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A Toxicological Detection Method of Trace Water Samples Based on Caenorhabditis elegans

A Caenorhabditis elegans and detection method technology, which is applied in the field of toxicological detection of trace water samples based on Caenorhabditis elegans, can solve the problems of high cost, complicated detection process, and long time consumption, and achieve small water sample volume and simple cultivation , the effect of detection step optimization

Inactive Publication Date: 2017-03-01
AGRI ENG INST ANHUI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The purpose of the present invention is to solve the deficiencies in the prior art about the detection method for the individual-level toxicological effect evaluation of environmental water samples and the related evaluation methods for DNA damage and oxidative stress, and to provide a method for quickly detecting the toxicological effect of environmental water samples
By using different strains of Caenorhabditis elegans as the research object and using different staining reagents, the reproductive toxicology, oxidative stress and stress, and DNA damage of environmental water samples can be quickly detected, which solves the previous toxicological evaluation process of environmental water samples Time-consuming, high cost and complicated detection process

Method used

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  • A Toxicological Detection Method of Trace Water Samples Based on Caenorhabditis elegans
  • A Toxicological Detection Method of Trace Water Samples Based on Caenorhabditis elegans
  • A Toxicological Detection Method of Trace Water Samples Based on Caenorhabditis elegans

Examples

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Embodiment 1

[0034] Example 1: Method for detecting apoptosis of gonad cells in trace water samples based on Caenorhabditis elegans

[0035] This example describes a method for the detection of gonad cell apoptosis in trace water samples based on Caenorhabditis elegans, the steps of which include the sterilization treatment of environmental water samples, the cultivation process of Caenorhabditis elegans, and the exposure of biological samples of environmental water samples. Process, fluorescence microscopy and differential interference contrast microscopy for toxicological determination procedures.

[0036] Step 1. Sterilization of environmental water samples

[0037] Use 2 pieces of 0.22 μm microporous filter membranes, place them in the middle of the filter of the corresponding size, sterilize the assembled filter at 121 degrees Celsius for 20 minutes, and then filter the environmental water samples;

[0038] Step 2, the cultivation process of Caenorhabditis elegans

[0039] Wild-type C...

Embodiment 2

[0044] Example 2: Method for detecting oxidative stress level in trace water samples based on Caenorhabditis elegans

[0045] The basic method of this example is the same as that of Example 1, except that in step 2, the staining reagent of the nematode is replaced with 10 μg / mL CM-DCFDA, and in step 4, the fluorescence intensity in the nematode is observed with a fluorescence microscope and image proplus software is used to carry out Analysis of fluorescence intensity.

Embodiment 3

[0046] Example 3: Method for detection of oxidative stress in trace water samples based on Caenorhabditis elegans

[0047] The basic method of this embodiment is the same as that of Example 1, except that the strain of the nematode is replaced by CF1553 , this strain is based on wild-type nematodes, and the green fluorescent protein is coupled to the promoter of SOD-3, so that the intensity of green fluorescent protein is equal to the expression level of SOD-3. SOD-3 is an inhibitor of oxidative free radicals in nematodes, and the level of oxidative stress in nematodes can be detected by observing the expression of SOD-3. The difference is that in step 4, the fluorescence intensity in the nematode is observed with a fluorescence microscope and the analysis of the fluorescence intensity is performed with Image pro plus software.

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Abstract

The invention discloses a Caenorhabditis elegans based micro water sample toxicology detection method, belonging to the technical field of the environmental toxicology detection on water samples. The method mainly comprises the steps of carrying out sterilization treatment on an environment water sample, culturing nematode, exposing a biological sample of the environment water sample, and performing toxicology detection on the water sample. The Caenorhabditis elegans based micro water sample toxicology detection method is characterized in that the complexity effect of a multi-kind polyvalent mixture in the environment water sample on a living body is evaluated comprehensively, the required water sample amount is low, the operation is simple, and the method can be used for solving the problems that the waste liquid amount in an existing pollutant toxicologic study is high and can be treated difficultly and secondary pollution can be caused, overcoming the experimental error caused by change of the concentration of substances in the water sample due to water evaporation, achieving rapid detection of the toxicology of the living body in the water sample, solving the problems that the existing water sample toxicology detection time is long, the cost is high and the like, and improving the accuracy and the precision of the environmental water sample evaluation.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of environmental toxicology detection, and more specifically relates to a trace water sample toxicological detection method based on Caenorhabditis elegans. [0003] Background technique: [0004] Environmental Toxicology is a discipline that uses toxicological methods to study the impact and potential harm of harmful substances in the environment on humans, other species, and the ecological environment from the perspective of medicine and biology. Through the research on the biological effect, action mechanism and damage type of environmental pollutants, the environmental health standards of the pollutants are determined, and the scientific basis for the environmental treatment methods and processes of the pollutants is provided. [0005] This discipline mainly evaluates the damage and mechanism of action of pollutants by observing and testing the toxicological effects of environmental pollutants on ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/18G01N33/48
Inventor 郭肖颖施六林王磊
Owner AGRI ENG INST ANHUI ACADEMY OF AGRI SCI
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