RESUMEN
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.
Asunto(s)
Infecciones por Birnaviridae , Pollos , Virus de la Enfermedad Infecciosa de la Bolsa , Filogenia , Enfermedades de las Aves de Corral , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Animales , Egipto/epidemiología , Infecciones por Birnaviridae/veterinaria , Infecciones por Birnaviridae/virología , Infecciones por Birnaviridae/epidemiología , Enfermedades de las Aves de Corral/virología , Enfermedades de las Aves de Corral/epidemiología , Pollos/virología , Brotes de Enfermedades/veterinaria , Secuencia de Aminoácidos , Vacunas Virales/inmunología , Vacunación/veterinaria , Proteínas Estructurales Virales/genética , Virulencia , Variación GenéticaRESUMEN
Infectious bursal disease (IBD) is one of the most important immunosuppressive diseases of young chickens, causing considerable economic losses to the poultry industry. More than 30 years ago, an antigenic variant (av) pathotype of the IBD virus (IBDV) was reported to originate in, and subsequently spread among, poultry farms in the USA. Recently, a novel avIBDV lineage was identified in China and was shown to exhibit clear differences in its pathogenicity as well as molecular characteristics compared with the previously isolated variant strains. In this study, we conducted a passive surveillance of chicken carcasses submitted to our research division from June-December 2019, and detected the IBDV strains by reverse transcription PCR. Five avIBDV strains were isolated, and their pathogenicity was determined by necropsy and molecular analysis. Additionally, a coinfection field case involving an avIBDV strain and a very virulent IBDV (vvIBDV) strain was identified. Multiple sequence alignment and phylogenetic analysis of partial viral protein 1 (VP1) and hypervariable region (hv) VP2 genes revealed that those strains originated from two different avIBDV lineages. The co-occurrence of two sub-groups of avIBDVs in South Korea confirms for the first time the evolution of antigenic variant IBDV strains, and highlights the urgency for the development of new strategies for IBDV intervention in South Korea.RESEARCH HIGHLIGHTS Five avIBDV strains were identified in South Korea by passive surveillance test in 2019.A coinfection between two IBDV strains from different genogroups was reported in a field case.By phylogenetic analysis, Korean avIBDVs belonged to two distinct lineages of antigenic variant genogroup.
Asunto(s)
Variación Antigénica/genética , Infecciones por Birnaviridae/veterinaria , Pollos/virología , Virus de la Enfermedad Infecciosa de la Bolsa/inmunología , Enfermedades de las Aves de Corral/virología , Proteínas Estructurales Virales/genética , Animales , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/patología , Infecciones por Birnaviridae/virología , Monitoreo Epidemiológico , Genotipo , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/crecimiento & desarrollo , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Filogenia , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/patología , República de Corea/epidemiologíaRESUMEN
Recently, multiple spillover events between domesticated poultry and wild birds have been reported for several avian viruses. This phenomenon highlights the importance of the livestock-wildlife interface in the possible emergence of novel viruses. The aim of the current study was to investigate the potential spillover and epidemiological links of infectious bursal disease virus (IBDV) between wild birds and domestic poultry. To this end, twenty-eight cloacal swabs were collected from four species of free-living Egyptian wild birds (i.e. mallard duck, bean goose, white-fronted goose and black-billed magpie). Genetic and phylogenetic analysis of three positive isolates revealed that the IBDV/USC-1/2019 strain clustered with previously reported very virulent IBDV (vvIBDV) Egyptian isolates. Interestingly, two other wild bird-origin isolates (i.e. IBDV/USC-2/2019 and IBDV/USC-3/2019) grouped with a vaccine strain that is being used in commercial poultry. In conclusion, our results revealed the molecular detection of vaccine and vvIBDV-like strains in Egyptian wild birds and highlighted the potential role of wild birds in IBDV epidemiology in disease-endemic regions.
Asunto(s)
Animales Salvajes/virología , Enfermedades de las Aves , Aves/virología , Infecciones por Birnaviridae , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Animales , Enfermedades de las Aves/epidemiología , Enfermedades de las Aves/virología , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/virología , Egipto , Aves de Corral/virología , Enfermedades de las Aves de Corral/virologíaRESUMEN
Infectious bursal disease virus (IBDV), which infects young chickens, is one of the most important pathogens that harm the poultry industry. Evaluation of the immune status of birds before and after vaccination is of great importance for controlling the disease caused by this virus. Therefore, the development of low-cost and easy-to-manufacture test systems for IBDV antibody detection remains an urgent issue. In this study, three expression systems (bacteria, yeast, and human cells) were used to produce recombinant VP3 protein of IBDV. VP3 is a group-specific antigen and hence may be a good candidate for use in diagnostic tests. Comparison of the antigenic properties of the obtained polypeptides showed that the titres of antibodies raised in chickens against bacteria- or human-cell-derived recombinant VP3 were high, whereas the antibody level against yeast-derived recombinant VP3 was low. The results of an enzyme-linked immunosorbent assay (ELISA) of sera from IBDV-infected chickens demonstrated that the recombinant VP3 produced in E. coli would be the best choice for use in test systems.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/inmunología , Péptidos/inmunología , Enfermedades de las Aves de Corral/virología , Proteínas Estructurales Virales/inmunología , Animales , Anticuerpos Antivirales/inmunología , Infecciones por Birnaviridae/virología , Pollos , Ensayo de Inmunoadsorción Enzimática , Mapeo Epitopo , Escherichia coli/genética , Escherichia coli/metabolismo , Virus de la Enfermedad Infecciosa de la Bolsa/química , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Péptidos/química , Péptidos/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Estructurales Virales/química , Proteínas Estructurales Virales/genéticaRESUMEN
Infectious bursal disease virus (IBDV), the etiological agent of infectious bursal disease (IBD), is a variable RNA virus of Avibirnavirus. Some artificially attenuated vaccine strains of IBDV can adapt to cell culture of chicken embryo fibroblast (CEF) cell or its immortalized cell line DF1 in vitro while wild-type IBDV cannot. In this study, for the first time, a naturally occurring cell-adapted classic strain (genogroup 1) of IBDV named IBD17JL01 was identified in China. Animal experiments showed that IBD17JL01 could severely damage the central immune organ of infected chickens. Sequence analysis of the full-length genome revealed the peculiar molecular characteristics of IBD17JL01 with a few amino acid substitutions that might be involved in cell-tropism, antigenicity, and virulence of IBDV. Identification of this novel strain is beneficial to our understanding of the complexity of the epidemiology of IBDV. And the expansion of viral cell-tropism might increase the potential risk of the reassortment of different IBDVs including the live vaccines.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Pollos/virología , Genoma Viral , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Tropismo Viral , Adaptación Biológica , Sustitución de Aminoácidos , Animales , Técnicas de Cultivo de Célula , Línea Celular , China , Fibroblastos/virología , Genotipo , Filogenia , Enfermedades de las Aves de Corral/virología , VirulenciaRESUMEN
Infectious bursal disease virus (IBDV) is an immunosuppressive pathogen of poultry causing great economic losses to the poultry industry. In this study, the IBDV antibodies were detected in captured free-living wild birds in Zaria, Nigeria. One hundred and fifty free-living wild birds, comprising 30 birds each of 5 different species, were sampled over a period of 9 months. Blood samples were collected from each bird, and harvested sera were tested for IBDV antibodies using enzyme-linked immunosorbent assay. Results indicated IBDV seroprevalences in speckled pigeon (6.67%) and cattle egret (3.33%). In conclusion, the detection of IBDV antibodies in free-living wild birds in this study is indicative of previous natural exposure of these birds to the virus. These species of wild birds could therefore serve as carriers of these viruses and, consequently, transmit these viruses to chickens.
Asunto(s)
Enfermedades de las Aves/epidemiología , Infecciones por Birnaviridae/veterinaria , Columbidae , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Pájaros Cantores , Animales , Animales Salvajes , Anticuerpos Antivirales/sangre , Enfermedades de las Aves/virología , Aves , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/virología , Nigeria/epidemiología , Prevalencia , Estudios SeroepidemiológicosRESUMEN
Infectious bursal disease (IBD) is an immunosuppressive viral disease of chickens, associated with severe economic losses and major threats to poultry production worldwide. Disease prevention programs rely on unequivocal identification of the pathogen, as well as vaccination programs. This study developed a sensitive, one-step, real-time, quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay using a hydrolysis probe system for infectious bursal disease virus (IBDV, VP1 gene) detection and quantification, which was compared to other routinely used diagnostic methods. The assay successfully detected IBD reference viruses and field isolates. The absence of cross-reactivity was detected with negative samples or with other avian viruses in the analytical specificity test. The detection limit of this assay was 70 RNA copies. RT-qPCR was more sensitive in the detection of serially diluted IBDV isolates compared to virus isolation. For clinical samples, the sensitivity and specificity values of RT-qPCR compared to enzyme-linked immunosorbent assay (ELISA) were 97.5% and 100%, respectively, and compared to histopathology, these values were 100% and 93.94%, respectively. RT-qPCR can provide a simple and reliable assay for IBDV surveillance programs and for evaluation of control strategies.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Proteínas de la Cápside/genética , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Enfermedades de las Aves de Corral/diagnóstico , Animales , Infecciones por Birnaviridae/diagnóstico , Infecciones por Birnaviridae/virología , Pollos , Pruebas Diagnósticas de Rutina/métodos , Pruebas Diagnósticas de Rutina/normas , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Técnicas de Diagnóstico Molecular , Enfermedades de las Aves de Corral/virología , Reacción en Cadena en Tiempo Real de la Polimerasa , Estándares de Referencia , Sensibilidad y Especificidad , Virulencia/genéticaRESUMEN
Infectious bursal disease virus (IBDV) is the aetiological agent of a highly contagious chicken immunodeficiency disorder known as Gumboro disease, which cause severe economic loses to the poultry worldwide. The emergence of very virulent IBDV strains (vvIBDV) during the late 80s resulted in drastic changes to the epidemiology of IBDV with a dramatic increase in the mortality of the animals affected. Molecular studies determined that the emergence of the vvIBDV was a consequence of a genomic reorganization of IBDV known as reassortant event by which the virus combined two emergent genetic background vvIBDV for segment A and vvIBDV for segment B. In the current study, a retrospective analysis was conducted, and samples collected during acute outbreaks of Gumboro disease in Poland during 1992-2015 were submitted to sequencing and further molecular and phylogenetic analyses. The results obtained not only revealed a high genetic diversity for Polish IBDV strains but a new population of IBDV was identified. These novel reassortant strains with a unique genetic background contain the segment A from very virulent strains and segment B from an unidentified source, phylogenetically segregated and classified as 'transitional lineage'. The results obtained also showed the presence of this new lineage in Finland, evidencing the expansion of this new genomic reorganized viral strain in Europe representing an additional threat to the global situation of IBDV.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Brotes de Enfermedades/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Enfermedades de las Aves de Corral/epidemiología , Virus Reordenados/aislamiento & purificación , Enfermedad Aguda , Animales , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/virología , Pollos/virología , Finlandia/epidemiología , Genoma Viral , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Polonia/epidemiología , Enfermedades de las Aves de Corral/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Virus Reordenados/genética , Estudios RetrospectivosRESUMEN
Recently, a new genotype of infectious bursal disease virus (IBDV), named ITA, was detected in IBD-vaccinated Italian broilers. Genome characterization revealed ITA to be a genetically different IBDV, belonging to genogroup 6 according to a recently proposed IBDV classification. The currently available clinical data do not allow any definition of the degree of pathogenicity of the ITA-IBDV isolates. In the present study, a pathogenicity trial was conducted by the oral inoculation of specific-pathogen-free (SPF) chickens. Birds were housed in poultry isolators and inoculated at 35 days of age with an ITA-IBDV isolate (35 birds) or a strain belonging to the G1a genogroup as a comparison (35 birds). Control birds (25 birds) were contextually mock-inoculated with sterile water. Birds were observed daily for clinical signs and at 0, 7, 14, 21 and 28 days post-inoculation (dpi) were bled for IBDV antibody detection. At 2, 4, 7, 14, 21 and 28 dpi, five birds from each of the inoculated groups, and three from the control group, were euthanized and subjected to a post-mortem examination; the bursa:body weight and thymus:body weight ratios were calculated. Microscopic lesions of the bursa and thymus were scored on the basis of lymphoid necrosis and/or depletion or cortex atrophy, respectively. Both viruses induced a subclinical course of disease, as neither clinical signs nor mortality were recorded during the study, even in the presence of typical IBDV gross and microscopic lesions. Bursal damage, measured by the bursa:body weight ratio, was more noticeable and precocious after ITA-IBDV inoculation. Histopathology scores of the bursa, indicative of rapid lymphoid depletion, confirmed the aggressiveness of the ITA-IBDV strain in this organ. This study showed that, although the ITA-IBDV strain tested causes infection with a subclinical course, it induces severe damage to lymphoid tissues. Therefore, its circulation in birds might be a threat for the poultry industry and may jeopardize the success of the production cycle.
Asunto(s)
Anticuerpos Antivirales/sangre , Infecciones por Birnaviridae/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Enfermedades de las Aves de Corral/virología , Vacunación/veterinaria , Animales , Infecciones por Birnaviridae/virología , Bolsa de Fabricio/patología , Bolsa de Fabricio/virología , Pollos , Genotipo , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Italia/epidemiología , Tejido Linfoide/patología , Tejido Linfoide/virología , Organismos Libres de Patógenos Específicos , VirulenciaRESUMEN
Between July and September 2017, samples collected from six unvaccinated chickens in Namibia were shown to be positive for infectious bursal disease virus (IBDV) by RT-PCR. Partial sequence and phylogenetic analysis of the VP1 and VP2 genes from six viruses revealed that they all belong to the very virulent pathotype (Genogroup 3) and are genetically very similar to IBDVs identified in neighbouring Zambia. This is the first molecular characterisation of IBDV in Namibia and has implications on the control and management of the disease in the country.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Pollos , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Enfermedades de las Aves de Corral/epidemiología , Animales , Infecciones por Birnaviridae/epidemiología , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Namibia/epidemiología , Filogenia , Enfermedades de las Aves de Corral/virología , Análisis de Secuencia de ADN/veterinariaRESUMEN
Fowl adenovirus serotype 4 (FAdV-4) is the pathogenic agent of hydropericardium hepatitis syndrome (HHS) in chickens and ducks, which has caused huge economic losses for the Chinese poultry industry since 2015. In order to objectively determine the prevalence and co-infection status of the virus in Shandong province in China, we analyzed a total of 679 clinical cases of chickens and ducks from 36 farms in the province. The results showed that the FAdV-4 infection rate was 65.2% (443/679), and the rate in breeder ducks was almost two-fold higher than that in breeder chickens (68.57% vs. 34.30%). Notably, co-infection by H9N2 avian influenza virus, infectious bursal disease virus, and/or chicken infectious anemia virus was very common in the 443 FAdV-4-positive cases. Furthermore, phylogenetic analysis of the hexon genes of four Shandong FAdV-4 isolates revealed that these strains clustered into Indian reference strains, indicating that the Shandong FAdV-4 strains might have originated in India. These findings provide the first data on the prevalence and co-infection status of FAdV-4 in Shandong province, which may serve as a foundation for the prevention of FAdV-4 in the field.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Coinfección/veterinaria , Adenovirus A Aviar , Enfermedades de las Aves de Corral/epidemiología , Adenoviridae/genética , Adenoviridae/inmunología , Adenoviridae/aislamiento & purificación , Infecciones por Adenoviridae/epidemiología , Animales , Proteínas de la Cápside/genética , Virus de la Anemia del Pollo/aislamiento & purificación , Pollos/virología , China/epidemiología , Coinfección/epidemiología , Coinfección/virología , Patos/virología , Adenovirus A Aviar/genética , Adenovirus A Aviar/inmunología , Adenovirus A Aviar/aislamiento & purificación , Genes Virales , Hepatitis Animal/epidemiología , Hepatitis Animal/virología , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Subtipo H9N2 del Virus de la Influenza A/aislamiento & purificación , Filogenia , Filogeografía , Aves de Corral/virología , Enfermedades de las Aves de Corral/virología , Prevalencia , SerogrupoRESUMEN
Bursa tissue samples from a pullet flock in New York State that was experiencing immune suppression related disease were sent to our laboratory in 2018. A very virulent infectious bursal disease virus (vvIBDV) was identified in those samples through molecular and pathogenicity studies and designated 1/chicken/USA/1054NY/18. Phylogenetic analyses of the hypervariable VP2 nucleotide sequence region indicated that this strain belonged to genogroup 3 which comprises the vvIBDV. Partial sequence data of the VP1 gene indicated this virus also had a VP1 typical of vvIBDV. While vvIBDV have previously been identified in the United States in California and Washington State, the 1054NY vvIBDV was most closely related to isolates from Ethiopia, suggesting it is a new introduction into the U.S. The 1054NY vvIBDV was used to challenge four-week old specific-pathogen-free (SPF) layer chicks where it caused 100% morbidity and 68.7% mortality within 4 days. Upon necropsy, gross pathological findings in infected SPF birds included small yellowish coloured bursas, some with haemorrhages on the serosal and mucosal surfaces. Microscopic lesions included inflammation, severe lymphocyte necrosis, atrophy of the follicles and follicular depletion of lymphocytes. RESEARCH HIGHLIGHTS A very virulent infectious bursal disease virus (vvIBDV) was detected in a pullet flock in New York state, USA. Nucleotide sequence analysis of the vvIBDV VP2 gene indicates it is not related to previous US vvIBDV isolates and appears to be a new introduction into the US. The New York vvIBDV caused 100% morbidity and 68.7% mortality in four-week-old specific-pathogen-free chicks.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Enfermedades de las Aves de Corral/virología , Animales , Infecciones por Birnaviridae/diagnóstico , Infecciones por Birnaviridae/patología , Infecciones por Birnaviridae/virología , Pollos , Femenino , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , New York , Filogenia , Enfermedades de las Aves de Corral/diagnóstico , Enfermedades de las Aves de Corral/patología , Organismos Libres de Patógenos Específicos , Proteínas Estructurales Virales/genética , VirulenciaRESUMEN
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.
Asunto(s)
Infecciones por Birnaviridae/virología , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Secuencia de Aminoácidos , Sustitución de Aminoácidos/genética , Animales , Variación Antigénica/genética , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/veterinaria , Pollos/virología , Brotes de Enfermedades , Genotipo , Filogenia , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/virología , Vietnam/epidemiología , Proteínas Estructurales Virales/genéticaRESUMEN
BACKGROUND: Lates calcarifer, known as seabass in Asia and barramundi in Australia, is a widely farmed species internationally and in Southeast Asia and any disease outbreak will have a great economic impact on the aquaculture industry. Through disease investigation of Asian seabass from a coastal fish farm in 2015 in Singapore, a novel birnavirus named Lates calcarifer Birnavirus (LCBV) was detected and we sought to isolate and characterize the virus through molecular and biochemical methods. METHODS: In order to propagate the novel birnavirus LCBV, the virus was inoculated into the Bluegill Fry (BF-2) cell line and similar clinical signs of disease were reproduced in an experimental fish challenge study using the virus isolate. Virus morphology was visualized using transmission electron microscopy (TEM). Biochemical analysis using chloroform and 5-Bromo-2'-deoxyuridine (BUDR) sensitivity assays were employed to characterize the virus. Next-Generation Sequencing (NGS) was also used to obtain the virus genome for genetic and phylogenetic analyses. RESULTS: The LCBV-infected BF-2 cell line showed cytopathic effects such as rounding and granulation of cells, localized cell death and detachment of cells observed at 3 to 5 days' post-infection. The propagated virus, when injected intra-peritoneally into naïve Asian seabass under experimental conditions, induced lesions similar to fish naturally infected with LCBV. Morphology of LCBV, visualized under TEM, revealed icosahedral particles around 50 nm in diameter. Chloroform and BUDR sensitivity assays confirmed the virus to be a non-enveloped RNA virus. Further genome analysis using NGS identified the virus to be a birnavirus with two genome segments. Phylogenetic analyses revealed that LCBV is more closely related to the Blosnavirus genus than to the Aquabirnavirus genus within the Birnaviridae family. CONCLUSIONS: These findings revealed the presence of a novel birnavirus that could be linked to the disease observed in the Asian seabass from the coastal fish farms in Singapore. This calls for more studies on disease transmission and enhanced surveillance programs to be carried out to understand pathogenicity and epidemiology of this novel virus. The gene sequences data obtained from the study can also pave way to the development of PCR-based diagnostic test methods that will enable quick and specific identification of the virus in future disease investigations.
Asunto(s)
Lubina/virología , Enfermedades de los Peces/virología , Genoma Viral , Virus de la Enfermedad Infecciosa de la Bolsa/clasificación , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Animales , Acuicultura , Línea Celular , Secuenciación de Nucleótidos de Alto Rendimiento , Virus de la Enfermedad Infecciosa de la Bolsa/ultraestructura , Microscopía Electrónica de Transmisión , Filogenia , Reacción en Cadena de la Polimerasa , SingapurRESUMEN
Infectious bursal disease (IBD) is one of the most important immunosuppressive diseases that seriously threaten poultry farming and food safety worldwide. The variant strain of infectious bursal disease virus (IBDV) has been greatly neglected for more than 30 years. Recently, the subclinical infection of suspected IBD, causing considerable economic losses, occurred in the main chicken-farming regions of China. Through RT-PCR, sequencing, and phylogenic analyses, novel variant IBDVs were first identified in six provinces of eastern China. Immunological detection further confirmed the antigenic variation of the Chinese variant IBDVs. The Chinese IBDV variants were obviously different from the American IBDV variants, with less than a 97.7% (VP1) or 98.7% (VP2) amino acid sequence identity. Animal experiments further confirmed the serious threat of the variant IBDVs to chickens, demonstrating irreversible damage to the central immune organ, obvious immunosuppression, and growth retardation. This study not only identified the pandemic nature of the novel variant IBDVs for the first time but also discovered the distinct molecular epidemiological characteristics of these viruses, which will contribute more to the control of the disease.
Asunto(s)
Variación Antigénica , Infecciones por Birnaviridae/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Enfermedades de las Aves de Corral/epidemiología , Animales , Infecciones por Birnaviridae/epidemiología , Pollos/virología , China/epidemiología , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Filogenia , Enfermedades de las Aves de Corral/virología , ARN Viral/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ADN , Proteínas Estructurales Virales/genéticaRESUMEN
Fowl adenovirus (FAdV), which causes the high-impact diseases such as inclusion body hepatitis and hepatitis-hydropericardium syndrome, is of major concern to the poultry industry internationally. This study was carried out in direct response to mortality rates of up to 75% in commercial broiler flocks in Trinidad, West Indies. Symptoms in 3- to 8-week-old broilers and 13- to 18-week-old pullets pointed to infection with an immunosuppressive viral pathogen. The objectives of the study were to determine whether the infectious agent FAdV, along with other viral pathogens, was responsible for the clinical disease, and to obtain information on the serotypes of FAdV that were infecting the birds. Tissue samples from clinically affected birds from eight different farms were tested for chicken infectious anaemia virus (CIAV) and infectious bursal disease virus (IBDV) by real-time reverse transcription polymerase chain reaction (PCR) and for FAdV by conventional PCR. The birds tested positive for FAdV and CIAV, but negative for IBDV. The gene corresponding to the L1 loop of the hexon protein for FAdV was amplified and sequenced. Phylogenetic analysis of seven FAdV strains inferred that four serotypes were likely to be circulating in the chickens. Well supported genetic relatedness was observed for serotype 8a (97.8%), 8b (97.8%), 9 (95.8%) and 11 (98.8%-99.5%). This is the first published report from Trinidad and Tobago on the presence and circulation of pathogenic FAdV strains, in combination with CIAV, in poultry. The data demonstrate a possible need for the introduction of serotype-specific vaccines against FAdV, as well as vaccines against CIAV, in broilers in the region and emphasize the importance of maintaining high levels of biosecurity on farms to prevent the spread of these potentially devastating viruses between farms.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Adenoviridae/aislamiento & purificación , Virus de la Anemia del Pollo/aislamiento & purificación , Pollos/virología , Infecciones por Circoviridae/veterinaria , Enfermedades de las Aves de Corral/virología , Adenoviridae/genética , Adenoviridae/inmunología , Infecciones por Adenoviridae/epidemiología , Infecciones por Adenoviridae/virología , Animales , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/veterinaria , Infecciones por Birnaviridae/virología , Virus de la Anemia del Pollo/genética , Virus de la Anemia del Pollo/inmunología , Infecciones por Circoviridae/epidemiología , Infecciones por Circoviridae/virología , Coinfección/veterinaria , Femenino , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/inmunología , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Filogenia , Enfermedades de las Aves de Corral/epidemiología , Serogrupo , Trinidad y Tobago/epidemiologíaAsunto(s)
Infecciones por Birnaviridae/veterinaria , Brotes de Enfermedades/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Virus Reordenados/genética , Animales , Infecciones por Birnaviridae/mortalidad , Genoma Viral , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Pakistán , Filogenia , Enfermedades de las Aves de Corral/virología , Reacción en Cadena en Tiempo Real de la Polimerasa , Virus Reordenados/aislamiento & purificación , Virus Reordenados/patogenicidad , Análisis de Secuencia de ADNRESUMEN
Infectious bursal disease virus (IBDV) is the causative agent of a highly contagious immunosuppressive disease affecting young chickens. The recently described "distinct IBDV" (dIBDV) genetic lineage encompasses a group of worldwide distributed strains that share conserved genetic characteristics in both genome segments making them unique within IBDV strains. Phenotypic characterization of these strains is scarce and limited to Asiatic and European strains collected more than 15 years ago. The present study aimed to assess the complete and comprehensive phenotypic characterization of a recently collected South American dIBDV strain (1/chicken/URY/1302/16). Genetic analyses of both partial genome segments confirmed that this strain belongs to the dIBDV genetic lineage and that it is not a reassortant. Antigenic analysis with monoclonal antibodies indicated that this strain has a particular antigenic profile, similar to that obtained in a dIBDV strain from Europe (80/GA), which differs from those previously found in the traditional classic, variant and very virulent strains. Chickens infected with the South American dIBDV strain showed subclinical infections but had a marked bursal atrophy. Further analysis using Newcastle disease virus-immunized chickens, previously infected with the South American and European dIBDV strains, demonstrated their severe immunosuppressive effect. These results indicate that dIBDV strains currently circulating in South America can severely impair the immune system of chickens, consequently affecting the local poultry industry. Our study provides new insights into the characteristics and variability of this global genetic lineage and is valuable to determine whether specific control measures are required for the dIBDV lineage. Research Highlights A South American strain of the dIBDV lineage was phenotypically characterized. The strain produced subclinical infections with a marked bursal atrophy. Infected chickens were severely immunosuppressed. The dIBDV strains are antigenically divergent from other IBDV lineages.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Pollos/virología , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/inmunología , Enfermedades de las Aves de Corral/virología , Animales , Infecciones por Birnaviridae/inmunología , Infecciones por Birnaviridae/virología , Pollos/inmunología , Genotipo , Inmunogenicidad Vacunal , Terapia de Inmunosupresión/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Fenotipo , Enfermedades de las Aves de Corral/inmunología , VirulenciaRESUMEN
BACKGROUND: Infectious 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. METHODOLOGY: IBDV 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. RESULTS: The 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. CONCLUSION: During 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.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Enfermedades de las Aves de Corral/epidemiología , Aves de Corral , Animales , Infecciones por Birnaviridae/epidemiología , Demografía , India/epidemiología , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , Enfermedades de las Aves de Corral/virología , ARN Viral/genéticaRESUMEN
Infectious bursal disease virus is a causative agent of one of the most important disease which causes frequent tragic disaster in the poultry industry all over the world. Therefore, in the present study a new fluorescence resonance energy transfer-based technique was developed to detect VP2 gene of infectious bursal disease virus using two oligonucleotide probes labeled with quantum dots and rhodamine- immobilized gold nanoparticles (AuNPs-Rh). Quantum dots labeled with an amino-modified first oligonucleotide, and AuNPs-Rh labeled with thiol-modified second oligonucleotides were added to the DNA targets upon which hybridization occurred. In the presence of target the AuNPs-Rh will be located in the vicinity of the quantum dots and leads to the fluorescence resonance energy transfer to be occurred and subsequently the fluorescence intensity of quantum dots was stimulated. The immobilization of rhodamine to the surface of AuNPs increased the fluorescence intensity of rhodamine. The maximum fluorescence resonance energy transfer efficiency for the developed sensor is monitored at a quantum dots-PA/AuNPs-Rh-PT molar ratio of 1:10. Moreover, the feasibility of the developed nanobiosensor was demonstrated by the detection of a synthetic 49-mer nucleotide derived from infectious bursal disease virus and the limit of detection was estimated as 3â¯×â¯10-8â¯M. The developed DNA detection scheme is a simple, rapid and efficient technique which does not need excessive washing and separation steps.