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: 2009-12-09
SHENYANG INST OF APPLIED 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, zeo

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) Bacterial 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 acclimatization of degrading bacteria: Inoculate 2g of bacteria samples in 100ml enriched medium containing dichlorvos and mix well for transfer culture. 10% of the culture solution was inoculated in 100ml enriched medium containing dichlorvos gradually increasing by volume, and the dichlorvos was gradually increased by 100mg / L each time, 4 times. During the transfer process, dichlorvos was 100, 200, 300, and 400 mg / L, respectively.

[0022] The composition of enrichment medium in described step 1) is: 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, 100mg of dichlorvos respectively, add distilled water to 1000ml, adjust pH to 7.0-.5.

[0023] 2) Separation and screening: take the bact...

Embodiment 2

[0026] Determination of degrading activity of degrading strains:

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

[0028] 2) Determination of degradation rate: the acetone collected above was measured by gas chromatography, and the gas chromatography 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 tempera...

Embodiment 3

[0032] The difference from Example 1 is that:

[0033]1) Enrichment and domestication of degrading bacteria: Inoculate 3 g of bacteria samples into 120 mL enriched medium containing dichlorvos for transfer culture, each transfer culture condition is 32 ° C, 200 r / min shaker culture for 5 days, co-transferred Connect 5 times, and the transfer amount is 8% of the culture solution per volume percentage, inoculated in 120ml enriched medium containing dichlorvos gradually increasing for use; wherein the first addition of dichlorvos is 150mg / L, and then gradually increases by 150mg / L, Dichlorvos was 150, 300, 450, 600, and 750 mg / L during the 5 transfers.

[0034] 2) Separation and screening: take the bacterial liquid acclimation liquid cultured in step 1), dilute 10 5 After that, it was coated on beef extract peptone solid plate, and incubated at a constant temperature of 32°C for 3-4 days;

[0035] 3) Re-screening of degrading bacteria: The strains grown in step 2) are streaked ...

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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 biodegradable pesticide residues, in particular to a screening method for degrading strains using dichlorvos as a substrate. Background technique [0002] Dichlorvos, also known as DDVP, a kind of organophosphorus pesticide, 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 with the advantages of high efficiency, low toxicity, broad spectrum, low dosage and low price. It is currently one of the main insecticides widely used in vegetables, fruit trees and various farm crops. . Dichlorvos decomposes slowly in aqueous solution, accelerates decomposition in case of alkali, is thermally stable, and is corrosive to iron. It is poisonous to humans and animals, highly toxic to fish...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
Inventor 张惠文蔡颖慧张晓黎苏振成李旭
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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