[0047] Preparation of media and reagents
[0048] Basic medium: potassium dihydrogen phosphate 3.0g, magnesium sulfate heptahydrate 0.1g, ammonium nitrate 2.0g, anhydrous CaCl 2 Mix 0.01g, 1.5g of dipotassium hydrogen phosphate, and 0.01g of disodium ethylenediaminetetraacetate, add to 1000mL of filtered aquaculture water, adjust the pH to 7.6-7.8, and make it after autoclaving (121°C, 15min);
[0049] Nutrient medium: 10.0g of peptone, 3.0g of beef powder, 5.0g of sodium chloride, and 1.0g of glucose are mixed, added to 1000mL of filtered culture water, adjusted to pH 7.6-7.8, and prepared after high-pressure steam sterilization (121°C, 15min);
[0050] Enrichment culture solution: add malachite green drug to the nutrient medium, so that the concentration of malachite green drug is 5 mg / L;
[0051] Malachite green degradation liquid medium: add malachite green to 99% basal medium and 1% nutr...
Embodiment 2
[0057] Embodiment 2: Identification of malachite green drug-degrading bacterial strain
[0058] The bacterial strain D3 that above-mentioned embodiment 1 obtains carries out morphological characteristic and molecular biological identification, and the bacterium colony morphology of this bacterial strain is as follows figure 1 As shown, the scanning electron microscope pictures of the strains are as follows figure 2 and image 3 shown;
[0059] The main biological characteristics of the strain are:
[0060] Gram stain negative, bacilli; ornithine decarboxylase test, lysine decarboxylase test, urease test, arginine double hydrolysis test, gelatin liquefaction test negative; mannitol test, sorbitol test positive; glucose can be used , lactose, sucrose; the strain grows in a fishery breeding environment, the optimum growth temperature is 30°C, and the optimum growth pH is 7.6-7.8. On solid medium, the colony is round, white and translucent;
[0061] The strain was identified ...
Embodiment 3
[0062] Embodiment 3: Degradation test of malachite green degrading bacteria to malachite green in fishery water body
[0063] The bacterial strain D3 that embodiment 1 separates, purifies obtains with bacterial quantity OD 600nm =0.2 was inoculated into 100mL of 250mL Erlenmeyer flasks containing fishery water with a malachite green drug concentration of 5mg / L, compared with fishery water without bacteria, cultured on a constant temperature shaker at 30°C and 160rpm for 36h. After the cultivation, the culture solution was centrifuged at 7000rpm for 3 minutes, 100 μL of the solution was taken, diluted into 10 mL of water, 300 μL of deuterated malachite green and deuterated recessive malachite green with a concentration of 0.2 μg / mL were added, and 1 mL of the sample was passed through the PRS solid phase. Extraction column, PRS column after activation (2mL acetonitrile, 2mL water), sample loading, rinsing (2mL water, 2mL acetonitrile), and finally eluted with 3mL acetonitrile:a...
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