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Heterotrophic nitrification-aerobic denitrification bacterium resistant to low carbon and high nitrogen and application thereof

A technology of aerobic denitrifying bacteria and heterotrophic nitrification, applied in the field of environmental microorganisms, can solve the problems of increased cost and high ammonia nitrogen tolerance concentration, and achieve the effects of widespread promotion, fast growth of bacteria and high removal rate

Pending Publication Date: 2021-09-21
重庆理享生态修复研究院有限公司 +1
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In recent years, some researchers have discovered heterotrophic nitrification-aerobic denitrification bacteria from the environment. This type of microorganism can simultaneously perform nitrification and denitrification under aerobic conditions, and has a fast growth rate and a high ammonia nitrogen tolerance concentration. The currently reported heterotrophic nitrification-aerobic denitrification bacteria require high C / N in the denitrification process, and an external carbon source is required under low C / N conditions, which increases the cost

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  • Heterotrophic nitrification-aerobic denitrification bacterium resistant to low carbon and high nitrogen and application thereof
  • Heterotrophic nitrification-aerobic denitrification bacterium resistant to low carbon and high nitrogen and application thereof
  • Heterotrophic nitrification-aerobic denitrification bacterium resistant to low carbon and high nitrogen and application thereof

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Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1, strain enrichment and optimization

[0027] 1) Enrichment culture: Take 2ml of biogas slurry water sample from Chongqing Muan pig farm in Banan District, Chongqing, inoculate it into a 250ml Erlenmeyer flask containing 100ml of LB medium, and put it into a shaker set at 30°C and 160r / min. After culturing for 72 hours, inoculate at 2% into a 250ml Erlenmeyer flask also equipped with 100ml LB medium, and subculture twice to obtain the enriched bacterial solution.

[0028] 2) Subculture and domestication: Take 2ml of the enriched bacteria liquid, inoculate it into a 250ml Erlenmeyer flask containing 100ml of heterotrophic nitrification, aerobic and denitrification selective medium, put it into a shaker set at 30°C, 160r / min, and test every 24h OD in culture flask 600 , NH 4 + -N, TN, subcultured three times to obtain a mixed flora with stable denitrification effect.

[0029] 3) Isolation of pure bacteria: use cooled sterile distilled water to carry out 10...

Embodiment 2

[0031] Embodiment 2, strain identification

[0032] Sichuan Qingke Company was entrusted to complete the 16SrDNA sequencing of the screened strains. After sequencing, the 16S rDNA sequence of the strain was obtained. According to the result, Blast comparison and identification was carried out in NCBI, and the strain with similar homology to the target gene sequence was selected, and the phylogenetic tree was constructed by Neighbor-Joining method with MEGA 7.0 software.

[0033] The identification results showed that the bacteria belonged to Gram-negative bacteria, the colony diameter was 2-4mm, aerobic, the surface was moist, shiny and opaque. The sequencing of the 16S rDNA of Shinella sp.CY-9 is shown in SEQ ID NO.1, and the base sequence was compared with the GenBank nucleic acid sequence database and found to be the same as that of Shinella sp. strains W G48, ADC-10, The homology of C72 is 100%, and the phylogenetic tree of Shinella sp.CY-9 can be seen figure 2 .

Embodiment 3

[0034] Example 3. Degradation effect of Shinella sp.CY-9 on ammonia nitrogen and TN under different C / N conditions

[0035] Prepare heterotrophic nitrification, aerobic and denitrification selection medium with C / N between 1-15 respectively, in triplicate, and sterilize the above culture at 121°C, and inoculate 2ml Shen Bacteria Shinella sp.CY-9 bacteria liquid, cover the rubber stopper and shake well, put it into the shaker and set it at 30°C, 160 rpm to cultivate, and measure the concentration of the bacteria liquid every 24h (OD 600 ), ammonia nitrogen (NH 4 + -N), total nitrogen (TN) concentration. The detection method of ammonia nitrogen is Nessler's reagent spectrophotometry, and the cuvette and wavelength are 20mm glass cuvette and 420nm respectively. The reagents used in the detection process were Nessler's reagent and potassium sodium tartrate. The detection method of total nitrogen is the alkaline potassium persulfate spectrophotometer method, the cuvette and opt...

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Abstract

The invention discloses a heterotrophic nitrification-aerobic denitrification bacterium resistant to low carbon and high nitrogenand application thereof. The strain is preserved in the China Center for Type Culture Collection, the preservation number is CCTCC NO: M 2021681, and the strain is named as Shinella sp.CY-9. The strain can remove ammonia nitrogen, total nitrogen and chemical oxygen demand under low-carbon and high-nitrogen conditions, has a heterotrophic nitrification-aerobic denitrification function, can maintain high activity under a low C / N condition, can be used for biological treatment of municipal sewage, culture biogas slurry, high-speed service area wastewater, decentralized sewage and the like, and is expected to make a new breakthrough in the field of biological nitrogen removal in an extreme environment.

Description

technical field [0001] The invention relates to the technical field of environmental microorganisms, in particular to low-carbon and high-nitrogen resistant heterotrophic nitrifying-aerobic denitrifying bacteria, and also relates to the application of low-carbon and high nitrogen-resistant heterotrophic nitrifying-aerobic denitrifying bacteria. Background technique [0002] Excessive nitrogen-containing compounds in the water body will cause eutrophication of the water body, causing algae and other plankton to multiply rapidly, the dissolved oxygen in the water body will decrease, the water quality will deteriorate, and fish and other organisms will die in large numbers, resulting in deterioration of water quality and seriously endangering ecological balance and human health. . The biological denitrification method is widely used in the treatment of ammonia nitrogen wastewater due to its economical, high efficiency and no secondary pollution. Traditional biological denitrif...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/30C12R1/01C02F101/16
CPCC02F3/302C02F2101/16Y02W10/10
Inventor 赵天涛陈沛沛刘毫张丽杰张福攀谭娜
Owner 重庆理享生态修复研究院有限公司
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