Bacillus subtilis N2 with ammonia nitrogen degradation function and application thereof

A Bacillus subtilis, ammonia nitrogen technology, applied in the field of microorganisms, can solve the problems of lack of scientific and rigorous technical guidance, unstable quality, long starting time, etc., and achieve the effects of good application prospects, wide tolerance and strong adaptability

Active Publication Date: 2019-02-19
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since the early 1980s, my country has begun to apply micro-ecological preparations to aquaculture. The most important ones are nitrifying bacteria and denitrifying bacteria. Most of them are photoautotrophic bacteria, and their existence and function start for a long time. Unstable quality, difficult to become dominant bacte...

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  • Bacillus subtilis N2 with ammonia nitrogen degradation function and application thereof
  • Bacillus subtilis N2 with ammonia nitrogen degradation function and application thereof
  • Bacillus subtilis N2 with ammonia nitrogen degradation function and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Enrichment and screening of Bacillus subtilis N2 with the function of reducing ammonia nitrogen

[0022] Sample source: Litopenaeus vannamei (Litopenaeus vannamei) cultured water body in the breeding house of South China Sea Institute of Oceanology, Chinese Academy of Sciences.

[0023] The specific implementation steps are as follows: under sterile conditions, inoculate the prawn culture water body into a 250mL Erlenmeyer flask containing 100mL liquid screening medium with an inoculum size of 1% v / v, place it in a shaker at 200rpm and culture at 30°C for 48h ; Take 100 μL of the culture solution and spread it on the solid screening medium, cultivate it in a 30°C incubator for 24 hours, pick a single colony, and inoculate it again in the liquid screening medium for pure culture, after 36 hours of pure culture, take the culture solution and dilute it 10 times The ammonia nitrogen content in the culture solution was measured, and the strain with the best effect ...

Embodiment 2

[0027] The identification of embodiment 2 bacillus subtilis N2 bacterial classification

[0028] 1. Morphological observation

[0029] The pure culture strains were inoculated on the surface of 2216E solid medium according to the plate streaking method, and cultured in a 30°C incubator for 24 hours to observe the morphological characteristics of the colonies. See the photo of N2 strain colony figure 1 , the colony is dot-shaped, opaque white, with smooth edges and raised colonies.

[0030] The 2216E solid medium is prepared by adding agar powder with a mass fraction of 1.5% to the 2216E liquid medium components and mixing. The 2216E liquid medium contains 5g of peptone per liter, 1g of yeast extract powder, 0.1g of ferric citrate, 19.45g of sodium chloride, 5.98g of magnesium chloride, 3.24g of sodium sulfate, 1.8g of calcium chloride, and 0.55g of potassium chloride , sodium carbonate 0.16g, potassium bromide 0.08g, strontium chloride 0.034g, boric acid 0.022g, sodium sili...

Embodiment 3

[0040] Example 3 Exploration of Bacillus subtilis N2 Optimum Growth Conditions

[0041] The effects of different pH, temperature and NaCl mass fraction (salinity) on the growth of Bacillus subtilis N2 were studied respectively. Among them, the different temperatures are: 20°C, 25°C, 30°C, 37°C, 43°C; the different pH values ​​are: 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0; the different NaCl mass fractions are: 0 %, 1%, 2%, 3%, 4%, 5%.

[0042] In the above experiments, except for the investigation factors, the fermentation conditions were all to inoculate the Bacillus subtilis N2 culture solution in the LB fermentation medium with an inoculum size of 1% (v / v), and after cultivating for 24 hours at 30°C and 200rpm, samples were taken to measure each Conditional OD 600 Value, the LB fermentation medium without inoculation was used as blank control. Take culture temperature, pH value, NaCl mass fraction as abscissa, OD 600 Values ​​plot the respective curves on the ordinate. see r...

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Abstract

The invention discloses a Bacillus subtilis N2 with an ammonia nitrogen degradation function and application thereof. The Bacillus subtilis N2 is isolated from a shrimp culture water body. Studies confirm that the Bacillus subtilis N2 can quickly and effectively reduce the ammonia nitrogen content in the water body, and do not convert ammonia nitrogen into nitrite to cause stress or damage to a cultured object again. In addition, the Bacillus subtilis N2 and artemia gnotobiote tests show that at higher concentrations, a strain does not produce toxic effects on farmed animals, and has certain probiotic effects; the strain is more tolerant, a salinity and the environmental pH have little effect on bacterial growth, and regardless of being isolated from the freshwater or the seawater and theacid or the alkali environment, the strain N2 can grow well and function efficiently; it is of great significance in the marine aquaculture industry. The Bacillus subtilis N2 is inoculated into the aquaculture water body, the water quality is stable, the cultured object grows healthily, the adaptability is strong, and the safety is high. The strain can be used as a good strain of a novel microecological preparation and have a good application prospect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bacillus subtilis N2 with the function of degrading ammonia nitrogen and its application. Background technique [0002] With the expansion of aquaculture scale and the improvement of aquaculture density, problems such as environmental pollution and economic constraints have become increasingly serious, which has greatly hindered the development of aquaculture. Especially in the intensive aquaculture system, in pursuit of output and economic benefits, a large amount of feed, fertilizer and drug inputs and the deposition of metabolites in cultured fish and shrimps lead to serious endogenous pollution in ponds, deterioration of the breeding ecological environment, residual bait and After the feces are decomposed by microorganisms, a large amount of ammonia nitrogen is produced. High-concentration ammonia nitrogen is highly toxic to the breeding objects, often leading to lar...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/125C02F101/16C02F103/20
CPCC02F3/34C02F2101/16C02F2103/20C12N1/205C12R2001/125
Inventor 陈偿丁雄祺谢媚杨艺滢方建相管云雁云龙
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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