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Method for detecting toxicity of aflatoxin B1

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

Active Publication Date: 2017-12-12
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology uses tiny creatures called Zebrafixia (a type of bacterium) that are found naturally on seafood around 4000 years ago. These small organisms have been shown to be able to detect harmful chemicals like pesticides or carbamates from food sources such as peanuts during production processes. They also help researchers study how these substances affect their reproductive health by observing them under different conditions.

Problems solved by technology

This patents describes different methods used during testing for detecting chemical substances called flutaxes, such as arboviruses like yellow fever virus (YFV), bacteria like Bacillaria spp., fungi-like organisms, amoeba spirochetes, nematodes, yeast cells, protozoan insects, seafood fish, shellfish, mice, dogs, horseshoe shoe feet, whales, birds, rodents, pollen, worms, plants, algae, microorganism, and other similar species. However, these techniques require expensive laboratory equipment with trained operators who may lack confidence due to their subjectivity.

Method used

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  • Method for detecting toxicity of aflatoxin B1
  • Method for detecting toxicity of aflatoxin B1
  • Method for detecting toxicity of aflatoxin B1

Examples

Experimental program
Comparison scheme
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

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Claims

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Application Information

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Owner WUHAN POLYTECHNIC UNIVERSITY
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