Method for monitoring benzo(a)pyrene pollution of seawater by utilizing fish blood cells

A blood cell and seawater technology, which is applied in the detection of toxic organic pollutants in seawater, can solve problems such as difficult promotion, cumbersome chemical monitoring methods, and inability to truly reflect the organic pollution of surface seawater, and achieve the effect of convenient preparation and simple operation

Inactive Publication Date: 2011-06-08
张振冬
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the actual monitoring process of marine toxic organic pollutants, chemical monitoring methods are relatively cumbersome, and the higher the detection sensitivity, the more expensive the equipment required, so it ...

Method used

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  • Method for monitoring benzo(a)pyrene pollution of seawater by utilizing fish blood cells
  • Method for monitoring benzo(a)pyrene pollution of seawater by utilizing fish blood cells

Examples

Experimental program
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Embodiment 1

[0022] 1. Centrifuge and wash Staphylococcus aureus 3 times with 0.1mol / L PBS buffer, and adjust the concentration to 2×10 8 CFU / mL bacterial suspension, stored at 4°C for later use.

[0023] 2. Take 50 μL of Staphylococcus aureus suspension and mix it with the red blood cell suspension of flounder cultured in normal seawater and benzo-a-pyrene-containing seawater respectively in equal volumes. The ratio of cell number is about 20:1. After mixing gently, Incubate at 20°C for 30 min.

[0024] 3. Gently shake and mix every 10 minutes during the incubation to prevent red blood cells and bacterial cells from settling.

[0025] 4. After the incubation is complete, stop the reaction on ice. Add 20 μL of pre-cooled 0.25% glutaraldehyde solution to each tube, and fix at 4°C for 15 min. Take an appropriate amount of incubation solution to make a smear, dry it and fix it with methanol for 2 min.

[0026] 5. Add an appropriate amount of Wright's staining solution to each smear and st...

Embodiment 2

[0031] A 3L glass beaker was selected as the experimental container, and a total volume of 2L of spiked seawater was added, and 5 experimental fish were placed in each beaker, and 2 parallels were used for each spiked concentration. Benzoapyrene concentration gradient: 0.49 μg / L, 0.98 μg / L, 1.96 μg / L, 3.92 μg / L, 7.84 μg / L. The experimental period was 14 days, the temperature was (20±2)°C, and the standard solution was changed every 24 hours to measure the change of the immune adhesion rate of blood cells.

[0032] 1. Use a 2.5mL sterile syringe to collect blood from the tail vein of the experimental fish, mix it with Arbori anticoagulant and flounder peripheral blood 1:1, and store it in a refrigerator at 4°C for later use

[0033] 2. Centrifuge and wash Staphylococcus aureus 3 times with 0.1mol / L PBS buffer, and adjust the concentration to 2×10 8 CFU / mL bacterial suspension, stored at 4°C for later use

[0034] 3. Take 50 μL of Staphylococcus aureus suspension and mix it wi...

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Abstract

The invention provides a method for monitoring benzo(a)pyrene in seawater by utilizing fish blood cells. The method comprises the following steps: taking 50mu L of staphylococcus aureus suspension obtained through treatment of buffer solution on ice and mixing the staphylococcus aureus suspension with isovolumetric paralichthys olivaceus erythrocyte suspension exposed in the seawater containing benzo(a)pyrene and incubating the mixture at the constant temperature of 20 DEG C for 30 minutes, wherein the ratio of the cell number is 20:1; shaking the mixture to mix the mixture uniformly every 10 minutes to prevent precipitates; terminating reaction after completion of incubation, adding 20mu L of 0.25% of precooled glutaraldehyde solution to each tube, immobilizing the tube in a refrigerator at the temperature of 4 DEG C for 15 minutes, taking incubation solution to prepare smears and immobilizing the smears with methanol after airing the smears; dying each smear with Wright's dye solution for 3 minutes, washing the smear with distilled water until the smear becomes colorless, carrying out observation under an oil lens after airing the smear and computing the erythrocyte C3b receptor rosette rate; and forming a rosette by adhering two or more staphylococcus aurei to each erythrocyte. The detection method is completed in one hour and whether the seawater is polluted by benzo(a)pyrene can be better monitored and analyzed by the method.

Description

technical field [0001] The invention belongs to the invention and relates to a method for detecting toxic organic pollutants in seawater based on changes in the immune adhesion activity of fish blood cells. Background technique [0002] In recent years, with the advancement of science and technology and the development of industrial and agricultural production, more and more toxic organic pollutants (including petroleum hydrocarbons, polychlorinated biphenyls, organochlorine pesticides, polycyclic aromatic hydrocarbons, tributyltin, etc.) Inshore seawater, sediments and marine organisms are commonly detected, and the problem of organic pollution has become increasingly prominent, which has become a new focus of attention of scientific circles and governments around the world. Studies have shown that toxic organic pollutants are highly toxic, easily dissolved in the biological organic phase, highly enriched in marine organisms, destroy the genetic material of marine organisms...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N15/10
Inventor 张振冬闫启仑米盛景王睿睿王立俊
Owner 张振冬
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