Method for evaluating developmental toxicity of environmental pollutants by utilizing lengths of egg yolk extension parts of Danio rerio

A technology for environmental pollutants and extensions, applied in the field of environmental detection technology and toxicity research, can solve the problems of unpredictable developmental toxicity risks, high pollutant exposure doses, and large dose differences.

Pending Publication Date: 2020-07-31
HUAIYIN TEACHERS COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Zebrafish, as a test object for the developmental toxicity of environmental pollutants, evaluates the presence or absence and strength of developmental toxicity mainly through lethality and teratogenicity at the individual level, which inevitably brings about the following problems: 1) The lethal effect requires a higher 2) The toxic effect of growth retardation cannot be evaluated; 3) The evaluation value of the minimum effective dose of pollutants is too high, resulting in the lower exposure dose. Potential developmental toxicity risk is difficult to predict
However, there is no report on the development of zebrafish yolk extension as an evaluation index of the toxicity of environmental pollutants

Method used

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  • Method for evaluating developmental toxicity of environmental pollutants by utilizing lengths of egg yolk extension parts of Danio rerio
  • Method for evaluating developmental toxicity of environmental pollutants by utilizing lengths of egg yolk extension parts of Danio rerio
  • Method for evaluating developmental toxicity of environmental pollutants by utilizing lengths of egg yolk extension parts of Danio rerio

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preparation example Construction

[0035] The method for preparing fertilized eggs preferably includes the following steps: separately placing male and female zebrafish at the age of 5 to 6 months in a culture tank with a constant temperature circulating water system for feeding, wherein the culture water in the culture tank is aerated and heated The water temperature in the culture tank is 27.5-28.5°C, the dissolved oxygen in the water is greater than 6.0mg / L, the pH of the water body is 7.0-8.0, and the light cycle of 14h light / dark 10h is maintained; 1 day before spawning, according to male and female The zebrafish was moved to the spawning box at a ratio of 1:1; the spawning box was placed in an artificial climate box at 27.5-28.5°C overnight, and the light was turned on at 6:30-7:30 the next morning, so that the female and male The fish mate naturally, and after about 15 minutes, the fertilized eggs are collected, placed in a plate, and the feces and impurities are cleaned with a rubber dropper. Select emb...

Embodiment 1

[0052] A method for evaluating the developmental toxicity of environmental pollutants by using the length of zebrafish yolk extension, comprising the following steps:

[0053] Breeding of zebrafish: The zebrafish is raised in a culture tank with a constant temperature circulating water system, wherein the culture water in the culture tank is tap water heated by aeration, and the water temperature in the culture tank is 28±0.5°C. The dissolved oxygen is greater than 6.0mg / L, the pH of the water is 7.0-8.0, and the photoperiod of 14h light / 10h dark is maintained. Feed freshly hatched Artemia every day from 7:00 to 9:00, 14:00 to 15:00 and 19:00 to 20:00.

[0054] Obtaining fertilized eggs: Separate male and female zebrafish at the age of 5 to 6 months and raise them in a culture tank with a constant temperature circulating water system, wherein the culture water in the culture tank is aerated and heated tap water. The water temperature is 28±0.5°C, the dissolved oxygen in the w...

Embodiment 2

[0070] Evaluation of the developmental toxicity of 2,2',4,4'-tetrabromodiphenyl ether using the length of yolk extension in zebrafish embryos

[0071] Environmental pollutants: organic pollutant 2,2’,4,4’-tetrabromodiphenyl ether (BDE-47).

[0072] Preparation method: Dissolve 25mg BDE-47 in 5mL dimethyl sulfoxide (DMSO), add 495mL embryo culture solution to make BDE-47 mother solution (50mg / L); take 100mL BDE-47 mother solution, add 900mL embryo culture solution , made into BDE-47 stock solution (5mg / L); BDE-47 stock solution was diluted with embryo culture medium to a concentration of 1.25 and 0.625 mg / L.

[0073]The developmental toxicity of BDE-47 was tested by the method of Example 1. Embryos in the experimental group were cultured in 0.625 or 1.25 mg / LBDE-47 solution, and a control group was established, and the embryos in the control group were cultured in the control solution. In the blank control group of the present invention, it is preferable to add culture soluti...

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Abstract

The invention provides a method for evaluating the developmental toxicity of environmental pollutants by utilizing the lengths of egg yolk extension parts of Danio rerio, and belongs to the technicalfield of environmental detection technology and toxicity research. The method comprises the steps of exposing Danio rerio embryos developed to 4-5 hpf to the environmental pollutants or a control culture solution, and counting the lengths of the egg yolk extension parts of the embryos, the ratio of the lengths of the egg yolk extension parts to the total length of egg yolk spheres and the extension parts or the ratio of the lengths of the egg yolk extension parts to the lengths of the embryos when the Danio rerio embryos are developed to 60-84 hpf; and when any one of the lengths of the yolk extension parts, the ratio of the lengths of the yolk extension parts to the total length of the yolk spheres and the extension parts, and the ratio of the lengths of the yolk extension parts to the lengths of the embryos of an environmental pollutant group is obviously different from that of a control group, indicating that the environmental pollutants have developmental toxicity. Compared with egg yolk sphere development and embryo development, the development of the yolk extension parts is more sensitive to the toxicity of the environmental pollutants, the developmental retardation toxicityeffect of lower-dose environmental pollutants can be effectively evaluated, and meanwhile, the method is simple, convenient and rapid to operate and high in detection sensitivity.

Description

technical field [0001] The invention belongs to the technical fields of environmental detection technology and toxicity research, and in particular relates to a method for evaluating the developmental toxicity of environmental pollutants by using the length of zebrafish yolk extension. Background technique [0002] Environmental pollution has become an important problem facing the world today, and environmental pollutants are potentially harmful to organisms and humans. The embryonic development of vertebrates is a finely regulated process of cell growth and reproduction, organ and system morphogenesis. Developmental organisms are particularly sensitive to the hazards of environmental pollutants. Developmental toxicants can interfere with the developmental process of embryos and affect their normal development, manifested as embryonic lethality, teratogenicity, growth retardation, and incomplete or abnormal functions. Generally speaking, developmental toxicants can initial...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
CPCG01N33/5014G01N33/5044
Inventor 庄娟潘正军马心如张丽娅
Owner HUAIYIN TEACHERS COLLEGE
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