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Acinetobacter calcoaceticus T32 capable of metabolizing furazolidone and application thereof

A technology of furazolidone and calcium acetate, which is applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problem of less furazolidone, etc., and achieve the effect of fast degradation speed, simple operation steps and low cost

Active Publication Date: 2011-09-14
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, compared with the detection and monitoring of furazolidone, there are few research reports on how to remove furazolidone from the environment and organisms

Method used

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  • Acinetobacter calcoaceticus T32 capable of metabolizing furazolidone and application thereof
  • Acinetobacter calcoaceticus T32 capable of metabolizing furazolidone and application thereof
  • Acinetobacter calcoaceticus T32 capable of metabolizing furazolidone and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Acquisition of Acinetobacter calcium acetate T32 16S rDNA:

[0013] The Acinetobacter calcium acetate T32 was registered and preserved in the General Microbiology Center (CGMCC) of the China Microbial Culture Collection Management Committee; the preservation date was November 06, 2010; the serial number was CGMCC No. 4311. Collect the stable Acinetobacter calcium acetate T32, and extract the genomic DNA of the bacteria. The specific steps are:

[0014] (1) The T32 strain cultured overnight at 30°C was inoculated into a test tube containing 10ml LB medium at an inoculum of 1% (V / V), and the OD of the bacteria 600 When ≈1.0, take 1ml of bacteria liquid in a 1.5ml centrifuge tube and centrifuge at 10,000g for 1min to collect the bacteria.

[0015] (2) Add 1ml of 0.9% NaCl to wash the precipitate and centrifuge at 10,000g for 3min.

[0016] (3) Discard the supernatant, resuspend the pellet with 450μl TE buffer, and add 50μl 20% SDS, gently invert the centrifuge tube upside down to...

Embodiment 2

[0024] The method of using Acinetobacter calcium acetate T32 to treat a solution containing 5ppm furazolidone:

[0025] (1) Microbial strain: Acinetobacter calcium acetate T32 CGMCC No. 4311 was selected.

[0026] (2) Seed culture: select a ring of colonies from the plate and insert it into LB medium, the volume of the shake flask is 30%, the culture temperature is 28-30°C, the culture speed is 180-200 rpm / min, and the culture time is 10-16h. The formula of LB medium is: tryptone, 10g, yeast extract, 5g, sodium chloride, 10g, distilled water, 1L.

[0027] (3) Acclimation culture: Inoculate the T32 in step (2) in an inorganic salt peptone medium at an inoculum of 1% (V / V), and cultivate overnight at 28°C for 10-16 hours. The formula of the inorganic salt peptone culture medium is: Na 2 HPO 4 ·12H 2 O 3g / L, KH 2 PO 4 1g / L, NaCl 3g / L, MgSO 4 0.3g / L, Tryptone 2.5g / L; adjust pH 6.0-8.0.

[0028] (4) T32 degrades furazolidone: the T32 cell in step (3) is set to 1×10 at the initial concen...

Embodiment 3

[0033] The method of using Acinetobacter calcium acetate T32 to treat a solution containing 10ppm furazolidone:

[0034] (1) Microbial strain: Acinetobacter calcium acetate T32 CGMCC No. 4311 is selected;

[0035] (2) Seed culture: Connect the T32 stored in the refrigerator at -80℃ to 10ml of LB medium, shake the bottle with 30% liquid, culture temperature 28-30℃, culture speed 180-200rpm / min, culture time For 10-16h. The formula of LB medium is: tryptone, 10g, yeast extract, 5g, sodium chloride, 10g, distilled water, 1L.

[0036] (3) Acclimation culture: Inoculate the T32 in step (2) with 1% of the inoculum (V / V) in an inorganic salt peptone medium, and cultivate overnight at 28°C for 10-16 hours. The formula of the inorganic salt peptone culture medium is: Na 2 HPO 4 ·12H 2 O 3g / L, KH 2 PO 4 1g / L, NaCl 3g / L, MgSO 4 0.3g / L, Tryptone 2.5g / L; adjust pH 6.0-8.0.

[0037] (4) T32 degrades furazolidone: the T32 cell in step (3) is set to 1×10 at the initial concentration 8 CFU mL -1 I...

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Abstract

The invention relates to the field of treating environment by using microorganisms, in particular to acinetobacter calcoaceticus T32 capable of metabolizing furazolidone, and application thereof. The acinetobacter calcoaceticus T32 has been registered and collected in the China General Microbiological Culture Collection (CGMCC) on sixth, November, 2010; and the collection number is CGMCC No.4311. The acinetobacter calcoaceticus T32 can be used as bactericide for degrading the furazolidone. The furazolidone in the environment is removed by the metabolic process of T32, and the acinetobacter calcoaceticus T32 has the advantages of high degradation speed, low cost, convenience of operation and the like.

Description

Technical field [0001] The invention relates to the field of environmental treatment using microorganisms, in particular to Acinetobacter calcium acetate capable of metabolizing furazolidone and its application. Background technique [0002] Furazolidone is also known as furazolidone, and its molecular formula is C 8 H 7 N 3 O 5 , Is a synthetic broad-spectrum antibacterial drug with the basic structure of 5-nitrofuran. Because furazolidone has the advantages of cheap price and fast drug effect, it is commonly used to prevent and control infections caused by bacteria and original organisms such as cholera, coccidiosis, swine enteritis and turkey blackhead. It also has certain applications in aquaculture. However, studies have shown that furazolidone can cause genetic mutations in experimental animals and induce cancer. The residue of furazolidone in the aquaculture environment causes chronic poisoning of fish and shrimp, which seriously endangers the healthy cultivation of fish....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A62D3/02C12R1/01A62D101/28
Inventor 陈令新张卫卫牛宗亮刘萍徐冬雪
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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