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Spring water monoucleosis for degrading alflatoxin B1 and ochratoxin A and application of spring water mononucleosis

A kind of spring monas, aflatoxin technology, applied in the direction of bacteria, microbe-based methods, biochemical equipment and methods, etc., can solve the problems of no substantial detoxification significance, rare reports, etc.

Active Publication Date: 2016-01-20
CHACHA FOOD CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the virus-free strains reported so far have the following two common problems in practical application: First, the detoxification mechanism of some virus-free strains belongs to physical adsorption, not substantial biodegradation, and is adsorbed on the cell wall of the bacteria detoxification may occur under different physical and chemical environments in animals or humans, and has no substantial detoxification significance; second, the detoxification rate of the reported strains to toxins is mostly measured at a higher concentration of 100 μg / kg or more. There are few reports on the actual detoxification ability of low-concentration aspergillus toxoids (less than 20 μg / kg) in complex matrices such as food raw materials and feedstuffs

Method used

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  • Spring water monoucleosis for degrading alflatoxin B1 and ochratoxin A and application of spring water mononucleosis
  • Spring water monoucleosis for degrading alflatoxin B1 and ochratoxin A and application of spring water mononucleosis
  • Spring water monoucleosis for degrading alflatoxin B1 and ochratoxin A and application of spring water mononucleosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Screening, identification and cultivation of aflatoxin B1 and ochratoxin A degrading bacteria

[0023] 1. Screening of strains

[0024] The samples were collected from soil heavily contaminated by polycyclic aromatic hydrocarbons (near oil refineries and auto repair shops) and moldy foods heavily contaminated by mycotoxins. Take 0.5 g of the collected sample and add it to 150 mL of enrichment medium (the content of OTA and AFB1 are each about 20 μg / L), and enrich and culture in a constant temperature incubator at 30° C. for 7 days. After the first enrichment culture, take 5mL enrichment culture solution and inoculate 150mL fresh OTA and AFB1 enrichment medium, increase the concentration of OTA and AFB1 to 30μg / L, and continue enrichment culture under the same conditions 7 day. Complete the third enrichment culture experiment with the same enrichment method, and increase the concentration of OTA and AFB1 to 50μg / L. After the enrichment culture is completed, the e...

Embodiment 2

[0034] Example 2 Detecting the degradation characteristics of strain CW282 by high performance liquid chromatography

[0035] Degradation dynamic detection of degrading strain: The degrading strain CW282 was continuously activated on the most suitable solid medium for 2 generations, inoculated into 4mL liquid medium and cultivated overnight to obtain fresh bacterial liquid. Inoculate 50μL of fresh bacterial solution into 4mL OTA test medium (containing 20.0μg / LOTA) and AFB1 test medium (containing 20.0μg / LAFB1) respectively, and culture the inoculated test tube with shaking for 0h, 12h, 24h and 48h, each degraded The experiment is set to 3 replicates. E.coli K12 was used as a negative control strain. After the incubation, mix well, centrifuge at 8000r / min for 10 minutes, and collect the supernatant and the bacterial pellet.

[0036] The degraded supernatant was passed through the OTA immunoaffinity column and the AFB1 immunoaffinity column, respectively, and eluted with methanol....

Embodiment 3

[0041] Example 3 Application of strain CW282 in detoxification treatment of corn soybean meal feed

[0042] 1. Experimental materials

[0043] Strain activation medium I: Tryptone 17.0g / L, Soy peptone 3.0g / L, Glucose 2.5g / L, NaCl5.0g / L, K 2 HPO 4 2.5g / L, agar 20.0g / L.

[0044] Strain activation medium II: peptone 5.0g / L, beef extract 30.0g / L, NaCl5.0g / L, pH7.0-7.2.

[0045] The above-mentioned medium was autoclaved at 120°C for 15 minutes, and the experimental feed was corn soybean meal type diet.

[0046] 2. Experimental method

[0047] Add an appropriate amount of OTA and AFB1 standard stock solution to 50mL of phosphate buffer, and immediately pour it into 100g of the crushed feed sample after mixing. Stir evenly to make the final concentration of OTA and AFB1 reach 40.0μg / kg. Dry in a cool, ventilated place for later use. Continuously activate the tested strains on the solid activation medium I for 2 generations, inoculate the activated strains in 150 mL of liquid activation medium...

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Abstract

The invention provides spring water monoucleosis for degrading alflatoxin B1 and ochratoxin A and application of the spring water mononucleosis and particularly provides a strain of difunctional efficient degrading bacteria (Silanimonas sp.) CW282 and application thereof in degrading low-pollution-concentration alflatoxin B1 and ochratoxin A. Compared with existing aflatoxin degrading bacteria, the strain CW282 can achieve excellent degrading effects under the condition of low-concentration toxin pollution. Under the condition of liquid fermentation, in a fermentation culture solution containing alflatoxin B1 and ochratoxin A with the final concentration being 20 micrograms per liter, the degrading rate of the strain CW282 on ochratoxin A is 70.7% in 24 h and reaches 95.6% in 48 h; the degrading rate of the strain CW282 on alflatoxin B1 is 86.9% in 24 h and reaches 91.3% in 48 h. When the strain CW282 is used for treating fodder (the final concentration is 20 micrograms per kg) polluted by toxins, the degrading rate on ochratoxin A is 53.2% in 48 h, and the degrading rate on alflatoxin B1 is 58.3% in 48 h. The strain CW282 has substantive application value and significance on the application aspects of food and feed biological detoxification.

Description

Technical field [0001] The present invention relates to the field of microbiology and biodegradation, in particular to a spring water monas which can efficiently degrade aflatoxin B1 and ochratoxin A and its application. Background technique [0002] Aspergillus mycotoxins are mainly secondary metabolites produced by Aspergillus flavus, Aspergillus parasitica and Aspergillus ochraceus. More than 20 structural analogues have been isolated and identified, among which aflatoxin B1 (AFB1) and ochratoxin A (OTA) is the most toxic and has the most widespread pollution in agricultural production and food industry. In 1993, aflatoxin was designated by the World Health Organization (WHO) Cancer Research Institute as a Class I carcinogen. Because of its strong carcinogenic, teratogenic and mutagenic properties, it has gradually become the focus of public health and scientific research. Aflatoxin and ochratoxin contamination of grains and oil crops has become a global problem. In animal hu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/01
Inventor 周育王旭姜楠韦朝领
Owner CHACHA FOOD CO LTD
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