A norfloxacin-degrading Acinetobacter calcoacetate nor‑36 and its application

A technology of NOR-36 and norfloxacin, which is applied in norfloxacin to degrade Acinetobacter calcoaceticus NOR-36 and its application field, can solve the problems affecting the development of animal husbandry and aquaculture industry, the impact of environmental microecology, etc. Achieve the effect of improving the quality and safety of the water environment, good removal effect, and protecting the ecological environment

Active Publication Date: 2017-12-08
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, antibiotics are refractory to biodegradable substances. Once antibiotic pollution is caused, it is easy to accumulate in the environment, induce drug-resistant strains, and have a serious impact on the environmental microecology.
[0003] Fluoroquinolones have been widely used in my country's animal husbandry and aquaculture industry, but in use, it was found that fluoroquinolones will produce a series of adverse reactions, such as skin allergies and photosensitivity reactions, cartilage toxicity, circulatory system reactions, central nervous Systemic (CNS) reactions, gastrointestinal reactions, etc., have directly affected the development of animal husbandry and aquaculture

Method used

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  • A norfloxacin-degrading Acinetobacter calcoacetate nor‑36 and its application
  • A norfloxacin-degrading Acinetobacter calcoacetate nor‑36 and its application
  • A norfloxacin-degrading Acinetobacter calcoacetate nor‑36 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Norfloxacin degrades the screening and isolation of Acinetobacter calcoacetate NOR-36

[0026] Get the activated sludge 3.0ml that obtains from the wastewater treatment tank of a certain pesticide factory production workshop that produces norfloxacin and add 100ml norfloxacin concentration and be in the LB medium containing piperazine of 50mg / L, its formula is: Contain 5g / L piperazine, NaCl10.00g / L, peptone 10.00g / L, yeast powder 5.00g / L, pH 7.0, shake culture at 160r / min, 30℃, inoculate with 3% every 7 days The amount was transferred to fresh inorganic salt medium, and transferred 5 times in a row.

[0027] Take 1.0ml of the enriched bacteria solution obtained above, add 9.0ml of sterile water, and make 10 -1 The rich solution, and then draw 1.0ml prepared 10 -1 The enrichment solution was added to 9.0ml sterile water, mixed thoroughly to make 10 -2 The enrichment solution, and so on, carry out gradient dilution on the enrichment solution. The dilution so...

Embodiment 2

[0029] Example 2 Shake Flask Degradation Experiment of Acinetobacter Calcoacetate NOR-36

[0030] 2.1 The effect of inoculum size on the degradation efficiency of NOR-36

[0031]In the inorganic salt medium containing 500ug / L norfloxacin, the formula is: every liter contains 500ug / L piperazine, 1.50 gK 2 HPO 4 , 0.50g KH 2 PO 4 , 0.20g MgSO 4 ×7H 2 O, 1.00g NaCl, 1.00g (NH 4 ) 2 SO 4 , pH 7.0, inoculate NOR-36 in the culture medium according to the inoculum amount of 1%, 2%, 3%, 4%, 5%, shake culture at 30°C, and take samples for measurement after 72 hours. The result is given by figure 2 It can be seen that when 1% is inoculated, the degradation rate is 91%. With the increase of the inoculum amount, the degradation rate of norfloxacin increases slightly. Considering the reasons of bacterium growth and economy, the inoculum amount of 1% is adopted The requirement can be met.

[0032] 2.2 The effect of pH on the degradation efficiency of NOR-36

[0033] The pH valu...

Embodiment 3

[0035] Embodiment 3 Containing the preparation of calcium acetate Acinetobacter NOR-36 bacterial agent

[0036] The original species of norfloxacin-degrading calcoacetic acid Acinetobacter NOR-36 screened and isolated in Example 1 was activated on a petri dish, and its degradation performance was measured, and it was inoculated on the inclined surface of the test tube for subsequent use. The test tube was inoculated in a 1000ml shake flask containing 200ml of LB medium containing piperazine. The formula of LB medium was: piperazine 5g / L, yeast extract 5.00g / L, peptone 10.00g / L, NaCl10.00g / L, pH 7.0, constant temperature shaking culture to logarithmic phase, ready to inoculate seed tanks. The seed tank is 500 liters, the feeding amount is 400 liters, the medium formula is: piperazine 5g / L, glucose 8g / L, (NH 4 ) 2 SO 4 1g / L, K 2 HPO 4 2g / L, MgSO 4 0.5g / L, NaCl1g / L%, CaCO 3 0.5g / L, yeast extract 2g / L, pH 7.2-7.5. After the feeding is completed, 121°C high-pressure dam...

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Abstract

The invention provides a degrading bacterial agent for eliminating norfloxacin residues in waste water, which belongs to the technical field of biological high technology. The strain used was Gram-negative bacteria NOR‑36, which was identified as Acinetobacter . calcoaceticus. It was deposited in the General Microbiology Center of China Microbial Cultures Preservation Management Committee (CGMCC for short, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Zip code 100101), on February 9, 2015, with a deposit number CGMCC NO.10540, identified as Acinetobacter calcoaceticus. The main biological characteristics are negative Gram stain, smooth, short rod-shaped colonies, positive for contact oxidase, and negative for nitrate reductase. The direct application of degrading bacteria products can reduce the residual amount of norfloxacin in water by more than 90%.

Description

technical field [0001] The invention belongs to the technical field of biological high technology, and in particular relates to a norfloxacin-degrading Acinetobacter calcoacetate NOR-36 and its application. Background technique [0002] Veterinary antibiotics refer to antibiotics used in animal feed at sub-therapeutic doses to protect animal health, promote animal growth and production, and improve feed utilization. Antibiotics themselves are harmless to the human body, but if used in excess, pathogenic bacteria will develop drug resistance, causing antibiotics to fail. The excessive use of antibiotics has caused drug resistance and drug residues in animal products to become serious problems that cannot be ignored, especially the destruction of animal immune functions. At the same time, antibiotics are refractory to biodegradable substances. Once antibiotic pollution is caused, they are easy to accumulate in the environment, induce drug-resistant strains, and have a serious...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/38
CPCC02F3/34C02F2101/38C12N1/20C12N1/205C12R2001/01
Inventor 蔡天明杜雨昕陈立伟杨倩蔡舒
Owner NANJING AGRICULTURAL UNIVERSITY
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