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A strain of Sphingomonas lpn080 assimilating isoammonia and its microbial preparation and application

A technology of sphingomonas and microorganism preparations, applied in the directions of microorganisms, microorganisms, microorganism-based methods, etc., can solve the problems of no application examples, lack of reports on heterotrophic nitrous assimilating bacteria, etc.

Active Publication Date: 2020-06-12
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is still a lack of reports and research on heterotrophic nitrous acid assimilating bacteria, and there are no examples of application in aquaculture

Method used

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  • A strain of Sphingomonas lpn080 assimilating isoammonia and its microbial preparation and application
  • A strain of Sphingomonas lpn080 assimilating isoammonia and its microbial preparation and application
  • A strain of Sphingomonas lpn080 assimilating isoammonia and its microbial preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Enrichment and isolation of Sphingomonas LPN080

[0022] Enrichment and isolation medium (EIM): 0.5g ammonium sulfate, 1g peptone, 6g glucose, 6g sucrose, 0.5g yeast extract, NaH 2 PO 4 0.5g, 1L of sand-filtered seawater, adjust the pH to 7.0, and sterilize by high-pressure steam.

[0023] Sample source: Mixed samples of prawn breeding water and pond bottom mud in Dianbai District, Maoming City, Guangdong Province.

[0024] The specific implementation steps are as follows: under sterile conditions, filter 135ml of the sample into a sterilized Erlenmeyer flask, add 10×EIM medium (the formula of 10×EIM medium is: ammonium sulfate 5g, peptone 10g, glucose 60g, sucrose 60g, yeast extract 5g, NaH 2 PO 4 5g, 1L of sand-filtered seawater, adjusted to pH 7.0, autoclaved) 15ml, cultured at 30°C for 48h in a shaker at 150rpm, to obtain the enrichment solution. Take 100 μl of the enrichment solution and spread it on the above-mentioned EIM solid medium, culture i...

Embodiment 2

[0026] Example 2: Molecular Identification of Sphingomonas LPN080

[0027] Take the strain LNP080 bacterial liquid and centrifuge, discard the supernatant, extract the genomic DNA from the bacterial cell pellet, use the genomic DNA as a template, and use 27F / 1492R as a primer to amplify 16SrDNA (doi:10.1128 / AEM.02272-07), and sequence, Its nucleotide sequence is shown in SEQ ID NO.1. The sequence was compared by Blast search and identified as a bacterium of the genus Sphingomonas (Sphingomonassp.), named Sphingomonas sp. LPN080.

Embodiment 3

[0028] Embodiment 3: the removal effect of sphingomonas (Sphingomonas sp.) LPN080 to the ammonia nitrogen of aquaculture water

[0029] Take aquaculture water samples and use ammonium sulfate to adjust the ammonia nitrogen concentration to about 8mg / L, add glucose to a final concentration of 1g / L, and after sterilization, divide 150ml into sterilized 250ml shaker flasks. Experimental groups (E1, E2, E3) added 500 μl of LPN080 bacterial solution (medium is EIM medium) cultured overnight at 30°C on a shaker at 200 rpm, and the control group (C1, C2, C3) added the same amount of EIM medium, 30 Cultivate on a shaker at ℃ (rotating speed 200rpm), and take water samples at 0h, 3h, 6h, 9h, 12h, 24h, 36h, and 48h. Utilize Nessler's reagent spectrophotometer to measure ammonia nitrogen content at different time points, the results are shown in figure 1 .

[0030] Depend on figure 1 It can be seen that when the concentration of ammonia nitrogen in the initial culture water sample rea...

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Abstract

The invention discloses a strain of sphingomonas LPN080 assimilating isoammonia and its microbial preparation and application. The strain was isolated from the prawn breeding environment in Maoming City, Guangdong Province. It has a strong ability to degrade ammonia nitrogen. It was identified as Sphingomonas sp. by molecular identification. Sphingomonas LPN080 can pass heterotrophic The form rapidly absorbs and assimilates ammonia nitrogen, so it is different from autotrophic nitrifying bacteria and heterotrophic nitrifying bacteria. In addition, Sphingomonas LPN080 can also significantly compete and inhibit Vibrio in aquaculture water. The application of Sphingomonas LPN080 in aquaculture water can significantly and quickly reduce the content of ammonia nitrogen in aquaculture water, which is safe for aquaculture animals.

Description

Technical field: [0001] The invention belongs to the technical field of aquaculture technology and ecological environment protection, and in particular relates to a strain of sphingomonas LPN080 assimilating isoammonia and its microbial preparation and application. Background technique: [0002] In aquaculture, a large amount of residual bait, feces and their metabolites in the aquaculture pond continue to increase. These substances are decomposed by microorganisms to produce a large amount of ammonia nitrogen and nitrite, both of which are toxic to farmed animals. Excessive ammonia nitrogen and nitrite It will also cause a large number of algae in the aquaculture water to explode, and the algae will die to produce toxins after the outbreak, which will also cause the death of aquaculture animals in large quantities, and at the same time, pathogenic microorganisms will also breed in large numbers. Therefore, controlling the concentration of ammonia nitrogen and nitrite in aqu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34A61K35/74A61P31/04C12R1/01C02F101/16C02F103/20
CPCA61K2035/11A61P31/04C02F3/34C02F2101/16C02F2103/20C12N1/20C12N1/205C12R2001/01
Inventor 罗鹏云龙王青柏李颖颖胡超群田雨顺
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI