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Fluoroquinolone antibiotic degrading bacterium and application thereof in compost

A technology of fluoroquinolones and antibiotics, applied in the field of microorganisms, can solve the problems of resistance gene pollution and poor effect

Active Publication Date: 2021-08-27
INST OF SOIL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are few reports on the degradation of antibiotics by microbial agents in composting, mainly because the high temperature generated by composting can inhibit the activity of antibiotic-degrading bacteria, resulting in poor results in practical applications.
In addition, the continued existence of antibiotics will also cause contamination of resistance genes, while antibiotic-degrading strains themselves carry resistance genes, and the strains may also have the risk of resistance gene contamination in application

Method used

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  • Fluoroquinolone antibiotic degrading bacterium and application thereof in compost
  • Fluoroquinolone antibiotic degrading bacterium and application thereof in compost
  • Fluoroquinolone antibiotic degrading bacterium and application thereof in compost

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preparation example Construction

[0039] The present invention also provides a preparation method of the above-mentioned fermented bacterial agent, comprising the following steps:

[0040] The above bacterial strains are inoculated in LB medium and cultured for 24 to 40 hours to obtain the fermentation bacterial agent.

[0041] In the present invention, the temperature of the culture is preferably 20-40°C, more preferably 20-25°C; the culture method preferably further includes: activating in LB liquid medium for 12h, and then activating to the logarithmic phase The bacterium solution was transferred to fresh LB liquid medium again according to the inoculum amount of 1% (v / v), and expanded for 24 hours. The present invention has no special requirements on the source of the LB medium, which is prepared by using commercially available products or preparation methods well known to those skilled in the art.

[0042] The present invention also provides the application of the above-mentioned bacterial strain or the ...

Embodiment 1

[0053] Isolation, purification and identification of strains

[0054] The present invention uses the preparation of substratum as follows:

[0055] Inorganic salt medium: K 2 HPO 4 5.8g, KH 2 PO 4 4.5g, (NH 4 ) 2 SO 4 2.0g, MgCl 2 0.16g, CaCl 2 0.02g, Na 2 MoO 4 2H 2 O 0.0024g, KNO 3 0.0012g, FeCl 3 0.0018g, MnCl 2 2H 2 O 0.0015g, add ultrapure water to 1000mL, adjust pH to 7.0-7.5.

[0056] Separation medium: add 20g agar powder per liter of inorganic salt medium.

[0057] LB liquid medium: tryptone 10.0g, yeast extract 5.0g, sodium chloride 10.0g, add ultrapure water to 1000mL, adjust pH to 7.0-7.5.

[0058] LB solid medium: Add 20g of agar powder per liter of LB liquid medium.

[0059] Acclimatization, isolation and purification of strains:

[0060] First, take about 5g of compost samples (organic fertilizer strips from Jiangxi Zhenghe Ecological Agriculture Co., Ltd.) and add them to inorganic salt medium containing 5 mg / L ciprofloxacin, and put them in ...

Embodiment 2

[0069] Study on Degradation Effect of Brucella WXX-3 on Ciprofloxacin

[0070] Expanded culture of Brucella WXX-3

[0071] First the WXX-3 bacterial strain obtained by screening in Example 1 was activated in LB liquid medium for 12h, then the bacterium liquid activated to the logarithmic phase was transferred to fresh LB liquid medium again according to 1% (volume concentration) inoculum , the number of bacteria reached 2×10 after expanding the culture for 24 hours 10 CFU / mL to obtain the fermentation agent.

[0072] Degradation effect of WXX-3 strain on ciprofloxacin in liquid environment

[0073] 2 treatments are set: 1) CFC: LB liquid medium containing 20mg / L ciprofloxacin; 2) CFC+WXX-3: the bacterial strain after the expansion culture is inoculated to contain 20mg by 1% (volume concentration) inoculum / L ciprofloxacin in LB broth.

[0074] Wrap the reaction bottle with tinfoil to avoid light, and place it in a constant temperature incubator at 30°C and 180r / min for inc...

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Abstract

The invention relates to the technical field of microorganisms, in particular to a Brucella WXX-3 strain and application of the Brucella WXX-3 strain in degradation of fluoroquinolone antibiotics. The preservation number of the strain provided by the invention is CGMCC NO.22259. The WXX-3 strain provided by the invention can efficiently degrade fluoroquinolone antibiotics, and according to the embodiment, the highest degradation rate of ciprofloxacin reaches 44.7% after the strain is added into an LB liquid culture medium containing 20mg / L ciprofloxacin and is cultured in a dark place; besides, the strain is inoculated into livestock and poultry manure for propagation expanding for 8-10 days, then conditioning agents such as wood chips and straw are added for conventional composting for 30 days, and the degradation rate of ciprofloxacin can reach 85.7%; meanwhile, compost prepared from the strain can degrade antibiotic resistance genes, so that the safety of compost products is improved, and the strain is suitable for large-scale popularization and application.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain of fluoroquinolone antibiotic-degrading bacteria and its application in composting. Background technique [0002] In recent years, the risk of antibiotic and resistance gene contamination has attracted more and more attention. By 2030, the consumption of antibiotics in the global livestock and poultry industry will reach 1 million tons. However, most antibiotics are difficult to be absorbed by animal intestines, and about 30% to 90% of antibiotics will be excreted with animal feces and urine. The survey found that antibiotics were detected in animal feces, wastewater, manure and soil, and the concentration could reach mg / kg level. Residues of antibiotics will also lead to the enrichment and spread of resistant bacteria and resistant genes in the environment, increasing the pressure on the ecological environment and the risk of human health. [0003] Composting ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F17/20C12R1/01
CPCC12N1/20C05F17/20Y02W30/40
Inventor 王兴祥张静岳政府周志高丁昌峰
Owner INST OF SOIL SCI CHINESE ACAD OF SCI
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