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Master the technique of inoculation of Newcastle disease virus in chicken embryo allantoic cavity.
Second, equipment
1. The chicken embryo should use the fertilized egg of the healthy chicken group. The chicken embryo inoculated with the chicken should have no maternal antibody, so as not to affect the proliferation of the virus in the embryo. The embryos used in the experiment mostly used live embryos of 9-10 days old.
2. Inoculation material Newcastle disease I vaccine
3. Various equipment incubators, egg cartons, egg racks, punchers, 1ml tuberculin syringes, 41/2 needles, tweezers, 3% iodine, 75% alcohol cotton balls, pencils, alcohol lamps, scissors , ophthalmic tweezers, sterilized vaccine bottles, sterile droppers and paraffin wax.
Third, content and method
(1) Preparation before vaccination
1. Preparation of chicken embryos
In this experiment, the chicken embryos of 9-10 days old were selected, and the air chamber was drawn with a pencil when the eggs were photographed. In the area where the embryos were active and the blood vessels were obvious below the air chamber, the gap between the blood vessels was marked as the inoculation. The parts; wash the eggshell with 0, 1% chlorhexidine and wipe it dry, then disinfect the inoculation site with 2% iodine, wipe it again with 75% alcohol, and punch the hole in the inoculation, the gas chamber is placed upright On the egg rack.
2. Preparation of virus inoculation materials
The virus material to be isolated and cultured should be sterile. Therefore, for the purpose of sterility of the material, 100-500 IU of penicillin and streptomycin can be added per ml of inoculant, and it can be set to 1 hour at room temperature or 12-24 in the refrigerator. Hourly processed. Or sterilized by a filter.
(ii) Vaccination
Intracavitary injection of chorioallantoic: Newcastle disease virus material was aspirated with a 1 ml syringe with a 6-gauge needle. The needle was inserted into the chicken embryo from the punched hole to 0.5-1 cm, and 0, 1 ml (compared 0 and 5 feathers). ), add zinc oxide tape to seal, add melted paraffin seal, put it back into the incubator for hatching, turn the egg twice a day, look once, and die within 24 hours.
(three) harvest
Newcastle disease I virus material can be harvested 24 to 48 hours after inoculation. Chicken embryos should be stored in a 4 degree refrigerator for 4 hours or overnight before harvest to avoid rupture of blood vessels during dissection. Iodine disinfection of eggshells, use tweezers to remove the egg shell and allantoic membrane, use the suction head to absorb the allantoic fluid in the bottle, collect 5-8ml, sterilize and store frozen.
First, the purpose of the request