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Method for evaluating ecotoxicological impact of microplastics

A technology of toxicology and microplastics, applied in the field of high-throughput sequencing, can solve problems such as the analysis of the intestinal flora of artemia that has not been applied, and achieve the effect of short experimental period, simple operation and rapid detection

Pending Publication Date: 2020-02-04
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this technology has not been applied to the analysis of the intestinal flora of Artemia; there is no report on the use of Illumina MiSeq sequencing technology to analyze the intestinal flora of Artemia to evaluate the ecotoxicological impact of microplastics

Method used

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  • Method for evaluating ecotoxicological impact of microplastics
  • Method for evaluating ecotoxicological impact of microplastics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A method for evaluating the ecotoxicological impact of microplastics, comprising the following steps:

[0033] (1) Artemia exposure experiment:

[0034] Absorb mature Artemia and put it into artificial seawater as the blank group, put into the suspension of polyethylene microspheres with a concentration of 0.13g / L as the experimental group 1 (PE group), put into the polystyrene microspheres with a concentration of 0.13g / L The suspension is the experimental group 2 (PS group), and each group is equipped with 6 parallel sample beakers, and each beaker draws 50 mature artemia individuals, feeds 30ml of chlorella every other day, and cultivates them under the conditions of 28°C and light intensity of 1500lx 9 days;

[0035] The solvent of the plastic microsphere suspension is artificial sea water;

[0036] (2) Collect intestines:

[0037] Draw Artemia in the blank group and the experimental group, dissect the Artemia in the ultra-clean bench, and collect the Artemia inte...

Embodiment 2

[0060] (1) Artemia exposure experiment:

[0061] Absorb mature Artemia and put it into artificial seawater as the blank group, put it into the polyethylene microsphere suspension with a concentration of 0.1g / L as the experimental group 1, and put it into the polystyrene microsphere suspension with a concentration of 0.1g / L as the experimental group. Group 2, set up 6 parallel sample beakers in each group, absorb 50 mature artemia individuals in each beaker, feed 30ml chlorella every other day, and cultivate for 10 days under the condition of 25°C and light intensity of 2000lx;

[0062] The solvent of the plastic microsphere suspension is artificial sea water;

[0063] (2) Collect intestines:

[0064] Draw Artemia in the blank group and the experimental group, dissect the Artemia in the ultra-clean bench, and collect the Artemia intestinal tract;

[0065] (3) DNA extraction and PCR amplification:

[0066] according to Soil Kit Instructions Extract the total intestinal DNA ...

Embodiment 3

[0081] (1) Artemia exposure experiment:

[0082] Absorb mature Artemia and put it into artificial seawater as the blank group, put it into the polyethylene microsphere suspension with a concentration of 0.15g / L as the experimental group 1, and put it into the polystyrene microsphere suspension with a concentration of 0.15g / L as the experimental group. Group 2, set up 6 parallel sample beakers in each group, absorb 50 mature artemia individuals in each beaker, feed 30ml of chlorella every other day, and cultivate for 8 days under the condition of 30°C and light intensity of 1500lx;

[0083] The solvent of the plastic microsphere suspension is artificial seawater;

[0084] (2) Collect intestines:

[0085] Absorb the Artemia in the blank group and the experimental group, dissect the Artemia in the ultra-clean bench, and collect the Artemia intestine;

[0086] (3) DNA extraction and PCR amplification:

[0087] according to Soil Kit Instructions Extract the total intestinal DN...

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Abstract

The invention discloses a method for evaluating the ecotoxicological impact of microplastics. The method includes the following steps that (1) artemia exposure experiment is performed; (2) intestinaltract collecting is performed; (3) DNA extraction and PCR amplification are performed; (4) Illumina Miseq sequencing is performed; (5) data processing is performed; (6) data analysis is performed, specifically, cluster analysis of the processed data finds that the main phylums in each group at the phylum level are Actinobacteria and Proteobacteria; and (7) ecotoxicology evaluation is performed, specifically, the microplastics have a certain toxic effect on organisms. The method is simple in operation, high in accuracy and short in experimental period, and can rapidly detect intestinal microorganisms. The method can quickly and accurately evaluate the ecotoxicological impact of the microplastics.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing, and relates to a method for evaluating the ecotoxicological impact of microplastics. Background technique [0002] Microplastics (MPs) usually refer to plastic fragments, particles or fibers with a diameter or length of less than 5 mm, mainly derived from leaked raw materials in the plastic industry, debris formed in the ocean by large plastic waste, and plastics in various daily necessities. Microbead additives, etc. Due to the special properties of MPs, such as larger specific surface area, ability to absorb more pollutants, and easier entry into organisms, the impact on the marine environment is far greater than that of large plastic garbage. Due to the small particle size, MPs are easily absorbed by marine organisms. ingested and passed through the food chain. At present, the pollution status and ecological risks of MPs have attracted great attention from scholars at home...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12Q1/689C12Q1/04A01K67/033
CPCC12Q1/6869C12Q1/689A01K67/033C12Q2600/142C12Q2531/113C12Q2535/122C12Q2537/165
Inventor 刘宪华陈宏伟白璐任俊
Owner TIANJIN UNIV
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