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Biological denitrifier and application thereof

A technology of denitrification and biological activity, applied in the field of microorganisms, can solve the problems of long treatment time, low efficiency, lack of tolerance to high-concentration ammonium salts, etc., and achieve good denitrification effect

Active Publication Date: 2010-10-06
南京曜动节能环保科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the Bacillus subtilis that have been reported so far have problems such as weak removal ability of ammonia nitrogen, lack of tolerance to high concentrations of ammonium salts, long processing time, and low efficiency.

Method used

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  • Biological denitrifier and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Ammonia nitrogen removal ability of CPU BS-1 for simulated ammonium-containing wastewater

[0045] 1. Determination of ammonia nitrogen in medium by indophenol blue method

[0046] 1.1 Ammonium standard solution preparation:

[0047] Precisely measure 0.2358g of ammonium sulfate dried to constant weight at 105°C, dissolve in an appropriate amount of double-distilled water, transfer to a 100mL volumetric flask, and then set the volume to the mark. That is, the mother liquor containing 500 μg / mL of ammonium. Accurately dilute 10 times before use.

[0048] 1.2 Reagent A:

[0049] Accurately weigh 5.0000g of phenol and 0.0290g of ferrous sodium ferricyanide (dihydrate) and dissolve in double distilled water, dilute to 500mL, and store in a brown bottle at 4°C.

[0050] 1.3 Reagent B:

[0051] Weigh 2.50g of sodium hydroxide, 2.0g of trisodium citrate and 3.5mL of sodium hypochlorite, dissolve them in double distilled water, dilute to 500mL, and store in a brown bottle ...

Embodiment 2

[0064] Preparation of freeze-dried powder and determination of viable bacteria concentration

[0065] 1. Preparation of freeze-dried powder

[0066]Add 15% (m / v) skimmed milk powder to the fermentation broth of Bacillus subtilis CPU BS-1, pre-freeze in the cold trap of a lyophilizer for 7h-8h, and dry in a vacuum in a lyophilizer. The first stage is -30°C for 240min (4h), the second stage is -10°C for 600min (10h), the third stage is 0°C for 960min (16h), and the fourth stage is 4°C for 360min (6h).

[0067] 2. Determination of the concentration of live bacteria in freeze-dried powder

[0068] The lyophilized powder was used to determine the concentration of viable bacteria, and diluted with starch to an appropriate concentration.

[0069] The bacteria count reporting rules should select the average bacterial colony count between 30 and 300 as the basis for the bacteria count report (taking two significant figures).

[0070] 2.1 When the number of colonies in only one dilut...

Embodiment 3

[0076] Large-scale cultivation of Bacillus subtilis CPU BS-1 in fermenter

[0077] Two rings of bacterial lawn were taken from the agar slant and put into a 250ml Erlenmeyer flask with 50ml of seed medium, and cultured at 37°C and 200rpm on a shaker for 12h. Inoculate according to 5% inoculum amount, and transfer 500ml of seed culture solution to a 10L fermenter equipped with 9.5L of fermentation medium. According to the research result of shake flask experiment, determine the culture condition of optimum fermentor: 37 ℃, stirring speed 100rpm, ventilation rate 0.07vvm, inoculation amount 5%, fermentation medium composition (g / L): glucose 160g, corn steep liquor 40g, urea 20g, soybean cake powder 5g, calcium carbonate 5g, the initial pH was adjusted to 7.2, and the strain grew into a plateau after 24h. After culturing for 24 hours, a higher concentration of bacteria can be harvested, which is conducive to large-scale use in practical applications. Growth curve see attached ...

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Abstract

The invention relates to the field of microorganisms, in particular to a biological denitrifier and application thereof. The biological denitrifier is a strain of bacillus subtilis, named bacillus subtilis CPU BS-1, and preserved in China General Microbiological Culture Collection Center in March, 17th, 2010 with the preservation number of CGMCC No.3668. The invention also discloses a method for culturing bacteria with denitrification biological activity and application thereof in degrading nitrogen in a water body.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a bacterium with denitrification biological activity, a method for cultivating the bacterium, preparation of a biological denitrification agent with the bacterium as the main component, and its application in degrading nitrogen in water . Background technique [0002] Nitrogen cycle is one of the most important components of ecosystem material cycle in nature. However, the use of nitrogen fertilizers and the development of high-density aquaculture have caused serious environmental problems while bringing huge economic benefits. In aquaculture, eutrophication frequently occurs due to excessive nitrogen and phosphorus content. In addition to deteriorating water quality and destroying the biological balance, ammonia nitrogen and nitrite nitrogen are highly toxic to farmed animals. Especially ammonia nitrogen (NH 4 + -N), it can directly or indirectly affect the growth, reproduction...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/125C02F101/16
Inventor 贾源宾王涛和金周洪秀娟窦洁周长林
Owner 南京曜动节能环保科技有限公司
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