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A kind of aerobic denitrifying bacteria and its application

A technology of aerobic denitrifying bacteria and Pseudomonas, applied in the directions of bacteria, chemical instruments and methods, biochemical equipment and methods, etc.

Active Publication Date: 2022-06-14
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, while high-density farming satisfies people's demand for aquatic products, it also causes serious harm to the farming environment and the surrounding ecological environment due to factors such as excessive input of feed and frequent use of antibacterial agents.
On the one hand, the deterioration of the aquaculture water itself, such as the accumulation of excessive bait, feces and secretions, increases the concentration of ammonium nitrogen and nitrite nitrogen in the water, endangering the healthy growth of aquaculture animals, affecting the production of aquaculture, and improving the quality of aquaculture. cost; on the other hand, the random discharge of untreated aquaculture wastewater has also caused serious pollution to the water environment around the aquaculture

Method used

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  • A kind of aerobic denitrifying bacteria and its application
  • A kind of aerobic denitrifying bacteria and its application
  • A kind of aerobic denitrifying bacteria and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Isolation, purification and preservation of Pseudomonas JM22B6

[0029] The samples were collected from the pond water of mixed culture of bighead carp and Penaeus vannamei in Guanghai Town breeding base (N21°56′31″; E112°46′16″) of Nianfeng Company, Taishan City, Jiangmen City, Guangdong Province. Breeding water sample 1mL, gradient dilution 10 -1 , 10 -2 , 10 -3 and 10 -4 After that, take 200 μL of 10 -2 、10 -3 and 10 -4 The diluted solution was spread on the selective medium used for the isolation and screening of denitrification bacteria, and cultured in a 30°C incubator. Observe the colony morphology with the naked eye, pick single colonies with differences in colony morphology, streak and purify them separately, inoculate the purified colonies into 5mL R2A liquid culture concentration, culture at 30°C and 180rpm, and store them in glycerol tubes for subsequent storage, thus obtaining Strain JM22B6.

[0030] The selective medium formula is as ...

Embodiment 2

[0033] Embodiment 2: Physiological and biochemical characteristics of Pseudomonas JM22B6

[0034] like figure 1 and figure 2 As shown, the strain JM22B6 is a Gram-negative bacterium. When the strain was cultured on NA medium for 48 hours, its colony morphological characteristics were: beige, irregular edges, slightly raised, smooth and opaque surface. The morphological characteristics of the cells observed by transmission electron microscope are: the size is (0.3×2.4) μm, long rod-shaped, with a single extreme flagella.

[0035] The strain JM22B6 was inoculated into NB medium with NaCl concentration of 0, 0.5%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0% and 10.0%, respectively, Cultivate at 30°C, 180rpm. Strain JM22B6 was inoculated into NB medium with pHs of 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0 and 11.0 respectively, and cultivated at 30°C and 180rpm. The strain JM22B6 was inoculated on NA medium, and cultured in incubators at 10°C, 15°C, 20°C...

Embodiment 3

[0038] Example 3: 16S rRNA sequence analysis of Pseudomonas JM22B6

[0039] HiPure Bacterial DNA Extraction Kit (Guangzhou Meiji Biotechnology Co., Ltd.) was used to extract the genomic DNA of strain JM22B6, and the bacterial 16S rRNA gene was amplified with universal primers 27F / 1492R (27F: 5'-AGAGTTTGATCATGGCTCAG-3, 1492R: 5'- TACGGTTACCTTGTTACGACTT-3') was added to obtain the PCR product, and sent to Suzhou Jinweizhi Biotechnology Co., Ltd. for sequence sequencing. The sequence is shown in SEQ ID NO.1. The sequence result is homologous to the 16S rRNA sequence in the EzBioCloud website database Comparative analysis, the results showed that the strain JM22B6 and the strain Pseudomonas mendocina NBRC 14162 T The 16S rRNA gene sequence similarity was the highest at 98.5%, which was less than the threshold value of 98.7% at the bacterial species level. Based on the analysis of the above results, the strain JM22B6 of the present invention was preliminarily identified as a new s...

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Abstract

The invention discloses an aerobic denitrifying bacteria and application thereof. The preservation number of Pseudomonas JM22B6 of the present invention is: GDMCC No: 61212. According to comprehensive analysis of morphology, physiology and biochemistry, 16SrRNA sequence and genome sequence, the strain of the present invention is a new species of Pseudomonas. This strain is a new type of aerobic denitrifying bacteria, which can efficiently remove nitrate nitrogen, nitrite nitrogen and ammonium nitrogen in water, that is, after 12 hours of cultivation, the removal efficiency of nitrite nitrogen and ammonium nitrogen Both were 100%; the removal rate of nitrate nitrogen was 100% after culturing for 24 hours. In addition, the strain 22B6 has a good denitrification effect in the range of nitrite nitrogen concentration from 14 to 280 mg / L. Therefore, Pseudomonas JM22B6 has important application potential in the denitrification treatment of wastewater polluted by nitrogen such as nitrate nitrogen, nitrite nitrogen and ammonium nitrogen, especially wastewater polluted by various concentrations of nitrite nitrogen.

Description

technical field [0001] The invention belongs to the technical field of microbes and environmental engineering, and in particular relates to an aerobic denitrifying bacterium and its application. Background technique [0002] With the rapid development of the domestic aquaculture industry, the high-density farming model is more and more popular. However, while high-density farming meets people's demand for aquatic products, it also causes serious harm to the breeding environment and surrounding ecological environment due to factors such as excessive feed input and frequent use of antibacterial agents. On the one hand, the deterioration of the aquaculture water itself, such as the accumulation of excessive bait, feces and secretions, increases the concentration of ammonium nitrogen and nitrite nitrogen in the water, endangering the healthy growth of aquaculture animals, affecting the production of aquaculture, and improving the quality of aquaculture. cost; on the other hand,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/38C02F101/16
CPCC12N1/20C02F3/34C02F2101/16C02F2101/163C02F2101/166
Inventor 张明霞肖波涛臧毅朱红惠吴清平李安章陈猛徐帅帅
Owner SOUTH CHINA UNIV OF TECH
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