Cytopathic effect was evident at 48-72 hours after inoculation of LMH and Vero cell cultures with the allantoic fluid
from dead embryos.
Biochemical profile of ovine amniotic and allantoic fluids
. Tamilnadu J.
As shown in Figure 2a, the virus sample prepared by centrifugation at sucrose chosen was found to be very pure compared to infected crude allantoic fluid
. The transmission electron microscopic observation (Jeol T-100) of the negative stained virus samples supported the purity of the viruses and gave information about the morphology of purified virus particles (Figure 2b).
In allantoic fluid
, lactic acid was only found after day 9 of incubation.
The 44th passaged virus was inoculated into five 9-11 days old embryonated SPF fowl eggs and allantoic fluids
were harvested after 5 days of post inoculation.
Spot Agglutination Test: A drop of allantoic fluid
was placed on a glass slide and mixed with an equal volume of 5% freshly prepared washed chicken RBC's (Tiwari et al, 1996)
A correlation coefficient was employed to diagnose relationship between volume of allantoic fluid
and initiation of calving.
Infectious allantoic fluid
([10.sup.6] [EID.sub.50]) generated using each type of reassortant was intranasally administrated to three chickens each.
The reference strains of influenza A virus (A/PR/8/34-H1N1 and A/Udorn/307/72-H3N2) procured from CDC, Atlanta, GA, USA, and the pandemic H1N1-2009 (A/Delhi/1/2009-H1N1) isolated at our laboratory, to serve as positive control, were propagated to sufficiently high titres in 10-days old embryonated chicken eggs and the virus-containing allantoic fluid
was harvested and stored in -80[degrees]C until use.
The allantoic fluid
, about 5 mL in quantity was aspirated from every embryonated egg in to a 10 mL pipette and poured in to a sterilized ampoule at - 20oC (Arshad et al., 1994).
(AF) from two eggs selected randomly was harvested at an hourly interval and stored at -70[degrees]C.