Sheep pox virus (SPV) is a member of the genus Capripoxvirus, of family Poxviridae, which affect sheep and goats causes significant economic losses. The present study was applied for isolation and identification of Sheep pox virus from clinically affected sheep during 2017 to 2019 in five Egyptian governorates,
The need to economic, sensitive and easily maintained cell cultures use for massive production of fowl pox vaccine was necessary. We adapted Fowl pox virus (FPV) Baudette strain on Primary chicken embryo fibroblast (CEF), Baby Grivet Monkey Kidney cell line (BGM) and African green monkey kidney (Vero) cell lines. The highest virus titers were 10 6.5 after the 9 th passage in CEF and 10 6.2 after the 12 th passage on BGM while Vero cells were abortive to the virus. BGM cells were selected for study cytopathic effect (CPE) and growth kinetic as it easily maintained than primary CEF cells. The characteristic CPE were rounding till 6 th passage with intracytoplasmic inclusion by the 7 th passage and syncytium formation appeared from the 9 th passage till the 13 th passage. The virus was highly cell associated for the first 84h post inoculation(PI) with a titer of 10 4.5 then a maximum titer was 10 6.2 in cell free portion after 120h PI. In conclusion, BGM were considered a new susceptible cell for growth of FPV with best harvesting time 120h PI to obtain a maximum titer for subsequent vaccine production.
Although primary lamb testis (LT) cell culture seemed to be sensitive and the most suitable for adaptation of recently isolated sheep poxvirus (SPV), giving rapid and complete Cytopathic Effect (CPE), these cells were difficult to be maintained as a monolayer cell line. Moreover, their primary and secondary cultures still have many of the detrimental, inherent characteristics and contaminating elements. Therefore, the use of more stable cell lines (Vero cells) becomes a necessity for an economic preparation of SPV seed vaccine. Three SPV isolates, which previously passaged and adapted in lamb testis cell culture (isolates with titer reaching to 6.5 Log10 TCID50 / ml), were chosen to be transferred and propagated on Vero cell line. With further studies on its growth curve and growth kinetics to identify the growth behavior of these isolates, followed by detection of the virus antigen in the infected Vero cells by indirect florescent antibody technique (IFAT) as the first step for antigenic identification of the isolated adapted virus. The chosen optimum multiplicity of infection (MOI) was (0.01) with the optimum virus harvestation time was 4 days post-inoculation (DPI) and virus titre was equal to 10 5.5 . Finally, we recommend the use of Vero cells as a continuous cell line for preparation of seed of SPV prepared from an Egyptian isolate.
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