Vibrio qinghaiensis Q67 based long-term microplate toxicity analyzing method of environmental pollutant

A technology for environmental pollutants, Vibrio qinghai, applied in biochemical equipment and methods, analysis by chemical reaction of materials, and determination/inspection of microorganisms, which can solve the problems of long action time and low concentration of emerging pollutants.

Inactive Publication Date: 2010-12-15
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The technical problem to be solved in the present invention is to focus on the shortcomings of the traditional pollutant luminescent bacteria toxicity detection method and the characteristics of low concentration and long action time of emerging pollutants in the environment, and emphasize the influence of time factors on toxicity on the basis of the traditional short-term toxicity detection method , to establish a long-term microplate toxicity analysis method for environmental pollutants based on Vibrio Qinghai Q67, which can effectively detect the toxicity of pollutants with low content in the environment, long-term existence and specific action sites on the tested organisms

Method used

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  • Vibrio qinghaiensis Q67 based long-term microplate toxicity analyzing method of environmental pollutant
  • Vibrio qinghaiensis Q67 based long-term microplate toxicity analyzing method of environmental pollutant
  • Vibrio qinghaiensis Q67 based long-term microplate toxicity analyzing method of environmental pollutant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] [Example 1] Carry out Wilbur's long-term toxicity assay with the toxicity analysis method of the present invention, comprising the following steps:

[0046] (1) Preparation and cultivation of Qinghai Vibrio Q67 common medium:

[0047] Weigh KH separately 2 PO 4 27.2 mg, Na 2 HPO 4 12H 2 O 71.6 mg, MgSO 4 ·7H 2 O 0.5g, MgCl 2 ·6H 2 O 1.22g, CaCl 2 66.0 mg, NaHCO 3 2.68g, NaCl 3.08g, yeast extract 5.0g, tryptone 5.0g, glycerin 3.0g, heated and dissolved in 1000mL distilled water, adjusted the pH value to 8.5±0.5 with 1mol / L NaOH, and sterilized at 121°C according to conventional sterilization Sterilize under high pressure steam for 20 minutes and use after cooling. All of the above media were purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd.

[0048] (2) Preparation of the solid plate culture medium of Vibrio Qinghai Q67:

[0049] Take 100mL of ordinary culture medium obtained in step (1), add 1.5-2.0g of agar, heat to dissolve until transparent...

Embodiment 2

[0069] [Example 2] Carry out the long-term toxicity determination of Promilone with the toxicity analysis method of the present invention

[0070] The remaining steps of the method are the same as in Example 1, except that the volume ratio in step (5) is 1:5, and the relative luminous intensity of the 100 μL microplate-loaded bacterial solution is 0.8×10 5 Relative luminescent unit; the highest inhibition rate reaches more than 80% in step (6), and the concentration of the sample stock solution is 2.22 × 10 -3 mol / L, dilution factor The raw data of Q67 luminescence intensity were measured according to the above method steps, and the concentration-effect data were obtained as shown in Table 2 after entering the preprocessing according to step (8). Select the Weibull function in the APTox software (software number 062731, registration number 2006SR15065) to carry out nonlinear least squares fitting to the concentration-effect data, obtain fitting parameters α=7.47, β=2.56, and...

Embodiment 3

[0073] [Example 3] Carrying out the long-term toxicity determination of mecitrione with the toxicity analysis method of the present invention

[0074] The remaining steps of the method are the same as in Example 1, except that the volume ratio in step (5) is 1:3, and the relative luminous intensity of the 100 μL microplate-loaded bacterial solution is 1.2×10 5 Relative luminescence unit; in step (6), the concentration of the sample stock solution is 3.56×10 -3 mol / L, dilution factor

[0075] The raw data of Q67 luminescence intensity were measured according to the steps of the above method, and the concentration-effect data were obtained by preprocessing according to step (8), as shown in Table 3. Select the Logit function in the APTox software (software number 062731, registration number 2006SR15065) to carry out nonlinear least squares fitting to the concentration-effect data, obtain fitting parameters α=9.09, β=2.92, and the square of the correlation coefficient is 0.968...

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Abstract

The invention discloses a vibrio qinghaiensis Q67 based long-term microplate toxicity analyzing method of environmental pollutants. The long-term microplate toxicity analyzing method is established by using the vibrio qinghaiensis Q67 as a tested organism and introducing time factors into the toxicity concentration-effect relationship of the traditional compound on the basis of a microplate toxicity analyzing method. The vibrio qinghaiensis Q67 based long-term microplate toxicity analyzing method of the environmental pollutants is characterized by adopting a specific microplate design and sampling scheme and a microplate culture medium suitable for the growth of the vibrio qinghaiensis Q67, measuring the long-term toxicity effect of the environmental pollutants on the vibrio qinghaiensis Q67 and repeatedly measures the toxicity of the same pollutant through multiple plates to ensure the statistical significance of an experimental result. By measuring the long-term toxicity of a part of the environmental pollutants on the vibrio qinghaiensis Q67, compared with short-term toxicity, the long-term toxicity of the tested environmental pollutants is discovered to be obviously higher than the short-term toxicity of the tested environmental pollutants, wherein the difference of the long-term toxicity and the short-term toxicity of antibiotics is most significant. The invention can actually reflect the toxicity of a site compound having specific action on compounds and more reasonably evaluate the toxicity of the pollutants.

Description

technical field [0001] The invention relates to the field of toxicity detection of environmental pollutants, in particular to a method for analyzing the long-term toxicity of environmental pollutants to Qinghai Vibrio Q67. Background technique [0002] With the continuous development of modern industry, environmental pollution is becoming more and more serious, especially the pollution of chemicals. The water body environment is the final collection place of various pollutants. The intervention of a large number of chemicals with adverse ecological effects affects the normal function of the water body, seriously destroys the ecological balance, and directly or indirectly threatens human health. In order to protect water resources and aquatic ecosystems and control pollution, it is necessary to monitor and evaluate pollutants in the water environment, and establish water quality evaluation standards based on chemical analysis and biological detection. [0003] Chemical analy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76C12Q1/02
Inventor 朱祥伟刘树深葛会林张丽芬
Owner TONGJI UNIV
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