RESUMO
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.
Análisis de las secuencias genéticas y de la patogenicidad del virus de la enfermedad infecciosa de la bolsa de pollos en Egipto durante los años 20172021. La circulación continua del virus de la enfermedad infecciosa de la bolsa (IBDV) en Egipto, a pesar del uso de varias vacunas, continua siendo un problema serio que requiere la detección continua de este virus. En el presente estudio, se realizó una prueba de transcripción reversa y reacción en cadena de la polimerasa en tiempo real de 100 parvadas enfermas de pollos durante los años 20172021 y reveló la presencia de virus muy virulentos (vvIBDV) en el 67% de las parvadas, otros tipos diferentes a los muy virulentos en el 11%, y una mezcla de virus muy virulentos y otros tiposen un 4% de las parvadas. Se enviaron veintinueve aislados del virus de la enfermedad infecciosa de la bolsa para la secuenciación parcial de la región hipervariable de la proteína viral 2 (VP2-HVR), y se confirmó que 27 aislados pertenecían al genogrupo A3 (vvIBDV) con una similitud del 96.3% al 98.5% con el genogrupo A3 global (vvIBDV) y de 88.9% a 97% de similitud con las cepas vacunales del genogrupo A1. Los dos aislamientos restantes no resultaron ser muy virulentos y mostraron un 91.1% y un 100% de identidad con las cepas clásicas del genogrupo A1, respectivamente. Además, la secuencia y el análisis filogenético de la proteina VP1 (aminoácidos 33-254) de dos aislados seleccionados de genogrupo A3, 5/2017 y 98/2021, los agruparon como cepas B2, similares a virus muy virulentos, con alta similitud (99.5%) con cuatro aislamientos de Egipto, con similitud de 99% con aislados chinos y europeos, y de 97.7% con aislados muy virulentos chinos y polacos. La infección experimental de pollos de engorde comerciales con dos aislados muy virulentos tipo A3B2 (5/2017 y 98/2021) no mostró mortalidad a pesar de las lesiones tisulares típicas, los cambios histopatológicos claros y la fuerte respuesta de anticuerpos por ELISA. El aislado 98/2021 fue más patógeno, según lo confirmado por histopatología, mientras que el aislado 5/2017 indujo una respuesta serológica más fuerte. En conclusión, las cepas muy virulentas (A3B2) con dos sustituciones de aminoácidos (aa) en la proteina VP1 como V141I y V234I, así como en VP2 tales como Y220F y G254S, todavía circulan en Egipto.
Assuntos
Infecções por Birnaviridae , Galinhas , Vírus da Doença Infecciosa da Bursa , Filogenia , Doenças das Aves Domésticas , Animais , Vírus da Doença Infecciosa da Bursa/genética , Vírus da Doença Infecciosa da Bursa/patogenicidade , Infecções por Birnaviridae/veterinária , Infecções por Birnaviridae/virologia , Infecções por Birnaviridae/epidemiologia , Doenças das Aves Domésticas/virologia , Doenças das Aves Domésticas/epidemiologia , Egito/epidemiologia , VirulênciaRESUMO
Infectious bursal disease (IBD) represents a greatly transmissible viral disease found worldwide, causing significant health and production challenges in young chickens. The aim of this research was to assess the immune reaction induced by different vaccines targeting IBD. These vaccines included recombinant (Vac1; HVT-IBD vector), immune complex (Vac2; Bursa-Plex®), and intermediate plus (Vac3; Bursine plus) IBD vaccines. Our assessment relied on serological and histopathological analyses, as well as the pattern of immune-related cytokine expression in the bursal tissue. The vaccinated groups, along with a control positive (CP) group, were subjected to a vvIBDV challenge on their 28th day of life, while the control negative (CN) group received a mock vaccination with PBS. Our study revealed that Vac1 resulted in the most favorable growth performance, as well as maintained normal liver and kidney function, mitigating the impact of IBDV infection. Serological analysis using VP2 ELISA kits indicated that Vac1 induced the strongest immunological response among all vaccines. Histopathological examination demonstrated that Vac1 caused minimal lymphoid depletion observed in the lymphoid organs, followed by Vac2. Analysis of cytokine expression profiles showed significant upregulation in all vaccinated groups, particularly Vac1, during the pre-challenge period. Following IBDV infection, Vac1 resulted in a noteworthy increase in the expression of IL2 and IFN-γ, Vac2 showed a significant upregulation in TNF-α and granzyme, and both Vac1 and Vac3 exhibited increased levels of IL1ß and IL10. In conclusion, our study suggests that the various vaccines triggered immune responses against IBD through both humoral and cell-mediated immunity. However, recombinant followed by immune complex vaccines appeared to induce more robust immunity while also being safer for broiler chickens in contrast to the intermediate plus vaccine.