Trichothecene toxin biodegradation agent and preparation method thereof

A technology of trichothecenes and biodegradants, applied in the field of trichothecenes biodegradants and their preparation, to achieve high detoxification efficiency, strong specificity, and high efficiency

Active Publication Date: 2017-06-23
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 that efficiently degrade trichothecenes in feed, optimizing and producing biodegradants that efficiently degrade trichothecenes are important for improving feed utilization and ensuring the safety and economy of animal husbandry. The improvement of efficiency is of great significance

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The screening of embodiment 1 Devosella ANSB714 strain

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

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

[0034] Wherein, the formula of 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 screening of samples: add 100 μl of deoxynivalenol (100 μg / mL) to each seed liquid, use the scr...

Embodiment 2

[0049] The cultivation method of embodiment 2 Devosella ANSB714

[0050] Get Devosella ANSB714 seed liquid 0.5-5ml (being 0.25%, 0.5%, 1%, 2%, 5% and 10% inoculum size), be inoculated in the 50-100ml medium and carry out shaking flask fermentation culture, fermentation The temperature is 16-37°C, the pH value is 6.0-9.0, the rotation speed is 100-200r / min, and the fermentation time is 20-26h. Wherein, the shake flask fermentation medium consists of the following components: tryptone 8-12g, yeast extract 2-5g, sodium chloride 8-12g, water 800-1200mL, pH value 6.0-9.0. Preferably, the shake flask fermentation medium consists of the following components: 10 g of tryptone, 5 g of yeast extract, 10 g of sodium chloride and 1000 mL of distilled water, with a pH value of 7.0. Shake flask fermentation conditions are: fermentation temperature 16-37°C, fermentation time 20-26h, rotation speed 100-200r / min. Preferably, the shaking flask fermentation conditions are: 26.5° C., 120 r / min ...

Embodiment 3

[0052] Application of Example 3 Devosella ANSB714 in the Degradation of Trichothecenes

[0053] Detect the absorbance value at 600nm wavelength of the De Vosella ANSB714 shake flask culture fermentation broth in Example 2, and use the plate count method to detect the number of viable bacteria in the fermentation. At the same time, 980 μl of fermentation broth was drawn and reacted with 20 μl of trichothecenes (NIV, VER and DON) (5000 μg / mL); the control group added 20 μl of trichothecenes (NIV, VER and DON) (5000 μg / mL); the pH value of the reaction system was adjusted to 7.0, and the treatment group and the control group were reacted at 37° C. respectively. Take 0.3mL of the reaction solution and mix it with methanol at a ratio of 1:1, let it stand for more than half an hour, centrifuge at 10,000 rpm for 5min, take the supernatant, pass it through a 0.22μm organic membrane, and measure it on the machine. Determine the remaining amount of toxin, calculate the toxin degradatio...

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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 fields of microbiology and feed additives, in particular to a trichothecene toxin biodegradant and a preparation method thereof. Background technique [0002] Contamination of grain by trichothecenes poses a great threat to human and livestock health. Such toxins are mainly produced by Fusarium, Cephalosporium, Lacquerella, Staphylocera, Trichoderma and other molds, including nearly 150 compounds, which can be divided into four subgroups, of which Class A and Class B are the most important. Type A trichothecenes include: T-2 toxin, HT-2 toxin, verrucarin (VER), fusaric acid (neosolaniol, NEO) and diacetoxy saralenol (DAS); Class B trichothecenes include: deoxynivalenol (DON) and its 3-acetyl or 15-acetyl derivatives, nivalenol (NIV) and fusarenone -X (Fusarenon-X, FX). The chemical structures of the trichothecene mycotoxins produced by different Fusarium species are different, so they can be distinguished according to th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K20/10A23L5/20C12R1/01
Inventor 赵丽红计成马秋刚李笑樱
Owner CHINA AGRI UNIV
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