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Relative quantitative method of tubifex in sludge reduction technology and application of relative quantitative method

A technology of sludge reduction and relative quantification, which is applied in biological sludge treatment, microbial measurement/inspection, biochemical equipment and methods, etc. It can solve the problems of difficult separation and quantification, and worms mixed in various sundries

Active Publication Date: 2020-09-18
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in production practice, worms are often mixed in various sundries, or intertwined with fibrous fillers, making it difficult to separate and quantify

Method used

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  • Relative quantitative method of tubifex in sludge reduction technology and application of relative quantitative method

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preparation example Construction

[0051] 2. Preparation of the concentration solution of the gene fragments of the order of Trichomonas

[0052] 2.1 Ordinary PCR

[0053] The DNA sample of the worm was extracted as a template, and the target fragment was amplified by ordinary PCR using forward and reverse primers, and a large number of target fragments of the worm DNA were amplified. A total of 5 groups (parallel samples) were used for PCR amplification using a 20 μL system, and a negative control with sterile water instead of the template was added at the same time. The primer information and 20 μL system of the PCR working part are shown in Table 1 and Table 2 below.

[0054] Table 1 Information of the primers for amplification of the target fragment of the wormwood DNA

[0055] Forward primer information 5’-AGC TCG TAG TTG GAT CTC-3’ Reverse Primer Information 5'-CTG CTT TGA GCA CTC TAA-3'

[0056] Table 2 Components of 20 μL system for PCR of wormwood DNA samples

[0057] ...

Embodiment 1

[0108] Example 1 Comparative analysis of the content of worms in biochemical pools of different sewage treatment plants

[0109] Using the relative quantitative detection method of the worm of the present invention, the operation steps are as follows:

[0110] 1) During the maintenance of the biochemical pools of sewage treatment plant A and sewage treatment plant B (both located in Guangzhou, Guangdong Province), the sediments in different areas of the aerobic zone of the biochemical pools were collected respectively, and mixed to form mixed sediment A and mixed sediment B.

[0111] 2) Another sediment sample was taken, from which worms with strong activity and bright red color were selected, rinsed with enzyme-free water, and 20 mg of worm tissue was separated and collected, and 20 μL of DNA solution was extracted using a mollusk tissue DNA extraction kit.

[0112] 3) Using the forward and reverse primers (Table 1) of the target fragment of the worm DNA, a total of 5 groups of...

Embodiment 2

[0129] Example 2 Contrastive analysis of the content of worms in different wetland sediments

[0130] Using the relative quantitative detection method of the worm of the present invention, the operation steps are as follows:

[0131] 1) In Constructed Wetland A and Natural Wetland B (both located in Guangzhou City, Guangdong Province), sediment collectors were used to collect sediment from different areas of the wetland, and they were mixed to form Constructed Wetland Sediment A and Natural Wetland Sediment B.

[0132] 2) Buy a small amount of worms from the market (Guangzhou City, Guangdong Province), select worms with strong activity and bright red color, wash them with enzyme-free water, separate and collect 20 mg of worms tissue, and use a mollusk tissue DNA extraction kit, Extract to obtain 20 μL DNA solution.

[0133] 3) Using the forward and reverse primers (Table 1) of the target fragment of the worm DNA, a total of 5 groups of 20 μL systems (Table 2) were used for co...

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Abstract

The invention discloses a relative quantitative method of tubifex in sludge reduction technology and an application of the relative quantitative method. According to the relative quantitative method of tubifex, copy numbers of tubifex DNA in samples of different groups of objects to be tested can be quickly obtain by the simple operation of "sample mixing and drying + fluorescence quantitative PCR+ marking formula calculation", the relative amount of tubifex biomass in different groups of objects to be tested is judged by direct comparison of the copy numbers. The problem that quantitative work of the tubifex is difficult to finish by conventional methods such as counting and weighing at present is solved, and the objectives of comparative evaluation of sludge reduction effects of different sludge reduction reactors or judgment of the growth status of the tubifex in a single sludge reduction reactor are achieved. The method has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of sewage sludge treatment, and in particular relates to a method for relative quantification of worms in sludge reduction technology and an application thereof. Background technique [0002] Most urban sewage treatment plants adopt the activated sludge method, which has the disadvantage of large surplus sludge production. Excess sludge contains a large amount of harmful substances such as refractory organic pollutants, heavy metals, and pathogenic microorganisms. It is difficult to dispose of and costly, and secondary pollution is easily generated during the treatment process. It has become a huge problem that plagues urban development and environmental protection. problem. [0003] In order to reduce the excess sludge production in the sewage treatment process, researchers have proposed a series of sludge reduction processes based on micro-metazoan predation sludge. In the relevant sludge reduction proce...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6851C02F11/02
CPCC12Q1/6888C12Q1/6851C02F11/02C12Q2531/113C12Q2545/114C12Q2563/107
Inventor 黄少斌杜至力
Owner SOUTH CHINA UNIV OF TECH
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