A kind of detection method of aflatoxin b1 toxicity

An aflatoxin and detection method technology, applied in the field of aflatoxin B1 toxicity detection, can solve the problems of low single litter size, high research cost, long animal reproduction cycle, etc., achieves easy observation, reduced test cost, The effect of improving efficiency

Active Publication Date: 2018-06-12
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In order to carry out the toxicity research of AFB1, researchers mostly use rats, hamsters, pigs, dairy cows and poultry as the test objects. The research costs are high, the animal reproduction cycle is long, and the single litter size is low, so it is difficult to obtain homogeneous test individuals. , cannot visually display the pathological changes of living tissues and organs, so it is not suitable as an ideal method for AFB1 toxicity detection and research

Method used

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  • A kind of detection method of aflatoxin b1 toxicity
  • A kind of detection method of aflatoxin b1 toxicity
  • A kind of detection method of aflatoxin b1 toxicity

Examples

Experimental program
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Effect test

Embodiment 1

[0076] Zebrafish feeding and management methods are as follows:

[0077] (1) Breed zebrafish in a constant temperature circulating water system. The water used for breeding is tap water heated by aeration. Heating rods are used to heat the water tank to maintain the water temperature at 28±1°C, the dissolved oxygen is greater than 5.0mg / L, and the pH is 7.0± 0.2, light alternately (light 14h / dark 10h). Among them, the feeding bait for adult fish is small tropical fish feed (purchased from Beijing Crazy Aquatic Plant Company) and Artemia worm, and they were fed at 7:00~9:00, 12:00~15:00 and 18:00~20:00 every day. bait; the feeding bait for juvenile fish is paramecium, which is fed once a day.

[0078] (2) Breeding method of Artemia: Add 800mL of water, 3.2g of salt and a little worm into the hatching bottle, put the whole device into the bucket, use a heating rod to heat the water to keep the water temperature at 28±1°C; Inflate the airnia to keep the Artemia eggs in a state ...

Embodiment 2

[0081] Zebrafish embryos were selected as follows:

[0082] (1) At 9:00 the night before egg collection, move zebrafish (female to male ratio 1:3) from the culture tank to the spawning box with a fishing net, and put the spawning box into an artificial climate box (RTOP1000B, Zhejiang Top Instrument Co., Ltd.) overnight, the temperature was controlled at 27±1°C, and the light was turned on at 9:00 the next morning to allow the female and male fish to mate naturally, and fertilized eggs were collected 15 minutes later;

[0083] (2) Place the fertilized eggs in a plate, use a plastic dropper to clean up the feces and impurities, and culture them in an artificial climate box for 3 hours, the temperature is controlled at 27±1°C, and use a dissecting mirror to select the eggs that develop normally 3 hours after fertilization Gastrula.

Embodiment 3

[0085] The preparation of the culture solution with different AFB1 concentrations is as follows:

[0086] (1) Configuration of DMSO mother solution: Take 10 mL of DMSO (analytical pure) in a 4°C refrigerator and put it into a test tube, wrap the test tube with tin foil, store it at room temperature, and use it as a test DMSO mother solution;

[0087] (2) Configuration of DMSO stock solution: pipette 0.5 mL of DMSO mother solution into a centrifuge tube, add 49.5 mL of aquaculture water, and configure 50 mL of DMSO solution with a volume concentration of 1% of DMSO;

[0088] (3) Configuration of AFB1 mother liquor: Take 1mg of AFB1 pure product (99.9%), add 1mL of DMSO mother liquor, place the mixture in an ultrasonic breaker (20-25kHz) for crushing treatment for 10min, shake well, and make AFB1 concentration AFB1 stock solution of 3200μM / L;

[0089] (4) Configuration of AFB1 stock solution: use a pipette to draw 100 μL of AFB1 mother solution into a centrifuge tube, add cultu...

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Abstract

The invention discloses a method for detecting the toxicity of aflatoxin B1. The method comprises the following steps: preparing culture solutions with different aflatoxin B1 concentrations; culturing zebra fish embryos by virtue of the culture solutions with different aflatoxin B1 concentrations for 24-120 hours; and observing the death rate of the zebra fish embryos and the hatching rate, malformation rate, areas of heart cysts and yolk sacs and areas of fish larvae of young zebra fish hatched from the zebra fish embryos, calculating a heart cyst / larva area ratio and yolk sac / larva area ratio, and judging the toxicity effect of the aflatoxin B1 to the zebra fish embryos. According to the method, the aflatoxin B1 is detected by taking zebra fishes as an experiment object, so that a large number of experimental animals with good homogeneity can be rapidly obtained, the experiment cost is lowered, the toxic symptom can be easily observed, the pathological change of each tissue of the fish body can be visually observed in real time, and the detection efficiency, visuality and accuracy of the aflatoxin B1 are improved.

Description

technical field [0001] The invention relates to the technical field of toxicology detection, in particular to a method for detecting the toxicity of aflatoxin B1. Background technique [0002] Aflatoxins (AFs) are secondary metabolites mainly produced by Aspergillus flavus and A. parasiticus in the genus Aspergillus after contaminating grains and grain products. The harmfulness of aflatoxin is that it can damage the liver of human body and animals, and can lead to liver cancer or even death in severe cases. It has been listed as a class I carcinogen by the International Agency for Research on Cancer. According to literature reports, more than 20 types of AFs have been identified, among which aflatoxin B1 (AFB1) is the most common in naturally contaminated food, and its toxicity and carcinogenicity are also the strongest. [0003] Studies have shown that exposure to aflatoxin B1 can cause multiple systemic toxic effects in humans and animals, including digestive system toxic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/50
CPCG01N33/5014
Inventor 熊江林吴灵英王锐刘玉兰周华林
Owner WUHAN POLYTECHNIC UNIVERSITY
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