Bacillius firmus GY-49, screening method and applications thereof

A technology of Bacillus firmus and GY-49, which is applied in the direction of chemical instruments and methods, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of high cost of oxygenation, waste of resources, low treatment efficiency, etc., and achieve detoxification Stable nitrogen effect, lower COD, and high treatment efficiency

Active Publication Date: 2015-03-11
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the process of aquaculture wastewater treatment, anaerobic bacterial agents are used to remove nitrogen and aerobic bacterial agents are used to remove COD. The death of aerobic bacteria, anaerobic bacteria can not denitrify under aerobic conditions, effectively remove nitrogen and the cost of oxygenation is high, due to the difference in growth and metabolic environment, the two bacteria can not be used at the same time
As a result, it leads to waste of resources, increased cost, and low treatment efficiency in the process of aquaculture wastewater treatment.

Method used

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  • Bacillius firmus GY-49, screening method and applications thereof
  • Bacillius firmus GY-49, screening method and applications thereof
  • Bacillius firmus GY-49, screening method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Material preparation

[0026] (1). Bacterial source

[0027] Bacterial source: the soil at a depth of 10-15 cm below the surface of the vegetable field in Wanshou Village, Qixia District, Nanjing City, Jiangsu Province.

[0028] (2). Medium

[0029] (1) Beef extract peptone medium: 5.0 g beef extract, 10.0 g peptone, 5.0 g KCl, MgSO 4 ·7H 2 O 2.5 g, dilute to 1 L with water, and adjust the pH to 7.0~7.2.

[0030] (2) LB medium: 10.0 g peptone, 5.0 g yeast powder, 10.0 g NaCl, dilute to 1 L with water, and adjust the pH to 7.0~7.2.

[0031] (3) Denitrification medium: NaNO 3 0.6 g, trisodium citrate 4.1 g, MgSO 4 ·7H 2 O 0.1 g, KH 2 PO 4 1.5 g, 1 mL of trace element solution, dilute to 1 L with water, adjust pH to 7.0-7.5.

[0032] Trace element solution: Na 2 EDTA 63.7 g, ZnSO 4 2.2 g, CaCl 2 5.5 g, MnCl 2 4H 2 O 5.1 g, FeSO 4 ·7H 2 O 5.0 g, Na 2 MoO 4 4H 2 O 1.1 g, CuSO 4 ·5H 2 O 1.6 g, CoCl 2 ·6H 2 O 1.6 g, dilute to 1 L with water, adju...

Embodiment 2

[0042] Example 2: Identification of strains.

[0043] 1. Morphological, physiological and biochemical characteristics of Bacillus firmus GY-49.

[0044] (1) Bacillus firmus Bacillus firmus. Morphological characteristics of GY-49 colonies ( figure 1 )

[0045] Strain GY-49 can be seen after being cultured on LB medium for 2 days, and the colony shape is round, white, with neat edges, rough surface, dry, and protrusions.

[0046] (2) Bacillus firmus Bacillus firmus. Physiological and biochemical characteristics of GY-49

[0047]

[0048] Note: + means positive, - means negative.

[0049] 2. Determination of the 16S rDNA sequence of strain GY-49

[0050] (1) Extraction of bacterial total DNA

[0051] Pick a single colony and activate it on an LB plate first, then transfer it to 5 mL LB liquid medium, culture it to the logarithmic phase overnight with shaking, centrifuge at 8000 rpm for 2 min, collect the bacterial liquid, and wash it with STE for 2 to 3 times; use 0....

Embodiment 3

[0099] Embodiment 3: bacterial strain of the present invention adds KNO with beef extract peptone medium 3 To simulate the anaerobic denitrification and COD removal of high-nitrogen and high-COD aquaculture wastewater, the steps are as follows:

[0100] 1. The Bacillus firmus ( Bacillus firmus. ) GY-49 was inoculated onto the slant of sterilized LB medium, and activated at 30°C for 24-36h; the activated strain was inoculated in beef extract peptone liquid medium, and 100ml of medium was filled in a 250ml Erlenmeyer flask. The medium is beef extract 5.0 g, peptone 10.0 g, NaCl 10.0 g, KCl 5.0 g, MgSO 4 ·7H 2 O 2.5 g, distilled water 1 L, pH7.0-7.5, 30°C, 120rpm shaking culture for 24-36h;

[0101] 2. Centrifuge the bacterial solution obtained in step 1 at 5000rpm for 3 minutes to collect the bacterial cells, and transfer to the KNO-containing 3 Cultured in the beef extract peptone medium for 48 hours, the bacterial agent must be applied; containing KNO 3 The formula of be...

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Abstract

The invention discloses bacillius firmus GY-49. The classification name is Bacillus firmus GY-49, the preservation number is CCTCC No.M2014347, and the bacterium strain has been preserved in the China Center for Type Culture Collection (CCTCC) on July 21st, 2014. The invention further discloses a screening method of bacillius firmus GY-49. The invention also discloses an application of bacillius firmus GY-49 in wastewater treatment under an anoxic condition. The invention also discloses an application of bacillius firmus GY-49 in an anaerobic treatment of wastewater with high nitrogen and COD concentrations. The invention further discloses an application bacterium inoculant. The provided bacterium strain can sustain high concentration nitrates and COD, has an efficient denitrifying performance and COD and Cr removing performance under a completely anoxic condition, and is capable of effectively removing nitrates (nitrogen) and reducing the COD content.

Description

technical field [0001] The invention belongs to the field of environmental application microorganisms, and in particular relates to a bacillus firmus GY-49 and its screening method and application. Background technique [0002] With the development of aquaculture industry, the removal of high-concentration nitrogen and COD in aquaculture wastewater has become an important part of the pollution prevention and control of aquaculture industry. The biological denitrification and COD removal process has been widely used because of its good treatment effect, low treatment cost, and convenient operation and management. [0003] However, most biological denitrification and COD removal cannot be carried out simultaneously. The main reason is that the biological denitrification process mostly performs denitrification under anaerobic conditions to convert nitrogenous substances into N 2 emissions, while the removal of COD generally relies on the aerobic respiration of microorganisms...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04C02F3/34C12R1/07
CPCC02F3/34C02F2103/20C12Q1/04C12N1/205C12R2001/07
Inventor 胡南高远高浩峰王鹏胤杨立志
Owner NANJING TECH UNIV
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