Biological enhanced functional flora analysis method based on high-flux sequence testing

A bioaugmentation and analysis method technology, applied in the field of microbial bioaugmentation treatment, can solve the problems of key factors such as core flora and characteristic flora information missing system flora structure and treatment effect, low throughput of analysis methods, cumbersome operations, etc. , to achieve the effect of ensuring repeatability, broad application prospects, and easy and flexible operation

Active Publication Date: 2018-09-04
NANJING UNIV
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Problems solved by technology

[0005] Purpose of the invention: The present invention aims at the lack of key core flora and characteristic flora information in the biointensive treatment system of pharmaceutical wastewater, and the key factors affecting the system flora structure and treatment effect are not yet clear, and the existing analysis methods have low throughput and cumbersome operations , heavy workload, high cost, etc., the present invention aims to realize the transparency of the "black box" of the pharmaceutical wastewater biointensive treatment system, and adopts a bioinformatics analysis method based on a new generation of high-throughput sequencing to provide a biological A method for in-depth analysis of functional flora in enhanced treatment systems

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  • Biological enhanced functional flora analysis method based on high-flux sequence testing
  • Biological enhanced functional flora analysis method based on high-flux sequence testing
  • Biological enhanced functional flora analysis method based on high-flux sequence testing

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Embodiment 1

[0035](1) Take the returning sludge from the secondary settling tank of the urban domestic sewage treatment plant, first filter the debris through the filter, and inoculate it into the MBR reactor, the concentration of the inoculated sludge is 3500-4500mg / L; the MBR The operating conditions are: the hydraulic retention time (HRT) is 14 hours, the dissolved oxygen is 3-6mg / L, the pH value is 7-8, and the temperature is normal temperature (25±2°C); continuously increase the load in the influent, first gradually Increase the load of COD (from 200mg / L to 1000mg / L) and ammonia nitrogen (from 20mg / L to 250mg / L) in the influent, and then continuously increase the influent concentration of penicillin in each stage (from 100mg / L to 1000mg / L) At the same time, reduce the concentration of glucose in the influent water (from 1000mg / L to 0mg / L), until the carbon source is completely replaced by penicillin, monitor the COD and ammonia nitrogen concentration in the influent and effluent water...

Embodiment 2

[0043] (1) Take the returning sludge from the secondary settling tank of the urban domestic sewage treatment plant, first filter the debris through the filter, and inoculate it into the MBR reactor, the concentration of the inoculated sludge is 3500-4500mg / L; the MBR The operating conditions are as follows, the hydraulic retention time (HRT) is 14 hours, the dissolved oxygen is 3-6mg / L, the pH value is 7-8, and the temperature is normal temperature (25±2°C); continuously increase the load in the influent, first gradually Increase the load of COD (from 200mg / L to 1000mg / L) and ammonia nitrogen (from 20mg / L to 250mg / L) in the influent, and then continuously increase the influent salinity (from 1% to 4%) at each stage, each Monitor the concentration of COD and ammonia nitrogen in the influent and effluent during the operation stage, and properly discharge the sludge after running for a period of time to maintain the sludge concentration in the reactor at 4000mg / L-5000mg / L, so as t...

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Abstract

The invention discloses a biological enhanced functional flora analysis method based on high-flux sequence testing. The method comprises the following operation steps of using a membrane bioreactor; performing functional microbe group domestication by continuously improving the load in inlet water; collecting sludge after the stable operation in each operation stage of the membrane bioreactor; performing centrifugal precipitation so as to enrich microbes; extracting microbe total DNA on sludge samples in each operation stage; performing target fragment PCR amplification; purifying the amplification products; identifying the amplification product quality; mixing different sample PCR amplification products according to the equal mass; performing fragment size and quantitative detection on amixed library; performing high-flux sequence testing; finally performing original data quality control and biological information analysis. The analysis method provided by the invention can be used for systemically analyzing the structure features of the microbe flora in the waste water biochemical treatment process from the ecological aspect, determining the core function flora in the waste watertreatment system and identifying the key water quality parameters influencing the microbe flora structures.

Description

technical field [0001] The invention relates to a method for analyzing bioaugmented functional flora based on high-throughput sequencing, and belongs to the technical field of microbial bioaugmentation treatment. Background technique [0002] The environmental pollution caused by pharmaceutical wastewater is becoming more and more serious. Typical pharmaceutical wastewater such as antibiotics, vitamins, and pesticides has caused serious pollution to the human living environment. At present, the biological method is still the mainstream technology for pharmaceutical wastewater treatment. However, due to the large fluctuations in water quality and quantity of pharmaceutical wastewater, it is easy to affect the biological treatment effect. Previous studies mostly focused on the determination of the treatment process and its parameters, but little is known about the key core flora and characteristic flora information in the pharmaceutical wastewater treatment system, and the ke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C12Q1/04
CPCC12Q1/6869C12Q2531/113C12Q2535/122C12Q2537/165
Inventor 黄开龙任洪强张徐祥叶林孙浩浩赵福正
Owner NANJING UNIV
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