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ABSTRACT: Migratory wild birds can carry various pathogens, such as influenza A virus, which can spread to globally and cause disease outbreaks and epidemics. Continuous epidemiological surveillance of migratory wild birds is of great significance for the early warning, prevention, and control of epidemics. To investigate the pathogen infection status of migratory wild birds in eastern China, fecal samples were collected from wetlands to conduct pathogen surveillance. The results showed that duck orthoreovirus (DRV) and goose parvovirus (GPV) nucleic acid were detected positive in the fecal samples collected from wild ducks, egrets, and swan. Phylogenetic analysis of the amplified viral genes reveals that the isolates were closely related to the prevalent strains in the regions involved in East Asian-Australasian (EAA) migratory flyway. Phylogenetic analysis of the amplified viral genes confirmed that they were closely related to circulating strains in the regions involved in the EAA migration pathway. The findings of this study have expanded the host range of the orthoreovirus and parvovirus, and revealed possible virus transmission between wild migratory birds and poultry.

Simple Summary: A genomic epidemiological study was conducted to describe the genetic diversity within turkey reoviruses circulating in Hungary. Phylogenetic analyses confirmed that turkey reoviruses comprise a well determined genetic clade; however, evidence shows occasional genetic interaction between turkey reoviruses and reoviruses of other avian species. A better understanding of the source and the route of reovirus infections in the field requires extended sampling of strains from additional geographic areas and from different phases of turkey production. Reovirus infections in turkeys are associated with arthritis and lameness. Viral genome sequence data are scarce, which makes an accurate description of the viral evolution and epidemiology difficult. In this study, we isolated and characterized turkey reoviruses from Hungary. The isolates were identified in 2016; these isolates were compared with earlier Hungarian turkey reovirus strains and turkey reoviruses isolated in the 2010s in the United States. Gene-wise sequence and phylogenetic analyses identified the cell-receptor binding protein and the main neutralization antigen, σC, to be the most conserved. The most genetically diverse gene was another surface antigen coding gene, μB. This gene was shown to undergo frequent reassortment among chicken and turkey origin reoviruses. Additional reassortment events were found primarily within members of the homologous turkey reovirus clade. Our data showed evidence for low variability among strains isolated from independent outbreaks, a finding that suggests a common source of turkey reoviruses in Hungarian turkey flocks. Given that commercial vaccines are not available, identification of the source of these founder virus strains would permit a more efficient prevention of disease outbreaks before young birds are settled to fattening facilities. [ABSTRACT FROM AUTHOR]

Simple Summary: Avian orthoreoviruses cause significant losses to the poultry industry worldwide. Reovirus-associated diseases in chickens are frequently reported even in vaccinated flocks due to the emergence and spread of novel strains. Herein we studied the genetic features of reovirus strains originating from parts of East-Central Europe. The isolated strains from the study area displayed large genetic variability and shared low genetic similarity with vaccine strains. These findings highlight the importance of immune pressure from vaccination in the epizootiology of reovirus infection in chickens. The concern that the vaccines currently used against Avian orthoreovirus (ARV) infections are less efficient in the field justifies the need for the close monitoring of circulating ARV strains. In this study, we collected necropsy samples from various chicken breeds and tested for ARV by virus isolation, RT-PCR assay and sequence analysis. ARVs were isolated from birds showing runting-stunting syndrome, uneven growth, lameness or increased mortality, with relative detection rates of 38%, 35%, 6% and 25%, respectively. Partial σC gene sequences were determined for nearly 90% of ARV isolates. The isolates could be classified into one of the major genetic clusters. Interestingly, cluster 2 and cluster 5 were isolated from vaccinated broiler breeders, while clusters 1 to 4 were isolated from unvaccinated broilers. The isolates shared less than 75% amino acid identities with the vaccine strains (range, 44.3–74.6%). This study reaffirms the global distribution of the major genetic clusters of ARVs in chicken. The diversity of ARV strains isolated from unvaccinated broilers was greater than those detected from vaccinated animals, however, the relative importance of passive and active immunity on the selection of novel strains in different chicken breeds needs to be better understood. [ABSTRACT FROM AUTHOR]

Avian orthoreovirus (ARV) is among the important viruses that cause drastic economic losses in the Egyptian poultry industry. Despite regular vaccination of breeder birds, a high prevalence of ARV infection in broilers has been noted in recent years. However, no reports have revealed the genetic and antigenic characteristics of Egyptian field ARV and vaccines used against it. Thus, this study was conducted to detect the molecular nature of emerging ARV strains in broiler chickens suffering fromarthritis and tenosynovitis in comparison to vaccine strains. Synovial fluid samples (n = 400) were collected from 40 commercial broiler flocks in the Gharbia governorate, Egypt, and then pooled to obtain 40 samples, which were then used to screen ARV using reverse transcriptase polymerase chain reaction (RT-PCR) with the partial amplification of ARV sigma C gene. The obtained RT-PCR products were then sequenced, and their nucleotide and deduced amino acid sequences were analyzed together with other ARV field and vaccine strains from GenBank. RT-PCR successfully amplified the predicted 940 bp PCR products from all tested samples. The phylogenetic tree revealed that the analyzed ARV strains were clustered into six genotypic clusters and six protein clusters, with high antigenic diversity between the genotypic clusters. Surprisingly, our isolates were genetically different from vaccine strains, which aligned in genotypic cluster I/protein cluster I, while our strains were aligned in genotypic cluster V/protein cluster V. More importantly, our strains were highly divergent fromvaccine strains used in Egypt, with 55.09-56.23% diversity. Sequence analysis using BioEdit software revealed high genetic and protein diversity between our isolates and vaccine strains (397/797 nucleotide substitutions and 148-149/265 amino acid substitutions). This high genetic diversity explains the vaccination failure and recurrent circulation of ARV in Egypt. The present data highlight the need to formulate a new effective vaccine from locally isolated ARV strains after a thorough screening of the molecular nature of circulating ARV in Egypt. [ABSTRACT FROM AUTHOR]

Avian orthoreovirus (ARV) is among the important viruses that cause drastic economic losses in the Egyptian poultry industry. Despite regular vaccination of breeder birds, a high prevalence of ARV infection in broilers has been noted in recent years. However, no reports have revealed the genetic and antigenic characteristics of Egyptian field ARV and vaccines used against it. Thus, this study was conducted to detect the molecular nature of emerging ARV strains in broiler chickens suffering from arthritis and tenosynovitis in comparison to vaccine strains. Synovial fluid samples (n = 400) were collected from 40 commercial broiler flocks in the Gharbia governorate, Egypt, and then pooled to obtain 40 samples, which were then used to screen ARV using reverse transcriptase polymerase chain reaction (RT-PCR) with the partial amplification of ARV sigma C gene. The obtained RT-PCR products were then sequenced, and their nucleotide and deduced amino acid sequences were analyzed together with other ARV field and vaccine strains from GenBank. RT-PCR successfully amplified the predicted 940 bp PCR products from all tested samples. The phylogenetic tree revealed that the analyzed ARV strains were clustered into six genotypic clusters and six protein clusters, with high antigenic diversity between the genotypic clusters. Surprisingly, our isolates were genetically different from vaccine strains, which aligned in genotypic cluster I/protein cluster I, while our strains were aligned in genotypic cluster V/protein cluster V. More importantly, our strains were highly divergent from vaccine strains used in Egypt, with 55.09–56.23% diversity. Sequence analysis using BioEdit software revealed high genetic and protein diversity between our isolates and vaccine strains (397/797 nucleotide substitutions and 148-149/265 amino acid substitutions). This high genetic diversity explains the vaccination failure and recurrent circulation of ARV in Egypt. The present data highlight the need to formulate a new effective vaccine from locally isolated ARV strains after a thorough screening of the molecular nature of circulating ARV in Egypt.

Reovirus infections in turkeys are associated with arthritis and lameness. Viral genome sequence data are scarce, which makes an accurate description of the viral evolution and epidemiology difficult. In this study, we isolated and characterized turkey reoviruses from Hungary. The isolates were identified in 2016; these isolates were compared with earlier Hungarian turkey reovirus strains and turkey reoviruses isolated in the 2010s in the United States. Gene-wise sequence and phylogenetic analyses identified the cell-receptor binding protein and the main neutralization antigen, σC, to be the most conserved. The most genetically diverse gene was another surface antigen coding gene, μB. This gene was shown to undergo frequent reassortment among chicken and turkey origin reoviruses. Additional reassortment events were found primarily within members of the homologous turkey reovirus clade. Our data showed evidence for low variability among strains isolated from independent outbreaks, a finding that suggests a common source of turkey reoviruses in Hungarian turkey flocks. Given that commercial vaccines are not available, identification of the source of these founder virus strains would permit a more efficient prevention of disease outbreaks before young birds are settled to fattening facilities.

The concern that the vaccines currently used against Avian orthoreovirus (ARV) infections are less efficient in the field justifies the need for the close monitoring of circulating ARV strains. In this study, we collected necropsy samples from various chicken breeds and tested for ARV by virus isolation, RT-PCR assay and sequence analysis. ARVs were isolated from birds showing runting-stunting syndrome, uneven growth, lameness or increased mortality, with relative detection rates of 38%, 35%, 6% and 25%, respectively. Partial σC gene sequences were determined for nearly 90% of ARV isolates. The isolates could be classified into one of the major genetic clusters. Interestingly, cluster 2 and cluster 5 were isolated from vaccinated broiler breeders, while clusters 1 to 4 were isolated from unvaccinated broilers. The isolates shared less than 75% amino acid identities with the vaccine strains (range, 44.3–74.6%). This study reaffirms the global distribution of the major genetic clusters of ARVs in chicken. The diversity of ARV strains isolated from unvaccinated broilers was greater than those detected from vaccinated animals, however, the relative importance of passive and active immunity on the selection of novel strains in different chicken breeds needs to be better understood.

Avian orthoreoviruses (ARV) are an emerging threat for the poultry industry, both in the United States (US) and globally. ARV infections in turkeys have been associated with arthritis, lameness and neurological disorders, and cost the US economy approximately USD 33 million per year. There is not a commercial vaccine available and the shortage of turkey ARV (TRV) genomic data hinders the efforts to explore the molecular epidemiology of this virus, although several studies suggest a close relationship between European TRVs and TRVs present in the US. This study shows a snapshot of the genomic diversity of Avian orthoreoviruses (ARV) circulating in Germany in the mid-2000s. Through a deep genomic analysis of 18 ARV isolates recovered from sick turkeys, we observed that co-infection was a common condition. 80% of the samples showed signs of a simultaneous infection with a TRV and a chicken ARV (CRV). We believe this is the first reported evidence of CRV and TRV naturally occurring co-infections in commercial turkeys. These co-infection events were identified due to the significant genomic diversity observed among ARV infecting various production bird species. Our phylogenetic analysis revealed a consistent host-associated ARV clustering, with three main clades: (i) a TRV clade, (ii) a CRV clade, and (iii) a Duck ARV (DRV)/Goose ARV (GRV) clade. Furthermore, our findings indicate that German TRVs have interacted with their European and American counterparts, suggesting active mobilization of the virus, likely through the commercial trading of live animals. However, we also consider the potential role of migratory birds in the international movement of ARV. • Our phylogenetic analyses demonstrate that ARVs share a high level of host-species specificity among the main production bird species. • This high host specificity allowed us to report, for the first time, naturally occurring co-infections of a chicken orthoreovirus and a turkey orthoreovirus. • Our results provide evidence of a transoceanic movement of ARVs during the early 2000s that could be explained by the commercial trading of live animals. • We cannot discard the possibility of migratory birds as the cause of the viral international movement. [ABSTRACT FROM AUTHOR]

Migratory wild birds can carry various pathogens, such as influenza A virus, which can spread to globally and cause disease outbreaks and epidemics. Continuous epidemiological surveillance of migratory wild birds is of great significance for the early warning, prevention, and control of epidemics. To investigate the pathogen infection status of migratory wild birds in eastern China, fecal samples were collected from wetlands to conduct pathogen surveillance. The results showed that duck orthoreovirus (DRV) and goose parvovirus (GPV) nucleic acid were detected positive in the fecal samples collected from wild ducks, egrets, and swan. Phylogenetic analysis of the amplified viral genes reveals that the isolates were closely related to the prevalent strains in the regions involved in East Asian-Australasian (EAA) migratory flyway. Phylogenetic analysis of the amplified viral genes confirmed that they were closely related to circulating strains in the regions involved in the EAA migration pathway. The findings of this study have expanded the host range of the orthoreovirus and parvovirus, and revealed possible virus transmission between wild migratory birds and poultry. [ABSTRACT FROM AUTHOR]

Outbreaks of avian orthoreovirus (ARV) infection with primary symptoms of arthritis/tenosynovitis syndrome have been occurring more frequently in broiler flocks in China in recent years. This study aimed to investigate the genetic characteristics of ARV field strains in broiler flocks exhibiting arthritis/tenosynovitis syndrome from 9 cities in Shandong province during 2015 to 2017. A total of 64 synovial and tendon samples were obtained from broilers with significant arthritis/tenosynovitis syndrome, and 21 ARV field strains were obtained. Phylogenetic analysis of the σC nt/aa sequences revealed that only 4 isolates were clustered in genotype I, including vaccine strains S1133, 1733, and most of the ARV field strains identified previously in China. Eleven and 6 ARV field isolates were identified in genotypes II and V, sharing 70.9 to 76.0% and 53.0 to 55.2% nt identities with the vaccine strains, respectively. Previous studies in China have not reported these 2 serotypes of field strains, and prevalence of these ARV variants may be increasing in Chinese broiler flocks. Results of this study suggest that large-scale investigation of epidemic ARV should be conducted to explore the genetic diversity of ARV field isolates in China.

In recent years, the emergence of avian orthoreovirus (ARV) has caused significant losses to the poultry industry worldwide. In this study, a novel ARV isolate, designated as AHZJ19, was isolated and identified from domestic chicken with viral arthritis syndrome in China. AHZJ19 can cause typical syncytial cytopathic effect in the chicken hepatocellular carcinoma cell line, LMH. High-throughput sequencing using Illumina technology revealed that the genome size of AHZJ19 is about 23,230 bp, which codes 12 major proteins. Phylogenetic tree analysis found that AHZJ19 was possibly originated from a recombination among Hungarian strains, North American strains, and Chinese strains based on the sequences of the 12 proteins. Notably, the σC protein of AHZJ19 shared only about 50% homology with that of the vaccine strains S1133 and 1733, which also significantly differed from other reported Chinese ARV strains. The isolation and molecular characteristics of AHZJ19 provided novel insights into the molecular epidemiology of ARV and laid the foundation for developing efficient strategies for control of ARV in China.

Common pathogens of intensive poultry farms, either parasitic or bacterial, such as Coccidiaor Salmonella, are well known and strictly controlled by veterinary management. This case study reports an unusual case of runting stunting syndrome (RSS) observed on a Sicilian poultry farm of broiler chickens during 2019. The investigation was carried out on five chickens which present delayed in body weight and growth performance. Animals showed also difficulty in deambulation and diarrhea. At necropsy, intestinal lesions were detected in three of the five clinical cases. Gut samples were collected and analyzed to identify potential pathogens responsible for the RSS. Presence of viruses was detected by using quantitative reverse transcription PCR (RT‑qPCR), while selected tissues were fixed and embedded in paraffin wax according to routine procedures. All histological sections were stained with hematoxylin‑eosin. RT‑qPCR successfully detected both Chicken astrovirus (CAstV) and Avian orthoreovirus (ARV). Histology evidenced severe specific lesions on the intestinal mucosa in liver and kidneys. Chicken astrovirus and Avian orthoreovirus RNA was also detected in cecal tonsils, kidney and liver, thus implying their possible primary role in inducing the disease. Further studies are needed to evaluate the role of other possible factors (low biosecurity measures, e.g.) and, most of all, the consequences in terms of economic losses and animal health impairment.

Avian orthoreovirus (ARV) infections of broiler flocks cause arthritis/tenosynovitis syndrome and significant economic losses. ARV variants were detected in the USA and Canada. Viral arthritis/tenosynovitis syndrome has occurred frequently in China in recent years. In this study, a variant ARV strain associated with viral arthritis/tenosynovitis syndrome was isolated from broilers and designated as LY383. Genomic sequence and phylogenetic analysis of the σC nucleic acid and amino acid sequences revealed that the isolate was closely related to ARV field strains Reo/PA/Layer/01224B/14, Reo/PA/Broiler/1551/13, GA/14602/2014, GA/13569/2013 and GA/13542/2013, in cluster V, but distinct from most Chinese field strains or commercial vaccine strains. Experimental challenge showed that the isolate could cause arthritis/tenosynovitis syndrome in broilers, which possessed a high level of maternal antibodies induced by commercial ARV vaccines (S1133, 1733 and T98). Furthermore, viral nucleic acid could be detected in cloacal swabs of all challenged birds throughout the entire test from 5 dpi onward. These results suggest that a novel ARV genotype emerges and might become prevalent in broiler flocks in China. RESEARCH HIGHLIGHTS A variant avian orthoreovirus was isolated from a vaccinated broiler flock in North China. The ARV field strain was distinct from previous China-origin ARV isolates and vaccine strains. The current commercial ARV vaccine could not provide effective protection of broilers against the field isolate infection. These findings indicated that variant ARV field strains might become frequent in broiler flocks in China and effective measures should be conducted to prevent and control the disease. [ABSTRACT FROM AUTHOR]

Avian reoviruses (ARVs) have a significant economic impact on the poultry industry, affecting commercial and backyard flocks. Spread feco-orally, or vertically, many do not cause morbidity, but pathogenic strains can contribute to several diseases, including tenosynovitis/arthritis, which is clinically the most significant. The last decade has seen a surge in cases in the US, and due to ongoing evolution, seven genotypic clusters have now been identified. Control efforts include strict biosecurity and vaccination with commercial and autogenous vaccines. Research priorities include improving understanding of pathogenesis and developing new vaccines guided by ongoing molecular and serologic surveillance.

Oncolytic viruses are cancer therapeutics with promising outcomes in pre-clinical and clinical settings. Animal viruses have the possibility to avoid pre-existing immunity in humans, while being safe and immunostimulatory. We isolated an avian orthoreovirus (ARV-PB1), and tested it against a panel of hepatocellular carcinoma cells.We found that ARV-PB1 replicated well and induced strong cytopathic effects. It was determined that one mechanism of cell death was through syncytia formation, resulting in apoptosis and induction of interferon stimulated genes (ISGs). As hepatitis C virus (HCV) is a major cause of hepatocellular carcinoma worldwide, we investigated the effect of ARV-PB1 against cells already infected with this virus. Both HCV replicon-containing and infected cells supported ARV-PB1 replication and underwent cytolysis. Finally, we generated in silico models to compare the structures of human reovirus- and ARV-PB1-derived S1 proteins, which are the primary targets of neutralizing antibodies. Tertiary alignments confirmed that ARV-PB1 differs from its human homolog, suggesting that immunity to human reoviruses would not be a barrier to its use. Therefore, ARV-PB1 can potentially expand the repertoire of oncolytic viruses for treatment of human hepatocellular carcinoma and other malignancies. [ABSTRACT FROM AUTHOR]

In recent years, the emerging avian orthoreovirus (ARV) strains that led to viral arthritis have attracted much attention from the chicken industry worldwide, due to the significant economic losses suffered. In mid-2020, with the assistance of next-generation sequencing technology, we achieved success in characterizing two divergent avian orthoreovirus (ARV) variants (0543/SDYT) and isolating them from the broiler tendons characterized by arthritis. As suggested by the genome characterization of the 0543/SDYT strains, they belong separately to clusters Ⅰ and Ⅳ. As revealed by sequence comparison, phylogenetic, and recombination analysis, for μA, μB, and σNS genes, considerable genetic divergences were also observed in the two new isolates. However, in the case of λA, λB, μNS, σA, and σB genes, very clear clustering patterns were observed for SDYT and 0543 field strains, respectively. In terms of the μA, μNS, p10, p17 genes of SDYT isolate and μNS, p17, σC, σA genes of 0543 isolate, the lower similarity was observed with NCBI stored sequences, with nt highest identity values below 90 %. In addition, there is an intra-fragmental recombination event in the M1 gene of the SDYT strain. In regard, the multiple segmental recombination and accumulation of point mutations play a role in the newly-emerging ARV strains. Not only did the isolates strain exhibit strong replication ability in vivo , but they also displayed strong arthritogenicity in broilers with low neutralizing maternal antibodies, indicating that maternal antibody treatment may not effectively reduce the oral infection of avian orthoreovirus. These findings suggest that it is necessary to develop a new strategy for enhanced effectiveness in preventing and controlling ARV infection. • We successfully isolated and identified two broiler-derived Avian orthoreovirus strains, namely SDYT and 0543. • Genetic characterization revealed that SDYT was an intra-segmental recombinant strain, classified as genotype IV, and 0543 was a genotype 1 variant, distinct from the S1133 vaccine strain. • We assessed pathogenicity in vivo by challenging S1133, SDYT, 0543 strains in chicks containing maternally derived antibodies (mainly from S11333 vaccinated breeders). [ABSTRACT FROM AUTHOR]

Avian reovirus (ARV) is a common pathogen in chickens and other birds causing a variety of clinical symptoms such as arthritis and tenosynovitis but also enteric and respiratory symptoms. A rapid method that detects as many ARV genotypes as possible, will contribute to the early identification and control of the virus infection that causes high economic damage to the poultry industry worldwide. In this study, a real-time reverse transcription polymerase chain reaction (RT-qPCR) assay for the detection of ARV was developed. The RT-qPCR detection threshold for ARV genomic RNA standard cases was 10 copies/µL. Reproducibility of the RT-qPCR was confirmed by intra- and inter-assays. When the nucleic acids of different ARV genotypes and other common avian pathogens (IBDV, AIV, NDV, and IBV) were subjected to that RT-qPCR test, only ARV samples tested positive while all other pathogens tested negative. Due to the simplicity, convenience, high sensitivity, and specificity of the assay, the probe-based RT-qPCR is proposed to be used as an alternative diagnostic assay for the detection of ARVs in veterinary diagnostic laboratories. • Developed a specific molecular-based technique for early detection of ARV through the RT-qPCR method. • The new primers targeted for the L1 segment were tested for their sensitivity and specificity. • The RT-qPCR analysis reveals that the developed method can detect up to 1 × 101 copies/µL. • The current RT-qPCR technique has a 100-fold increase in sensitivity over the traditional RT-PCR. • This assay will provide a platform for the rapid identification of ARVs in veterinary diagnostic laboratories. [ABSTRACT FROM AUTHOR]

Avian orthoreoviruses cause various diseases in wild birds and domesticated poultry. In this study we report the detection and genomic characterization of a partridge ( Perdix perdix ) origin reovirus strain, D1007/2008. The virus was isolated on cell culture from acute pneumonia and infra-orbital sinusitis. The 23,497 nucleotide long genome sequence was obtained by combined use of semiconductor and capillary sequencing. Sequence and phylogenetic analyses showed that the partridge reovirus strain was related to orthoreoviruses of gallinaceous birds. In fact, five (λB, λC, μB, σC, σNS) and one (σB) out of 10 genes clustered definitely with turkey or chicken origin orthoreoviruses, respectively, whereas in the λA, μA, μNS and σA phylogenies a more distant genetic relationship was observed. Our data indicate that the identified reovirus strain is composed of a mixture of chicken and turkey orthoreovirus related alleles. This finding implies that partridges may serve as natural reservoirs for orthoreoviruses of domesticated poultry. [ABSTRACT FROM AUTHOR]

Oncolytic viruses are cancer therapeutics with promising outcomes in pre-clinical and clinical settings. Animal viruses have the possibility to avoid pre-existing immunity in humans, while being safe and immunostimulatory. We isolated an avian orthoreovirus (ARV-PB1), and tested it against a panel of hepatocellular carcinoma cells. We found that ARV-PB1 replicated well and induced strong cytopathic effects. It was determined that one mechanism of cell death was through syncytia formation, resulting in apoptosis and induction of interferon stimulated genes (ISGs). As hepatitis C virus (HCV) is a major cause of hepatocellular carcinoma worldwide, we investigated the effect of ARV-PB1 against cells already infected with this virus. Both HCV replicon-containing and infected cells supported ARV-PB1 replication and underwent cytolysis. Finally, we generated in silico models to compare the structures of human reovirus- and ARV-PB1-derived S1 proteins, which are the primary targets of neutralizing antibodies. Tertiary alignments confirmed that ARV-PB1 differs from its human homolog, suggesting that immunity to human reoviruses would not be a barrier to its use. Therefore, ARV-PB1 can potentially expand the repertoire of oncolytic viruses for treatment of human hepatocellular carcinoma and other malignancies.

Highlights: [•] The full-length sequence and genetic traits of a classical MDRV isolates was determined and analyzed. [•] The classical MDRV isolates formed two distinct lineages (China and Europe lineages). [•] The classical MDRVs isolated in restricted geographical region are evolving by different and independent pathways. [ABSTRACT FROM AUTHOR]

Abstract: The complete sequence of a reovirus, strain 815-12 associated with necrotic focus formation in the liver and spleen of Muscovy ducklings in China, was determined and compared with sequences of other duck-, goose-, and chicken-origin reoviruses. The 815-12 genome comprised 22,969bp with 10 dsRNA segments ranging from 1125bp (S4) to 3958bp (L1), all of which (except S4) were almost identical in length to the cognate segments of other waterfowl and chicken isolates. Detailed analyses revealed that 815-12 and other waterfowl isolates contained the conserved 3′-terminal pentanucleotide sequence (UCAUC-3′) of the orthoreoviruses and 5′-terminal hexanucleotide sequence (5′-GCUUUU) of avian orthoreoviruses (ARVs), and conserved functional motifs previously identified in ARV proteins. Several notable differences, including organization of the polycistronic genome segments and genomic coding assignments of the S segments, existed between viruses represented by 815-12 and the waterfowl reoviruses emerging in China in recent years; the latter was somewhat similar to chicken isolates. Pairwise sequence comparisons demonstrated extensive sequence diversity among the various waterfowl isolates and between waterfowl and chicken isolates. Phylogenetic analyses identified two genetic groups for waterfowl reoviruses, and potential genetic reassortment of segment M2 between waterfowl and chicken reoviruses and segments encoding for λA, λB, μA, μNS and σA between waterfowl reoviruses. Taken together, it was suggested that common designation ARV-Wa should be used to represent ARV isolates from different waterfowl species and that the two ARV-Wa genotypes should be considered as two separate groups distinct from chicken isolates within the species Avian orthoreovirus. [Copyright &y& Elsevier]

The article presents a retrospective polymerase chain reaction (PCR) study of avian Orthoreovirus (ARV), chicken infectious anemia (CAV) and fowl Aviadenovirus (FAdV) genomes contamination in commercial poultry vaccines in Brazil. Based on results, vaccines produced in 1991-1998 from three manufacturers had contaminating genomes of the three viruses. The study shows that vaccines produced since 2001 tested negative for the genomes, which demonstrates the monitoring value of molecular methods.

• Using the NGS technology, we identified a recombinant cluster 2 ARV variant named PHC-2020-0545, isolated from tendons of broilers in China. • The isolate was genetically significantly distinct from known ARV strains in M1 and M3 genes. • Multiple segmental recombination, intra-segmental recombination and point mutations might contribute to the emergence of this isolate. • The isolate had a strong replication ability in 1-day-old broilers, and mainly affected the movement, digestion and metabolism of broilers. In mid-2020, using next-generation sequencing (NGS) technology, we identified a recombinant cluster 2 avian orthoreovirus (ARV) variant named PHC-2020−0545, isolated from tendons of 33-day-old broilers with leg swelling in China. Complete genomic sequencing and analyses demonstrated that the isolate was genetically significantly distinct from known ARV strains in M1 and M3 genes and its σC coding gene had an extremely high variability, compared with the identified ARV strains grouped into other genotyping cluster. Further analysis showed that many base substitutions were silent and non-silent substitutions are most likely to occur in the first positions of codons. Multiple segmental recombination, intra-segmental recombination and accumulation of point mutations might contribute to the emergence of this isolate. The PHC-2020−0545 strain had a strong replication ability in 1-day-old broilers, and mainly affected the movement, digestion and metabolism of broilers. In addition, the infection route of the isolate is related to its pathogenicity to broilers. Therefore, combined with its unique genetic characteristics and potential origin, we determined that the PHC-2020−0545 field strain is a novel recombinant ARV strain, which has certain reference value for the preparation and evaluation of new vaccines. [ABSTRACT FROM AUTHOR]

In recent years, the emergence of avian orthoreovirus (ARV) has caused significant losses to the poultry industry worldwide. In this study, a novel ARV isolate, designated as AHZJ19, was isolated and identified from domestic chicken with viral arthritis syndrome in China. AHZJ19 can cause typical syncytial cytopathic effect in the chicken hepatocellular carcinoma cell line, LMH. High-throughput sequencing using Illumina technology revealed that the genome size of AHZJ19 is about 23,230 bp, which codes 12 major proteins. Phylogenetic tree analysis found that AHZJ19 was possibly originated from a recombination among Hungarian strains, North American strains, and Chinese strains based on the sequences of the 12 proteins. Notably, the σC protein of AHZJ19 shared only about 50% homology with that of the vaccine strains S1133 and 1733, which also significantly differed from other reported Chinese ARV strains. The isolation and molecular characteristics of AHZJ19 provided novel insights into the molecular epidemiology of ARV and laid the foundation for developing efficient strategies for control of ARV in China., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhang, Li, Wang, Xie, Wan, Qin, Ye and Shao.)

The avian orthoreovirus (ARV) causes large economic losses to poultry industry around the whole world. ARV viral proteins can be classified into three sizes: λ (large), μ (medium) and σ (small). σC, one of the capsid protein of ARV, contains various specific neutralizing epitopes and can induce robust immune responses in infected chickens. An indirect enzyme-linked immunosorbent assay (I-ELISA) was established using the recombinant σC protein. The optimal dilutions of antigen, serum and conjugate goat anti-chicken IgY conjugate were 2.5 μg/ml, 1:80 and 1:4000, respectively. The optimal antigen coating condition was 4℃ for overnight. Hyperimmune serum of high-yielding hens of different age produced stronger reaction against σC. Chicken serum samples were randomly collected and total coincidence rate between commercial kit and σC-ARV-ELISA kit was 97.8%. Importantly, those discrepant serums were further verified using Western blot assay and three of them were determined to be positive. Thus, the method based on σC protein has higher sensitivity compared with the commercial kit and provide a rapid and simple method for serodiagnosis of ARV, contributing to convenient epidemiological monitoring of ARV., Competing Interests: Declaration of Competing Interest The authors declare no commercial of financial conflict of interest., (Copyright © 2021. Published by Elsevier B.V.)

• Whole genome sequences and phylogenetic analyses of novel pheasant orthoreoviruses are presented. • Reo/HUN/Pheasant/216/2015 is a reassortant strain showing genetic relationship with turkey, chicken and partridge reoviruses. • Strain D1996/2/1 is distantly related to any currently known avian origin orthoreoviruses. Avian reoviruses are well-known pathogens seriously affecting the productivity of poultry industry. Game birds represent a small segment of the agricultural sector and much remained to be learnt about factors affecting productivity. Here we show that reovirus infections might occur in pheasants and demonstrate that reoviruses of pheasants are of diverse origin. The complete or coding-complete genomic sequences of two Hungarian reovirus strains, D1996/2/1 and Reo/HUN/Pheasant/216/2015, have been determined in this study. The strain D1996/2/1 was isolated in 2012 from birds with gizzard erosion, whereas the other strain was isolated in 2015 from diarrheic pheasant poults. Phylogenetic analyses showed that none of the Hungarian isolates shared common origin with a pheasant reovirus detected recently in the United States. Additionally, we found that Reo/HUN/Pheasant/216/2015 is a multi-reassortant reovirus within the species Avian orthoreovirus that shared genetic relationship with turkey reoviruses (σC), partridge reoviruses (λA, σB), and chicken reoviruses (λB, λC, μA, σA, and σNS), in the respective gene phylogenies, whereas two genes (μB and μNS) did not reveal any possible common ancestors. The other isolate, D1996/2/1, was found to be distantly related to previously described reoviruses raising the possibility that it might represent a novel orthoreovirus species or a new genogroup within the newly accepted species, Neoavian orthoreovirus. The genetic diversity among pheasant reoviruses could raise challenges for virus classification as well as for development of molecular diagnostic tools and vaccine based prevention and control measures. [ABSTRACT FROM AUTHOR]