Growth of Infectious Bursal Disease Virus with Plaque Formation in Chick Embryo Fibroblast Cell Culture

Abstract

The WA69 isolant of infectious bursal disease virus (IBDV) induced cytopathic effects and plaque formation in chick embryo fibroblast (CEF) cultures after serial passages in embryonated eggs and then in CEF cultures. The plaque-forming agent was cloned (designated WA69 clone) and identified as IBDV on the basis of serologic response in inoculated birds and its antigenic relationship to other known IBDV isolants. The WA69 clone replicated rapidly in CEF cultures, reaching peak titers at 48 hours postinoculation, and the virus caused only minimal histologic lesions of the bursa when inoculated into 3-week-old chicks from a specific-pathogen-free flock. The growth of IBDV in CEF cultures with plaque formation may provide a simple in vitro system for virological and serological studies of IBDV.

Isolation, molecular characterization and pathogenicity studies of infectious bursal disease field virus isolates

“…Dead inoculated embryos with classical and vvIBDV strains showed typical lesions (severe congested liver and greenish coloration of the liver with mottling appearance, dwarfing embryo, splenomegally, hemorrhages on head, toes skin and along the feather tracts, edematous distension of the abdominal region and cerebral edema, while dead embryos from variant IBDV strains showed stunted growth, spleenomegally and liver necrosis. Previously mentioned results were recorded by Cho et al, (1979); McFerran et al, (1980); Lasher and Shane (1994); . showed morbidity rates of 70, 80 and 60%, respectively and the only detectable signs throughout the observation period were off food and lowered feed intake.…”

Propagation and Molecular Characterization of Bioreactor Adapted Very Virulent Infectious Bursal Disease Virus Isolates of Malaysia

“…However, this technique is not always effective, because some variant strains of the virus do not induce embryonic mortality [ 2 ], an infection index that CAM inoculation methods mostly depend on to indicate productive infection. Advances in IBD diagnosis made it possible for many IBD virus (IBDV) isolates to be adapted on primary avian cell cultures derived from chicken embryonic tissues from organs such as kidney (CEK), bursa (CEB), and muscles (CEF) [ 3 , 4 ]. These cells, however, were found to produce low virus titer, have limited cell growth, and may be a source of contamination with extraneous avian viruses [ 5 , 6 ].…”

Adaptation and Molecular Characterization of Two Malaysian Very Virulent Infectious Bursal Disease Virus Isolates Adapted in BGM-70 Cell Line

“…Traditional isolation of IBDV by CAM inoculation of 9- to 11-day-old SPF chicken embryos is expensive coupled with high risk of contamination especially for vaccine development and production [ 5 ]. Successful adaptation of IBDV isolates in chicken embryo fibroblast (CEF), chicken embryo kidneys, and chicken embryo bursas was reported [ 6 , 7 ], but the cells being of primary avian origin have limited lifespan, produce low virus titer, and may contain extraneous avian viruses that may contaminate vaccines developed using them [ 8 , 9 ]. Many continuous cell lines of mammalian origin were reported to support the growth of IBDV isolates including MA-104 [ 10 ], OK [ 11 ], BGM-70 [ 10 , 12 ], Vero cells [ 10 , 13 – 15 ], and RK-13 [ 14 , 16 ].…”