New bacillus subtilis strain, microecological preparation and application

A technology of Bacillus subtilis and microecological preparations, which is applied in the field of aquatic microecological preparations, can solve the problems of large differences in the effects of strains, etc., and achieve the effect of improving the quality of breeding and improving the economic benefits of breeding

Active Publication Date: 2016-03-30
BEIJING DABEINONG TECH GRP CO LTD +2
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many strains, the effects of the strains vary greatly, but few are actually effective

Method used

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  • New bacillus subtilis strain, microecological preparation and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Take 124 water samples and mud samples from fish ponds and shrimp ponds in Wuqing, Tianjin, Fujian, and Jiangsu. Take 1 mL of water samples or 1 g of mud samples in 9 mL of LB liquid medium in 18 ml test tubes, and treat them in a water bath at 85 ° C for 10 min.

[0018] The test tube after the high temperature treatment was placed under the aerobic condition of 30° C. and 180 rpm for constant temperature shaking culture for 16 hours, and then the enriched culture solution was obtained.

[0019] The enriched culture solution was serially diluted by the doubling dilution method, spread on LB solid medium, placed in a 30°C incubator for 20 hours, picked different single colonies grown on the plate, streaked, and purified Two times, live single colonies were obtained, and a total of 275 strains were obtained.

[0020] Use a toothpick to inoculate the single colony purified twice on a fresh hemolysis plate, and place it in a 30°C incubator to observe for 24h, 48h, and 72h....

Embodiment 2

[0026] Example 2 High-throughput determination of nitrogen-reducing function of strains, screening of efficient strains

[0027] In the ultra-clean bench, the 63 bacterial strains to be tested were inserted into the corresponding screening medium, and the final inoculation concentration was 5×10 5 CFU / mL, the number of repetitions is 3; culture in a shaker at 30°C, 180r / min, and culture for 48h; evenly sample into a 1.5mL centrifuge tube, centrifuge at 12000rpm for 3min, take 200μL of the supernatant and add it to a 96-well plate;

[0028] When measuring nitrite nitrogen, after adding 200 μL of the supernatant of the sample to be tested in a 96-well plate, add 20 μL of Griess reagent A and B to each well successively, and measure it colorimetrically at 550 nm; when measuring ammonium nitrogen, add Nessler's reagent 20 μL, colorimetric determination at 450nm;

[0029] There are two types of screening medium: ammonia nitrogen screening medium and nitrite nitrogen screening medi...

Embodiment 3

[0050] Comparison of H8-1 growth conditions under different salt concentration conditions in embodiment 3

[0051] Use water with salinities of 0‰, 3‰, and 15‰ to prepare LB solid medium, use a sterilized toothpick to plant H8-1 on plates with different salinities, and place them in a 30°C incubator for 16 hours. Observe the growth of colonies on the plate. After culturing, it was found that the colonies planted on the three plates with different salinities grew well, and the colony sizes after growth were relatively consistent, indicating that H8-1 had strong salt tolerance.

[0052] LB medium: tryptone: 10g / L; yeast extract 5g / L; sodium chloride 10g / L; agar powder 15g / L. Use a salinity meter to prepare water with a salinity of 0‰, 3‰, and 15‰, respectively, and prepare the culture medium.

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Abstract

The invention belongs to the technical field of aquatic-product microecological preparations, and concretely relates to a n new bacillus subtilis strain, a microecological preparation and application. The disclosed bacillus subtilis H8-1 possesses the preservation number of CGMCC No. 9356, is capable of reducing harmful substances such as ammonia nitrogen and nitrite-state nitrogen in water in a pollution-free residue-free way, improving culture quality and improving culture economic benefit, and possesses promotion value. The degradation rates of bacillus subtilis H8-1 on ammonia nitrogen and nitrite-state nitrogen both with the concentration of 2.5 mg / L are respectively 81.72% and 99.70%.

Description

technical field [0001] The invention belongs to the technical field of aquatic microecological preparations, and in particular relates to a new strain of bacillus subtilis, a microecological preparation and its application. Background technique [0002] In recent years, high-density and intensive aquaculture has gradually replaced the traditional free-range farming model, which has greatly improved the output and economic benefits of aquaculture. However, when people continuously increase the breeding density to pursue the breeding output, a series of common problems have also been caused. Excessive bait feeding reduces the utilization rate of the bait, resulting in an increase in organic matter in the water; increased fertilization, resulting in a sharp increase in nitrogen and phosphorus in the water; a large amount of feces and other organic matter pollute the aquaculture water body; the transparency of the water body decreases and the water quality deteriorates. Excessi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34A23K1/16C12R1/125C02F101/38
Inventor 胡婷马青山贠锦军何增国
Owner BEIJING DABEINONG TECH GRP CO LTD
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