Aspergillus fumigatus agar beads and preparation method thereof

A technology of Aspergillus fumigatus and its production method, applied in the direction of microorganism-based methods, biochemical equipment and methods, fungi, etc., to achieve the effects of no significant difference in feed intake and weight gain, convenient application, and high success rate

Inactive Publication Date: 2018-09-25
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are no reports on whether Aspergillus fumigatus can colonize the lungs through breathing, how long the colonization time is, and how to imitate the natural state to establish the colonization of Aspergillus fumigatus instead of an infection model. In addition, the

Method used

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  • Aspergillus fumigatus agar beads and preparation method thereof
  • Aspergillus fumigatus agar beads and preparation method thereof
  • Aspergillus fumigatus agar beads and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] (1) Preparation of Aspergillus fumigatus spore suspension

[0046] Aspergillus fumigatus (strain number CCCCMIDA1, purchased from the Institute of Dermatology and Pathology, Chinese Academy of Medical Sciences) was picked with a metal inoculation loop and inoculated on potato dextrose agar medium (PDA), and cultured at 37°C for 4 days. Wash the medium with phosphate buffer containing 0.1% Tween 80, and collect the spore suspension, 10000rpm, centrifuge for 3min, remove the supernatant, then resuspend with phosphate buffer containing 0.1% Tween 80, and finally use 8 Filter through a layer of sterile gauze. Count with a hemocytometer under a microscope, and adjust the concentration of the concentrated spore suspension to 1.0×10 9 CFU / ml.

[0047] (2) Preparation of Aspergillus fumigatus agar beads

[0048] Prepare 100ml of potato dextrose agar medium, sterilize at 121°C for 15min, and autoclave. Take 52°C (the choice of 52°C mainly considers two factors, one is that t...

Embodiment 2

[0051] Example 2: Construction of a chick respiratory tract model colonized with Aspergillus fumigatus

[0052] 1. Test animals:

[0053] 60 SPF chickens, 9 days old, were purchased from Jinan Spafasi Poultry Co., Ltd., and were raised in the animal room of the Animal Science and Technology College of Shandong Agricultural University. Aspergillus fumigatus agar beads (prepared in Example 1) were inoculated by intratracheal injection with a dose of 300 μl.

[0054] 2. Construction method:

[0055] Hold the 14-day-old chick, stand up and open its mouth, and fully display the position of its oropharynx and larynx under sufficient light. The operator uses a 1ml syringe to draw 300 μl of Aspergillus fumigatus agar beads (prepared in Example 1), and uses a No. 8 curved feeding needle. When the chick’s breathing throat is open, quickly insert the feeding needle into the trachea through the throat, and the insertion depth is Slowly inject 300 μl of Aspergillus fumigatus agar beads ...

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Abstract

Aspergillus fumigatus agar beads and a preparation method thereof are disclosed. Firstly, Aspergillus fumigatus spore suspension is prepared, and is mixed with a potato dextrose agar medium; the mixture is added into a container containing mineral oil and is stirred and centrifuged to form beads; then a sterile phosphate buffer solution is added for washing; centrifugation for oil removing is performed; beads the diameter of which is smaller than 280 [mu]m are screened; and the spore concentration in the agar beads is adjusted to 5*10<6> CFU/mL. The prepared Aspergillus fumigates-containing agar beads are applied for testing animals, compared with control groups, no abnormal clinical symptom is showed in the test, and food intake and weight gain differences are not significant. The agar beads achieve the objective of Aspergillus fumigates inplanting in the lung, are convenient to apply and free of adverse clinical manifestations, can be used for building a poultry Aspergillus fumigatesin-lung inplanting model, and provide a premise for further research of poultry aspergillosis and interactions with the body.

Description

technical field [0001] The invention relates to the technical field of animal experiment products, in particular to an Aspergillus fumigatus agar bead and a preparation method thereof. Background technique [0002] Aspergillus fumigatus (Aspergillus fumigatus) belongs to the genus Aspergillus and is the most common conditional pathogenic bacteria in molds. The particle size is about 2-3μm, and can enter the deep part of the respiratory tract and alveoli with breathing. [0003] In livestock and poultry farming, due to negligence in feeding and management, feed and litter are prone to mold and produce mold. Among them, Aspergillus fumigatus is the most common. The spores produced by it are dispersed in the air and can be inhaled by animals. Normal animals generally do not cause breathing after inhaling spores. Systemic infection, but in immunocompromised individuals, may cause aspergillosis characterized by lung and air sac infection, with chicks and geese being most suscept...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12R1/66
CPCC12N1/14
Inventor 蔡玉梅庄宏旭刘玉庆宋志刚刘传德
Owner SHANDONG AGRICULTURAL UNIVERSITY
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