1. There are various routes for inoculating eggs with viruses, including the allantoic, chorio-allantoic, yolk sac, and amniotic routes. The allantoic route through the air sac is most common. 2. For the chorio-allantoic route, two small holes are made - one in the air sac and another in the bloodless chorio-allantoic membrane. Air is passed through the first hole to create an artificial air sac space, and then the vaccine is injected. 3. For the yolk sac route, a longer needle is used to inject the virus seed through the air sac and into the deeper yolk sac area.

1. Various routes of egg inoculation are as follows:
1) Allantoic route A/C- Mostcommon route. Keeping the straight with
broad top end and pointed bottom, hole is drilled through air sac by
egg driller, passing the tubercular syringethrough air sac and .1ml
inoculated in allantoic cavity. Later the hole is sealed with WAX.
(Another site of hole can be lateral end of air sac)
2) Chorio- allantoic inoculation: DROP CAM: 2 holes are made , one
hole at air sac and other point is determined at the time of candling
– by slightly tilting the egg find the site of membrane whereno blood
vesselis presentand mark the point.by keep the egg in same tilted
position with a rubber bulb pass the air in firsthole created at the air
sac space, this will help to create a ARTIFICAL AIRSACSPACEatthe
second hole area in the blood less field of chorio-allantoic membrane,
seen as flow of somefluid fromthe second hole. Maintain egg in
same position pass the tubercular needle to inoculate the .1ml of
vaccine seed in the artificial air sac created. Seal the holes with wax
and keep the egg in horizontalposition so as to prevent the leak of
fluid fromthe egg.
3) Yolk sac inoculation: Keep the egg straight, mark the point on air sac
spaceand drill hole, through the air sac pass the 21-22 gauzeneedle
and .25mlof virus seed can be inoculated. Long needle is used as yolk
sac is deep. Seal the hole with wax.
4) Amniotic route: notroutinely performed, it’s guided by Candler.
5) Intra-venous route: For ARTHROPOD BORNEVIRUS VACCINE. Here
the egg shell is skillfully cracked at the site various thevessels streaks
are seen in the candling. Slowly remove the shell, this make the
vesselvisible, apply the paraffin jelly at the site to make the filed
more transparentand clear: for better visualization of vessels. Once
vesselfound the are inoculated.

2. After inoculating the egg, kept in the egg incubator at the specified
temperature and humidity. 24 hrs later the egg is examined by candling for the
meticulous inoculation of egg-checking the egg viability. Every day seen for the
changes which are proceeding. On 4-5 th day eggs are seen for the dead
embryo depicting the virus multiplication