Acinetobacter calcoaceticus NOR-36 for degradation of norfloxacin and application of acinetobacter calcoaceticus NOR-36

A technology of NOR-36 and norfloxacin, applied in the direction of bacteria, biological water/sewage treatment, microorganisms, etc., can solve problems affecting the development of animal husbandry and aquaculture, environmental micro-ecology, etc., to improve the quality and safety of water environment , Good removal effect, low production and use cost

Active Publication Date: 2015-08-26
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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  • Acinetobacter calcoaceticus NOR-36 for degradation of norfloxacin and application of acinetobacter calcoaceticus NOR-36
  • Acinetobacter calcoaceticus NOR-36 for degradation of norfloxacin and application of acinetobacter calcoaceticus NOR-36
  • Acinetobacter calcoaceticus NOR-36 for degradation of norfloxacin and application of acinetobacter calcoaceticus NOR-36

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, NaCl 10.00g / 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 s...

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, its formula is: every liter contains 500ug / L piperazine, 1.50 g K 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, NaCl 10.00g / 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, NaCl 1g / L%, CaCO 3 0.5g / L, yeast extract 2g / L, pH 7.2-7.5. After the feeding is completed, 121°C high-pressure d...

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Abstract

The invention provides a degrading strain capable of eliminating norfloxacin left in wastewater and belongs to the technical field of high techniques for biology. The strain is gram staining reaction negative bacteria NOR-36 and is identified as acinetobacter calcoaceticus. The acinetobacter calcoaceticus is preserved in CGMCC (China general microbiological culture collection center) in Microbiology Research Institute of China Science Academy in No.3, yard No.1, Beichen western road, Chaoyang district, Beijing, the postcode is 100101, the collection number is CGMCC No.10540, and the strain is identified as acinetobacter calcoaceticus. The main biological character is gram-negative, the bacterial colony is smooth, and the strain is short-rod-shaped and is positive when contacted with oxidase and negative when contacted with nitrate reductase. The degrading product can be directly applied and can reduce the amount of residual 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 serio...

Claims

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Application Information

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