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When the experimental data was analysed as a 23 full factorial design (testing two levels of virus, medium and passage number), the passage number was shown to have the most significant effect on polyhedra yield, while the virus medium interaction had the most important effect on polyhedra biological activity.
The level of the H5-specific ELISA antibodies was much higher in the group that received the H1N1 Nca9 ca virus as the first dose and the H5N1 HK03 ca virus as the second dose than the ferrets that previously received the H3N2 ca04 ca virus and those that did not receive any virus (medium), although the titer was lower than the ferrets that received 2 doses of the H5N1 HK03 ca vaccine (Fig. 4B).
After 4 h of infection, the virus medium was removed.
Like in case of the infection, the growth medium was withdrawn and new medium (virus medium without virus) was added.
Fresh cells were infected 3 times every third day by using virus medium collected from the previous infection.
After washing and addition of the virus medium (not containing virus in case of mock-infection) cells adapt to the new cultivation conditions.
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For this condition no growth was assumed because (1) mock-virus medium does not contain growth factors and (2) the cell monolayer is already confluent (like in the case of a viral infection).
The two swabs were then inserted into one vial containing 2 ml of virus transport medium (Gly medium).
These viruses were propagated in MRC-5 cells Virus maintenance medium (VMM) which consisted of H-MEM medium with 3% FBS, (Moregate, Australia).
Cells were infected in serum-free DMEM over night after which the virus containing medium was removed and replaced with normal medium.
Infection was performed by filtering the virus containing medium, addition of proteamine sulphate, and transferring of the virus containing medium to the target cells which were incubated further for 24 hrs.
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