A kind of identification method of in situ degrading bacteria of acetonitrile in waste water
A technology for degrading bacteria and acetonitrile, applied in the field of microbial identification, can solve the problems of low assimilation rate of nitrile compounds and adsorption of nitrile compounds
Active Publication Date: 2020-10-30
BEIJING NORMAL UNIVERSITY
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Problems solved by technology
There is no previous research on the use of SIP method to identify nitrile degrading bacteria. The main reasons include: low assimilation rate of nitriles, cross-feeding phenomenon, and strong adsorption of nitriles in environmental media.
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Embodiment 1
[0040] A method for identifying in situ degrading bacteria of acetonitrile in wastewater, comprising:
[0041] Step 1: Collect the activated sludge from the wastewater treatment plant for standby;
[0042] The activated sludge sample is taken from the aeration reaction stage of the reactor, and the reactor is running in a stable state during sampling; after sampling, the sample is transported to the laboratory in time for storage at 4°C for future use; among them, the nitrile content of the activated sludge sample is less than 0.02 mg / L.
[0043] Step 2: Add to the activated sludge collected in step 1 15 N-labeled acetonitrile for acclimation culture;
[0044] Cultivate about 50mL of raw activated sludge in a 250mL beaker, and then add the final concentration of about 55mg / L 15 N labeled acetonitrile. Step 3: Join every once in a while 15 N-labeled acetonitrile for sequential batch culture;
[0045] Take about 10mL samples for analysis on the 3rd, 6th, 12th, 18th and 24th ...
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The invention relates to a method for identifying in-situ degrading bacteria of acetonitrile in wastewater, and belongs to the technical field of microorganism identification. The method comprises thefollowing steps: step 1, collecting activated sludge of a wastewater treatment plant for later use; step 2, adding 15N-labeled acetonitrile into the activated sludge collected in the step 1, and carrying out acclimatization culture; step 3, adding the 15N-labeled acetonitrile at set intervals, and carrying out sequencing batch culture; step 4, performing activated sludge DNA extraction through centrifugal separation; step 5, carrying out ultra-high-speed centrifugation on the extracted DNA obtained in the step 4; step 6, determining the abundance of 16S rRNA microorganisms in each layer of DNA of the 15N-labeled acetonitrile by using quantitative PCR; step 7, determining the community structure of the microorganisms; and step 8, identifying the active acetonitrile degrading bacteria in the SIP treatment by comparing enrichment ratios. According to the method disclosed by the invention, various microorganisms which are not reported yet are identified to be related to acetonitrile degradation, so that the diversity of acetonitrile degrading bacterium communities is enriched.
Description
technical field [0001] The invention relates to the technical field of microbial identification, and specifically provides an identification method for in-situ degradation bacteria of acetonitrile in wastewater. Background technique [0002] With the rapid development of my country's industry, the content of nitriles in industrial waste water is also increasing day by day. my country's "Comprehensive Wastewater Discharge Standard" (GB 8978-1996) stipulates that the maximum allowable discharge concentration of total cyanides in industrial waste water: a The standards for grade two and grade two are both 0.5mg / L, and the standard for grade three is 1.0mg / L. Therefore, it is of great practical value to study the treatment method of nitrile-containing wastewater to effectively control the concentration of nitrile in wastewater and reduce its impact on the environment and human body. [0003] At this stage, chemical and biological methods are the main methods for treating nitrile...
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IPC IPC(8): C12Q1/6851C12Q1/06
CPCC12Q1/6851C12Q2531/113C12Q2535/122C12Q2563/107
Inventor 孙寓姣尹萌郑丹阳
Owner BEIJING NORMAL UNIVERSITY