Abstract
Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 (also known as B00/81) and UPM190 (also known as UPM04/190) isolated from local IBD outbreaks in 2000 and 2004, respectively, were separately passaged for 12 consecutive times in 11-day-old specific pathogen free (SPF) chicken embryonated eggs (CEE) via the chorioallantoic membrane (CAM) route. The CEE passage 8 (EP8) isolates were passaged once in BGM-70 cell line yielding UPM0081EP8BGMP1 and UPM190EP8BGMP1, while the EP12 isolates were passaged 15 times in BGM-70 cell line yielding UPM0081EP12BGMP15 and UPM190EP12BGMP15 using T25 tissue culture flask. These isolates were all propagated once in bioreactor using cytodex 1 as microcarrier at 3 g per liter (3 g/L) yielding UPM0081EP8BGMP1BP1, UPM190EP8BGMP1BP1, UPM0081EP12BGMP15BP1, and UPM190EP12BGMP15BP1 isolates. The viruses were harvested at 3 days after inoculation, following the appearance of cytopathic effects (CPE) characterized by detachment from the microcarrier using standard protocol and filtered using 0.2 μm syringe filter. The filtrates were positive for IBDV by RT-PCR and immunofluorescence. Sequence and phylogenetic tree analysis indicated that the isolates were of the vvIBDV strains and were not different from the flask propagated parental viruses.
Highlights
Infectious bursal disease virus (IBDV), the causative agent of infectious bursal disease (IBD), was isolated through inoculation of suspected samples into 9 to 11day-old chicken embryonated eggs (CEE) via chorioallantoic membrane (CAM) route [1]
BGM-70 cell line was selected for the propagation of local Malaysian infectious bursal disease virus (IBDV) isolates in a table top bioreactor to evaluate the ability to support the growth of the viruses in the bioreactor
The CEE adapted UPM0081EP8BGMP1 passaged once in BGM-70 cells and propagated in the bioreactor developed cytopathic effects (CPE) between 48 and 72 hours that was characterized by cell detachment from the microcarrier
Summary
Infectious bursal disease virus (IBDV), the causative agent of infectious bursal disease (IBD), was isolated through inoculation of suspected samples into 9 to 11day-old CEE via CAM route [1]. 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, were found to produce low virus titer, have limited cell growth, and may be a source of contamination with extraneous avian viruses [5, 6]. The cell line BGM-70 derived from baby grivet monkey was reported to support the growth and rapidly attenuate classical, variant, and very virulent IBDV strains and with high viral titer [6, 15, 18] The ability of this continuous cell line to yield higher viral titers is a valuable characteristic that makes growing viruses in them advantageous over primary cell cultures apart from convenience. This is due to the high titer obtained when the cell line was used to propagate the virus in a conventional flask culture, confirming the earlier reports of its potential [14, 18, 20]; Abdel-Alim et al, 2003; [15]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
