A penta-dog inactivated cell culture vaccine was prepared to protect dogs againstcanine distemper virus, canine parvovirus, canine adenovirus1, 2 and rabies virus.The potency of this vaccine was compared with that of single inactivated vaccinesprepared against each disease, in different groups of susceptible dogs. It was foundthat the protective dose of penta-dog vaccine (2ml) including the protective amounts of the five viral proteins resulted in full protection of vaccinated dogs against the challenge with virulent strain of the used viruses showing no antagonizing effect between each other with and no adverse postvaccinal reaction. So, the prepared inactivated cell culture penta-dog vaccine is a safe and potent vaccine for dogs which resulted in saving time, cost, and effort stress factors on animals and providing good immune statues.

The present study was designed in a trial to use serum replacement instead of thenewborn calf serum in propagation of BHK-21 cell cultures with subsequent reducingthe cost of foot and mouth disease (FMD) vaccine production. Two batches of BHK-21 cell culture were prepared where the medium of the first batch was supplemented with newborn calf serum while the medium of the second batch was supplemented with serum replacement. FMD virus was propagated 7 passages using BHK-21 cell culture. Both virus titration and complement fixation titer (CF) revealed that propagation of FMD virus in cell cultures supplemented with newborn calf serum yields a titre higher than that in case of cells supplemented with serum replacement. Also two batches of FMD inactivated vaccine were prepared from the virus propagated in the two-mentioned cell culture batches. Two groups of susceptible calves were vaccinated with these vaccines. Both of virus neutralization test (VNT) and enzyme linked immunosorbent assay (ELISA) revealed that higher antibody levels were induced in calves vaccinated with the vaccine prepared from cells supplemented with calf serum than those vaccinated with vaccine prepared from cells grown with serum replacement. BHK-21 cell culture supplement with newborn calf serum is most susceptible for FMD virus propagation yielding higher titer of the virus. Moreover, the growth pattern of the used cell culture was much better when the newborn calf serum was used.

Avian coronavirus (AvCoV) infects a range of tissues in chickens and several other avian species. Although the virus can be isolated in chicken embryos, only a few strains of the 6 genotypes/33 lineages can grow in cell lines, with the Beaudette strain (GI-1 lineage) being the most used for in vitro studies. Considering the differences between cell lines and chicken embryos as habitats for AvCoV, this study aimed to assess the diversity of the genes coding for the nonstructural protein 3 (nsp3) and spike envelope protein (S) after serial passages in BHK-21 and Vero cells. After 14 passages of an embryo-adapted Beaudette strain, the virus loads fluctuated in both cell lines, with the highest loads being 8.72 log genome copies/µL for Vero and 6.36 log genome copies/µL for BHK-21 cells. No polymorphisms were found for nsp3; regarding S, not only aa substitutions (Vero: 8th passage A150S, and 14th S150A; BHK-21: 4th S53F, 8th F53Y, and 8th S95R), but also minor variants could be detected on chromatograms with fluctuating intensities. As the regions of these aa substitutions are within the receptor-binding domain of S, it can be speculated that differences in cell receptors between Vero and BHK-21 cells and the speed of cell death led to the selection of different dominant strains, while the stability of nsp3 supports its function as a protease involved in AvCoV replication. In conclusion, AvCoV quasispecies evolution is influenced by the biological model under consideration, and a gradual transition is seen for minor and major variants.

The current study was conducted to determine to what extent human platelet lysate (HPL) can be used to enhance the growth of three cell lines used for propagation of Bovine Ephemeral Fever Virus (BEFV) and Rabies virus (RV) for vaccine production. HPL; Fetal Calf Serum (FCS) and Newborn Calf Serum (NBCS) were used as supplements to the used cell culture medium (Minimum Essential Medium “MEM”) for passaging of three cell lines including baby hamster kidney (BHK-21); African green monkey kidney (VERO) and Madin Derby canine kidney (MDCK). Through three successive cell culture passages; for each cell line; it was found that HPL supported growth and propagation of BHK-21, VERO and  MDCK cell lines without altering their morphologic and functional characteristics. It has been accepted as a suitable alternative to FBS and NBCS, and it can be used in vaccinal production.