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Aerobic denitrifying bacillus subtilis preparation and preparation method thereof

A Bacillus subtilis, aerobic denitrification technology, applied in biochemical equipment and methods, chemical instruments and methods, microorganism-based methods, etc., can solve problems such as poor biological denitrification technology, and achieve healthy and sustainable development. , the effect of easy cultivation and mild reaction conditions

Inactive Publication Date: 2015-11-11
GUANGDONG BIDE BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most domestic aquaculture adopts a highly intensive aquaculture model. Sufficient oxygen is the key to ensure the success of high-density aquaculture. During the high-density aquaculture process of aquaculture, it must be continuously aerated to ensure a certain amount of dissolved oxygen in the water. However, the traditional Most of the denitrifying bacteria are anaerobic bacteria, and denitrification can only be carried out under anaerobic or anoxic conditions. Under the condition of sufficient oxygen, its ability to degrade nitrate nitrogen and nitrite nitrogen will be greatly inhibited. This has caused the problem that the current biological denitrification technology is not effective in the aquaculture environment.

Method used

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  • Aerobic denitrifying bacillus subtilis preparation and preparation method thereof
  • Aerobic denitrifying bacillus subtilis preparation and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Pick a loop of bacteria from the freeze-dried seeds of Bacillus subtilis GIM1.7152 and inoculate it into LB agar test tube slant medium, and cultivate at 37°C for 16 hours; transfer the test tube slant seeds to LB liquid medium at 37°C Shake culture for 20 hours to form a seed culture solution with a concentration of 2.0×10 8 cfu / mL.

[0028] The seed culture solution was transferred to a 30L seed tank equipped with a primary fermentation medium at an inoculum of 3% (volume ratio) for fermentation culture, the filling volume was 70%, the fermentation temperature was 37±1℃, the stirring rate was 200rpm, and the aeration rate 0.8~0.9V / V·min, dissolved oxygen concentration 80~90%, take samples every 2h after inoculation to measure the absorbance at 600nm, use the uninoculated medium as a blank control to make the growth curve of Bacillus subtilis, such as figure 1 As shown, it can be seen from the curve that the number of Bacillus subtilis increases sharply in 8-12 hours, indi...

Embodiment 2

[0058] Pick a loop of bacteria from the freeze-dried seeds of Bacillus subtilis GIM1.7152 and inoculate it into LB agar test tube slant medium, and cultivate at 37°C for 20 hours; transfer the slant seeds of the test tube to LB liquid medium at 37°C Shake culture for about 25 hours to form a seed culture solution with a concentration of 1.1×10 9 cfu / mL.

[0059] The seed culture solution was transferred to a 30L seed tank with a primary fermentation medium at an inoculum of 4% (volume ratio) for fermentation culture, the filling volume was 70%, the fermentation temperature was 37±1°C, the stirring rate was 300rpm, and the aeration rate 0.8-0.9V / V·min, dissolved oxygen concentration of 80-90%, fermentation time of 10h, made into first-class seed liquid. Transfer the first-level seed liquid to a 300L fermentor equipped with a second-level fermentation medium at an inoculum of 6% (volume ratio) for further expansion. The filling amount is 70%, the fermentation temperature is 37±1°C,...

Embodiment 3

[0066] Pick a loop of bacteria from the freeze-dried seeds of Bacillus subtilis GIM1.7152 and inoculate it into LB agar test tube slant medium, and culture at 37°C for 24 hours; transfer the test tube slant seeds to LB liquid medium at 37°C Shake culture for about 30 hours to form a seed culture solution with a concentration of 2.0×10 9 cfu / mL.

[0067] The seed culture solution was transferred to a 30L seed tank equipped with a first-level fermentation medium at a 5% (volume ratio) inoculum for fermentation culture, the filling volume was 70%, the fermentation temperature was 37±1℃, the stirring rate was 350rpm, and the aeration rate 0.8-0.9V / V·min, dissolved oxygen concentration 80-90%, fermentation time is 14h, the bacteria have reached the logarithmic growth phase after testing, and the first-level seed liquid is made. Transfer the first-level seed liquid to a 300L fermentor equipped with a second-level fermentation medium at an inoculum of 10% (volume ratio) to further expan...

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Abstract

The invention discloses an aerobic denitrifying bacillus subtilis preparation and a preparation method thereof. According to the invention, the aerobic denitrifying bacillus subtilis preparation with high spore content and capable of highly actively degrading nitrite nitrogen is cultured by adopting a preparation process easy in realizing industrial production and comprising the following steps: by utilizing the modern biological fermentation technology, carrying out liquid-solid combined fermentation, liquid high-density fermentation, solid fermentation, process material supplement, low-temperature drying, and the like. Each gram of the microbial preparation contains 20-25 billion living bacteria, the spore ratio is greater than 80%, and the infectious bacteria ratio is 0.5-1%. When used as a modifier as aquatic water, the aerobic denitrifying bacillus subtilis preparation can effectively reduce the content of nitrites in the aquatic water, so that the water quality is improved, and the demand for the healthy sustainable development of the aquaculture industry is guaranteed.

Description

Technical field [0001] The invention belongs to the technical field of aquaculture water quality regulation and control, and specifically relates to an aerobic denitrification Bacillus subtilis preparation and a preparation method thereof. Background technique [0002] With the rapid development of intensive aquaculture industry, the excretion of farmed animals and excess bait have caused the content of ammonia nitrogen and nitrite in the water body to seriously exceed the standard, and the water quality has deteriorated, which has led to frequent diseases and severely affected the health of the aquaculture industry. sustainable development. The denitrification of microorganisms plays a very important role in improving water quality, especially reducing nitrite. Denitrifying bacteria are a kind of microorganisms that can use nitrite as a nitrogen source and organic carbon as a carbon source, and can reproduce themselves. At present, most domestic aquaculture adopts highly inten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/125
Inventor 许国焕张丽程炜轩魏逸峰陈亚剑李志朋谭文俊梁建庆张嘉辉
Owner GUANGDONG BIDE BIO TECH
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