Search Results (71)

Infectious bursal disease virus (IBDV) is one of the most important immunosuppressive viruses in poultry, causing the global spread of infectious bursal disease (IBD). It poses a significant threat to the healthy development of the poultry industry. Vaccination is an effective approach for controlling IBDV infection. Therefore, reliable immune monitoring for IBDV is critical for maintaining poultry health. The enzyme-linked immunosorbent assay (ELISA) is a common technique used to detect specific antibodies in clinical serum testing and for the serological evaluation of IBDV vaccines. Among the currently available and under development IBDV vaccines, IBD VP2 subunit-based vaccines account for a considerable proportion. These vaccines stimulate the production of antibodies that are specific only to VP2. However, most IBDV antibody ELISA kits approved for use have applied the whole virus as the coating antigen, which does not adequately meet the diverse requirements for IBDV detection across different conditions. This study utilized a prokaryotic expression system to express the VP2 protein of the IBDV epidemic strain, assembling it into virus-like particles to be used as coating antigens. This approach enabled the establishment of an indirect ELISA method for detecting IBDV VP2 antibody (VP2-ELISA). The optimal coated antigen concentration was determined to be 2.5 μg/mL, with overnight coating at 4 °C; sealing with 5% skim milk at 37 °C for 4 h; serum dilution at 1:500 with incubation at 37 °C for 30 min; secondary antibody dilution at 1:4000 with incubation at 37 °C for 40 min; and then incubation with the substrate solution 3,3′,5,5′-tetramethylbenzidine at room temperature for 20 min. The criterion for interpreting the detection results was OD_450nm ≥ 0.111 indicates IBDV antibody positivity, while OD_450nm < 0.111 indicates negativity. The established VP2-ELISA can specifically detect IBDV-positive sera at the lowest serum dilution of 1:6400, with intra- and inter-batch coefficients of variation of <2%. This indicates that the VP2-ELISA exhibits good specificity, sensitivity, and stability. Detection experiments using 20 laboratory-immunized chicken serum samples and 273 clinical serum samples demonstrated that the results of VP2-ELISA were consistent with those of commercial ELISA kits coated with whole virus. In summary, the VP2-ELISA developed in this study offers advantages in immune response detection for IBD VP2 subunit-based vaccines and is appropriate for evaluating the efficacy of IBD vaccines and detecting clinical serum samples. Full article

The etiology of transmissible viral proventriculitis (TVP) of broiler chickens has been discussed since its initial recognition 40 years ago. Regardless of its low direct impact on mortality rate, it leads to high economic losses in the broiler industry through reduction of food conversion, weakening of birds, and their increased susceptibility to pathogens. The aim of the present study was to examine the potential presence of TVP on the broiler chicken farms in Bosnia and Herzegovina, to characterize microscopic lesions, and to investigate the viruses implicated in etiology of TVP by PCR-based methods. In total, 143 diseased broiler chickens from 16 farms in Bosnia and Herzegovina were euthanized and subjected to necropsy and subsequent histopathology of proventriculi. A representative number of proventriculi samples (n = 50) that exhibited histopathologic changes were processed for molecular detection of chicken proventricular necrosis virus (CPNV), girovirus (GyV3), chicken anemia virus (CAV), and infectious bursal disease virus (IBDV) by PCR-based methods. In addition, samples of bursa of Fabricius (n = 39) and spleen (n = 50) were tested for IBDV. Histopathology revealed changes consistent with TVP in 39.8% (57/143) and LP (lymphocytic proventriculitis) in 2.1% (3/143) of samples. All 50 proventricular samples showed positivity to CPNV with Ct values ranging between 18 and 26. GyV3 was detected in eight samples (16%), with Ct values ranging from 11.1 to 27.5. The presence of CAV was more prominent (38%), with 19 positive broiler chickens (Ct ranging from 9.6 to 35.6). Pooled samples of spleen, bursa, and proventriculi from three farms were positive for IBDV. The obtained results represent the first documented data on TVP and the first record of CPNV and GyV3 presence in broiler farms from Bosnia and Herzegovina. Full article

Infectious bursal disease (IBD) is a major immunosuppressive disease affecting young chickens, and causes significant economic losses to the poultry industry. This work represents the first pathogenicity assessment of Moroccan very virulent IBD virus. Molecular characterization and sequence analysis of this isolate previously identified specific substitutions, including seven amino acid substitutions in segment A, and I472L and E688D in segment B, specific and unique to Moroccan vvIBDV strains. Two chicken lines, broiler and specific-pathogen-free (SPF) chickens, were inoculated via the occulonasal route with 0.2 mL of the 10⁵EID₅₀ /mL viral solution of the IB19 vvIBDV strain at 29 days of age. Experimental monitoring was carried out for 10 days post-challenge (dpc). Clinical signs started on the second dpc, with peak severity observed between 3 and 6 dpc. The total mortality rate reached 10% in broilers (group G1) and 93% in SPF chickens (G3). Macroscopic lesions in G1 broilers included marked hypertrophy of the bursa of Fabricius (BF), followed by very pronounced atrophy, while macroscopic examinations of deceased SPF birds (G3) revealed very hemorrhagic BF with a black cherry appearance in 80% of dead birds. The mean Bursa/Body Index (BBI) of challenged broilers (G1) showed a decrease of 46% compared to the control group (G2), indicating bursal atrophy. Microscopic lesions in the BF consisted mainly of inflammation, with severe lymphoid depletion of the follicles in challenged G3 SPF birds. This in vivo study of Moroccan vvIBDV demonstrated a distinctive virulence profile, and confirmed its classification as a very virulent strain with substantial disease-causing potential. It is crucial to obtain comprehensive knowledge of the prevalence, emergence, pathogenicity, and control of Moroccan IBDV strains. Full article

Infectious bursal disease virus (IBDV) is an important pathogen affecting the poultry industry worldwide. IBDV serotype 1, including classical virulent strains (cvIBDV), variant strains (varIBDV), and very virulent strains (vvIBDV), is pathogenic for chickens. IBDV mainly infects immature B-lymphocytes in the bursa of Fabricius, weakening the humoral immune response and leading to secondary infections and increased morbidity and mortality. The Laboratory of Avian Pathology received ten live 8-week-old pullets from a laying hen operation experiencing increased mortality, prostration, diarrhea, and sudden death. Upon necropsy, the affected birds presented swollen, hemorrhagic, and edematous bursa of Fabricius, as well as hemorrhage in the breast and thigh muscles. RT-PCR confirmed that the samples from the bursa of Fabricius were positive for IBDV. Phylogenetic analysis of the VP1 and VP2 gene nucleotide sequences classified the strain, isolated in embryonated chicken eggs, as the A3B5 genotype. Amino acid sequence analysis of the VP2 hypervariable region revealed the presence of amino acid residues commonly found in vvIBDV. Additional studies are required to investigate the epidemiological situation of this genotype in Chile and to evaluate current vaccination plans and their effectiveness against new variants. Full article

Mycoplasma, reticuloendotheliosis virus (REV), avian leukosis virus (ALV), chicken infectious anemia virus (CIAV), bovine polyomavirus (BPV), bovine viral diarrhea virus (BVDV), and porcine circovirus (PCV) are considered common contaminants in live veterinary vaccines against Newcastle disease virus (NDV), fowlpox virus (FPV), infectious bursal disease virus (IBDV), classical swine fever virus (CSFV), pseudorabies virus (PRV), and porcine reproductive and respiratory syndrome virus (PRRSV). In the past five years, Getah virus (GETV), an arbovirus affecting many farming mammals, was reported as a new contaminant in live PRRSV vaccines in two previous studies, which arouses our considerable interest. Therefore, in this paper, we aim to analyze and discuss the source, biological hazard, and genomic characteristics of these contaminating GETV strains further. Full article

Novel antigenic variant strains of the infectious bursal disease virus (IBDV) classified into genogroup A2d have been found in the western part of Japan since 2017. Novel antigenic variant IBDVs now occur in higher frequencies in poultry houses and have been detected in the eastern part of Japan, indicating the spread of IBDVs despite the usual IBDV vaccination. We isolated a novel antigenic variant IBDV, designated as the B2977CE2C3 strain. The B2977CE2C3 strain had two genogroup A2d specific amino acids—lysine and isoleucine, at 221 and 252 aa—along with the other genogroup A2 common amino acids in the projection domains of the VP2 protein corresponding to the virus-neutralizing epitopes and viral pathogenicity. Experimental infection of the B2977CE2C3 strain did not produce any apparent clinical signs in the specific-pathogen-free chickens during the observation period (21 days), but atrophy of the bursa of Fabricius (BF) was apparent. The mean BF to the body weight ratio was 0.35 in negative control chickens at 21 days post-infection (pi) but 0.06 in the B2977CE2C3 infected group. An extremely high copy number of the IBDV genome (>10⁸ copies/µL) was observed in the BF at 3 days pi, while a high copy number of the IBDV genome (>10⁶ copies/µL) was observed in the thymus, spleen cecal tonsil, and bone marrow even though macroscopic lesions were not apparent in these organs. Full article

Infectious bursal disease (IBD) continues to threaten poultry production globally, with highly virulent strains circulating in many parts of Africa. In this study, molecular characterization was performed on a circulating infectious bursal disease virus (IBDV) strain from an outbreak in a layer flock in Ghana. Layer chicks presented for necropsy had markedly enlarged and hemorrhagic bursae of Fabricius, with necrotic foci and catarrhal exudate on the serosal surface. Histopathology of the bursa of Fabricius revealed scattered to effacing hemorrhages on the plicae, extensive necrosis with expansion of the stroma between the follicles, and depletion of lymphocytes within the interfollicular epithelium. Reverse transcription polymerase chain reaction (RT-PCR) and subsequent sequencing of the VP2 gene showed the presence of IBDV in formalin-fixed paraffin-embedded tissues. A phylogenetic analysis compared 62 other IBDV sequences from different parts of the world and placed the Ghanaian IBDV in genogroup 3 (vvIBDV), closely related to IBDV from Nigeria. In comparison to reference vvIBDV, there were amino acid substitutions at positions 252, 254, and 300. To the best of our knowledge, this is the first report in which an IBDV from a disease outbreak in Ghana has been sequenced and compared with other IBDVs in a phylogenetic analysis. Full article

Infectious bursal disease (IBD) is among the most impactful immunosuppressive diseases of poultry. Its agent, infectious bursal disease virus (IBDV), is prone to both mutation and reassortment, resulting in a remarkable variability. Traditionally, IBDV characterization relies on antigenicity and pathogenicity assessment, but multiple phylogenetic classifications have been recently proposed, whose implementation in molecular surveys helps generating informative and standardized epidemiological data. In the present study, the Algerian IBDV scenario was assessed based on the novel classification guidelines by sequencing portions of both genome segments. Seventy pools of bursal samples were collected in 2022–2023 in 11 districts of Northern Algeria, mostly from broiler flocks. Out of 55 (78.6%) positive flocks, 40 (57.1%) were infected by field strains, which were characterized as very virulent strains (genotype A3B2) and phylogenetically related to previously reported Algerian strains. Significant differences in the percentage of field infections were observed between vaccinated (25/52, 46.2%) and unvaccinated (14/17, 82.3%) groups, and also between birds immunized with live intermediate (13/20, 65.0%) and intermediate plus (10/28, 35.7%) vaccines. Nonetheless, the number of field strain detections suggests a high infectious pressure and the inadequacy of current vaccination efforts, demanding a reevaluation of control measures coupled with attentive monitoring activities. Full article

In the realm of poultry production, viral superinfections pose significant challenges, causing substantial economic losses worldwide. Among these, avian leukosis virus subgroup J (ALV-J) and infectious bursal disease virus (IBDV) are particularly concerning as they frequently lead to superinfections in chicken, further exacerbating production losses and health complications. Our previous research delved into the pathogenicity and immunosuppressive effects of these superinfections through in vitro and in vivo analyses. Yet, the underlying key genes and pathways governing this phenomenon remained elusive. In this study, we randomly selected three chickens at 21 days post infection from each treatment group (ALV-J, IBDV, ALV-J+IBDV, and control group) to collect the bursa of Fabricius samples for full transcriptome analysis. Utilizing these data, we constructed a comprehensive circRNA/lncRNA-miRNA-mRNA network which elucidated both synergistic and specific activations during the superinfection. Notably, three pivotal genes (FILIP1L, DCX, and MYPN) were pinpointed in datasets reflecting synergistic activations. Conversely, four other genes (STAP, HKR6, XKR4, and TLR5) emerged in datasets associated with specific activations. Further exploration revealed diverse significant GO terms and pathways associated with both synergistic and distinct activation processes. These ceRNA network and core genes potentially wield substantial influence over the synergistic or specific activation of tumorigenesis and pathogenesis induced by ALV-J and IBDV. These findings could help develop targeted therapies and improve disease control in poultry, reducing economic losses. Full article

Duck Tembusu virus (DTMUV), duck hepatitis virus (DHV), Muscovy duck reovirus (MDRV), and Muscovy duck parvovirus (MDPV) represent four emergent infectious diseases impacting waterfowl, which can be challenging to differentiate due to overlapping clinical signs. In response to this, we have developed a one-step multiplex real-time fluorescence quantitative reverse transcription PCR (qRT-PCR) assay, capable of simultaneously detecting DTMUV, DHV, MDRV, and MDPV. This method exhibits high specificity, avoiding cross-reactivity with other viruses such as Fowl adenoviruses (FADV), infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), Haemophilus paragallinarum (Hpg), duck circovirus (DUCV), goose astrovirus (GoAstV), and mycoplasma gallisepticum (MG). The limit of detection (LOD) established for DTMUV, DHV, MDRV, and MDPV was determined to be 27 copies/μL. In the repeatability test, the intra-assay and inter-assay coefficients of variation (CVs) of the recombinant plasmid standard were less than 2%. Utilizing this method, we analyzed 326 clinical specimens sourced from Guangxi over the period spanning October 2021 through December 2023, yielding promising and precise outcomes. The qRT-PCR method established herein exhibits commendable specificity, sensitivity, and repeatability. Furthermore, it boasts a high clinical detection rate, making it a highly effective tool for diagnosing these pathogenic agents in waterfowl. Full article

Background: Infectious bursal disease (IBD) is a highly infectious disease of chicken found in all latitudes, due to the very high resistance to environmental conditions and commonly used disinfectants of the IBD virus (IBDV). Methods: This study was conducted to evaluate three IBD vaccination protocols in broiler chicken in terms of their effectiveness (clinical observations and production performance of the flock), estimation of serological baseline values (with the use of two different commercial ELISA kits) and the degree of progression of BF lesions (histopathological lesion score (HLS)) after vaccination. The three protocols were (I) single vaccination using an intermediate plus vaccine, (II) double vaccination with an intermediate vaccine and (III) double vaccination with an intermediate plus vaccine. Results: Birds on farms vaccinated with protocol II were characterized by the lowest antibody titers in both ELISA tests and the lowest average HLS. The highest IDEXX titers were obtained in birds vaccinated with protocol III, while in the BIOCHEK test the highest titers were obtained for birds vaccinated with protocol I. Protocols I and III were characterized by similar HLSs. Birds vaccinated with protocols I and III had higher immune uniformity. Conclusions: The estimated serological baseline values and the degree of HLSs presents a clear picture of the differences between the different vaccination protocols and allows their adaptation for different farms depending on the current IBD epidemiological situation. Full article

Infectious bronchitis virus (IBV) causes infectious bronchitis in chicken, an acute, highly contagious respiratory infection. Because of genetic mutations and recombination, IBV forms many subtypes, which makes it difficult to treat the disease and apply commercial vaccines. Therefore, to detect IBV in time and stop the virus from spreading, a novel and convenient IBV detection technology based on reverse transcription recombinase-aided amplification (RT-RAA) was established in this study. According to the S1 gene of IBV CH I–V and Mass genotypes and S1 gene of IBV CH VI genotype, a set of optimal primers were designed and selected to establish a real-time dual fluorescence RT-RAA method. The lowest detection line was 10 copies/μL of RNA molecules and the method exhibited no cross-reactivity with avian reticuloendotheliosis virus (REV), infectious bursal disease virus (IBDV), avian leukosis virus (ALV), Newcastle disease virus (NDV), chicken infectious anemia virus (CIAV), infectious laryngotracheitis virus (ILTV), Marek’s disease virus (MDV), and H9N2 avian influenza virus (H9N2), demonstrating high specificity. When compared to qPCR detection results, our method achieved a sensitivity of 96.67%, a specificity of 90%, and a Kappa value of 0.87 for the IBV CH I–V and Mass genotypes, and achieved a sensitivity of 100%, a specificity of 97.73%, and a Kappa value of 0.91 for the IBV CH VI genotype. Full article

Infectious bursal disease virus (IBDV) is a significant burden for poultry production and market due to both direct disease and induced immunosuppression. In the present study, the expression of different cytokines in the bursa of Fabricius and thymus was evaluated during a 28-day-long experimental infection with two strains classified in the G1a (Classical) and G6 (ITA) genogroups. Although both strains significantly affected and modulated the expression of different molecules, the G6 strain seemed to induce a delayed immune response or suppress it more promptly. A recovery in the expression of several mediators was observed in the G1a-infected group at the end of the study, but not in the G6 one, further supporting a more persistent immunosuppression. This evidence fits with the higher replication level previously reported for the G6 and with the clinical outcome, as this genotype, although subclinical, has often been considered more immunosuppressive. However, unlike other studies focused on shorter time periods after infection, the patterns observed in this paper were highly variable and complex, depending on the strain, tissue, and time point, and characterized by a non-negligible within-group variability. Besides confirming the strain/genogroup effect on immune system modulation, the present study suggests the usefulness of longer monitoring activities after experimental infection to better understand the complex patterns and interactions with the host response. Full article

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). Full article

Infectious bursal disease (IBD) is an immunosuppressive disease causing significant damage to the poultry industry worldwide. Its etiological agent is infectious bursal disease virus (IBDV), a highly resistant RNA virus whose genetic variability considerably affects disease manifestation, diagnosis and control, primarily pursued by vaccination. In Egypt, very virulent strains (genotype A3B2), responsible for typical IBD signs and lesions and high mortality, have historically prevailed. The present molecular survey, however, suggests that a major epidemiological shift might be occurring in the country. Out of twenty-four samples collected in twelve governorates in 2022–2023, seven tested positive for IBDV. Two of them were A3B2 strains related to other very virulent Egyptian isolates, whereas the remaining five were novel variant IBDVs (A2dB1b), reported for the first time outside of Eastern and Southern Asia. This emerging genotype spawned a large-scale epidemic in China during the 2010s, characterized by subclinical IBD with severe bursal atrophy and immunosuppression. Its spread to Egypt is even more alarming considering that, contrary to circulating IBDVs, the protection conferred by available commercial vaccines appears suboptimal. These findings are therefore crucial for guiding monitoring and control efforts and helping to track the spread of novel variant IBDVs, possibly limiting their impact. Full article