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Evidence of genetic drift and reassortment in infectious bursal disease virus and emergence of outbreaks in poultry farms in India.

ABSTRACT: Recurrent outbreaks of infectious bursal disease (IBD) have become a burning problem to the poultry industry worldwide. Here, we performed genetic analysis of IBD virus (IBDV) field isolates from recent outbreaks in various poultry farms in India. The sequence analysis of IBDV VP2 hypervariable region revealed amino acid pattern similar to that of very virulent (222A, 242I, 253Q, 256I, 272I, 279D, 284A, 294I, 299S and 330S) and intermediate plus virulent (222A, 242I, 253Q, 256I, 272T, 279N, 284A, 294I, 299S and 330S) type whereas analysis of VP1 revealed presence of sequence similar to that of very virulent (61I, 145T) and unique (61I, 141I, 143D, 145S) type in field isolates. Among the eight field isolates, two isolates contained very virulent type VP2 and unique type VP1, three contained intermediate plus virulent type VP2 and unique type VP1 whereas five contained both VP2 and VP1 of very virulent type. The phylogenetic analysis based on VP2 nucleotide sequence showed clustering of all eight isolates close to known very virulent strains whereas based on VP1, five isolates formed unique cluster and three isolates were placed close to very virulent strains. The isolates forming unique VP1 cluster showed highest similarity with classical virulent IBDVs suggesting their possible evolution from segment B of non-very virulent IBDVs. Interestingly, these five isolates were responsible for outbreaks in four different farms located at three different geographic locations in India. These observations indicates genetic reassortment between segment A and segment B from co-infecting IBDV strains leading to emergence of very virulent strains and their widespread prevalence in Indian poultry farms. The presence of 272I and 279D in VP2 protein of five field isolates may explain possible cause of Gumboro intermediate plus vaccine failure in prevention of the outbreaks. However, mortality caused by other three strains which are antigenically similar to VP1 of intermediate plus vaccine strains could not be explained and the possible role of their unique VP1 in enhancing the pathogenesis needs to be investigated further.

SUBMITTER: Patel AK

PROVIDER: S-EPMC4909000 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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Evidence of genetic drift and reassortment in infectious bursal disease virus and emergence of outbreaks in poultry farms in India.

Patel Amrutlal K AK Pandey Vinod C VC Pal Joy K JK

Virusdisease 20160211 2

Recurrent outbreaks of infectious bursal disease (IBD) have become a burning problem to the poultry industry worldwide. Here, we performed genetic analysis of IBD virus (IBDV) field isolates from recent outbreaks in various poultry farms in India. The sequence analysis of IBDV VP2 hypervariable region revealed amino acid pattern similar to that of very virulent (222A, 242I, 253Q, 256I, 272I, 279D, 284A, 294I, 299S and 330S) and intermediate plus virulent (222A, 242I, 253Q, 256I, 272T, 279N, 284A ...[more]

PMID: 27366767

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Project description:AimThe study was undertaken to isolate infectious bursal disease virus (IBDV) from clinical cases in broiler and cockerel flocks of Maharashtra state, India, and its molecular epidemiological investigation.Materials and methodsThe morbid bursal tissues were collected from flocks suspected for IBD. The samples were subjected for virus adaptation in primary chicken embryo fibroblast (CEF) cells followed by confirmation by reverse transcription polymerase chain reaction (RT-PCR) for partial VP2 sequence and phylogenetic analysis.ResultsThe isolation of IBDV from field samples took seven blind passages for adaptation in CEF. The cytopathic effects included rounding, aggregation, vacuolation, and detachment of the cells. The RT-PCR showed amplification of 627 bp amplicon specific to the primers for VP2 gene fragment which confirmed successful adaptation and isolation of IBDV using CEF. The nucleotide and deduced amino acids based on phylogeny clustered the current isolate in a distinct clade with classical virulent and antigenic variants. It showed divergence from very virulent (vv) and vaccine strains of Indian origin. The isolate showed unique amino acid substitution at A329V as compared to all other IBDVs. The variation in key amino acids was reported at A222, I242, Q249, Q253, A256, T270, N279, T284, I286, L294, N299, and V329. It shared conserved amino acids at position A222, I242, and Q253 as reported in vvIBDV isolates. However, the amino acids reported at position T270, N279, T284, L294, and N299 are conserved in classic, antigenic variant and attenuated strains of IBDV. The amino acids at positions N279 and T284 indicated that the isolate has key amino acids for cell culture replication.ConclusionThe IBDV field isolate does not reveal the full nucleotide sequence signature of vvIBDV as well as vaccine strains. Hence, we can conclude that it might not belong to vvIBDVs of Indian origin and the vaccine strain used in the region. This may be suggestive of the evolution of the IBDV in the field due to the coexistence of circulating field strains and live attenuated hot strains, resulting into morbidity and mortality, warranting the need for safer protective vaccines, and implementation of stringent biosecurity measures to minimize loss to farmers.

Project description:Vaccination is an essential component in controlling infectious bursal disease (IBD), however, there is a lack of information on the genetic characteristics of a recent infectious bursal disease virus (IBDV) that was isolated from IBD vaccinated commercial flocks in Malaysia. The present study investigated 11 IBDV isolates that were isolated from commercial poultry farms. The isolates were detected using reverse transcription-polymerase chain reaction (RT-PCR) targeting the hypervariable region (HVR) of VP2. Based on the HVR sequences, five isolates (IBS536/2017, IBS624/2017, UPM766/2018, UPM1056/2018, and UPM1432/2019) were selected for whole-genome sequencing using the MiSeq platform. The nucleotide and amino acid (aa) sequences were compared with the previously characterized IBDV strains. Deduced aa sequences of VP2HVR revealed seven isolates with 94-99% aa identity to very virulent strains (genogroup 3), two isolates with 97-100% aa identity to variant strains (genogroup 2), and two strains with 100% identity to the vaccine strain (genogroup 1) of IBDV. The phylogenetic analysis also showed that the isolates formed clusters with the respective genogroups. The characteristic motifs 222T, 249K, 286I, and 318D are typical of the variant strain and were observed for UPM1219/2019 and UPM1432/2019. In comparison, very virulent residues such as 222A, 249Q, 286T, and 318G were found for the vvIBDV, except for the UPM1056/2018 strain with a A222T substitution. In addition, the isolate has aa substitutions such as D213N, G254D, S315T, S317R, and A321E that are not commonly found in previously reported vvIBDV strains. Unlike the other vvIBDVs characterized in this study, UPM766/2018 lacks the MLSL aa residues in VP5. The aa tripeptides 145/146/147 (TDN) of VP1 were conserved for the vvIBDV, while a different motif, NED, was observed for the Malaysian variant strain. The phylogenetic tree showed that the IBDV variant clustered with the American and Chinese variant viruses and are highly comparable to the novel Chinese variants, with 99.9% identity. Based on the sequences and phylogenetic analyses, this is the first identification of an IBDV variant being reported in Malaysia. Further research is required to determine the pathogenicity of the IBDV variant and the protective efficacy of the current IBD vaccines being used against the virus.

Project description:The growth of the poultry industry in Nigeria is constrained by major poultry diseases, despite the implementation of vaccination programs. This study aimed to assess the level of protection against Newcastle disease (ND), infectious bursal disease (IBD), and avian infectious bronchitis (IB) afforded by current vaccination schedules and characterize the circulating virus strains in commercial poultry flocks in Nigeria. A cross-sectional study was conducted on 44 commercial poultry farms in Oyo and Kano states of Nigeria. Serum and tissue samples and data on flock, clinical and vaccination records were collected on each farm. Farms were classified as being protected or not protected against ND, IBD and IB based on a defined criterion. Real-time reverse transcription polymerase chain reaction (rRT-PCR) testing was performed for each target virus on tissue samples and positive samples were sequenced. A total of 15/44 (34.1%), 35/44 (79.5%), and 1/44 (2.3%) farms were considered to be protected against ND, IBD, and IB, respectively, at the time of sampling. NDV RNA was detected on 7/44 (15.9%) farms and sequences obtained from 3/7 farms were characterized as the lentogenic strain. Infectious bursal disease virus (IBDV) RNA was detected on 16/44 (36.4%) farms tested; very virulent (vv) IBDV and non-virulent (nv) IBDV strains were both detected in 3/16 (18.8%) positive samples. Sequences of IBDV isolates were either clustered with a group of genotype 3 virulent IBDV strains or were related to vaccine strains MB and D78 strains. IBV RNA was detected on 36/44 (81.8%) farms, with variant02, Massachusetts, 4/91, and Q1 variants detected. Sequences of IBV isolates were either clustered with the vaccines strains Massachusetts M41 and H120 or were most closely related to the D274-like strains or a clade of sequences reported in Nigeria and Niger in 2006 and 2007. This study revealed that most study farms in Oyo and Kano states did not have adequate protective antibody titers against IBV and NDV and were therefore at risk of field challenge. Infectious bursal disease virus and IBV RNA were detected on farms with a history of vaccination suggesting potential vaccination failure, or that the vaccine strains used mismatch with the circulating strains and are therefore not protective.

Project description:The poultry sector contributes four percent to the national GDP of Nepal. However, this sector is under threat with periodic outbreaks of Avian Influenza (AI) subtypes H5 and H9 since 2009. This has been both a public health threat and an economic issue. Since the past few years, outbreaks of AI subtype H9 have caused huge economic losses in major poultry producing areas of Nepal. However, the risk factors associated with these outbreaks have not been assessed. A retrospective case-control study was conducted from April 2018 to May 2019 to understand the risk factors associated with AI subtype H9 outbreaks in Kathmandu valley. Out of 100 farms selected, 50 were "case" farms, confirmed positive to H9 at Central Veterinary Laboratory, Kathmandu, and another 50 farms were "control" farms, matched for farm size and locality within a radius of three km from the case farm. Each farm was visited to collect information using a semi-structured questionnaire. Twelve potential risk factors were included in the questionnaire under the broad categories: birds and farm characteristics, and management and biosecurity status of the farms. Univariable and multivariable logistic regression analysis was conducted and corresponding odds ratios were calculated. Risk factors, associated with AI subtype H9 outbreaks in Kathmandu valley, identified in the final multivariable model were: "farms that have flock size greater than median flock size of study farms (>1500)" (OR = 4.41, 95% CI: 1.53-12.71, p = 0.006), "farms that did not apply rules to wear boots for visitors inside the farms" (OR = 4.32, 95% CI: 1.52-12.29, p = 0.006) and "other commercial farms located within one km periphery" (OR = 10, 95% CI: 1.8-50, p = 0.007). This study showed that outbreaks of AI subtype H9 in Kathmandu valley were associated with a higher population of birds in the farm, poor management practices, and weak biosecurity measures in poultry farms. We suggest improving management practices and increase biosecurity in the farms to reduce incidences of AI subtype H9 outbreaks in Kathmandu valley.

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Evidence of genetic drift and reassortment in infectious bursal disease virus and emergence of outbreaks in poultry farms in India.

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Evidence of genetic drift and reassortment in infectious bursal disease virus and emergence of outbreaks in poultry farms in India.

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