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Bacillus sp. NB-1 and culture method and application thereof

A technology of bacillus and culture, applied in the field of microorganisms, can solve the problems of high inoculation amount, low degradation treatment efficiency, low ammonia nitrogen and nitrite degradation effect, and achieve the effect of improving emission reduction

Active Publication Date: 2020-12-22
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current microbial preparation products for degrading ammonia nitrogen and nitrite generally have problems such as high production and application costs, low degradation treatment efficiency, and poor stability of treatment effects.
[0003] There are existing Chinese published patents, the patent numbers are: CN200610011604.1, CN201810782000.X, CN201811325758.7 and CN201910309817.X, all of which disclose different types of Bacillus, but there are the following problems: 1. High inoculation amount; 2. One type Bacillus strains can only effectively degrade one of ammonia nitrogen and nitrite nitrogen, and cannot effectively degrade both of them at the same time. 3. The degradation effects of ammonia nitrogen and nitrite nitrogen are low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, the separation and screening of bacillus NB-1

[0023] Sampling: Cut 10g of nylon filaments of biofilm water purification grid with sterile scissors from the water body of the eel breeding pond in the eel culture workshop of Jimei University Fisheries Experimental Field, Xiamen City, Xiamen City, Fujian Province, and put them into a container. In a blue cap bottle with 100mL of sterile water, the bacterial suspension was obtained after ultrasonic vibration for 30 minutes.

[0024] Enrichment: Add 10% of the bacterial suspension into sterile enrichment medium, culture at 30°C, 180r / min constant temperature shaking, add 2mL of 5% sodium nitrite solution every day, and enrich for 1 week.

[0025] Enrichment medium (g / L): glucose 5, sodium chloride 0.1, nitrite 0.2, magnesium sulfate 0.02, potassium dihydrogen phosphate 4, dipotassium hydrogen phosphate 6, prepared with single distilled water, pH 7.4, extinguished at 121°C Bacteria 30min.

[0026] Separation...

Embodiment 2

[0030] Embodiment 2, identification of bacillus NB-1

[0031]Pick a single colony of the above-mentioned preserved strains, inoculate them in LB medium at 30°C, shake and culture at 170 r / min for 12 hours, collect the bacteria by centrifugation, and use the bacterial DNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.) Genomic DNA of the strain was extracted, using universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') (shown in SEQ ID No:1), and 1492R (5'-GGTTACCTTGTTACGACTT-3') (shown in SEQ ID No:2 ), PCR amplification of 16S rRNA gene was carried out, and the PCR reaction conditions were: 95°C pre-denaturation for 5 min, 30 cycles (95°C for 1 min, 53°C for 1 min, 72°C for 1.5 min), and extension at 72°C for 10 min. A gel recovery kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.) was used to purify the target fragment of about 1.5 kb, and the purified PCR product was sent to (Shanghai Meiji Biomedical Technology Co., Ltd.) for sequencing.

[0032] LB ...

Embodiment 3

[0035] Example 3, Ammonia nitrogen and nitrite degradation rate and safety detection in culture water of Bacillus NB-1

[0036] Pick a single colony of strain NB-1 and inoculate it in sterilized LB medium, and shake it at 170 r / min for 24 hours at 30°C. to 4×10 4 The inoculum volume of cfu / mL was added to contain 1m 3 In the plastic barrels of the cultured American eel in the water body, the control group does not add bacteria, and a double parallel control experiment is established. The weight of the cultured American eel in each barrel of the treatment group and the control group is 11.2kg, and the average size is 140g / tail. During the test, at 0h, 24h Collect water samples from each tank at 168h and 168h respectively. After the water samples are centrifuged, the supernatant is taken to measure the concentration of ammonia nitrogen and nitrite nitrogen, and the degradation rate is calculated. Both the treatment group and the control group were fed and managed at a feeding...

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Abstract

The invention provides Bacillus sp. NB-1 and a culture method and application thereof. The Bacillus sp. NB-1 is Bacillus sp. NB-1, is preserved in the China Center for Type Culture Collection on July13 , 2020, the address is Wuhan University, Wuhan Province, China, and the preservation number is CCTCC NO: M2020304; the Bacillus sp. NB-1 is cultured for 24h at 30 DEG C by using an LB liquid culture medium; and the Bacillus sp. NB-1 is used for degrading the concentration of ammonia nitrogen and nitrite in aquaculture water and aquaculture tail water. The Bacillus sp. NB-1 can obviously reduce the concentration of ammonia nitrogen and nitrite in the aquaculture water and the aquaculture tail water, is harmless to people and cultured animals, does not generate secondary pollution, and can be applied to improvement of aquaculture environment and aquaculture tail water emission reduction in large-scale production.

Description

【Technical field】 [0001] The invention relates to the technical field of microorganisms, in particular to a bacillus NB-1 capable of degrading ammonia nitrogen and nitrite, and a culture method and application thereof. 【Background technique】 [0002] my country is a big aquaculture country, and its aquaculture production accounts for about 68% of the world's aquaculture production, of which pond aquaculture production accounts for about 45%. During the breeding process, due to feeding a large amount of feed, the accumulation of nitrogen elements in the water is also one of the main reasons for the deterioration of the breeding water body. Among them, ammonia nitrogen and nitrite are the most serious water quality physical and chemical factors that endanger the quality of aquaculture water and farmed animals. Animals are highly toxic. Ammonia nitrogen will affect the enzyme reaction and the stability of the cell membrane in the cultured animals, and it will also affect the t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/07C02F103/20C02F101/16
CPCC12N1/20C02F3/34C02F2101/16C02F2101/166C02F2103/20C12R2001/07C12N1/205
Inventor 江兴龙郭少鹏
Owner JIMEI UNIV
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