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Compound strains, inoculants and their applications

A technology of bacterial strains and bacterial agents, applied in the field of bacterial agents and their applications, and compound bacterial strains, can solve the problems of large amount of sludge and high cost, and achieve the effect of efficient denitrification treatment

Active Publication Date: 2022-07-08
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In view of this, the main purpose of the present invention is to propose a composite bacterial strain, bacterial agent and its application, the composite bacterial strain can directional transform nitrate and nitrite, realize efficient denitrification treatment, and can solve the problem of low C / N livestock and poultry The problems of high cost and large amount of sludge in the process of denitrification of biogas slurry

Method used

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  • Compound strains, inoculants and their applications
  • Compound strains, inoculants and their applications
  • Compound strains, inoculants and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] This example is the screening and purification of Enterobacter C1-3 strain and Pantoea A1 strain:

[0036] 1. Build a microbial strain library:

[0037] 1.1. Use The SPIN Kit for Soil (MP Biomedicals, CA, USA) is used to extract DNA from the fully mixed and freeze-dried activated sludge samples. The activated sludge samples are from Jinjing, Institute of Subtropical Ecology, Chinese Academy of Sciences. Activated sludge deposits in the livestock and poultry biogas slurry disposal device of the livestock and poultry breeding wastewater treatment system of the experimental station; and F27 and 1492R were used as primers, and the extracted DNA was used as a template for PCR amplification, and the 50ul amplification system was 2 × Taq PCR Master Mix 25ul, F27 1ul, 1492R 1ul, DNA template 2ul, ddH 2O21ul. The amplification procedure was pre-denaturation at 94°C for 1 min, denaturation at 94°C for 30s, annealing at 57°C for 20s, extension at 72°C for 30s, and finally exte...

Embodiment 2

[0067] The present embodiment is a bacterial preparation, the bacterial preparation includes Pantoea A1 strain and Enterobacter C1-3 strain, wherein the number ratio of Pantoea A1 strain and Enterobacter C1-3 strain is 1:1.

[0068] The above-mentioned microbial inoculum is prepared by the following method:

[0069]Flush argon gas into the serum bottle to provide anaerobic conditions for anaerobic denitrifying microorganisms, inject Enterobacter C1-3 strain and Pantoea A1 strain at a ratio of 1:1 into 4 ml of sterile distilled water and use a sterile syringe Inject it into a serum bottle containing 40ml of LB liquid medium, and after culturing for 12h, collect the culture medium in the logarithmic growth phase, and centrifuge it in a centrifuge at 5000 rpm for 5 minutes, then rinse it with sterilized 1×PBS for 3 times to remove residues LB medium, and then resuspend the bacteria with 1×PBS to obtain the above bacterial agent.

experiment example 1

[0071] This experimental example is a study on the biotransformation of nitrate by Enterobacter C1-3 strain

[0072] Flush argon gas into the serum bottle to provide anaerobic conditions for anaerobic denitrifying microorganisms, inject the Enterobacter C1-3 strain into 4 ml of sterile distilled water, and use a sterile syringe to inject serum with 40 ml of LB liquid medium After culturing for 12 h, collect the culture medium in the logarithmic growth phase and centrifuge it in a centrifuge at 5000 rpm for 5 minutes, then rinse it with sterilized 1×PBS three times to remove the residual LB medium, and then use 1×PBS To resuspend the bacteria, the OD of the resuspended bacterial solution 600 1.42 and inoculated into sterilized DM1 (KNO 3 As the only nitrogen source, the concentration is 30mg / L), and cultivated at 150rpm and 30°C for 72h, and each strain was set up in three parallels;

[0073] The nitrate conversion results are as image 3 shown, by image 3 It can be seen t...

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Abstract

A composite bacterial strain, bacterial agent and application thereof, the composite bacterial strain is composed of Enterobacter C1-3 with preservation number CGMCC No.20581 obtained by separating and screening activated sludge and Pantoea A1 with preservation number CGMCC No.20582, Also provided is the application of the composite strain or the bacterial agent containing the composite strain in denitrification treatment, and a high-efficiency denitrification treatment effect is achieved. The composite strain denitrification system constructed by the invention has good application value in nitrate (biogas slurry) denitrification.

Description

technical field [0001] The invention relates to the technical field of denitrification, in particular to a compound strain, a bacterial agent and an application thereof. Background technique [0002] With the comprehensive control of point source pollution in urban areas, China's intensive livestock and poultry breeding has gradually become prominent, and livestock and poultry breeding wastewater contains a large amount of chemical oxygen demand (COD) and ammonia nitrogen; according to the characteristics of high organic matter in livestock and poultry wastewater, anaerobic Oxygen fermentation is used to obtain biogas with higher value, thereby realizing energy conversion. The anaerobic fermentation stage realizes the conversion of organic matter to CH 4 However, the removal of ammonia nitrogen (ammonia nitrogen ≈ 500 mg / L) is not obvious, so the COD / TN (total nitrogen) ratio of biogas slurry is significantly reduced. At present, the COD / TN ratio of biogas slurry for livest...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C02F101/16C12R1/01
CPCC12N1/20C02F3/34C02F2101/163Y02W10/10
Inventor 庄绪亮徐圣君徐立娜左佳靓张小寒姜参参杨东敏白志辉李瑞杨凡
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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