Viral Vaccine: Eggs Drilling, Virus Inoculation and Vaccine Harvesting
Before proceeding to the inoculation process, the eggs have to be drilled to make a tiny hole in order to let the syringe penetrate the egg shell. A small electric rotary (driller) is used to drill a tiny hole on the drilling point that was marked during the marking process. The hole is made by slowly touch the rotary part of the driller on the drilling point. This step has to be done very carefully without breaking the inner membrane of the eggs.
Immediately after the eggs are drilled, the eggs will be transfered to the clean room (Egg Laboratory) for inoculation process through the passbox system. Optimal volume of 0.2ml inoculums is injected per egg. The syringe was thrust down vertically through the hole on the shell at 45° and the inoculum is discharged into the allantoic cavity. The hole is immediately sealed with melted paraffin wax and petroleum jelly to prevent contamination. Then the eggs are incubated for a few days depending on the virus used. After the incubation, the eggs have to be chilled in cold room (+4oC) for 24 hours in order to harden the blood vessel. This step must be done because it can make the process harvesting easier.
The harvesting step is to collect the allantoic fluid of the eggs. The eggs first have to be swabbed with iodine containing alcohol around the sealed tiny hole in order to further sterilize the outer surface of the eggs. Then, the egg’s shell is broken with forceps according to the air sack marking. The allantoic will be harvested into the sterilized bottle of 500ml. 10ml of the antibiotic will be added in the bottle and 3ml allantoic fluid from each bottle will be taken out for sterility test. All the bottles will then send to the cold room at +4oC.
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