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Method for screening and evaluating genetic toxicity of industrial wastewater with complex ingredients

A technology for industrial wastewater and genotoxicity, applied in biochemical equipment and methods, analytical materials, microbe determination/inspection, etc., to achieve the effect of sensitive promotion and easy promotion

Inactive Publication Date: 2013-06-05
HUAINAN NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem solved by the present invention is based on the Hormesis effect and significance analysis of the actual DNA damage level of faba bean root tip cells induced by complex component industrial wastewater to identify and evaluate the genotoxicity of complex component industrial wastewater

Method used

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  • Method for screening and evaluating genetic toxicity of industrial wastewater with complex ingredients
  • Method for screening and evaluating genetic toxicity of industrial wastewater with complex ingredients
  • Method for screening and evaluating genetic toxicity of industrial wastewater with complex ingredients

Examples

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Embodiment 1

[0051] The inventor applied the above-mentioned inventive method to detect the genotoxicity of faba bean root tip cells induced by sewage from China Resources Snow Breweries (Changzhou) Co., Ltd., and evaluated the toxicity of raw sewage accordingly. Firstly, the raw wastewater should be diluted with tap water into gradient solutions: 1×, 2×, 4× and 8×. After the root tips were exposed to each dilution solution for 2 weeks, the root tips of each treatment group were cut and divided into two groups. One group was filtered with deionized water to separate nuclei and subjected to SCGE electrophoresis. The results showed that, compared with the tap water control group, the comet tail length and tail moment did not increase significantly in the undiluted 1× treatment group. Comet tail length and tail moment increased significantly after 2- and 4-fold dilutions. When diluted to 8 times, tail length and tail moment increased but not significantly ( figure 1 and figure 2 ). The ...

Embodiment 2

[0053] The inventor tested the toxicity of wastewater from Changzhou Laosan Group Co., Ltd. by using the above-mentioned inventive method. Seedlings were poisoned and died after root tip suspension culture of broad bean was diluted 1-5 times in wastewater solution. The root tip was exposed to 10-160 times of wastewater dilution solution, and after 2 weeks, the root tip was vacuum filtered with 100 μg / mL proteinase K solution, and the root tip cell nucleus was separated and subjected to SCGE electrophoresis. The results showed that 10-80 times of wastewater dilution solutions induced a significant increase in comet tail length and tail moment ( Figure 5 and Figure 6 ). Accordingly, it is believed that the raw wastewater is highly toxic.

Embodiment 3

[0055] The inventor also detected and evaluated the toxicity changes of the wastewater from Wujin Shuanghui Environmental Engineering Co., Ltd. before and after treatment by applying the above-mentioned inventive method. The seedlings wilted and died soon after the broad bean root tip suspension culture was diluted 1-10 times with the influent (before treatment)-and effluent (after treatment) solutions. After the seedlings were cultured in 20-320 times diluted water for 2 weeks, the root tip was vacuum filtered with 100 μg / mL proteinase K solution, and SCGE electrophoresis was performed. It was found that the water dilution of 20-160 times induced a significant increase in the tail length and tail moment of the comet, but no significant change was seen after the dilution of 320 times. At the same time, the 20-320-fold diluted effluent induced a significant increase in the comet tail length and tail moment. Based on this, it is believed that the company's wastewater was highly...

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Abstract

The invention relates to a method for screening and evaluating genetic toxicity of industrial wastewater with complex ingredients. Gemmiparous broad bean seeds are cultivated in a suspension manner in to-be-detected wastewater solution prepared with tap water by means of gradient dilution; after two weeks, root tips are cut down and placed in proteinase K solution, and vacuumizing is carried out. Root tip cell nucleuses are separated, and the single-cell gel electrophoresis (SCGE) technique is used for separating DNA fragments. A fluorescence microscope is used for shooting comet pictures. A cometscore image-analysis system is used for measuring the tail length and the tail moment of the comet. Then, an SPASS program is used for statistic analysis. The method promotes proteinase K to penetrate into root tip meristem cell nucleuses by means of vacuumizing, accelerates degradation of DNA crosslinked protein, thus removes retardation of DNA-protein cross-linking on DNA fragments, and facilitates revelation and accurate evaluation of the true level of NDA breakage and damage inducted by the to-be-detected wastewater.

Description

technical field [0001] The invention relates to a method for screening and evaluating the genotoxicity of industrial wastewater with complex components, specifically a Hormesis dose-effect evaluation of the genotoxicity of the polluting medium based on the dose-effect of Hormesis on the true DNA damage level induced by industrial wastewater with complex components Technology. Background technique [0002] In recent years, with the rapid development of industry, industrial wastewater has been continuously produced, and its pollution components have become increasingly complex. Because it is impossible to analyze all the pollution components in complex pollution media by chemical analysis methods, it is even more impossible to accurately predict and evaluate the potential biological toxicity induced by long-term exposure to complex pollutants. In recent years, environmental science and ecotoxicology workers have begun to monitor and evaluate environmental pollution from the r...

Claims

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Application Information

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IPC IPC(8): G01N27/447C12Q1/68
Inventor 汪承润卢韫徐向华姜传军王勤英李志啸
Owner HUAINAN NORMAL UNIV
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