Trichothecene mycotoxin biodegradation agent and preparation method thereof

A technology of trichothecenes and biodegradants, which is applied in the field of trichothecenes biodegradants and their preparation, and achieves the effects of simple operation, high detoxification efficiency and high efficiency

Active Publication Date: 2014-12-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although some experiments have confirmed that animal intestinal microorganisms can degrade trichothecenes, so far, there are almost no biodegradants that can effectively remove trichothecenes in feed by using these microorganisms. Therefore, looking for new bacterial strains

Method used

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  • Trichothecene mycotoxin biodegradation agent and preparation method thereof
  • Trichothecene mycotoxin biodegradation agent and preparation method thereof
  • Trichothecene mycotoxin biodegradation agent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Screening of Devosella ANSB714 Strain

[0032] Strain screening: screening of strains from animal intestinal contents, moldy feed, moldy food and natural environment (such as soil).

[0033] Seed solution preparation: Mix the animal intestinal contents, moldy feed, moldy food and samples obtained in the natural environment with twice the volume of sterile water, place them in a sterilized 18mm×180mm test tube, shake overnight; 0.1 mL of the supernatant was mixed with 0.8 mL of the screening medium, and cultured in a 30°C incubator with shaking for 12 hours to obtain a seed solution.

[0034] Among them, the formula of the screening medium is: NaH 2 PO 4 0.25g, NH 4 NO 3 1.0g, MgSO 4 .·7H 2 O 0.25g, FeSO 4 0.001g, 17g agar, 2.0g glucose, add distilled water to 1000mL, pH 7.0, steam sterilization at 121°C for 20min.

[0035] Preliminary sample screening: add 100μl of vomitin (100μg / mL) to various sub-liquids, use the non-infected vomitin-containing screening medium as a...

Embodiment 2

[0049] Example 2 Cultivation method of Devosella ANSB714

[0050] Take 0.5-5ml of Devosella ANSB714 seed liquid (ie 0.25%, 0.5%, 1%, 2%, 5% and 10% inoculum), inoculate it in 50-100ml medium for shaking flask fermentation culture, and fermentation The temperature is 16-37°C, the pH value is 6.0-9.0, the speed is 100-200r / min, and the fermentation time is 20-26h. Among them, the shake flask fermentation medium consists of the following components: tryptone 8-12g, yeast extract 2-5g, sodium chloride 8-12g and water 800-1200mL, pH 6.0-9.0. Preferably, the shake flask fermentation medium is composed of the following components: tryptone 10g, yeast extract 5g, sodium chloride 10g and distilled water 1000mL, pH 7.0. The fermentation conditions of the shake flask are: fermentation temperature 16-37°C, fermentation time 20-26h, rotating speed 100-200r / min. Preferably, the shake flask fermentation conditions are: 26.5°C, rotation speed 120r / min, fermentation 24h, and pH value 7.0 during...

Embodiment 3

[0052] Example 3 Application of Devosella ANSB714 in the degradation of trichothecenes

[0053] The absorbance at 600nm of the fermentation broth of Devosella ANSB714 shake flask culture in Example 2 was detected, and the number of viable bacteria in the fermentation was detected by the plate counting method. At the same time, draw 980μl of fermentation broth and react with 20μl of trichothecenes (NIV, VER and DON) (5000μg / mL); the control group is 980μl of fermentation medium and add 20μl of trichothecenes (NIV, VER) And DON) (5000μg / mL); the pH of the reaction system was adjusted to 7.0, and the treatment group and the control group were reacted at 37°C. Pipette 0.3 mL of the reaction solution and methanol 1:1 and mix, let stand for more than half an hour, centrifuge at 10,000 rpm for 5 min, take the supernatant, pass through a 0.22 μm organic membrane, and measure on the machine. Determine the remaining amount of toxins and calculate the toxin degradation rate ( Figure 1-3 ...

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Abstract

The invention provides a feed trichothecene mycotoxin biodegradation agent. Trichothecene mycotoxin comprises but is not limited to nivalenol, verrucarine and deoxynivalenol. By adopting the preparation method provided by the invention, a Devosia sp. ANSB714 for safely and efficiently degrading feed trichothecene mycotoxin is separated and screened out from a natural resource for the first time, and biological characteristics and toxin degradation characteristics of the bacterium are studied. The invention also provides a culture method of the bacterium, a method for preparing a biodegradation agent by using the bacterium, and a method for applying the biodegradation agent to degradation of the trichothecene mycotoxin in a feed. After the ANSB714 reacts with a moldy feed for 24 hours, the degradation rate of the trichothecene mycotoxin in the feed can reach 98%. The Devosia sp. ANSB714 in the invention has high activity, strong specificity and mild effect during degradation of the trichothecene mycotoxin, cannot damage nutritional ingredients in the feed, and improves the utilization ratio of the feed at the same time, so that the safety in production of animal husbandry is ensured and the economic benefit is improved.

Description

Technical field [0001] The invention relates to the field of microbiology and feed additives, in particular to a trichothecenes toxin biodegradation agent and a preparation method thereof. Background technique [0002] The contamination of grains by trichothecenes toxins poses a huge threat to the health of people and livestock. Such toxins are mainly produced by Fusarium, Cephalosporium, Myrothecium, Stachys, Trichoderma and other molds, including nearly 150 compounds, which can be divided into four subclasses, of which Class A and Class B are the most important. Class A trichothecenes include: T-2 toxin, HT-2 toxin, verrucarin (VER), fusaric acid (neosolaniol, NEO) and diacetoxy stubbyenol (DAS); Class B trichothecenes include: Fusarium deoxyquinol (DON) and its 3-acetyl or 15-acetyl derivatives, Fusarium nivalenol (NIV) and Fusarium ketone -X (Fusarenon-X, FX). The trichothecenes produced by different Fusarium species have different chemical structures, so they can be disti...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K1/16C12R1/01
Inventor 赵丽红计成马秋刚李笑樱
Owner CHINA AGRI UNIV
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