Bacillus subtilis BY7 and application thereof in degrading residual feeds and ammonia nitrogen

A Bacillus subtilis and ammonia nitrogen technology, applied in the field of microorganisms, can solve the problems of high ammonia nitrogen content in water, affecting the growth of cultured aquatic animals, endangering the lives of cultured animals, etc., and achieves the effect of reducing ammonia nitrogen content, improving ammonia nitrogen pollution and high safety.

Active Publication Date: 2016-08-24
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, since nitrogen-containing organic matter such as residual bait and feces are decomposed, ammonia nitrogen (NH 4 + The form of -N) is released into the water body, resulting in excessively high ammonia nitrogen content in the water body, affecting the growth of aquaculture animals, and even endangering the life of aquaculture animals. Therefore, it is of great significance to screen microbial strains that can reduce ammonia nitrogen in water bodies and apply them to aquaculture production

Method used

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  • Bacillus subtilis BY7 and application thereof in degrading residual feeds and ammonia nitrogen
  • Bacillus subtilis BY7 and application thereof in degrading residual feeds and ammonia nitrogen
  • Bacillus subtilis BY7 and application thereof in degrading residual feeds and ammonia nitrogen

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0034] Example 1 Isolation, purification and identification of Bacillus subtilis BY7

[0035] 1. Separation and purification of bacteria

[0036] 1. Enrichment of protein and ammonia nitrogen strains

[0037] Take 100mL of water samples from the tilapia culture pond, freshwater white pomfret culture pond, black snakehead culture pond, carp culture pond, and golden crucian carp culture pond in the hot spring aquaculture experiment field in Daiyue District, and mix the water samples from each culture pond Take 5mL and inoculate it into 100mL enriched medium (add NH to basal medium 4 Cl, adjust the concentration of ammonia nitrogen to 50mg / L to obtain the enrichment medium) in the Erlenmeyer flask, 37 ° C, 200r / min shaking culture for 1 day; take 1 mL of the culture solution after 1 day of cultivation and re-inoculate in 100mL enrichment medium , so repeated enrichment culture for 6 generations.

[0038] 2. Screening and isolation of protein and ammonia nitrogen strains

[00...

example 2

[0062] The research of example 2 Bacillus subtilis BY7 growth conditions

[0063] 1. The effect of C / N on the growth of strains

[0064] 1. Pick a single clone of Bacillus subtilis BY7, inoculate it in 10mL basal medium, and culture it with shaking at 37°C and 200r / min for 12h (make the OD of the bacterial solution in the culture system 600 value reaches 1.0), centrifuged at 4000rpm for 10min, collected the bacteria, and washed 3 times with sterile saline.

[0065] 2. Resuspend the above-mentioned bacteria with sterile normal saline, inoculate 2 mL of the bacterial suspension into 9 conical flasks containing 100 mL of colorless medium (pH 6.0), adjust each conical flask to culture Concentrations of sodium citrate and ammonium chloride in the base, so that the C / N in the 9 Erlenmeyer flask mediums were 1, 3, 4, 5, 6, 7, 9 and 11, and then 37 ° C, 200r / min Shaking culture, sampling once every 2h to determine the OD of the bacterial solution 600 Value, find after cultivating 1...

example 3

[0073] Example 3 Bacillus subtilis BY7 protease activity analysis

[0074] 1. Drawing of standard curve

[0075] 1. Prepare various concentrations of tyrosine solutions according to Table 3

[0076] Table 3 prepares various concentrations of tyrosine solutions

[0077] pipe number

1

2

3

4

5

6

Distilled water (mL)

10

8

6

4

2

0

100μg / mL Tyrosine (mL)

0

2

4

6

8

10

Tyrosine final concentration (μg / mL)

0

20

40

60

80

100

[0078] 2. Take 6 test tubes, labeled No. 1-6, add 0-10 mL of tyrosine at a concentration of 100 μg / mL respectively to prepare a solution with a final concentration of tyrosine as described in Table 3, and then add 0.4 mol / mL of tyrosine to each test tube. L of sodium carbonate solution 5mL and Folin's reagent 1mL. After fully mixing with a mixer, put it in a constant temperature water bath at 37°C to react for 20 minutes.

[0079] 3. Take No. 1 tube as ...

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Abstract

The invention relates to bacillus subtilis BY7 and an application thereof in degrading residual feeds and ammonia nitrogen. The bacillus subtilis BY7 is preserved in China Center for Type Culture Collection on September 29, 2015, with preservation number of CCTCC M 2015585; and the 16s rNDA of the bacillus subtilis BY7 is as shown in SEQ ID NO.1. The bacillus subtilis BY7 or a BY7 suspension can degrade ammonia nitrogen in a contaminated water body or an aquaculture water body and residual feeds in fee. The bacillus subtilis provided by the invention is applicable to the following aspects: (1) the bacillus subtilis, with a strong ammonia nitrogen degrading capacity, can be used for improving an ammonia-nitrogen pollution problem in the water body and for reducing an ammonia-nitrogen content in an aquaculture pond; and (2) the bacillus subtilis, which is capable of degrading protein in the residual feeds of the feed, can be used for degrading the residual feeds in the aquaculture pond and organic matters in the bottom of the pond. The bacillus subtilis disclosed by the invention has a broad application prospect in aquaculture ponds at high temperature in the summer season.

Description

1. Technical field [0001] The invention relates to bacillus subtilis BY7 and its application for degrading residual bait and ammonia nitrogen, and belongs to the technical field of microorganisms. 2. Background technology [0002] In aquaculture ponds, nitrogen-containing organic matter such as residual bait and manure at the bottom of the pond caused by a large amount of feeding is difficult to be completely oxidized and decomposed by natural microorganisms, and the remaining organic matter accumulates at the bottom of the pond over time, polluting the bottom and water quality of the pond. Therefore, it is very important to artificially apply microbial strains that can degrade a large amount of residual bait to accelerate the decomposition of organic matter at the bottom of the pond. In addition, since nitrogen-containing organic matter such as residual bait and feces are decomposed, ammonia nitrogen (NH 4 + The form of -N) is released into the water body, resulting in ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34A01K61/00C12R1/125
CPCA01K61/00C02F3/341C12N1/205C12R2001/125Y02A40/81
Inventor 季相山王慧王晓云赵燕陈红菊刘洪军
Owner SHANDONG AGRICULTURAL UNIVERSITY
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