Bacillus cereus and application thereof

A Bacillus cereus, straight rod technology is applied in the field of microorganisms to achieve the effects of maintaining the ecological balance of water bodies, good hydrophilicity, and simple culturing and fermentation methods

Active Publication Date: 2013-12-04
无锡亚克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of Bacillus for harmful algae control ...

Method used

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  • Bacillus cereus and application thereof
  • Bacillus cereus and application thereof
  • Bacillus cereus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 strain isolation

[0040] The inventor collected soil samples from Lhasa, Tibet, ground the soil into powder, put it into 45 ml of normal saline, and oscillated on a shaker for 30 minutes. Let it stand for clarification, pipette 100 μl of the supernatant with a sterile pipette tip, add it to 900 μl sterile water, and dilute to 10 with sterile water -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 A total of six concentration gradients. Put the diluted solution in a metal bath at 80°C and take it out after 10 minutes. Cultivation: Take 200 μl of each concentration dilution and spread it on LB solid medium to make a plate, place it upside down in an incubator, and cultivate overnight at 37°C, try to pick a single colony with different shapes in the liquid LB medium, shake it at 37°C Shake the culture overnight, then purify by streaking the plate until a pure culture is obtained. The isolated strain NMSW16 was identified (detailed below) as Bacillus cereus, and...

Embodiment 2

[0041] Example 2 strain identification

[0042] A. Morphological identification:

[0043] The isolated strain NMSW16 was observed under a microscope (1000 times), the cells were straight rods, produced spherical or elliptical spores, aerobic or facultative anaerobic, moved with perinatal flagella, no capsule; Staining identified it as Gram-positive bacteria (see figure 1 ).

[0044] B. Molecular identification:

[0045] 1. Use a sterile toothpick to pick a single colony and inoculate it into LB medium, culture at 37°C and 200 rpm for 24 hours with shaking, and use the conventional bacterial genomic DNA extraction method to extract genomic DNA as a template for PCR reaction.

[0046] 2. Amplify bacterial 16S rDNA and gyrB gene by PCR method

[0047] The primers were synthesized by Nanjing GenScript Biotechnology Co., Ltd., and the components of the PCR system (buffer, enzyme, dNTP, etc.) were purchased from Treasure Bioengineering (Dalian) Co., Ltd. (TaKaRa Company).

[00...

Embodiment 3

[0067] Example 3 Inhibitory effect of Bacillus cereus NMSW16 on the growth and reproduction of Microcystis aeruginosa

[0068] Pick a single colony with a sterile toothpick and inoculate it into LB medium (tryptone 10g / L, yeast extract 5g / L and sodium chloride 10g / L), culture at 37°C, 200rpm shaking for 24h; transfer to Landy culture base (every 1L contains 20g of glucose, 5g of L-glutamic acid, MgSO 4 0.5g, KCl0.5g, KH 2 PO 4 1g, FeSO 4 ·6H 2 O0.15mg, MnSO 4 ·H 2 O5mg, CuSO 4 ·5H 2 O0.16mg, L-phenylalanine 2mg, yeast extract 1g, pH7.0), at 30 ℃, 200rpm shaking culture fermentation for 38h.

[0069] Add 150 μl of fermentation broth to 30 ml of Microcystis aeruginosa FACHB-942 (from the Algae Collection Center of the Institute of Hydrobiology, Chinese Academy of Sciences) in the logarithmic growth phase, and then place it in a light intensity of 2500lx, a temperature of 28°C, Light-to-dark ratio 16h: 8h light incubator for static culture, shake regularly twice a day. ...

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Abstract

The invention discloses bacillus cereus and application thereof. The bacillus cereus has a very strong algal inhibiting effect, and particularly has a very strong inhibiting effect on microcystis aeruginosa, and the algal inhibition rate after a seven-day process is more than 90%. The bacillus cereus NMSW16 is simple in culture and fermentation, can be cultured in a large scale, can generate fermentation liquor through fermenting, and can be used for governing water bloom and restoring water eutrophication.

Description

technical field [0001] The invention relates to the field of microbes, in particular to a Bacillus cereus which has a high-efficiency inhibitory effect on harmful algae and its application in controlling algal blooms and restoring eutrophication of water bodies. Background technique [0002] With the acceleration of urbanization and the continuous development of industry and agriculture, a large amount of urban domestic sewage rich in nutrients such as nitrogen and phosphorus, industrial and agricultural wastewater flows into lakes, estuaries and other slow-flowing water bodies, causing excess nutrients in the water body and causing excessive algal blooms. Reproduction, leading to the decline of dissolved oxygen in water, the deterioration of water quality, the phenomenon of mass death of fish and other aquatic organisms, that is, the phenomenon of eutrophication in water body, which is called algae bloom in fresh water and red tide in ocean. There are many lakes in China, m...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P1/04C02F3/34C12R1/085
Inventor 高学文吴黎明张万宽伍辉军高林白子金
Owner 无锡亚克生物科技有限公司
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