Virulent IBDV Research Articles

Rapid identification, pathotyping and quantification of infectious bursal disease virus by high-resolution melting curve quantitative reverse transcription PCR analysis: An innovative technology well-suited for real-time large-scale epidemiological surveillance

With the virus continuing to evolve, very virulent IBDV (vvIBDV) and novel variant IBDV (nvIBDV) have become the predominant epidemic strains in China, exacerbated by the widespread use of attenuated vaccine strains (attIBDV), making a complex infection situation of IBDV in the field. Therefore, developing a rapid and accurate high-resolution melting curve quantitative reverse transcription PCR (HRM-qRT-PCR) for the identification and pathotyping of IBDV is crucial for clinical monitoring and disease control. Extensive data analysis and genome-screening of the three dominant IBDV pathotypes identified a specific region (nucleotides 2450–2603 in segment A) with distinct GC content as the detection target. Experimental testing of HRM-qRT-PCR revealed distinct melting curves and high sensitivity, with the detection limits of 61.2 copies/μL, 61.1 copies/μL and 67.5 copies/μL for vvIBDV, nvIBDV and attIBDV, respectively. The method exhibited excellent specificity, with no inter-genotypes cross-reactivity among the three pathotypes and no reactivity to other common avian pathogens. Applied to samples with double and triple co-infections of different IBDV pathotypes, the method displayed specific melting peaks corresponding to the viruses present in the samples, with an accuracy rate of 100 %. This method precisely identifies and differentiates all the single or co-infected samples, generating distinct peaks corresponding to the Tm values of each virus pathotype in traditional melting curve plots. Furthermore, the method overcomes the limitations of traditional pathotyping methods, requiring only one reaction to achieve rapid viral pathotyping and facilitating quantitative analysis of viruses within the samples. This study introduces an innovative HRM-qRT-PCR method, offering new technology to rapid and accurate identification, pathotyping and quantification of vvIBDV, nvIBDV, and attIBDV. With strong discriminatory power, user-friendliness and a short processing time, this method is highly attractive for the rapid IBDV pathotyping in real-time large-scale epidemiological surveillance during outbreaks.

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, Molecular, and Histopathological Patterns of a Novel Variant of Infectious Bursal Disease Virus in Chicken Flocks in Egypt.

After an extended period of detecting classical virulent, attenuated, and very virulent IBDV, a novel variant (nVarIBDV) was confirmed in Egypt in this study in 18, IBD vaccinated, chicken flocks aged 19-49 days. Partial sequence of viral protein 2 (VP2) [219 aa, 147-366, resembling 657 bp] of two obtained isolates (nos. 3 and 4) revealed nVarIBDV (genotype A2d) and OR682618 and OR682619 GenBank accession numbers were obtained. Phylogenetic analysis revealed that both nVarIBDV isolates were closely related to nVarIBDV strains (A2d) circulating in China, exhibiting 100% identity to SD-2020 and 99.5-98.1% similarity to ZD-2018-1, QZ, GX and SG19 strains, respectively. Similarity to USA variant strains, belonging to genotypes A2b (9109), A2c (GLS) and A2a (variant E), respectively, was 95.5-92.6%. Also, the VP2 hypervariable region in those two, A2d, isolates revealed greater similarities to Faragher 52/70 (Vaxxitek®) at 90.4% and to an Indian strain (Ventri-Plus®) and V217 (Xtreme®) at 89.7% and 86-88.9% in other vaccines. Histopathological examination of both the bursa of Fabricius and spleen collected from diseased chickens in flock no. 18 revealed severe atrophy. In conclusion, further studies are required to investigate the epidemiological situation of this novel genotype across the country, and to assess various vaccine protections against nVarIBDV. Additionally, vaccination of breeders with inactivated IBD vaccines including this nVarIBDV is essential to obtain specific maternal antibodies in their broilers.

The Over-40-Years-Epidemic of Infectious Bursal Disease Virus in China.

Infectious bursal disease (IBD) is an acute, highly contagious, immunosuppressive disease of chickens caused by the virus (IBDV), which critically threatens the development of the global chicken industry and causes huge economic losses. As a large country in the poultry industry, the epidemic history of IBDV in China for more than 40 years has been briefly discussed and summarized for the first time in this report. The first classic strain of IBDV appeared in China in the late 1970s. In the late 1980s and early 1990s, the very virulent IBDV (vvIBDV) rapidly swept across the entirety of China, threatening the healthy development of the poultry industry for more than 30 years. Variants of IBDV, after long-term latent circulation with the accumulation of mutations since the early 1990s, suddenly reappeared as novel variant strains (nVarIBDV) in China in the mid-2010s. Currently, there is a coexistence of various IBDV genotypes; the newly emerging nVarIBDV of A2dB1 and persistently circulating vvIBDV of A3B3 are the two predominant epidemic strains endangering the poultry industry. Continuous epidemiological testing and the development of new prevention and control agents are important and require more attention. This report is of great significance to scientific cognition and the comprehensive prevention and control of the IBDV epidemic.

Residues 318 and 323 in capsid protein are involved in immune circumvention of the atypical epizootic infection of infectious bursal disease virus.

Recently, atypical infectious bursal disease (IBD) caused by a novel variant infectious bursal disease virus (varIBDV) suddenly appeared in immunized chicken flocks in East Asia and led to serious economic losses. The epizootic varIBDV can partly circumvent the immune protection of the existing vaccines against the persistently circulating very virulent IBDV (vvIBDV), but its mechanism is still unknown. This study proved that the neutralizing titer of vvIBDV antiserum to the epizootic varIBDV reduced by 7.0 log2, and the neutralizing titer of the epizootic varIBDV antiserum to vvIBDV reduced by 3.2 log2. In addition, one monoclonal antibody (MAb) 2-5C-6F had good neutralizing activity against vvIBDV but could not well recognize the epizootic varIBDV. The epitope of the MAb 2-5C-6F was identified, and two mutations of G318D and D323Q of capsid protein VP2 occurred in the epizootic varIBDV compared to vvIBDV. Subsequently, the indirect immunofluorescence assay based on serial mutants of VP2 protein verified that residue mutations 318 and 323 influenced the recognition of the epizootic varIBDV and vvIBDV by the MAb 2-5C-6F, which was further confirmed by the serial rescued mutated virus. The following cross-neutralizing assay directed by MAb showed residue mutations 318 and 323 also affected the neutralization of the virus. Further data also showed that the mutations of residues 318 and 323 of VP2 significantly affected the neutralization of the IBDV by antiserum, which might be deeply involved in the immune circumvention of the epizootic varIBDV in the vaccinated flock. This study is significant for the comprehensive prevention and control of the emerging varIBDV.

Protective Efficacy of IBDV Winterfield H-2512 and SYZA-26 Immune-complex Vaccines against Recent Egyptian Very Virulent IBDV in Commercial Broiler Chickens

Protective Efficacy of IBDV Winterfield H-2512 and SYZA-26 Immune-complex Vaccines against Recent Egyptian Very Virulent IBDV in Commercial Broiler Chickens

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

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.

Gene cloning, protein expression and examination of biological activity of chicken CD40L

To obtain chicken CD40L protein, the cDNA was prepared from chicken splenic cells and used as a template to clone and amplify CD40L by PCR. The target gene was cloned into pFastBac vector to construct a pFastBac-chCD40L donor plasmid. Recombinant plasmid was transformed into DH10Bac and recombinant Bacmid-chCD40L was obtained. The Bacmid-chCD40L plasmid was transfected into sf9 insect cells to obtain His-chCD40L protein. In addition, the target gene was cloned into pQM01 vector to construct a pQM01-chCD40L plasmid, recombinant plasmid was transfected into HEK 293T cells to obtain Strep-chCD40L protein. The chCD40L protein was purified by affinity chromatography, and the concentration of purified chCD40L protein was determined to be 0.01 mg/mL. Primary cells were isolated from the bursal tissue of 3-week old SPF chickens, and the chCD40L protein was added to the culture medium to stimulate cells. The chCD40L could bind to CD40 on B cells as examined by Western blotting, indirect immunofluorescence assay and flow cytometry, suggesting that chCD40L protein is biologically active. We successfully obtained chicken CD40L protein of biological activity, which laid the foundation in the in vitro culture of primary B lymphocytes for the isolation and diagnosis of virulent IBDV.

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

BackgroundInfectious 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).ResultsIn 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.ConclusionsThe 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.

Development of a recombinant VP2 vaccine for the prevention of novel variant strains of infectious bursal disease virus

ABSTRACT Since 2017, novel variant strains of infectious bursal disease virus (nvIBDV) have been detected in China, while the current vaccines on the market against very virulent IBDV have limited protection against this subtype virus. In this context, a strain of the virus has been isolated, and sequencing alignment and bird regression experiments showed that the virus was IBDV, belonging to the nvIBDV subtype (and named IBDV FJ-1812). Furthermore, the Escherichia coli expression system was used to successfully express soluble nvIBDV rVP2, which is specifically recognized by an anti-IBDV standard serum and anti-nvIBDV positive serum, and could be assembled into 14 − 17 nm virus-like particles. Based on the purified nvIBDV rVP2, we developed an IBDV FJ-1812 VP2 VLP vaccine at a laboratory scale to evaluate protection by this vaccine; in addition, we also prepared an IBDV JZ 3/02 VP2 subunit vaccine targeting very virulent IBDV and evaluated its cross-protection against nvIBDV. Results of bird experiments showed that the nvIBDV rVP2 vaccine could induce high titres of specific antibodies, completely protect the bursa of Fabricius from viral infection, and provide 100% immune protection to SPF and Ross 308 broiler chickens. Furthermore, the IBDV JZ 3/02 VP2 subunit vaccine targeting very virulent IBDV could provide 60% protection for SPF chickens and 80% protection for Ross 308 broiler chickens. This report provides important technical supports for the prevention and control of nvIBDV in the future.

Efficacy of The Turkey Herpes virus-IBDV Vector Vaccine Against Recent Egyptian Very Virulent IBDV in Commercial Layers

The infectious bursal disease virus (IBDv) is widespread in poultry flocks all around the world. During the period between 2016- 2017, 22 IBD outbreaks were investigated in El-Minoufiya Governorate in different chicken sectors (11 broilers, 7 commercial layer pullets and 4 native baladi varieties). The disease occurred at 23-42 days of age. The mortality ranged between 3-9 % in commercial broilers farms, 5.8-12% in native breed varieties and 5-10 % in commercial layer replacement pullets. Two hundred one-day-old commercial male layer chicks was used for assessment of protection obtained after vaccination with HVT-IBDV vector vaccine Vaxxitek against challenged with vvIBDV local field isolate Lay./Men.Egypt/17 at 46th-day of age. Maternal derived antibodies (MDA) was followed up by ELISA to determine MA waning and the suitable age of chicks for IBDV vaccination and/or challenge. Chicks divided into 4 groups of (50 birds/ of each), G1 (vaccinated with HVT-IBD vector vaccine at 1 day old of age and non-challeneged) and G2 (vaccinated with HVT-IBDV at one-day of age and challenged with vvIBDV local field isolate Lay./Men.Egypt/017 at 46-day of age), G3 (non- vac-cinated and non-challenged) and G4 (non-vaccinated and challenged chickens). Our results revealed that the mortality percentage of commercial male layer chickens chal-lenged with recent vvIBDV local field isolate lay./Men.Egypt/017 was (0%) in G2, while in G4 was (90%), at 7-days post challenge. The highest bursa body ratio observed was (5.32 and 5.31) in non- vaccinated and non-challenged birds (G3) and birds vac-cinated with the VAXXITEK HVT-IBDv vaccine (G1). Conclusion, the recombinant VAXXITEK HVT-IBDv vaccine has provided protection for commercial male layer chicks against challenge with recent vvIBDv isolate. Vaccination with vector vaccine in en-demic areas with vvIBDV, protect against mortality, partially against bursal atrophy and decreases immunosuppressive effect of vvIBDV.

Genetic point mutation inducing antigenic drift in hypervariable region of a very virulent IBDV isolate in chickens in Egypt during 2014-2016

Genetic point mutation inducing antigenic drift in hypervariable region of a very virulent IBDV isolate in chickens in Egypt during 2014-2016

Molecular characterization of field isolates of infectious bursal disease virus from three decades, 1987-2018, reveals a distinct genotypic subgroup in Vietnam.

The complete nucleotide sequence of the viral protein 2 (VP2) ORF was determined for 26 Vietnamese infectious bursal disease isolates collected from clinical outbreaks in vaccinated flocks from 1987 to 2018 and two commercial vaccine specimens. These sequences were compared for molecular classification with 42 reference strains representing all four main classes of serotype 1, including very virulent (vvIBDV), classical (cvIBDV), antigenic variant (avIBDV) and attenuated (atIBDV) strains, and serotype 2 strains. Amino acids at nine key positions in the VP2-HVR in 20 Vietnamese isolates, A222, I242, Q253, I256, D279, A284, I294, S299, A329, which are typical of the vvIBDV class, were found to be identical in all of the isolates. Eighteen of these isolates had a unique change at residue 212 (D212N) located in the PAB loop. Phylogenetic analysis revealed a distinct lineage/subclade with strong nodal support (96%) that included recent Chinese IBDV strains that were distinct from typical vvIBDVs. Six isolates contained the amino acid substitutions P222, V242, Q253, V256, D279, A284, I294, N299, A329, which are present in two vaccine strains derived from strain 2512 and these isolates were also closely related to the classical virulent STC strain. Data from this study show that there is considerable genetic diversity among vvIBDVs, which vary according to geographic region. Antigenic drift and differences in genetic characteristics between virulent strains and IBDV vaccine strains may be the cause of vaccine failure. Better antigenic matching of vaccines to the strains circulating in Vietnam is therefore required.

Phylogenetic analysis of Infectious Bursal Disease viruses according to newly proposed model of classification into geno-groups

BackgroundInfectious bursal disease virus (IBDV) is the causative agent of Infectious Bursal Disease (IBD), the disease causes immunosuppression which leads to secondary infections among rearing poultry flocks. Characterization of the virus is important for its control and eradication. The circulating IBDVs are classified on the basis of their antigenic and pathogenic properties. The virus is categorised as classical, variant and very virulent IBDV (vvIBDV). IBDV is a non-envelop, icosahedral double stranded virus. Viral protein 2 (VP2) is the major structural protein of capsid that determines the host-pathogen relationship. The aim of this study was to characterise the IBD virus of Pak-Asian region. MethodologyIBDV suspected flocks were examined in Punjab, Pakistan from 2014–2018. Two hundred and fifty samples were collected with complete history of the disease. The suspected samples were collected from broiler, layer and rural poultry farms. RNA was extracted and hyper-variable region of VP2 gene was amplified using specific primers. Nucleotide sequence of the VP2 gene was determined and its Amino Acid sequence was deduced. Moreover, phylogenetic analysis was also performed. ResultsThe current classifications based in a hyper-variable region of the capsid protein VP2 (hvVP2), classification of IBDVs is split into newly proposed geno-groups according to Jackwood group. Among these prevailing, some IBDVs are limited geographically whereas, others are reported cosmopolitan. Genetic alterations are continuously playing role in evolution of new strains of the virus. ConclusionDuring this study it was found that isolates of IBDV fall in first three geno-groups. Most of the geno-groups are prevalent around the world, whereas the mutated and re-assorted ones are confined in particular areas of the globe.

©ПАТОМОРФОЛОГІЧНІ ЗМІНИ У ЛІМФОЇДНІЙ ТКАНИНІ БУРСИ ФАБРІЦІУСА КУРЕЙ, ВИКЛИКАНІ РІЗНИМИ ВАКЦИННИМИ ТА ПОЛЬОВИМИ ШТАМАМИ ВІРУСУ ХВОРОБИ ГАМБОРО ВИЯВЛЕНИМИ В УКРАЇНІ

The infectious bursal disease virus was detected in the Gumboro рrovince of the United States in 1962 and since then has become widespread throughout the world. Gumboro disease is characterized by severe immunosuppression in affected birds, and often mortality. Since 1986, very virulent strains of IBDV appeared in central Europe, and caused up to 70% mortality of chickens. Vaccination became the main method of protection against the disease. It is important to differentiate the post-infectious and post-vaccination effects of the virus on the bursa of the Fabricius of birds to control the vaccination process. Therefore, the aim of the study was to detect the effects of various vaccine and very virulent IBDV strains on the bursa of Fabricius of chickens. For this purpose, histological examination of bursas of vaccinated and infected birds was carried out. Samples were taken from different poultry farms in Ukraine. In results of the study we observed positive correlation between degree of depletion of the cloacal bursa and the residual virulence of the vaccine strains. Also, we detected very severe (up to 100%) depletion of bursa follicles caused by very virulent strains. The results of the work can be further used for the analysis of the post-vaccination reaction in chickens during preventive measures in places of industrial bird production.

Preparation Of Inactivated Infectious Bursal Disease Virus Vaccine From Classical Strain And Local Variant Isolates

This study was conducted to assess an inactivated vaccine prepared from field of Infectious Bursal Disease (IBD) local variant and virulent isolates and a classical strain to improve the full control over the problem of IBD in Egypt. This experiment applied on 21 day old SPF chicks divided into six groups, the first group was vaccinated with inactivated vaccine containing (local variant isolate, classical bursa vacc strain and virulent isolate), the second group was vaccinated with inactivated vaccine containing (classical bursa vacc strain and variant isolate), the third group was vaccinated with classical bursa vacc vaccine, and the group 4 vaccinated with variant isolate vaccine while last two groups were left as a positive and negative control groups. The immune response of chicks was evaluated in vitro by measuring serum neutralization test (SNT) and enzyme linked immunosorbent assay (ELISA) and in vivo by challenging birds with 103.5 EID50/ dose of virulent virus of infectious bursal disease virus (vvIBDV).The highest average antibody level was obtained at the 4th week post vaccination in chicks in group 1 vaccinated with (local variant isolate, classical bursa vacc and virulent isolate) vaccine, while the chicks in group 3 vaccinated with bursa vacc vaccine showed the lowest antibody level at the same week post vaccination using SNT and ELISA. The challenge of group 1, 2, 3 and 4 showed protection of 97.5%, 90%, 75% and 90% respectively using very virulent IBDV 28 day post vaccination with no clinical signs or lesions on examination. It was concluded that we can use an inactivated vaccine prepared from (classical strain and variant and virulent isolates) of Infectious Bursal Disease (IBD) virus as a method for control IBD disease in Egypt.

Gallid herpesvirus 3 SB-1 strain as a recombinant viral vector for poultry vaccination

Live herpesvirus-vectored vaccines are widely used in veterinary medicine to protect against many infectious diseases. In poultry, three strains of herpesvirus vaccines are used against Marek’s disease (MD). However, of these, only the herpesvirus of turkeys (HVT) has been successfully developed and used as a recombinant vaccine vector to induce protection against other avian viral diseases such as infectious bursal disease (IBD), Newcastle disease (ND) or avian influenza (AI). Although effective when administered individually, recombinant HVT vectors have limitations when combined in multivalent vaccines. Thus there is a need for developing additional viral vectors that could be combined with HVT in inducing protection against multiple avian diseases in multivalent vaccines. Gallid herpesvirus 3 (GaHV3) strain SB-1 is widely used by the poultry industry as bivalent vaccine in combination with HVT to exploit synergistic effects against MD. Here, we report the development and application of SB-1 as a vaccine vector to express the VP2 capsid antigen of IBD virus. A VP2 expression cassette was introduced into the SB-1 genome at three intergenic locations (UL3/UL4, UL10/UL11 and UL21/UL22) using recombineering methods on the full-length pSB-1 infectious clone of the virus. We show that the recombinant SB-1 vectors expressing VP2 induced neutralising antibody responses at levels comparable to that of commercial HVT-based VAXXITEKHVT+IBD vaccine. Birds vaccinated with the experimental recombinant SB-1 vaccine were protected against clinical disease after challenge with the very virulent UK661 IBDV isolate, demonstrating its value as an efficient viral vector for developing multivalent vaccines against avian diseases.

Gumboro disease (IBD)

Gumboro Disease (IBD) caused by Infectious Bursal Disease Virus is an immunosuppressive condition of chickens, which resulted in severe economical losses. Approximately 47 years after the first appearance of the disease, the changes in the form of presentation and pathogenicity of IBDV in the field has resulted in continual changes and adjustments of the vaccines used to control IBD. The original form of the disease, also known as classic or clinical IBD, was usually observed after the third week of life with high mortality, depending on factors, such as virulence of the strain of virus involved, age of the birds and maternal antibody status. The acute IBD is caused by the very virulent IBDV strains, characterized by high mortality rates in vaccinated chickens and the clinical signs and lesions are similar to those of the classic form. The subclinicalform of the disease is associated with the "variant" strains of IBDV and is characterized by low mortality and severe immunosuppression. Initially, the classic IBD was controlled by the use of mild vaccines produced in the early 1970s and from the late 1980s revealed absence of protection from wIBDV in vaccinated chickens. Another group of vaccinal strains, known as intermediate plus vaccine, has been used to control wIBDV strains. These vaccines multiply in birds, even in the presence of high maternal antibody titers. In the USA and other countries, the disease from the variant strains was controlled by intermediate strains as well as by inactivated vaccines that included variant strains, thus providing maternal antibodies against both the standard and variant strains. Until today, the IBD continues to pose an important threat to commercial poultry industry. The high resistence of IBDV to physical and chemical agents accounts for persistence of the virus in the outside environment, particularly on contaminated farms, despite disinfection. Nevertheless and not quite unexpected for the IBDV mutations in the genome, resulted in the emergence of antigenic variant strains in vaccinated flocks. The IBD requires heightened vigilance and the incidence and prevalence of the clinical and immunosuppressive forms must be evaluated more precisely.

Transcriptional profiles in bursal B-lymphoid DT40 cells infected with very virulent infectious bursal disease virus

BackgroundInfectious bursal disease virus (IBDV) causes a highly contagious, immunosuppressive disease in chickens. The virus mainly infects immature B lymphocytes in the bursa of Fabricius (BF). Chicken B cell line DT40, an avian leukosis virus-induced B cell line, supports very virulent IBDV (vvIBDV) infection in vitro and thereby serves as a good model for investigating the infection and pathogenesis of this virus. However, a transcriptome-wide understanding of the interaction between vvIBDV and B cells has not yet been achieved. This study aimed to employ time-course DNA microarrays to investigate gene expression patterns in DT40 cells after infection with vvIBDV strain LX.ResultsDT40 cells infected with vvIBDV exhibited alterations in the expression of many important host genes involved in signal transduction pathways, including MAPK signaling, PI3K/mTOR signaling, cell death and survival, BCR signaling, and antigen presentation. The changes in cellular mRNA levels identified by microarray analysis were confirmed for 8 selected genes using real-time reverse transcription-PCR. The upregulation of inflammatory cytokines and Toll-like receptors (TLRs) in the bursa of vvIBDV-infected chickens might involve excessive activation of the innate immune and inflammatory responses and contribute to tissue damage.ConclusionsThe present study is the first to provide a comprehensive differential transcriptional profile of cultured DT40 cells in response to vvIBDV infection and further extends our understanding of the molecular mechanisms underlying vvIBDV infection and pathogenesis.

English

In recent years very virulent (VV) IBDV strains and classical (CV) IBDV strains re-emerged and caused devastating outbreaks in different parts of the world. In this study, genetic evolution of fifteen IBDVs collected in Senegal in 1979, 1999, 2007, 2012, 2013 and 2014 was characterized to gain information for a better control of IBD. Following RT-PCR, nucleotide sequence of the VP2 hypervariable region was determined and compared with sequences available in GenBank. Phylogenetic analysis showed that the viruses diverged into two genotypes: Very virulent (VV) IBDV and classical virulent (CV) IBDV. The Senegalese field strains of the first genotype (VV) IBDV had 98.9 to 100% identity among themselves, whereas their identity with reported Nigerian (VV) IBDVs ranged between 96.7 and 99%. The close phylogenetic relationship of the Senegalese and Nigerian strains suggests that they likely derived from a common ancestor. In the phylogenetic tree, all the Senegalese (VV) IBDV strains belonged to the African very virulent types (VV2). The genotyping of senegalese field IBDV strains indicated that the majority of viruses circulating in Senegal are (VV) IBDVs and highlights a genetic stability. Key words: Infectious bursal disease, phylogenetic analysis, Senegal.