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

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Nafi’u Lawal ◽  
Mohd Hair-Bejo ◽  
Siti Suri Arshad ◽  
Abdul Rahman Omar ◽  
Aini Ideris
Keyword(s):  
Cell Line ◽  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Chorioallantoic Membrane ◽  
Infectious Bursal Disease ◽  
Phylogenetic Tree Analysis ◽  
Cytopathic Effects ◽  
Bursal Disease ◽  
Specific Pathogen ◽  
Syringe Filter

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.

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

2017 ◽  
Vol 2017 ◽  
pp. 1-19 ◽  
Author(s):  
Nafi’u Lawal ◽  
Mohd Hair-Bejo ◽  
Siti Suri Arshad ◽  
Abdul Rahman Omar ◽  
Aini Ideris
Keyword(s):  
Cell Line ◽  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Chorioallantoic Membrane ◽  
Infectious Bursal Disease ◽  
Single Mutation ◽  
Cytopathic Effects ◽  
Bursal Disease ◽  
Specific Pathogen

Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 109.50 TCID50/mL andlog109.80 TCID50/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogenetic analysis showed that these two isolates were of the vvIBDV. It appears that a single mutation of UPM190 and UPM0081 IBDV isolates at D279N could facilitate vvIBDV strain adaptability in CEE and BGM-70 cultures.

scholarly journals Growth and Replication of Infectious Bursal Disease Virus in the DF-1 Cell Line and Chicken Embryo Fibroblasts

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Kaliyaperumal Rekha ◽  
Chandran Sivasubramanian ◽  
Ill-Min Chung ◽  
Muthu Thiruvengadam
Keyword(s):  
Cell Line ◽  
Disease Virus ◽  
Chicken Embryo ◽  
Infectious Bursal Disease Virus ◽  
Infectious Bursal Disease ◽  
Population Doubling ◽  
Cytopathic Effects ◽  
Bursal Disease ◽  
Embryo Fibroblasts ◽  
Chicken Embryo Fibroblasts

Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chicks and leads to significant economic losses in the poultry industry. To determine a suitable cell line for IBDV infection, replication, and growth kinetics of the virus, DF-1 cells and chicken embryo fibroblasts (CEF) were used. The population doubling per day (Pd/D) was found to be higher in DF-1 as compared to CEF cells. A suitable time of infection (TOI) was established for increased production of virus and greater infectivity titers. The DF-1 and CEF cells were found to be susceptible to infection by producing marked cytopathic effects (CPEs), and the growth curves of IBDV in DF-1 and CEF cells were evaluated by infectivity assay using tissue culture infectious dose (TCID50). The cytopathic effects of the virus in DF-1 and CEF cells were found to be similar, but higher viral titers were detected in the DF-1 cells as compared to CEF. Thus the DF-1 cell line had a higher growth potential and infectivity, which will be of advantage in vaccine production.

scholarly journals Bursal immunopathology responses of specific-pathogen-free chickens and red jungle fowl infected with very virulent infectious bursal disease virus

2018 ◽  
Vol 163 (8) ◽  
pp. 2085-2097 ◽  
Author(s):  
Mohd Isa Farhanah ◽  
Abdul Rahaman Yasmin ◽  
Nguyen Phuc Khanh ◽  
Swee Keong Yeap ◽  
Mohd Hair-Bejo ◽  
...  
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Infectious Bursal Disease ◽  
Specific Pathogen Free ◽  
Red Jungle Fowl ◽  
Bursal Disease ◽  
Specific Pathogen

scholarly journals Identification of Chicken CD74 as a Novel Cellular Attachment Receptor for Infectious Bursal Disease Virus in Bursa B Lymphocytes

2019 ◽  
Vol 94 (2) ◽  
Author(s):  
Aijing Liu ◽  
Qing Pan ◽  
Yue Li ◽  
Nana Yan ◽  
Jing Wang ◽  
...  
Keyword(s):  
B Cells ◽  
Cell Line ◽  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
B Lymphocyte ◽  
Pivotal Role ◽  
Infectious Bursal Disease ◽  
Antigenic Peptides ◽  
Bursal Disease

ABSTRACT Infectious bursal disease virus (IBDV) is an important member of the Birnaviridae family, causing severe immunosuppressive disease in chickens. The major capsid protein VP2 is responsible for the binding of IBDV to the host cell and its cellular tropism. In order to find proteins that potentially interact with IBDV VP2, a liquid chromatography-mass spectrometry (LC-MS) assay was conducted, and the host chicken CD74 protein was identified. Here, we investigate the role of chicken CD74 in IBDV attachment. Coimmunoprecipitation assays indicated that the extracellular domain of CD74 interacted with the VP2 proteins of multiple IBDV strains. Knockdown and overexpression experiments showed that CD74 promotes viral infectivity. Confocal assays showed that CD74 overexpression allows the attachment of IBDV and subvirus-like particles (SVPs) to the cell surface of nonpermissive cells, and quantitative PCR (qPCR) analysis further confirmed the attachment function of CD74. Anti-CD74 antibody, soluble CD74, depletion of CD74 by small interfering RNA (siRNA), and CD74 knockdown in the IBDV-susceptible DT40 cell line significantly inhibited IBDV binding, suggesting a pivotal role of this protein in virus attachment. These findings demonstrate that CD74 is a novel important receptor for IBDV attachment to the chicken B lymphocyte cell line DT40. IMPORTANCE CD74 plays a pivotal role in the correct folding and functional stability of major histocompatibility complex class II (MHC-II) molecules and in the presentation of antigenic peptides, acting as a regulatory factor in the antigen presentation process. In our study, we demonstrate a novel role of CD74 during IBDV infection, showing that chicken CD74 plays a significant role in IBDV binding to target B cells by interacting with the viral VP2 protein. This is the first report demonstrating that CD74 is involved as a novel attachment receptor in the IBDV life cycle in target B cells, thus contributing new insight into host-pathogen interactions.

scholarly journals Replication of classical infectious bursal disease virus in the chicken embryo related cell line

2000 ◽  
Vol 29 (3) ◽  
pp. 213-217 ◽  
Author(s):  
T.C. Cardoso ◽  
P. Rahal ◽  
D. Pilz ◽  
M. C. B. Teixeira ◽  
C. W. Arns
Keyword(s):  
Cell Line ◽  
Disease Virus ◽  
Chicken Embryo ◽  
Infectious Bursal Disease Virus ◽  
Infectious Bursal Disease ◽  
Related Cell ◽  
Bursal Disease

Infectivity and propagation of attenuated infectious bursal disease virus in the chicken B-lymphocyte cell line DT40

2009 ◽  
Vol 154 (3) ◽  
pp. 513-517 ◽  
Author(s):  
Jun Luo ◽  
Gai-Ping Zhang ◽  
Jian-Ming Fan ◽  
Man Teng ◽  
Lei-Ming You ◽  
...  
Keyword(s):  
Cell Line ◽  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
B Lymphocyte ◽  
Infectious Bursal Disease ◽  
Bursal Disease ◽  
Lymphocyte Cell ◽  
Lymphocyte Cell Line

scholarly journals Pathogenicity and Effects on Lymphoid Organs of Recent Moroccan Very Virulent Infectious Bursal Disease Virus in Broilers and SPF Chickens

2021 ◽  
Author(s):  
Charifa DRISSI TOUZANI ◽  
Imane MAAROUFI ◽  
Siham FELLAHI ◽  
Ikhlass EL BERBRI ◽  
Fatima-zohra SIKHT ◽  
...  
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Clinical Signs ◽  
Total Mortality ◽  
Bursa Of Fabricius ◽  
Infectious Bursal Disease ◽  
Control Group ◽  
Bursal Disease ◽  
Specific Pathogen ◽  
And Control

Abstract The aim of the current study is to evaluate the pathogenicity of recent infectious bursal disease virus (IBDV) (1/chicken/Morocco/IB19/2017) genetically characterized as vvIBDV belonging to genogroup 3.Two chicken lines, broiler and specific-pathogen-free (SPF) chickens, were inoculated by occulonasal route with 0.2 ml of the 105EID50 /ml of viral solution of IB19 vvIBDV strain at 29 days of age. The experimental monitoring was carried out during 10 days post challenge (dpc). The clinical signs stared on day 2 pc with maximum severity observed between 3 and 6 dpc. The total mortality rate reached 10% in broilers (group G1) and 93% in SPF (G3). The macroscopic lesions in broilers G1 was a marked hypertrophy of the bursa of Fabricius (BF) with slight haemorrhage observed between 2 to 4 dpc, followed by very pronounced atrophy observed on the 5 dpc. The post-mortem examinations of dead SPF birds (G3) revealed on 3 dpc very haemorrhagic BF with black cherry appearance in 80 % of dead birds. The mean Bursa/Body Index (BBI) of challenged broilers (G1) showed a decrease of 46% on day 9 pc compared to broilers control group (G2) indicating bursal atrophy. The microscopic lesions found in the BF on 3 dpc consisted mainly of inflammation with severe lymphoid depletion of the follicles. The evaluation of recent vvIBDV outbreak is very important to understand its epidemiology and will contribute to the efficient prevention and control of IBD.

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