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Method for screening and degrading aflatoxin B1 bacteria

A technology of aflatoxin and bacteria, which is applied in the direction of bacteria, biochemical equipment and methods, and microbial measurement/inspection. It can solve the problems of health hazards, high risks, and heavy workload of operators, and achieve high screening efficiency. Inexpensive, highly accurate results

Active Publication Date: 2009-11-18
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detoxification studies of some other bacteria and fungi have shown that the degradation efficiency of most bacteria is not high, and the degradation efficiency is affected by the action time, pH value of the solution, the number of bacteria, the concentration of toxins, and the concentration of metal ions; so far there is no microbial screening method, can efficiently screen aflatoxin-degrading strains
[0003] Screening microorganisms capable of degrading toxins from nature is a time-consuming, laborious and highly dangerous task
Researchers at home and abroad mostly use toxin-containing medium for specific screening of microorganisms. The workload is huge and the efficiency is low. Long-term exposure to toxins is harmful to the health of operators, and toxins cause potential pollution to laboratories and surrounding environments.

Method used

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  • Method for screening and degrading aflatoxin B1 bacteria
  • Method for screening and degrading aflatoxin B1 bacteria
  • Method for screening and degrading aflatoxin B1 bacteria

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Embodiment 1 is used for the screening method of degradation aflatoxin B1 bacterium

[0039] 1) Preparation of medium: use coumarin as the only carbon source, add ammonium nitrate, calcium chloride, and agar to make a barren medium, adjust the pH to neutral; the medium formula is: 0.25gKH 2 PO 4 , 1.0 g NH 4 NO 3 , 0.25g CaCl 2 , 17g agar, 1.0g coumarin, pH7.0; add coumarin after steam sterilization at 121°C for 20min;

[0040] 2) Dilution of flora: Weigh 1g of various samples collected in nature, such as animal feces, moldy grain and feed, soil samples, rotten wood, etc., grind them, and place them in a sterilized 18mm×180mm test tube , add 9 mL of sterile water, seal with a cotton plug, shake overnight; dissolve with sterile water, enrich culture, and dilute;

[0041] 3) Coating of bacterial flora: draw 1 mL of supernatant, and serially dilute 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 , respectively take 0.2mL of each sample solution, and spread it on the flat plate...

Embodiment 2

[0056] Embodiment 2 is used for the screening method of degradation aflatoxin B1 bacterium

[0057] 1) Preparation of medium: use coumarin as the only carbon source, add ammonium nitrate, calcium chloride, and agar to make a poor medium, adjust the pH to be neutral; the medium formula is: 0.5g KH 2 PO 4 , 1 g NH 4 NO 3 , 0.5g CaCl 2 , 15g agar, 1g coumarin, pH 7.0; add coumarin after steam sterilization at 121°C for 20min;

[0058] All the other steps are basically the same as in Example 1.

[0059] 65 strains of bacteria were screened from 20 samples listed in Table 1 by this method. Through further re-screening, 8 strains with degradation activity were obtained.

Embodiment 3

[0060] Embodiment 3 is used for the screening method of degradation aflatoxin B1 bacterium

[0061] 1) Preparation of medium: use coumarin as the only carbon source, add ammonium nitrate, calcium chloride, and agar to make a poor medium, adjust the pH to neutral; the medium formula is: 0.2g KH 2 PO 4 , 0.5g NH 4 NO 3 , 1g CaCl 2 , 15g agar, 0.5g coumarin, pH 7.0; add coumarin after steam sterilization at 121°C for 20min;

[0062] All the other steps are basically the same as in Example 1.

[0063] 79 strains of bacteria were screened from 17 samples listed in Table 1 by this method. Through further re-screening, 14 strains with degradation activity were obtained.

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Abstract

The invention relates to a method for screening and degrading aflatoxin B1 bacteria, comprising the following steps: taking coumarin as the only carbon source for screening a culture medium; culturing a sample; finally, obtaining the aflatoxin B1 bacteria which can be degraded after screening. By adopting the method, the bacteria which can degrade toxins are screened without contact with the extremely toxic substance aflatoxin, so that the method has strong specificity, high screening efficiency, low price and good accuracy, and is applicable to screening and degrading the aflatoxin B1 bacteria on a large scale.

Description

technical field [0001] The invention relates to microorganism screening and purification, in particular to a method for screening highly efficient bacteria degrading aflatoxin B1. Background technique [0002] Since the discovery of aflatoxin in 1960, scientists have continued to study the prevention and detoxification methods of this toxoid. Most of the reports on screening microorganisms for detoxification from various samples in nature were concentrated in the past ten years. There are not many domestic studies, and only Liu Daling has screened out a fungal metabolite that can degrade toxins. According to foreign research reports, the detoxification strains screened out include lactic acid bacteria, bifidobacteria, yeast and white rot fungi, etc., as well as Flavobacterium aurantium, Rhodococcus, Pseudomonas, Rhizopus and so on. Studies on lactic acid bacteria, bifidobacteria and yeast have shown that most of the toxin removal efficiency is below 50%, and the most criti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04
Inventor 计成关舒马秋刚赵丽红王宁牛天贵梁志宏李俊霞
Owner CHINA AGRI UNIV