Virulent Infectious Bursal Disease Virus Research Articles

Construction and Efficacy of recombinant Newcastle disease virus co-expressing VP2 and VP3 proteins of Very Virulent Infectious Bursal Disease Virus

Construction and Efficacy of recombinant Newcastle disease virus co-expressing VP2 and VP3 proteins of Very Virulent Infectious Bursal Disease Virus

Very virulent infectious bursal disease virus infection triggered microscopic changes, apoptosis, and inflammatory cytokines imbalance in chicken spleen and thymus

ABSTRACT Infectious bursal disease virus (IBDV) can cause a highly contagious disease, resulting in severe damage to the immune system that causes immunosuppression in young chickens. Both spleen and thymus are important immune organs, which play a key role in eliciting protective immune responses. However, the effects of very virulent IBDV (vvIBDV) strain LJ-5 infection on chicken spleen and thymus are still unknown. In the present study, 3-week-old specific pathogen-free chickens were infected with vvIBDV for 1–5 days. The vvIBDV infection significantly increased the spleen index and decreased the thymus index. Microscopic analysis indicated necrosis, depletion of the lymphoid cells, and complete loss of structural integrity in spleen and thymus. Ultrastructural analysis displayed mitochondrial and nuclear damage, including mitochondrial cristae breaks, and deformation of nuclear membrane in vvIBDV-infected spleen and thymus tissues. Cytokine levels increased in the spleen and thymus after IBDV infection, promoting inflammation and causing an inflammatory imbalance. Moreover, the mRNA expression of apoptosis-related genes was significantly upregulated in the vvIBDV-infected group compared to the control group. Meanwhile, the mRNA expression of mitochondrial dynamics was altered in the spleen and thymus of vvIBDV-infected chickens. These results suggested that vvIBDV infection triggers an imbalance of inflammatory cytokines, and apoptosis in the spleen and thymus, resulting in immune injury in chickens. This study provides basic data for the further study of vvIBDV pathogenesis.

Epidemiological and molecular characteristics of Infectious bursal disease virus naturally infected in the broiler flocks in the Mekong Delta of Vietnam during 2015 and 2018

Infectious bursal disease is a highly contagious and economically devastating disease in the poultry industry worldwide caused by Infectious bursal disease virus (IBDV). However, the data on epidemiological and molecular characteristics of the IBDV outbreak in broiler flocks in the Mekong Delta of Vietnam is unclear. Herein, the epidemiological data of IBDV–positive flocks over a period of 2015–2018 were recorded and the hypervariable region of the VP2 gene of IBDV was amplified to analyze the local phylogeny. The current investigation showed that the overall morbidity and mortality rates of IBDV–positive flocks were 45% and 4.81%, respectively. The IBDV–positive birds occurred clinical signs and macroscopic findings involved with the very virulent (vv) IBDV outbreak. Epidemiological results revealed that IBDV was frequently infected in broiler flocks at 12–42 days, and birds belonging to Tau Vang and Binh Dinh breeds were more sensitive to IBDV. Also, the morbidity rate of IBDV was dramatically decreased in the open farming system. Flocks with complete vaccination significantly dropped morbidity in comparison with other groups. Regarding phylogenetic analysis, all identified IBDV sequences clustered in the same branch of vv phenotype and closely homology with prior strains circulated in Vietnam and other countries. These findings first indicated the epidemiological characteristics of the IBDV–positive broiler flocks in the Mekong Delta and highlighted the IBDV strain circulating in this region.

Emergence of the novel infectious bursal disease virus variant in vaccinated poultry flocks in Egypt

ABSTRACT Since the detection of antigenically atypical very virulent Infectious bursal disease viruses (vvIBDV) in Egypt in 1999, the country has been experiencing recurrent outbreaks with high mortality rates and typical gross lesions associated with typical vvIBDV. However, a significant change occurred in 2023, marked by a notable increase in reported subclinical IBDV cases. To evaluate the field situation, samples from 21 farms in 2023 and 18 farms from 2021 and 2022, all of which had experienced IBD outbreaks based on clinical diagnosis, were collected, and subjected to VP2-HVR sequencing. Phylogenetic analysis revealed that all samples collected in 2021 and 2022 clustered with classical virulent strains and vvIBDV. In 2023, one sample clustered with the Egyptian vvIBDV, another with classical virulent IBDV, and the rest with the novel variant IBDV (nVarIBDV) circulating in China. The alignment of deduced amino acid sequences for VP2 showed that all Egyptian classic virulent strains were identical to the Winterfield or Lukert strains, while vvIBDV strains exhibited two out of the three typical residues found in Egyptian vvIBDV, namely Y220F and G254S, but not A321T. Meanwhile, all Egyptian variant strains exhibited typical residues found in nVarIBDV. However, all Egyptian variants showed a mutation at position 321 (321V), which represents the most exposed part of the capsid and is known to have a massive impact on IBDV antigenicity, except for one sample that had 318G instead. This report highlights the emergence of a new variant IBDV in Egypt, clustered with the Chinese new variants, spreading subclinically in broiler farms across a wide geographic area. RESEARCH HIGHLIGHTS New variant IBDV which emerged in Egypt clustered with Chinese nVarIBDV. nVarIBDV spread subclinically across a wide geographic area. Mutation at 321 represents capsid's most exposed part, a defining feature. Antigenically modified vvIBDV still circulating in Egypt with typical lesions.

Isolation and molecular characterization of very virulent infectious bursal disease virus from Egypt

In Egypt, the poultry industry is plagued by the significant economic losses caused by the infectious bursal disease virus (IBDV). In the present study, a total of 30 diseased commercial chicken flocks aged 23–41 days old in three Egyptian governorates (Fayoum, Beni-Suef, and Minya) having symptoms consistent with IBDV were examined. The collected bursal samples (n=30, ten pooled bursae for each flock) were screened by RT-PCR to amplify the IBDV VP2 gene. Results revealed that 20 out of 30 samples were positive, as indicated by the amplification of 620 bp of the VP2 gene. The twenty positive samples were inoculated into specific pathogen-free embryonated chicken eggs (SPF-ECEs) via the chorioallantoic-membrane (CAM) route to isolate the suspected virus. Eight samples were successfully isolated and were subjected to a partial sequence of VP2 (targeting the HVR). The nucleotide sequences and phylogenetic analysis revealed that the examined isolates exhibited amino acids A222, I256, I294, and S299, which are extremely conserved among very virulent (vv)IBDV. The amino acid sequence analysis of the eight isolates revealed a high degree of similarity (97-100%) in between and 90-100% similarity compared to other Egyptian vvIBDV strains. In contrast, lower similarity was observed with attenuated IBDV vaccine strains D78, Lukert, and Bursavac (91-93%, 91-93%, and 92-94%, respectively). In conclusion, this study underscores the importance of ongoing surveillance of the IBDV situation in the field, as well as the need for further research to investigate the efficacy of current vaccination strategies to curb IBDV infection.

Protective effects of rabbit sacculus-derived antimicrobial peptides on SPF chicken against infection with very virulent infectious bursal disease virus

Previous studies here have demonstrated that the rabbit sacculus rotundus-derived antimicrobial peptides (RSRP) could alter the intestinal mucosal immune responses in specific-pathogen-free (SPF) chickens, however, the protective effects of RSRP on chickens against infection remain questionable. In the present study, eighty SPF chickens were randomly divided into five groups and challenged with very virulent infectious bursal disease virus (vvIBDV) to determine the protective effects and its underlying mechanism of RSRP. Histopathology examination found that vvIBDV-infection caused severe damage in the bursa of Fabricius, especially the bursal lymphoid follicles underwent severe necrosis, depletion, hemorrhage, and edema. Unexpectedly, RSRP intervention significantly reduced the necrosis and depletion of lymphoid follicles in the vvIBDV-infected chickens. Moreover, RSRP treatment significantly decreased the expression of Bax (P<0.01) as well as remarkably promoted the expression of Bcl-2 (P<0.01), concomitantly alleviated the excessive apoptosis in the immune organs such as the bursa of Fabricius during vvIBDV infection. Notably, consistent with our previous reports that increased mast cell activation and degranulation in the bursa after vvIBDV infection, RSRP administration considerably reduced the mast cell density and the expression of tryptase, a marker for activated mast cells. Collectively, the present study indicates that rabbit sacculus rotundus-derived antimicrobial peptides could effectively protect the major immune organs including the bursa of Fabricius from the damage caused by vvIBDV infection, which provides the possibility and a promising perspective for the future application of antimicrobial peptides for poultry production.

Genetic Sequence and Pathogenicity of Infectious Bursal Disease Virus in Chickens in Egypt During 2017-2021.

The continued circulation of infectious bursal disease virus (IBDV) in Egypt, despite the use of various vaccines, is a serious problem that requires continuous detection of IBDV. In the current study, real-time reverse transcriptase polymerase chain reaction testing of 100 diseased chicken flocks during 2017-2021 revealed the presence of very virulent IBDV (vvIBDV) in 67% of the flocks, non-vvIBDV in 11%, and a mixture of both vvIBDV and non-vvIBDV in 4%. Twenty-nine IBDV isolates were submitted for partial sequencing of the viral protein 2 hypervariable region (VP2-HVR), and 27 isolates were confirmed to be genogroup A3 (vvIBDV) with 96.3%-98.5% similarity to the global A3 (vvIBDV) and 88.9%-97% similarity to genogroup A1 vaccine strains. The remaining two isolates were non-vvIBDV and showed 91.1% and 100% identity with classical genogroup A1 strains, respectively. Furthermore, the sequence and phylogenetic analysis of VP1 (amino acids 33-254) of two selected isolates of A3, 5/2017 and 98/2021, clustered them as B2, vvIBDV-like, strains with high similarity (99.5%) to four Egyptian, 99% to Chinese and European, and 97.7% to Chinese and Polish vvIBDV isolates. Experimental infection of commercial broiler chickens with two vvIBDV-A3B2 isolates (5/2017 and 98/2021) showed no mortality despite typical tissue lesions, clear histopathological changes, and strong ELISA antibody response. Isolate 98/2021 was more pathogenic, as confirmed by histopathology, whereas isolate 5/2017 induced a stronger serological response. In conclusion, vvIBDV (A3B2) strains with two amino acid (aa) substitutions in VP1 as V141I and V234I as well as VP2 as Y220F and G254S are still circulating in Egypt.

Development of an improved reverse genetics system for avian metapneumovirus (aMPV): A novel vaccine vector protects against aMPV and infectious bursal disease virus

Avian metapneumovirus (aMPV), a paramyxovirus, causes acute respiratory diseases in turkeys and swollen head syndrome in chickens. This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase. Full-length cDNA of the LN16-A strain was constructed by assembling five cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme. Transfection of this plasmid, along with the supporting plasmids encoding the N, P, M2-1, and L proteins of LN16-A into BSR-T7/5 cells, resulted in the recovery of aMPV subtype B. To identify an effective insertion site, the enhanced green fluorescent protein (EGFP) gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As. The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites (between the leader and N genes or replacing the SH gene). To verify the availability of the site between G and L for foreign gene expression, the VP2 gene of very virulent infectious bursal disease virus (vvIBDV) was inserted into this site, and recombinant LN16-A (rLN16A-vvVP2) was successfully rescued. Single immunization of specific-pathogen-free chickens with rLN16A-vvVP2 induced high levels of neutralizing antibodies and provided 100% protection against the virulent aMPV subtype B and vvIBDV. Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.

The long noncoding RNA loc107053557 acts as a gga-miR-3530-5p sponge to suppress the replication of vvIBDV through regulating STAT1 expression

ABSTRACT Infectious bursal disease virus (IBDV) causes immunosuppression and high mortality in young chickens. Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) are important regulators during viral infection. However, detailed the regulatory mechanisms of lncRNA-miRNA-mRNA have not yet been described in IBDV infection. Here, we analysed the role of lncRNA53557/gga-miR-3530-5p/STAT1 axis in very virulent IBDV (vvIBDV) infection. Evidently upregulated expression of lncRNA53557 was observed in bursa of Fabricius and DT40 cells. Meanwhile, overexpression of lncRNA53557 promoted STAT1 expression and inhibited vvIBDV replication and vice versa, indicating that the upregulation of lncRNA53557 was part of the host antiviral defence. The subcellular fractionation assay confirmed that lncRNA53557 can be localized in the cytoplasm. Further, dual-luciferase reporter, RNA pulldown, FISH and RT-qPCR assays revealed that lncRNA53557 were directly bound to gga-miR-3530-5p and had a negative regulatory relationship between them. Subsequent mechanistic analysis showed that lncRNA53557 acted as a competing endogenous RNA (ceRNA) of gga-miR-3530-5p to relieve the repressive effect of gga-miR-3530-5p on its target STAT1, as well as Mx1, OASL, and ISG15, thereby suppressing vvIBDV replication. The study reveals that a network of enriched lncRNAs and lncRNA-associated ceRNA is involved in the regulation of IBDV infection, offering new insight into the mechanisms underlying IBDV-host interaction.

Development of a Highly Efficient CRISPR/Cas9-Mediated Herpesvirus of Turkey-Based Vaccine against Novel Variant Infectious Bursal Disease Virus.

Infectious bursal disease (IBD), caused by IBD virus (IBDV), is an extremely contagious immunosuppressive disease that causes major losses for the poultry industry worldwide. Recently, the novel variant IBDV (G2d) has been highly prevalent in Korea, but the current vaccines against this very virulent IBDV have limited efficacy against this novel variant. To develop a vaccine against this variant IBDV, a recombinant virus designated rHVT-VP2 was constructed by inserting the IBDV (G2d) VP2 gene into herpesvirus of turkeys (HVT) using CRISPR/Cas9 gene-editing technology. The PCR and sequencing results obtained showed that the recombinant virus rHVT-VP2 was successfully constructed. Vaccination with rHVT-VP2 generated IBDV-specific antibodies in specific pathogen-free chickens starting from 2 weeks post-immunization. Seven days after the challenge, the autopsy results showed that the bursa atrophy rates of the rHVT-VP2, HVT, vaccine A, and positive control groups were 0%, 100%, 60%, and 100%, respectively, and the BBIX values were 1.07 ± 0.22, 0.27 ± 0.05, 0.64 ± 0.33, and 0.32 ± 0.06, respectively. These results indicate that rHVT-VP2 can provide 100% protection against a challenge with the IBDV (G2d), whereas vaccine A only provides partial protection. In conclusion, vaccination with the recombinant virus rHVT-VP2 can provide chickens with effective protection against variant IBDV (G2d).

RNA-seq reveals role of cell-cycle regulating genes in the pathogenicity of a field very virulent infectious bursal disease virus.

Infectious bursal disease virus (IBDV) infection causes highly contagious and immunosuppressive disease in poultry. The thymus, serving as the primary organ for T cell maturation and differentiation, plays an important role in the pathogenicity of IBDV in the infected chickens. However, there are no reports on the molecular pathogenesis of IBDV in the thymus currently. The aim of the study was to elucidate the molecular mechanisms underlying the pathogenicity of a field very virulent (vv) IBDV strain NN1172 in the thymus of SPF chickens using integrative transcriptomic and proteomic analyses. Our results showed that a total of 4,972 Differentially expressed genes (DEGs) in the thymus of NN1172-infected chickens by transcriptomic analysis, with 2,796 up-regulated and 2,176 down-regulated. Meanwhile, the proteomic analysis identified 726 differentially expressed proteins (DEPs) in the infected thymus, with 289 up-regulated and 437 down-regulated. Overall, a total of 359 genes exhibited differentially expression at both mRNA and protein levels, with 134 consistently up-regulated and 198 genes consistently down-regulated, as confirmed through a comparison of the RNA-seq and the proteomic datasets. The gene ontology (GO) analysis unveiled the involvement of both DEGs and DEPs in diverse categories encompassing cellular components, biological processes, and molecular functions in the pathological changes in IBDV-infected thymus. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the host mainly displayed severely disruption of cell survival/repair, proliferation and metabolism pathway, meanwhile, the infection triggers antiviral immune activation with a potential emphasis on the MDA5 pathway. Network inference analysis identified seven core hub genes, which include CDK1, TYMS, MCM5, KIF11, CCNB2, MAD2L1, and MCM4. These genes are all associated with cell-cycle regulating pathway and are likely key mediators in the pathogenesis induced by NN1172 infection in the thymus. This study discovered dominant pathways and genes which enhanced our understanding of the molecular mechanisms underlying IBDV pathogenesis in the thymus.

Persistent Circulation of Very virulent Infectious Bursal Disease virus (IBDV) in Egypt: Phylogenetic analysis, Pathogenicity and Immunogenicity of an immune-complex vaccine

Persistent Circulation of Very virulent Infectious Bursal Disease virus (IBDV) in Egypt: Phylogenetic analysis, Pathogenicity and Immunogenicity of an immune-complex vaccine

Advancing IBDV diagnostics: a one-step multiplex real-time qRT-PCR for discriminating between vvIBDV and non-vvIBDV viruses, including the newly emerged IBDV variant.

The very virulent infectious bursal disease virus (vvIBDV) induces an acute, highly contagious and immunosuppressive disease in younger chicken causing massive economic losses globally. A major challenge in the field's clinical diagnosis is distinguishing gross lesions caused by vvIBDV from those induced by classic IBDV (cIBDV), commonly used as live attenuated vaccines. This study introduces a one-step multiplex real-time PCR assay designed to distinguish between vvIBDV and non-vvIBDV viruses. Via simultaneously targeting the VP2 sequence for vvIBDV detection and the VP1 sequence for non-vvIBDV identification, including classic, American variant and the recently emerged novel variant IBDV (nvarIBDV), the assay's specificity was validated against common avian viral diseases and nonspecific IBDV strains without any observed cross-reactions. It effectively differentiated between vvIBDV and non-vvIBDV field samples, including nvarIBDV, as confirmed by genotyping based on VP2 sequencing. The assay demonstrated a limit of detection ranging from 1.9×1010 to 103 DNA copies for vvIBDV-VP2, 9.2×1010 to 103 DNA copies for classic strains, and 1.2×1011 to 104 DNA copies for nvarIBDV in VP1 detection of non-vvIBDV. In conclusion, this study presents a specific, sensitive, and straight forward multiplex real-time PCR assay.

Protective effects of a novel chimeric virus–like particle vaccine against virulent NDV and IBDV challenge

Newcastle disease (ND) and infectious bursal disease (IBD) pose significant threats to the chicken industry, causing substantial economic losses. Currently, immunization through vaccination is the most effective strategy to prevent ND and IBD but currently used traditional vaccines, including inactivated or attenuated vaccines, face challenges in achieving a balance between immunogenicity and safety. To develop a green and efficient novel vaccine for ND and IBD, we developed a bivalent chimeric virus-like particle vaccine (ND-IBD cVLPs) displaying the ND virus (NDV) HN protein and the IBD virus (IBDV) VP2 protein based on the ND VLPs carrier platform and insect baculovirus expression system. This study aimed to evaluate the immunogenicity and protective efficacy of ND-IBD cVLPs in specific pathogen-free chickens. Chickens were immunized with 50 µg of purified ND-IBD cVLPs at 7 days old, boosted at 21 days old, and challenged at 42 days old. The results demonstrated that ND-IBD cVLPs stimulated highly effective hemagglutination inhibition antibody levels against NDV HN protein and enzyme-linked immunosorbent assay antibody levels against the IBDV VP2 protein. Furthermore, ND-IBD cVLPs provided complete protection against virulent NDV and IBDV challenges and mitigated pathological damage to the lung caused by NDV infection and the bursa of Fabricius caused by IBDV infection. These findings suggest that ND-IBD cVLPs hold promise as a safe and efficient novel vaccine candidate for the effective prevention of ND and IBD, extending the development of a foreign protein delivery platform of ND VLPs.

Continuous clinicopathological and molecular recognition of very virulent infectious bursal disease virus in commercial broiler chickens

Gumboro virus is one of the most dangerous immunosuppressant viruses that infect chickens and causes massive financial losses around the world. The purpose of the current study is to conduct a molecular survey of chicken farms for the infectious bursal disease virus (IBDV). On the basis of postmortem (PM) lesions, 125 bursal samples from 25 farms were collected from clinically diseased commercial chicken farms with increased mortality and suspected Gumboro virus infection. Pooled bursal samples from suspected IBD-vaccinated flocks were tested for IBDV by real-time polymerase chain reaction (RT-PCR). Fifteen out of 25 pooled specimens were found positive for IBDV, with a 60% detection rate, and confirmed positive for very virulent IBDV (vvIBDV). Nucleotide phylogenetic analysis of VP1 and VP2 genes was employed to compare the five chosen isolates with strains identified in various localities in 2022. The VP1 and VP2 genes are allotted into groups (I, II). The strains in our study were related to group II and it acquired a new mutation in the VP2 gene that clustered it into new subgroup B. By mutation analysis, the VP2 gene of all strains had a characteristic mutation to vvIBDV. It acquired new mutations in HVRs contrasted with HK64 in Y220F, A222T/V in all strains in our study, and Q221K that was found in IBD-EGY-AH5 and AH2 in the major hydrophilic region peak A in the loop PBC in addition to G254S in all strains in our study and Q249k that found in IBD-EGY-AH1 and AH3 in the minor hydrophilic region 1 in the loop PDE. These mutations are important in the virulency and antigenicity of the virus. The VP1 had 242E and 390L were characteristic of vvIBDV and KpnI restriction enzyme (777GGTAC/C782) in addition to a new mutation (F243Y and N383H) in tIBD-EGY-AH1 and AH4 strains. According to the current study, the strains were distinct from the vaccinal strain; they could be responsible for the most recent IBDV outbreaks observed in flocks instead of received vaccinations. The current study highlighted the importance of regular IBD monitoring to maintain up to date on the type of challenge viruses and the efficacy of commercial vaccinations.

Molecular characterization of the VP2 protein of BGM-70 adapted low passaged UPM190 infectious bursal disease virus isolate

The study evaluated the genetic profile of a low passaged BGM-70 adapted very virulent infectious bursal disease virus (vvIBDV) isolated in Malaysia in the year 2004 based on the VP2 hypervariable region. Embryonated chicken eggs were first used to propagate UPM190 12 times before being low passaged in BGM-70 cell line 7 times yielding UMP190BGMP7 whose VP2 nucleotide and amino acid sequences were determined and compared with reference sequences after Sanger sequencing of the low passage virus. The nucleotide and amino acid sequences of UPM190BGMP7 virus were similar with the sequences of viruses passaged in chicken embryonated eggs (CEE) but differed with higher BGM-70 passaged viruses. The E270 amino acid mutation associated with higher BGM-70 passaged viruses was absent in UPM190BGMP7. However, it has another amino acid mutation at position D279N that back-mutated on continuous BGM-70 cell propagation.

A Single Mutation of VP2 is Responsible for the Lethality and Antigenicity Differences between Novel Variant and Very Virulent IBDV Strains

Infectious bursal disease is an acute, immunosuppressive infectious disease in chickens caused by the infectious bursal disease virus (IBDV), which causes huge economic losses to the global poultry industry. Persistently circulating very virulent IBDV (vvIBDV) and newly emerging novel variant IBDV (nVarIBDV) are the two dominant epidemic strains of IBDV in East Asian countries such as China. Compared to lethal vvIBDV, nonlethal nVarIBDV has more insidious pathogenicity and can partially escape the immune protection of the existing vvIBDV vaccine, suggesting its potential adaptive survival strategy. However, the underlying molecular mechanism remains unknown. The viral capsid protein VP2 is closely related to cell tropism, virulence, and antigenic variation of IBDV. In this study, for the first time, we demonstrated that residue 279 of VP2 is responsible for the difference in pathogenicity between nVarIBDV and vvIBDV and that the D279N substitution reduces the lethality of vvIBDV from 70% to 0%. Moreover, a significant reduction in the viral load and inflammatory factor levels in the immune organs and blood of infected chickens may be important mechanisms for reducing the lethality of IBDV. Additionally, residue 279 was an important molecular basis for the antigenic differences between nVarIBDV and vvIBDV. D279N substitution reduced the neutralizing ability of vvIBDV antiserum against nVarIBDV by affecting the binding ability of antibodies and antigens. Our results indicate that nVarIBDV has an infection transmission strategy that facilitates its survival by hiding viral pathogenicity and escaping antiserum neutralization, which not only has significant implications for the systemic cognition of viral genetic evolution and pathogenesis but also provides new ideas for the comprehensive prevention and control of IBDV.

Efficacy of Fowlpox Virus Vector Vaccine Expressing VP2 and Chicken Interleukin-18 in the Protection against Infectious Bursal Disease Virus.

In mammals, the role of interleukin-18 (IL-18) in the immune response is to drive inflammatory and, normally therefore, anti-viral responses. IL-18 also shows promise as a vaccine adjuvant in mammals. Chicken IL-18 (chIL-18) has been cloned. The aim of this study was to investigate the potential of chIL-18 to act as a vaccine adjuvant in the context of a live recombinant Fowlpox virus vaccine (fpIBD1) against Infectious bursal disease virus (IBDV). fpIBD1 protects against mortality, but not against damage to the bursa of Fabricius caused by IBDV infection. The Fowlpox virus genome itself contains several candidate immunomodulatory genes, including potential IL-18 binding proteins (IL-18bp). We knocked out (Δ) the potential IL-18bp genes in fpIBD1 and inserted (::) the cDNA encoding chIL-18 into fpIBD1 in the non-essential ORF030, generating five new viral constructs -fpIBD1::chIL-18, fpIBD1ΔORF073, fpIBD1ΔORF073::chIL-18, fpIBD1ΔORF214, and fpIBD1ΔORF214::chIL-18. The subsequent protection from challenge with virulent IBDV, as measured by viral load and bursal damage, given by these altered fpIBD1 strains, was compared to that given by the original fpIBD1. Complete protection was provided following challenge with IBDV in chicken groups vaccinated with either fpIBDIΔ073::IL-18 or fpIBD1Δ214::IL-18, as no bursal damage nor IBDV was detected in the bursae of the birds. The results show that chIL-18 can act as an effective vaccine adjuvant by improving the fpIBD1 vaccine and providing complete protection against IBDV challenge.

MOLECULAR CHARACTERIZATION OF INFECTIOUS BURSAL DISEASE VIRUS ISOLATED FROM BROILER AND PULLET FLOCKS IN ALGERIA

Infectious bursal disease (IBD) or Gumboro disease is an acute, highly contagious viral disease of young chickens characterised by haemorrhagic syndrome, severe damage in the cloacal bursa, immunosuppression, and high mortality, generally at 3–6 weeks of age. The present study was performed to investigate the molecular characteristics and histopathological effects of infectious bursal disease virus (IBDV) isolated from broiler and pullet flocks in eastern and central Algeria. Fifty-five chickens collected from eleven broiler and pullet farms were investigated for IBD outbreaks over the period of 2019–2020. Only the birds with clinical signs and macroscopic lesions indicating IBD were selected for histopathological examination and molecular investigations using reverse transcription-polymerase chain reaction (RT-PCR) followed by sequencing. Except for Flock No4, all the birds in the study showed microscopic lesions of IBD. RT-PCR confirmed IBDV infection in samples from flocks No1, 2, 3, 9, 10, and 11. Using sequencing, a very virulent IBDV (vvIBDV) strain was detected in samples No10 and 11. The studied strains exhibited four conserved amino acids (222A, 256I, 294I, and 299S), characteristic of vvIBDV. According to the phylogenetic tree, the two strains in the study were closely related to previously isolated vvIBDVs and clustered together. This result may explain the frequent vaccine failure against IBD observed in broiler and pullet flocks in Algeria. Keywords: Molecular characterization, sequencing, vvIBDV, chickens, Algeria

Molecular epidemiology of infectious bursal disease virus in the Near East and Persian Gulf regions

ABSTRACT Infectious bursal disease virus (IBDV) is a ubiquitous immunosuppressive pathogen which is a severe burden to the poultry industry. Due to frequent mutation and reassortment events, its double-stranded, bi-segmented RNA genome displays a considerable heterogeneity, which in turn produces profound differences in antigenicity and pathogenicity. In recent years, the investigation of IBDV epidemiology has greatly benefited from the proposal of multiple phylogenetic classification systems which prompted the execution of molecular surveys all over the world, producing a wealth of standardized and easily sharable data. The present epidemiological survey was conducted in six Near East and Persian Gulf countries to characterize the field IBDVs circulating in the region. Ninety-three broiler flocks, immunized with different vaccination protocols, were investigated over a one-year period by collecting eight individual bursal samples at a time. Using molecular assays targeting both genome segments, field IBDVs belonging to four genotypes were detected from 42 flocks across all investigated countries. While traditional very virulent IBDVs (genotype A3B2) represented a minority, multiple atypical IBDV genotypes (A3B1, A4B1 and A6B1) were found to circulate in most of the region. Many of the detected strains were related to sequences from other continents, supporting the involvement of the Near and Middle East in complex networks of viral spreading. The obtained results, combined with the available literature, allowed us to update the epidemiological picture, hopefully favouring the identification of epidemiological threats and the planning of effective monitoring and control strategies. RESEARCH HIGHLIGHTS Different field IBDVs were found to circulate in the Near and Middle East. Multiple atypical genotypes (A3B1, A4B1, A6B1) were found to circulate extensively. Traditional very virulent IBDVs (A3B2) were a minority of the detected strains. Viral exchanges can be hypothesized between the region and different continents.