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Preservation method of anaerobic dehalogenated microorganisms and application of preservation method

A preservation method and microbial technology, which is applied in the field of water pollution control, can solve the problems of reduced activity of dehalogenated microorganisms, reduced efficiency of in-situ bioremediation, and difficulty in maintaining anaerobic conditions, so as to solve the problem of reduced dehalogenated microbial activity and reduced The effects of transportation cost and oxygen tolerance enhancement

Pending Publication Date: 2020-08-25
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

However, this transportation process is expensive and difficult to maintain strict anaerobic conditions, often resulting in greatly reduced degradation activity of dehalogenated microorganisms
On the other hand, organic chlorides higher than 0.5mM often have a certain toxicity inhibitory effect on dehalogenation microorganisms. However, in actual polluted sites, organic chlorine pollution problems much higher than this concentration or even in a non-dissolved state are very common, greatly reducing Efficiency of in situ bioremediation

Method used

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  • Preservation method of anaerobic dehalogenated microorganisms and application of preservation method
  • Preservation method of anaerobic dehalogenated microorganisms and application of preservation method
  • Preservation method of anaerobic dehalogenated microorganisms and application of preservation method

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Dehalococcoides mccartyi type strain 195 (species preservation number ATCC BAA-2266 T ) (hereinafter referred to as Dhc 195) as an example for immobilization.

[0046] S1. Cultivation of Dehalogenococcus: Inoculate Dhc 195 with 3% inoculum into a 160mL airtight serum bottle containing 100mL inorganic salt medium for culture, and the headspace is N 2 / CO 2 (80 / 20, v / v), and adding 5 mM sodium acetate as a carbon source and 10 mL hydrogen as an electron donor to the inorganic salt medium, and cultured at 30° C. in the dark.

[0047] The composition of the inorganic salt medium is: NaCl 1.0g / L, MgCl 2 ·6H 2 O 0.5g / L, KH 2 PO 4 0.2g / L, NH 4 Cl0.3g / L, KCl0.3g / L, CaCl 2 2H 2 O 0.015g / L, FeCl 2 4H 2 O 1.5mg / L, CoCl 2 ·6H 2 O 190μg / L, MnCl 2 4H 2 O 100μg / L, ZnCl 2 70μg / L, H 3 BO 3 6μg / L, Na 2 MoO 4 2H 2 O 36μg / L, NiCl 2 ·6H 2 O 24μg / L, CuCl 2 2H 2 O 2μg / L, Na 2 SeO 3 ·5H 2 O 6 μg / L, Na 2 WO 4 2H 2 O 8μg / L, resazurin indicator 0.025% (w / v), L-cys...

Embodiment 2

[0066] The method in Example 1 was used to immobilize the Dhc 195 bacteria, except that in step S3, no activated carbon was added, so as to realize the corresponding immobilization of the Dhc 195 bacteria.

[0067] The immobilized microspheres without activated carbon obtained in this example and the immobilized microspheres of Dhc 195 bacteria obtained in Example 1 were used for the degradation of trichlorethylene. The culture conditions, detection of degradation products and calculation of degradation rate were the same as in Example 1.

[0068] After testing, the degradation rates of trichlorethylene for immobilized microspheres without activated carbon and immobilized microspheres added with activated carbon were 95.3 μM d -1 and 129.9 μM d -1 , adding activated carbon can increase the degradation rate by 36.3%. It can be seen that due to the adsorption of activated carbon to microorganisms, the toxic effect of boric acid on microorganisms during the crosslinking process ...

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Abstract

The invention belongs to a water pollution treatment technology, and particularly relates to a preservation method of anaerobic dehalogenated microorganisms and application of the preservation method.The preparation method comprises the following steps: re-suspending dehalogenated microorganisms through an embedding medium solution, dropwise adding a re-suspension into a cross-linking agent, andperforming reaction for 10-20 minutes in an ice bath under an anaerobic condition; after the reaction, continuing crosslinking for 24 to 36h at 4 to 6 DEG C in an anaerobic environment; and after thecrosslinking, realizing the preservation of the anaerobic dehalogenated microorganisms. According to the invention, the anaerobic dehalogenated microorganisms are immobilized; through centrifugation,re-suspension, cross-linking balling and other operations under strict anaerobic conditions, an immobilized microbial agent is obtained while the microbial solution is concentrated and cells are wrapped, so that the tolerance degree of dehalogenated microorganisms to oxygen is remarkably improved, the problem that degradation activity is lost due to the fact that the dehalogenated microorganisms in a form of a culture solution are easily exposed to air during transportation is solved, and great convenience is provided for implementation of in-situ bioremediation engineering of a contaminated site.

Description

technical field [0001] The invention belongs to water pollution treatment technology, in particular to an anaerobic dehalogenation microbial preservation method and application thereof. Background technique [0002] The widespread use of organic chlorides as dry cleaning agents, pesticides, etc. worldwide has made them a large class of pollutants that are ubiquitous in groundwater. Organic chlorides have low solubility and strong mobility in soil, so they can easily infiltrate soil and pollute deep groundwater, forming heavy non-aqueous phase liquids (Dense non-aqueous phase liquids; DNAPLs). As a class of persistent organic pollutants, once organic chloride enters the aquifer, it will endanger the underground ecosystem and the safety of drinking water sources for a long time. Since the 1980s, studies have found that chlorinated olefins such as tetrachlorethylene have high toxicity, carcinogenic, teratogenic, and mutagenic effects on the human body, and trichlorethylene and...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10C12N11/084C12N11/04C02F3/34C12R1/01C02F101/36C02F103/06
CPCC12N11/14C12N11/10C12N11/084C12N11/04C02F3/34C02F2101/36C02F2103/06
Inventor 严俊王晶晶杨毅李秀颖
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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