Bacillus and application thereof in nitrogen removal and sulfur removal of culture water body

A technology of bacillus and breeding water, which is applied in the field of environmental microorganisms and environmental engineering, can solve the problems of high toxicity, and achieve the effects of low cost, strong tolerance and simple operation

Active Publication Date: 2020-05-05
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is generally believed that nitrogen and nitrate nitrogen in water bodies are harmless to aquatic animals, but higher concentrations of ammonia nitrogen and nitrite nitrogen are highly toxic to aquatic animals

Method used

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  • Bacillus and application thereof in nitrogen removal and sulfur removal of culture water body
  • Bacillus and application thereof in nitrogen removal and sulfur removal of culture water body
  • Bacillus and application thereof in nitrogen removal and sulfur removal of culture water body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: The isolation and screening of the Bacillus sp. SC16

[0041] The sample is the bottom sludge taken from aquaculture fish ponds. From the bottom sludge sample, 5 mL of the mud-water mixture is transferred to 95 mL of sterilized DM enrichment medium. Then put it into a 28°C constant temperature shaker at 200rpm for enrichment culture for 24h. Continuous enrichment culture was performed three times, and the third enrichment solution was diluted 10-fold and evenly spread on bromothymol blue (BTB) solid medium (agar 20g / L, see below for other components). After culturing in a constant temperature incubator at 28°C for 3 days, pick the strains with blue halos in the surrounding medium to LB medium. It was purified by streaking on the LB medium and saved by numbering. After preliminary screening, 19 strains were obtained, including one strain SC16.

[0042] The formula of DM enrichment medium is: KNO 3 0.5g / L, CH 3 COONa 2.0g / L, MgSO 4 ·7H 2 O 0.6g / L, CaCl 2 ·2H 2 O ...

Embodiment 2

[0046] Example 2: Morphological identification and phylogenetic tree analysis of strain SC16

[0047] (1) Morphological identification

[0048] Use an inoculation loop to pick the seed solution of the strain SC16 stored in Example 1 and streak it on the LB solid plate, and culture it at 28°C for 24 hours. Take a single colony for Gram staining, and observe under a microscope after staining. At the same time, a single colony was picked up in LB liquid medium for expansion culture. Take the LB solid plate, spread a layer of transparent sterile cellophane on the plate, dilute the above seed solution ten times with phosphate buffered saline (PBS), take the diluted solution and coat the plate on cellophane, and incubate at 28°C for 24h. The cellophane was sliced ​​(1cm×1cm), and it was fixed in 2.5% glutaraldehyde for 1.5h. Take the fixed samples and wash them with 30%, 50%, 70%, 90%, 100% absolute ethanol for 10 minutes. Take the eluted sample and vacuum dry for 2h. Take the dried...

Embodiment 3

[0058] Example 3: The aerobic denitrification performance experiment of Bacillus sp. SC16

[0059] The seed solution of Bacillus sp. SC16 stored in Example 2 was picked with an inoculating loop, streaked on the LB solid plate, and incubated at 28°C for 24 hours. Pick a loop of SC16 colonies from the LB plate into the LB liquid medium, culture it on a shaker at 28°C at 200rpm for 24h, wash the bacterial solution by centrifugation (6500rpm, 4°C) with PBS buffer, and inoculate at a ratio of 10% (volume ratio) To DM-1, DM-2 and DM-3 liquid medium (the nitrogen source in DM-1 is KNO 3 0.5g / L, the nitrogen source in DM-2 is NH 4 Cl 0.5g / L, the nitrogen source in DM-3 is NaNO 2 0.3g / L), shake culture at 28°C and 200rpm. Sampling every 12h to determine nitrate nitrogen Nitrite Ammonia nitrogen And bacteria density (OD 600 ), by analyzing the removal rate of the three nitrogen sources to judge the aerobic denitrification performance of the strain. by Figure 4 , Figure 5 with Imag...

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Abstract

The invention discloses a bacillus and an application thereof in nitrogen removal and sulfur removal of a culture water body, and belongs to the technical field of environmental microorganisms and environmental engineering. The strain is named as (Bacillus sp.)SC16, and has a preservation number of GDMCC No:60942. The strain is obtained by screening bottom mud of an aquaculture pond, do not have ecological safety risk when being applied to aquaculture water again, and accords with biological safety regulations. The strain has autotrophic and denitrification performance, can utilize inorganic carbon as a unique carbon source and utilize ammonia nitrogen, nitrate nitrogen or nitrite nitrogen as a unique nitrogen source for metabolism, and thus achieves the purpose of removing the nitrogen source; the strain has sulfur removal performance, can utilize sulfides as an electron donor to oxidize the sulfide to obtain energy; and meanwhile, the strain has strong tolerance to nitrite and has good growth. The strain can be prepared into a microecological preparation to be applied to nitrogen removal and sulfur removal treatment of a culture water body, has a wide application prospect, and has good social benefits.

Description

Technical field [0001] The invention belongs to the technical field of environmental microorganisms and environmental engineering, and relates to a bacillus with denitrification and sulfur removal functions and its application in denitrification and sulfur removal in aquaculture water. Background technique [0002] In recent years, my country's aquaculture industry has developed rapidly, and the scale of intensive aquaculture and high-density aquaculture has continued to expand. In order to pursue higher economic benefits, a large amount of feed will be put in. Concentrated feeding of bait will cause a large amount of residual bait, animal manure and dead animal carcasses to accumulate in the aquaculture water, resulting in the nitrogen content in the water exceeding the water quality standard, severely destroying the ecological environment of the aquaculture pond, and the decomposition of microorganisms will also produce hydrogen sulfide Gas, ultimately causing economic losses....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/07
CPCC02F3/34C12N1/20C12N1/205C12R2001/07
Inventor 张其中桑池国
Owner JINAN UNIVERSITY
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