scholarly journals Development of a Novel Assay Based on Plant-Produced Infectious Bursal Disease Virus VP3 for the Differentiation of Infected From Vaccinated Animals

2021 ◽  
Vol 12 ◽  
Author(s):  
Alessio Bortolami ◽  
Marcello Donini ◽  
Carla Marusic ◽  
Chiara Lico ◽  
Charifa Drissi Touzani ◽  
...  
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Severe Infection ◽  
Bursa Of Fabricius ◽  
Infectious Bursal Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Broiler Chicks ◽  
The Novel ◽  
Bursal Disease ◽  
Specific Pathogen

Infectious bursal disease virus is the causative agent of Gumboro disease, a severe infection that affects young chickens and is associated with lymphoid depletion in the bursa of Fabricius. Traditional containment strategies are based either on inactivated or live-attenuated vaccines. These approaches have several limitations such as residual virulence or low efficacy in the presence of maternally derived antibodies (MDA) but, most importantly, the impossibility to detect the occurrence of natural infections in vaccinated flocks. Therefore, the development of novel vaccination strategies allowing the differentiation of infected from vaccinated animals (DIVA) is a priority. Recently, commercial vectored and experimental subunit vaccines based on VP2 have been proved effective in protecting from clinical disease and posed the basis for the development of novel DIVA strategies. In this study, an engineered version of the VP3 protein of IBDV (His-VP3) was produced in plants, successfully purified from Nicotiana benthamiana leaves, and used to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-VP3 antibodies. The His-VP3 ELISA was validated with a panel of 180 reference sera and demonstrated to have 100% sensitivity (95% CI: 94.7–100.0) and 94.17% specificity (95% CI: 88.4–97.6). To evaluate the application of His-VP3 ELISA as a DIVA test, the novel assay was used to monitor, in combination with a commercial kit, detecting anti-VP2 antibodies, the immune response of chickens previously immunized with an inactivated IBDV vaccine, a recombinant Turkey herpes virus carrying the VP2 of IBDV (HVT-ND-IBD) or with plant-produced VP2 particles. The combined tests correctly identified the immune status of the vaccinated specific pathogen free white-leghorn chickens. Moreover, the His-VP3 ELISA correctly detected MDA against VP3 in commercial broiler chicks and showed that antibody titers fade with time, consistent with the natural decrease of maternally derived immunity. Finally, the novel assay, in combination with a VP2-specific ELISA, demonstrated its potential application as a DIVA test in chickens inoculated with VP2-based vaccines, being able to detect the seroconversion after challenge with a very virulent IBDV strain.

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.

scholarly journals The expression in plants of an engineered VP2 protein of Infectious Bursal Disease Virus induces formation of structurally heterogeneous particles that protect from a very virulent viral strain

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247134
Author(s):  
Carla Marusic ◽  
Charifa Drissi Touzani ◽  
Alessio Bortolami ◽  
Marcello Donini ◽  
Claudia Zanardello ◽  
...  
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Neutralizing Antibodies ◽  
Sucrose Density Gradient ◽  
Bursa Of Fabricius ◽  
Infectious Bursal Disease ◽  
Structural Protein ◽  
Vp2 Protein ◽  
Bursal Disease ◽  
Specific Pathogen

Infectious Bursal Disease Virus (IBDV), the etiological agent of Gumboro disease, causes mortality and immunosuppression in chickens and major losses to poultry industry worldwide. The IBDV major capsid protein VP2 is considered the best candidate for the production of novel subunit vaccines. This structural protein contains the major conformational epitopes responsible for the induction of IBDV neutralizing antibodies in chickens and has been demonstrated able to form supramolecular structures in yeast and insect cells. The aim of this study was to express an engineered version of the VP2 protein (His-pVP2) to verify its ability to self-assemble into virus-like particles in plants. The recombinant VP2 was transiently expressed by agroinfiltration in Nicotiana benthamiana and transmission electron microscopy of sucrose density gradient fractions revealed the presence of a mixed population of differently shaped particles ranging from spherical capsids, with a diameter between ~25 and ~70 nm, to tubular structures, with variable length (from 100 to 400 nm). The recombinant VP2-based particles when used for the intramuscular immunization of specific-pathogen-free chicks resulted able to induce the production of anti-IBDV specific antibodies at titers comparable to those induced by a commercial vaccine. Moreover, all the immunized birds survived to the challenge with a Moroccan very virulent IBDV strain with no major histomorphological alterations of the Bursa of Fabricius, similarly to what obtained with the commercial inactivated vaccine.

scholarly journals Isolation of Infectious Bursal Disease Virus Using Indigenous Chicken Embryos in Kenya

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
W. U. Mutinda ◽  
L. W. Njagi ◽  
P. N. Nyaga ◽  
L. C. Bebora ◽  
P. G. Mbuthia ◽  
...  
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Bursa Of Fabricius ◽  
Infectious Bursal Disease ◽  
Economic Activities ◽  
Indigenous Chicken ◽  
Chicken Embryos ◽  
Local Resources ◽  
Bursal Disease ◽  
Specific Pathogen

Infectious bursal disease virus (IBDV) isolates were recovered from outbreaks to initiate activities towards developing a local vaccine strain. Use of indigenous chicken embryos was exploited to determine their potential, promote utilization of local resources for research, and enhance household economic activities. Bursa of Fabricius (BFs) samples from outbreaks shown to be IBDV positive was homogenized and inoculated in 4-week-old specific pathogen-free (SPF) IBDV seronegative white leghorn chicks. The harvested virus was inoculated into 11-day-old indigenous chicken embryos that were IBDV seronegative and passaged serially three times after which they were inoculated into 4-week-old indigenous chicks to test for presence and virulence of propagated virus. Out of 153 BFs collected from outbreaks, 43.8% (67/153) were positive for IBDV antigen and 65.7% (44/67) caused disease in SPF chicks. The embryo mean mortalities were 88% on primary inoculation, 94% in 1st passage, 91% in 2nd passage, and 67% in 3rd passage. After the third passage in embryos all the 44 isolates were virulent in 4-week-old indigenous chicks. The results show that indigenous chicken embryos support growth of IBDV and can be used to propagate the virus as an alternative viral propagating tool for respective vaccine preparation.

scholarly journals STUDY THE EFFECT OF MYCOPLASMA CONTAMINATION OF EGGS USED IN VIRUS TITRATION AND EFFICACY OF SOME LIVE ATTENUATED POULTRY VIRAL VACCINES

2016 ◽  
Vol 10 (1) ◽  
pp. 216 ◽  
Author(s):  
Marwa Fathy ◽  
Mounir M. El-safty ◽  
Jakeen K. El-jakee ◽  
Howaida I. Abd-alla ◽  
Hala Mahmoud
Keyword(s):  
Disease Virus ◽  
Newcastle Disease ◽  
Disease Process ◽  
Mycoplasma Gallisepticum ◽  
Infectious Bursal Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infectious Bronchitis ◽  
Bursal Disease ◽  
Specific Pathogen ◽  
Viral Vaccines

ABSTRACTObjective: The study of Mycoplasma gallisepticum (MG) infection is needed, not only to understand the disease process but also to understand theinterference with the evaluation of some live viral poultry vaccines. This study aims to investigate the titration and potency of some live attenuatedpoultry viral vaccines; Newcastle disease, infectious bronchitis, infectious bursal disease, and Reo in both specific pathogen-free (SPF) embryonatedchicken eggs (ECEs) and chickens.Methods: Titration of live attenuated viral poultry vaccines in ECEs was carried out by dividing the inoculated eggs into four groups; the pre-,simultaneously-, post-, and non-MG contaminated. MG effect on the potency test was carried out using seventeen groups of SPF chickens (25 chicken/group) placed into separate isolators. Each live attenuated viral poultry vaccine was inoculated into 4 groups.Results: The highest titer of these vaccines that appeared in MG pre- contaminated ECEs were 1011, 107.5, 107.9, and 10, respectively. The lowest vaccinetiters that appeared in non-MG contaminated ECEs were 108, 106, 106.8, and 1067.5, respectively. Although the potency of these previous vaccines indicated thatthe highest antibodies titer that appeared in MG pre-infected vaccinated chickens were 7.5 log, 36 enzyme-linked immunosorbent assay unit (EU), and42 EU, respectively; the lowest antibodies titer that appeared in non-MG infected vaccinated chickens were 6.5 log22, 12 EU, 17 EU, and 10 EU, respectively.Conclusion: The present study findings underline the importance of using Mycoplasma -free eggs or chicken for the production of virus vaccines.Keywords: Mycoplasma gallisepticum, Newcastle disease virus, Infectious bronchitis virus, Infectious bursal disease virus, Reo virus, Chicken, Specificpathogen-free eggs.

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 Genome-wide analysis of differentially expressed mRNAs, lncRNAs, and circRNAs in chicken bursae of Fabricius during infection with very virulent infectious bursal disease virus

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Xuewei Huang ◽  
Junyan Zhang ◽  
Zengsu Liu ◽  
Meng Wang ◽  
Xiaolong Fan ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Expression Profiles ◽  
Circular Rna ◽  
Differentially Expressed ◽  
Bursa Of Fabricius ◽  
Infectious Bursal Disease ◽  
Non Coding Rna ◽  
Bursal Disease

Abstract Background Infectious bursal disease virus (IBDV) causes acute, highly contagious, immunosuppressive, and lethal infectious disease in young chickens and mainly infects the bursa of Fabricius (BF). To investigate interactions between IBDV and its host, RNA sequencing was applied to analyze the responses of the differentially expressed transcriptional profiles of BF infected by very virulent IBDV (vvIBDV). Results In total, 317 upregulated and 94 downregulated mRNAs were found to be significantly differentially expressed in infected chickens, compared to controls. Long non-coding RNA (lncRNA) and circular RNA (circRNA) alterations were identified in IBDV-infected chickens, and significantly different expression was observed in 272 lncRNAs and 143 circRNAs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed to assess the functions of significantly dysregulated genes, which showed that the JAK-STAT signaling pathway, the NOD-like receptor signaling pathway, and apoptosis may be activated by IBDV infection. We predicted interactions between differentially expressed genes and produced lncRNA-mRNA and circRNA-miRNA-mRNA regulator network. Conclusions The present study identified the expression profiles of mRNAs, lncRNAs, and circRNAs during vvIBDV infection and provides new insights into the pathogenesis of IBDV and antiviral immunity of the host.

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