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Method for analyzing sludge microbiological community configuration in urban sewage plant

A microbial community and urban sewage technology, applied in the field of microbial ecology, can solve problems such as difficulty in obtaining information about microbial communities, complex microbial diversity in sludge components, and inability to reflect the real situation of microorganisms, so as to avoid screening and enrichment, Avoid the loss of microbial information, the effect of simple separation and identification

Inactive Publication Date: 2005-02-23
TONGJI UNIV
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Problems solved by technology

However, this traditional method has many shortcomings: it cannot reflect the real situation of microorganisms in sludge samples from urban sewage plants with complex components; it is difficult to obtain relevant information on the structure and spatial distribution of microbial communities; Introduced, it is not very suitable for the sludge samples of urban sewage plants with complex components (for example, many microorganisms are oligotrophic in the natural ecological environment, and the total number of live bacteria in the sample is determined with nutrient-rich beef juice peptone in the laboratory, a large number The oligotrophic microorganisms are not suitable for growth, resulting in large errors in the measurement results); in general laboratories, bacteria are usually only cultured at 28°C when isolating and cultivating bacteria, and in addition to medium and high temperature bacteria, there are also low temperature bacteria in the sludge. Low temperature bacteria are neglected
Due to the complex composition of urban sewage sludge and rich microbial diversity

Method used

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  • Method for analyzing sludge microbiological community configuration in urban sewage plant
  • Method for analyzing sludge microbiological community configuration in urban sewage plant
  • Method for analyzing sludge microbiological community configuration in urban sewage plant

Examples

Experimental program
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Embodiment 1

[0026] Sampling and analysis of dewatered sludge from a sewage plant in Shanghai shows that the basic properties of the sludge are: moisture content 80%, water-soluble total nitrogen 3.0g / kgDM, total organic matter 550g / kgDM, total oil 30g / kg dry solids, Priority control pollutants were not detected, coliform value ≥ 24,000MPN / 100gDM. The microbial community structure of dewatered sludge was analyzed as follows:

[0027] Repeated extraction of DNA in the first step: Weigh 2g of sludge samples in a conical flask, add 13.5mL DNA extraction solution (100mmol / L Tris-HCl, 100mmol / L EDTA, 100mmol / L K2 HPO 4 , 100mmol / LKH 2 PO 4 , 1.5mol / L NaCl, 1% CTAB, pH8.0), then add 40μL lysozyme (20mg / mL) and 2-5 sterilized glass beads, shake at 37℃, 225r / min shaker for 20min, then add 1.5 mL of 20% active agent SDS, in a water bath at 65°C for 2 hours, gently shake a few times every 15 minutes, centrifuge the above extracted sample at 5000r / min for 10 minutes at room temperature, collect th...

Embodiment 2

[0032] The method of the invention can simultaneously analyze the sludge microbial population structure of multiple urban sewage plants to obtain their spatial dynamic changes, and provide good guidance and evaluation for the operation improvement of the urban sewage plant sludge treatment process. Sampling and analysis of the sludge at each treatment stage treated by a fast and high-efficiency composting process. The basic properties of the sludge are shown in Table 1:

[0033] Table 1. Basic properties of sludge in each treatment stage of composting process

[0034] No. Sampling characteristics Moisture content% VS%

[0035] 1 Last batch of return material 37 37.8

[0036] 2 Dewatered sludge (raw sludge) 81 10.5

[0037] After mixing with conditioner

[0038] 3 58 23.7

[0039] Material, processing day 1

[0040] Materials in high temperature period,

[0041] 4 42 40...

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Abstract

A method for analyzing sludge microbiocoenosis construction of municiple sewage plant relating PCR-DGGE technology analytical method for sludge microbiocoenosis construction of municiple sewage plant is disclose. It includes: DNA extracting, PCR amplifying, PCR product concentrating, DGGE electrophoretic separating to obtain DGGE map. It is characterized by two DNA extracting to obtain general DNA, taking 2-5g sludge sample, sludge DNA extracting by DNA extract, adding DNA extract etc. centrifugal collecting supernatant fluid in sediment, adding mixed liquor of chloroform and isopentanol, centrifugal pooling, DGGE electrophoretic separating by denaturing gradient 30%-60% polyacrylamide gel to obtain DGGE map. It achieves time saving and low cost.

Description

technical field [0001] The invention relates to a method for analyzing microbial community composition in urban sewage plant sludge by using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), which belongs to the field of microbial ecology. Background technique [0002] Municipal sewage plant sludge is an aggregate composed of bacteria gelatin formed by various microorganisms and organic and inorganic substances adsorbed on it. It contains a large number of bacteria, viruses, protozoa, parasite eggs and other microorganisms. The diversity of microorganisms Rich, complex microbial community composition. Analyzing the composition of microbial communities in sludge from municipal sewage treatment plants has great academic value and practical application prospects: it can not only obtain the spatial and temporal dynamic changes of microbial population structure in sludge through analysis, but also judge and predict the trend o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 赵建夫陈玲李竺王峰方萍傅以钢
Owner TONGJI UNIV
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