Some observations on the isolation and cultivation of avian reoviruses

Abstract

SUMMARYFor the cultivation of avian reoviruses a comparison was made of cell cultures of chick embryo fibroblasts, lungs, liver, kidney and chicken kidney and fertile eggs inoculated via the yolk sac, chorioallantoic membrane or allantoic cavity. Chick embryo liver was found to be the most sensitive cell culture method and the yolk sac the best inoculation route for eggs. Chicken kidney cells were second to embryonic liver cells, but peak titres of cell-bound and cell-free virus occurred later than in chick embryo liver.Using the plaque technique, minimal adsorption times for reovirus in embryonic liver and chicken kidney cells were found to be 30 and 45 min respectively. .Plaques produced by six different strains of reovirus in embryonic liver cells were larger and appeared sooner than in chicken kidney cells.

Adaptation and characteristics of replication of a strain of avian reovirus in vero cells

“…Avian reoviruses, however, have a predilection for primary cultures of chicken cells, particularly chicken kidney and chicken embryo liver cells (Guneratne et al, 1982), and these ceE types are commonly used for virus isolation and replication. There are a number of disadvantages in using these cells: due to the possible contamination with vertically transmitted avian reoviruses (Mustaffa-Babjee and Spradbrow, 1971;van der Heide and Kalbac, 1975) the cells need to be prepared from specific-pathogen-free (SPF) chickens or chicken embryos; due to the heterogenous nature of the cells and the apparent preference of avian reoviruses for cells of epithelial origin, consistent titration of infectious virus is difficult due to variation between batches of primary, cells, and early detection of a cytopathic effect (CPE) is difficult (Robertson and Wilcox, unpublished observations).…”

A survey of the viral flora of two commercial Pekin duck flocks

“…In contrast, EDS-76 virus produced haemagglutinins in both allantoic and cell culture fluids. CELi is a system which has been particularly conducive to isolation of reoviruses from chickens (Guneratne et al, 1982). However, in this study, only eight of the 28 reoviruses isolated were detected directly in CELi cells, and they were all from farm B.…”

Comparison of chick embryo liver and vero cell cultures for the isolation and growth of avian reoviruses

“…Primary chick embryo liver (CELi) cells were prepared from 14-day-old chicken embryos as previously described (Guneratne et al, 1982). For initial growth, Medium 199 contained 10% calf serum, while for maintenance of cultures, 5% calf serum was used (maintenance medium -MM).…”

“…For the isolation and cultivation of avian reoviruses, previous studies in this laboratory (Guneratne et al, 1982) have shown that chick embryo liver cell (CELi) cultures are superior to chick embryo fibroblasts (CEF), lung (CELu), kidney (CEK) or chicken kidney (CK) cultures, in that highest titres were reached and cytopathic effect (CPE) developed soonest in CELi cells. However, preparation of primary avian cell cultures is tedious and time-consuming, and growth of avian reoviruses in several mammalian continuous cell-lines has been examined.…”