A Simultaneous Nitrification and Denitrification Bacillus subtilis k8 and Its Application

A Bacillus subtilis and simultaneous nitrification technology, applied in the simultaneous nitrification and denitrification Bacillus subtilis K8 and its application fields, can solve the problems of single function and low efficiency, achieve high denitrification efficiency, high content of live bacteria, and improve utilization rate and the effect of nutrient content

Active Publication Date: 2022-03-01
天津市农业科学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a simultaneous nitrification and denitrification Bacillus subtilis K8 and its application, which solves the problems of low efficiency and single function in the prior art for removing nitrogen source pollutants in water bodies

Method used

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  • A Simultaneous Nitrification and Denitrification Bacillus subtilis k8 and Its Application
  • A Simultaneous Nitrification and Denitrification Bacillus subtilis k8 and Its Application
  • A Simultaneous Nitrification and Denitrification Bacillus subtilis k8 and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Isolation, screening and purification of a simultaneous nitrification and denitrification Bacillus subtilis K8

[0058] Take the activated sludge from the bottom of the Penaeus vannamei culture pond, dilute the sludge with sterile water at a ratio of 1:4, and treat it in a water bath at 80°C for 15 minutes, then streak on the plate, and further subculture different single colonies. The purified K8 strain was obtained after 5 subcultures.

Embodiment 2

[0059] Example 2 Morphological observation of a simultaneous nitrification and denitrification Bacillus subtilis K8 and sequence identification of 16s rRNA

[0060] The purified K8 strain was subjected to morphological observation and Gram staining observation to obtain the morphological characteristics of the K8 strain.

[0061] The DNA of the K8 strain was extracted and amplified by PCR. The amplified product was purified and then sequenced. The resulting sequence was analyzed by Blast, and a phylogenetic tree was constructed using the Neighbor-Joining software of MEGA 7.0. Phylogenetic tree results see figure 1 .

[0062] The results showed that the K8 strain was single-celled, rod-shaped, with a cell diameter of (0.7-0.8)×(3-4) μm, Gram staining was positive, spores were grown, the surface of the colony was rough and opaque, yellowish, and aerobic.

[0063] The length of the 16s rRNA sequence of the K8 strain is 1480bp, and the Blast analysis in the NCBI database shows t...

Embodiment 3

[0064] Example 3 Physiological and biochemical indicators of a simultaneous nitrification and denitrification Bacillus subtilis K8

[0065] The amylase activity of K8 strain was determined by 3,5-dinitrosalicylic acid method.

[0066] The protease activity of the K8 strain was determined by the azo-casein method.

[0067] The K8 strain was inoculated on LB medium with pH 3.0 and 4.0, cultured with shaking at 37°C for 24 hours, and the OD value of the K8 strain was detected.

[0068] The K8 strain was treated at 37° C. for 20 h at a bile salt concentration of 0.30%, and the survival rate of the K8 strain was detected.

[0069] The K8 strain was cultured in simulated gastric juice at 37°C for 240min, and the survival rate of the K8 strain was detected.

[0070] The results showed that the amylase activity of the K8 strain was 156.3U / mL, and the protease activity was 200.8U / mL. After the K8 strain was cultured on pH 3.0 and 4.0 media, the OD values ​​of the obtained strains we...

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Abstract

The invention relates to the technical field of agricultural microorganisms, in particular to a simultaneous nitrification and denitrification bacillus subtilis K8 and its application. The bacterial strain screened by the present invention is identified as Bacillus subtilis through physiological ecology analysis and molecular biology. The strain can grow under the condition of ammonia nitrogen, nitrite nitrogen or nitrate nitrogen as the only nitrogen source, and through synchronous nitrification and denitrification under heterotrophic and aerobic conditions, 100mg / L ammonia nitrogen, 100mg / L nitrate nitrogen The removal rate of nitrogen and 25mg / L nitrite nitrogen reaches 73.7%, 51.7% and 100% respectively, and does not accumulate inorganic nitrogen; the removal of 50mg / L ammonia nitrogen and 12.5mg / L nitrite nitrogen mixed nitrogen source in water The rate reaches 89.1% and 100% respectively, without nitrate accumulation; and it has a good control effect on cyanobacteria. The strain also has a strong ability to produce amylase and protease, and is resistant to acid and bile salts. It is compounded with yeast and lactic acid bacteria to make fermented feed, which can increase the content of amino acids and total protein in the feed.

Description

technical field [0001] The invention relates to the technical field of agricultural microorganisms, in particular to a simultaneous nitrification and denitrification bacillus subtilis K8 and its application. Background technique [0002] With the continuous improvement of the intensification of aquaculture, the accumulation of residual bait, excrement, and residues, coupled with the discharge of agricultural backwater, has resulted in serious pollution of the aquaculture water body. The most prominent is the high nitrogen content in the middle and late stages of aquaculture. Excessive nitrogen content in the water body will lead to eutrophication of the water body, algae multiply, consume dissolved oxygen in the water body, make the water smelly, and affect the survival of aquatic animals. However, how to economically and effectively remove nitrogen in aquaculture water and at the same time inhibit the production of cyanobacteria has always been a problem of concern to aquac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/16C02F3/34A23K10/18A23K10/12C12R1/125C12R1/225C12R1/74C02F101/16
CPCC12N1/20C12N1/16C02F3/34A23K10/18A23K10/12C02F2101/16C02F2101/163C02F2101/166C12R2001/125C12N1/205
Inventor 谢凤行张峰峰周可赵琼孙海波赵玉洁
Owner 天津市农业科学院
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