Method for quantitatively and rapidly screening active materials for inhibiting production of aflatoxin

A kind of aflatoxin and active substance technology, applied in the direction of biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problems of high cost, environmental pollution, human and animal health hazards, etc., and achieve reliable results and safe operation , observe the intuitive effect

Inactive Publication Date: 2015-01-21
HARBIN INST OF TECH AT WEIHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aflatoxins can widely contaminate food and feed, and are extremely harmful to human and animal health. At present, physical and chemical methods are mainly used to remove contaminated aflatoxins from food and feed, which is not only costly, but also unsatisfactory. And it will lead to the loss of nutrition. Therefore, inhibiting the production of aflatoxin in grain and feed has become the first choice for effective prevention and control of aflatoxin pollution.
[0003] The key to effectively inhibiting the production of aflatoxins is to screen out active substances that can efficiently inhibit the synthesis of aflatoxins. Since aflatoxins are extremely toxic fungal metabolites, they cannot be directly observed by the naked eye after they are produced. Relevant instruments and analysis must be used. It is not only cumbersome to detect the production of toxins, but also easy to cause environmental pollution and endanger the health of operators. Therefore, it is necessary to establish a trace, fast, safe and low-cost method to screen out active substances that can effectively inhibit aflatoxins. method is very important

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Screening of microbial fermentation products that inhibit aflatoxins

[0017] The cell-free fermentation supernatant stock solution containing microbial fermentation products was added to the 2-fold concentrated liquid medium at a volume ratio of 1:1 (supernatant stock solution: liquid medium) as the treatment group, and the control group used sterile water instead of microorganisms The cell-free fermentation supernatant stock solution of the fermentation product; after aseptic operation and mixing, take 200 microliters and add it to a micro-liquid incubator, and inoculate 5 microliters of Aspergillus parasitica DM strain into the micro-liquid incubator of the treatment group and the control group respectively. The conidia suspension was cultivated for 3 days in a constant temperature incubator at 28 degrees; the growth situation of the DM bacterial strain and the production of orange-red scatter-coated acid in the treatment group and the control group were vi...

Embodiment 2

[0019] Embodiment 2: Inhibit the screening of the chemical synthesis product of aflatoxin

[0020] The three chemically synthesized compounds, carbon-12, carbon-14 and carbon-16, were prepared with sterile water into solutions of appropriate concentration, added into the liquid medium aseptically, and prepared to contain drug concentrations of 0, 6.25, and 12.5 , 25, 50 and 100ppm liquid culture medium; get 500 microliters and join in the microincubator, inoculate 10 microliters of the conidia suspension of Aspergillus parasitica DM strain, cultivate 5 days in the constant temperature incubator of 28 degrees; Observe the growth situation of the DM bacterial strain in the micro liquid culture device of different drug concentrations and the production situation of the orange-red scatter-coated acid, put the micro liquid culture device into the centrifuge, centrifuge at 1000 rpm for 1 minute, and place the Put the mycelium into methanol / sodium hydroxide (volume to weight ratio: 9...

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Abstract

The invention relates to a method for quantitatively and rapidly screening active materials for inhibiting the production of aflatoxin, which comprises the following steps: adding a microbial fermentation product, a plant extract, a natural product, or a chemical synthetic product with certain mass or concentration into a liquid culture medium, taking 200-500 microliters of obtained liquid, and adding the taken liquid into a micro-liquid incubator; by taking a blank nutrient solution as a contrast, inoculating spore suspension of 5-10 microliters of aspergillus parasiticus DM strains, culturing for 3-6 days at the temperature of 28 DEG C, visually inspecting the mycelial growth of the DM strains and the production of orange norsolorinic acid, and centrifuging the micro-liquid incubator with a visually inspected inhibition effect for 1 minute at a rotating speed of 1000 rpm; weighing the fresh weight of a mycelium, putting the mycelium into a methanol/sodium hydroxide solution, extracting norsolorinic acid therein for 30-40 minutes, measuring substances at 560 nm, and comparing the measured values with a light absorption value of an extracting solution; calculating a detoxification rate and a bacteriostasis rate; and selecting the substances with an inhibition rate of 90-100% as active materials for inhibiting the production of aflatoxin. The method disclosed by the invention can realize the trace, intuitive, quick, safe, and low-cost screening of active materials for efficiently inhibiting the production of aflatoxin.

Description

technical field [0001] The invention relates to the technical fields of natural products, mycotoxin prevention and control, etc., and specifically relates to a method for quantitatively and rapidly screening active substances that inhibit aflatoxin production. Background technique [0002] Aflatoxin is a toxic secondary metabolite produced by fungi such as Aspergillus flavus and Aspergillus parasiticus. It is the most toxic mycotoxin found in nature. Its acute toxicity is more than 60 times that of arsenic. It is also highly carcinogenic and immune. Inhibitory and teratogenic, it is listed as a class of carcinogens by the International Agency for Cancer. Aflatoxins can widely contaminate food and feed, and are extremely harmful to human and animal health. At present, physical and chemical methods are mainly used to remove contaminated aflatoxins from food and feed, which is not only costly, but also unsatisfactory. And it will lead to the loss of nutrition. Therefore, inhib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/18
Inventor 闫培生王凯曹立新高秀君吴晗琪
Owner HARBIN INST OF TECH AT WEIHAI
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