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.
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).
Allantoic fluid (AF) from two eggs selected randomly was harvested at an hourly interval and stored at -70[degrees]C.