Screening method of degrading bacteria strains by taking DDVP as substrate

A screening method, the technology of dichlorvos, is applied in the field of screening of degrading strains, which can solve the problems of high price and secondary pollution of chemical treatment methods, and achieve the effect of simple screening method, good effect and good application potential

Active Publication Date: 2013-02-27
SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, a series of chemical methods are mainly adopted at home and abroad to degrade residual dichlorvos, zeolite loaded TiO2 photocatalysis, etc., but chemical treatment methods are expensive and easy to cause secondary pollution

Method used

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  • Screening method of degrading bacteria strains by taking DDVP as substrate
  • Screening method of degrading bacteria strains by taking DDVP as substrate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Screening of dichlorvos-degrading strains:

[0020] 1) Bacteria sample collection: collected from soil samples polluted by dichlorvos pesticides for a long time near Sujiatun Pesticide Factory in Shenyang City.

[0021] 2) Enrichment and domestication of degrading bacteria: Inoculate 2g of bacterial samples into 100ml enrichment medium containing dichlorvos and mix well for transfer culture. 1 day, a total of 4 transfers, the transfer amount was calculated by volume percentage, and 10% of each culture solution was inoculated in 100ml enrichment medium containing dichlorvos gradually increasing for use; wherein dichlorvos was gradually increased by 100mg / L each time, 4 times During the transfer process, the dichlorvos were 100, 200, 300, 400mg / L respectively.

[0022] The enrichment medium in the step 1) consists of: MgSO 4 ·7H 2 O 0.20g, (NH4) 2 SO 4 0.50g, KH 2 PO 4 0.50g, NaCl 1.0g, K 2 HPO 4 1.50g, dichlorvos 100mg respectively, add distilled water to 100...

Embodiment 2

[0026] Determination of degradative strain degradative activity:

[0027] 1) Pretreatment of the sample: take 4mL of the solution to be tested, centrifuge at 5000r / min for 8 minutes, take the supernatant, add 8mL of acetone, shake vigorously; then add 4g saturated sodium chloride and shake vigorously for 1min, and let it stand for 5min at room temperature to make The water phase and the acetone phase are separated, the upper layer of acetone is removed, and the upper layer of acetone is passed through an anhydrous sodium sulfate column at normal temperature and pressure to collect the outflowing acetone for gas chromatography determination.

[0028] 2) Determination of degradation rate: The acetone collected above was subjected to gas chromatographic determination, and the gas chromatographic conditions were: NPD detector; DB-1701P chromatographic column (30m×0.32mm×0.25μm); oxygen flow rate 60mL / min, hydrogen flow rate 2.3 mL / min, nitrogen flow rate 1.2ml / min, detector temper...

Embodiment 3

[0032] The difference from Example 1 is:

[0033]1) Enrichment and acclimatization of degrading bacteria: Inoculate 3g of bacterial samples into 120mL enrichment medium containing dichlorvos for transfer culture. Connect 5 times, the amount of transfer is calculated by volume percentage, and 8% of each culture solution is inoculated in the 120ml enrichment medium containing dichlorvos gradually increasing for use; wherein the first addition of dichlorvos is 150mg / L, and then gradually increased by 150mg / L, During the 5 transfers, the dichlorvos levels were 150, 300, 450, 600, and 750 mg / L respectively.

[0034] 2) Separation and screening: take the acclimatization liquid cultured in step 1) and dilute it by 10 5 Afterwards, spread on the beef extract peptone solid plate, and invert at 32°C for 3-4 days;

[0035] 3) Re-screening of degrading bacteria: the strains grown in step 2) were separated by streaking on a solid plate of selective inorganic salt medium containing dichlo...

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Abstract

The invention relates to a screening method of degrading bacteria strains by taking DDVP as a substrate, which belongs to the technical field of pesticide residue after biodegradation. The method comprises the following steps: the bacteria strain samples are inoculated in an enrichment medium containing 100-150 ml of DDVP for switchover culture, then the domesticated liquid cultured in the step 1 is taken and diluted for 10-10, then the diluent is coated on a beef extract-peptone solid flat plate, and then the bacteria strains are placed upside down at the constant temperature of 30-35 DEG C and cultured for 3-4 days; and the bacteria strains growing in the step 2 are streaked on a selective inorganic salt culture medium solid flat plate containing the DDVP as a sole carbon source and then cultivated for 4-5 days at the temperature of 30-35 DEG C and the growing bacteria strains are the degrading bacteria strains taking the DDVP as the sole carbon source. The provided screening method of the degrading bacteria strains by taking DDVP as the substrate can be used for screening the degrading bacteria strains for other organophosphorus insecticides and provides a certain theoretical guidance on the screening aspect of the pesticide residue degrading bacteria strains.

Description

technical field [0001] The invention relates to the technical field of biodegradation of pesticide residues, in particular to a screening method for degrading bacterial strains using dichlorvos as a substrate. Background technique [0002] Dichlorvos, also known as DDVP, a kind of organophosphorus insecticide, chemical name: 2,2-dichlorovinyl dimethyl phosphate (2,2-Dichlorovinyl dimethyl phosphate). Molecular formula C 4 h 7 Cl 2 o 4 P, the molecular weight is 220.98, and the chemical structural formula is: [0003] [0004] Dichlorvos is a broad-spectrum insecticide and acaricide. It has the advantages of high efficiency, low toxicity, broad spectrum, less dosage, and low price. It is currently one of the main insecticides widely used in vegetables, fruit trees and various farmland crops. . Dichlorvos decomposes slowly in aqueous solution, decomposes faster when encountering alkali, is stable to heat, and is corrosive to iron. Poisonous to humans and animals, high...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
Inventor 张惠文蔡颖慧张晓黎苏振成李旭
Owner SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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