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Degrading bacterium capable of efficiently degrading malachite green drug and application thereof

A malachite green, degrading bacteria technology, applied in microorganism-based methods, methods of using microorganisms, bacteria and other directions, can solve the problems of unfavorable quality of aquaculture ecosystems and aquatic products, and no public reports of microbial degrading bacteria, etc. Effects of malachite green drug residues, protection of fishery ecological environment, and easy operation

Active Publication Date: 2018-11-13
上海市水产研究所(上海市水产技术推广站)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the above-mentioned defects of the prior art, the technical problem to be solved by the present invention is that the existing malachite green drug residue in the fishery breeding environment is unfavorable to the breeding ecosystem and the quality of aquatic products, and the malachite green drug residue in the fishery breeding environment There are no public reports of microbial degrading bacteria

Method used

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  • Degrading bacterium capable of efficiently degrading malachite green drug and application thereof
  • Degrading bacterium capable of efficiently degrading malachite green drug and application thereof
  • Degrading bacterium capable of efficiently degrading malachite green drug and application thereof

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Experimental program
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Effect test

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 with 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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Abstract

The invention discloses a degrading bacterium capable of efficiently degrading a malachite green drug and application thereof. The degrading bacterium is a bacterial strain D3 of citrobacter sp., andis preserved in the China Center for Type Culture Collection on March 16, 2018 with a preservation number of CCTCC No. M 20181314. The bacterium has ability of degrading the malachite green drug in afish farming environment. The invention further discloses a preparing, separating and purifying method for the degrading bacterium capable of efficiently degrading the malachite green drug. The degrading bacterium capable of efficiently degrading the malachite green drug can safely, efficiently and quickly degrade the residual malachite green drug in the fish farming environment, and has a very good application prospect in fish ecological environment protection, farming environment drug residue pollution control and food safety guarantee.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a degrading bacterium capable of efficiently degrading malachite green medicine in a fishery breeding environment and an application thereof. Background technique [0002] Malachite green is widely used as an industrial stain and a cytochemical stain. In 1933, it was found that malachite green has antibacterial effect. After that, many countries began to widely use malachite green as a fungicide, mainly for killing parasites in aquaculture water environment and mold in fish eggs. Excessive malachite green metabolizes into colorless malachite green in aquatic animals, and malachite green and colorless malachite green tend to remain in aquatic animals for a long time, resulting in highly teratogenic, carcinogenic and mutagenic effects. In the process of using malachite green in violation of regulations, fish farming users often over-administer. These over-administered malachite green ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/36C02F3/34C12R1/01C02F103/20C02F101/38
CPCC02F3/34C02F2101/308C02F2101/38C02F2103/20C12N1/20C12N1/36C12N1/205C12R2001/01
Inventor 孙振中刘菁华张玉平张健陈琳刘云璐丁义
Owner 上海市水产研究所(上海市水产技术推广站)
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