Isolation of chicken astrovirus from specific pathogen-free chicken embryonated eggs

Núñez, L; Parra, Silvana Hipatia Santander; Mettifogo, Elena; Catroxo, Márcia Helena Braga; Astolfi-Ferreira, Claudete S.; Ferreira, Antônio José Piantino

doi:10.3382/ps/pev086

Cited by 23 publications

(29 citation statements)

References 31 publications

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“…This report is showing its presence alone or in concomitant infections during outbreaks and suggesting that they play an important role in the aetiology of enteric disease (Zsak et al 2013). These recent reports have led to the development of experimental models to recreate the disease and to try to determine its etiopathogenicity (Zsak et al 2013, Decaesstecker et al 1989, Nuñez et al 2015a). However, due to the complex nature of this disease, the exact aetiology remains unknown.…”

Section: Introductionmentioning

confidence: 79%

Detection of enteric viruses in pancreas and spleen of broilers with runting-stunting syndrome (RSS)

et al. 2016

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RESUMO.-[Detecção de vírus entéricos em pâncreas e baço de frangos com a síndrome de nanismo e raquitismo (RSS).]A doença entérica é um problema multifatorial em galinhas que causa alterações gastrointestinais, conversão alimentar elevada e deficiência de crescimento.Nos últimos anos, os vírus entéricos foram associados à doença entérica; casos reportados mostraram a infecção de um único vírus e também infecções concomitantes durante os surtos sugerindo a presença de múltiplos fatores etiológicos nas doenças entéricas. Este estudo mostra uma alta taxa de detecção dos vírus entéricos em amostras de pâncreas e baço de um surto de enterite e má-absorção em 16 lotes de frangos (n=80 frangos). O vírus de nefrite aviá-ria (ANV) foi o vírus mais detectado, estando presente em 75% dos lotes seguido pelo rotavírus aviário grupo A (ART--A) em 68,75% dos casos, e pelo astrovirus (CAstV) e parvovírus aviários (ChPV), ambos em 43,75% das amostras. Os vírus estavam presentes no pâncreas dos lotes positivos em percentuais elevados: 100% para ART-A e CAstV; 91,7% para ANV, e em 57,14% para ChPV. Em contraste, somente 16,7% e 57,14%, em amostras de intestino, foram positivos para ANV e CAstV, respectivamente. Reovírus aviário Enteric disease is a multifactorial problem in chickens, which causes gastrointestinal alterations, elevated feed conversions and impairment. In the last years, several enteric viruses were implicated in enteric disease; case reports have shown their presence alone or in concomitant infections during outbreaks and have suggested that they might be determining factors in the aetiology of enteric disease. This study shows high detection rates of enteric viruses in the pancreas and spleen in samples from an outbreak of enteritis and malabsorption in 16 chicken flocks (n=80 broilers). Avian nephritis virus (ANV) was the most ubiquitous virus, present in 75% of the flocks followed by avian rotavirus group A (ART-A) with 68.75%, and by chicken astrovirus (CAstV) and chicken parvovirus (ChPV) in 43.75% of samples. Viruses were present in the pancreas of positive flocks at extremely high rates: 100% for ART-A, 91.7% for ANV, 100% for CAstV and 57.14% for ChPV. By contrast, only 16.7% and 57.14% of intestine samples were positive for ANV and CAstV, respectively. Avian reovirus (AReo) and avian adenovirus group 1 (FAdV-1) were not detected. These results suggest that high viral detection rates in pancreas samples may be a result of viremia during enteric disease, with subsequent damage of the exocrine pancreas, leading to runting-stunting syndrome (RSS).INDEX TERMS: Broiler, enteric viruses, detection, pancreas, spleen, runting-stunting syndrome, RSS.

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Section: Introductionmentioning

confidence: 79%

Detection of enteric viruses in pancreas and spleen of broilers with runting-stunting syndrome (RSS)

et al. 2016

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“…Chicken astrovirus causes severe disease especially in young chickens (Bulbule et al 2013;Li et al 2013;Nunez et al 2016;Schultz-Cherry et al 2001;Smyth et al 2013;Todd et al 2009). Though culturing methods has been described for astroviruses (Baxendale and Mebatsion 2004;Nunez et al 2015), the isolation of virus is somewhat difficult due to its poor growth in the culture (Smyth et al 2010). Next generation sequencing technology is sophisticated as there is no need to isolate or culture the organism.…”

Section: Discussionmentioning

confidence: 99%

Complete genome sequence analysis of chicken astrovirus isolate from India

et al. 2016

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The consensus length of 7513 bp genome sequence of Indian isolate of chicken astrovirus was obtained after assembly of 14,121 high quality reads. The genome was comprised of 13 bp 5'-UTR, three open reading frames (ORFs) including ORF1a encoding serine protease, ORF1b encoding RNA dependent RNA polymerase (RdRp) and ORF2 encoding capsid protein, and 298 bp of 3'-UTR which harboured two corona virus stem loop II like "s2m" motifs and a poly A stretch of 19 nucleotides. The genetic analysis of CAstV/INDIA/ANAND/2016 suggested highest sequence similarity of 86.94% with the chicken astrovirus isolate CAstV/GA2011 followed by 84.76% with CAstV/4175 and 74.48%% with CAstV/Poland/G059/2014 isolates. The capsid structural protein of CAstV/INDIA/ANAND/2016 showed 84.67% similarity with chicken astrovirus isolate CAstV/GA2011, 81.06% with CAstV/4175 and 41.18% with CAstV/Poland/G059/2014 isolates. However, the capsid protein sequence showed high degree of sequence identity at nucleotide level (98.64-99.32%) and at amino acids level (97.74-98.69%) with reported sequences of Indian isolates suggesting their common origin and limited sequence divergence. The epitope analysis by SVMTriP identified two unique epitopes in our isolate, seven shared epitopes among Indian isolates and two shared epitopes among all isolates except Poland isolate which carried all distinct epitopes.

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“…The studied virus isolated in 7- and 14-day-old SPF-CEE confirmed to be infected based on the presence of pathological alterations in the embryos, detection of viral RNA, and visualization of viral particles. Viral isolation in SPF-CEE is an old technique that has been used for several years to propagate and isolate viral agents to permit the study of their pathogenicity and virus behavior or to make vaccines oriented to protect the chickens against diseases (Guy, 2008 ; Alemnesh et al, 2012 ; Nuñez et al, 2015 ). Several viruses have been isolated in SPF-CEE either using primary cultures cells from many organs or using cell lines, where some viruses grow better than in eggs or primary cultures (McNulty et al, 1979 ; Baxendale and Mebatsion, 2004 ).…”

Section: Discussionmentioning

confidence: 99%

“…The supernatant obtained from the embryo suspension described above was clarified and stained as described previously by Nunez et al ( 2015 ). The samples were examined by an electron microscope (Philips, Model EM 208, Amsterdam, Netherlands) to check for the presence of viral particles.…”

Section: Methodsmentioning

confidence: 99%

Isolation of avian nephritis virus from chickens showing enteric disorders

et al. 2018

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Runting-stunting syndrome (RSS) is one of the diseases associated with many detected viruses. In Brazil, there were reports of several enteric disease outbreaks in chickens in which avian nephritis virus (ANV) was detected; however, the role of ANV in the outbreaks and whether the virus was a causative agent of these cases of enteric diseases were not determined. The aim of this study was to isolate ANV in specific pathogen-free (SPF) chicken embryonated eggs (CEE) from the enteric contents of chickens showing signs of RSS. For this purpose, 22 samples of chicken enteric contents that were positive only for ANV were inoculated into 7 and 14-day-old SPF-CEE via the yolk sac route and incubated for 5 d, with a total of 3 passages. Virus isolation was confirmed by the presence of embryo injuries, detection of viral RNA by RT-PCR, and visualization of viral particles using electron microscopy. Therefore, the 7-day-old inoculated embryos showed dwarfism, gelatinous consistency, hemorrhage, and edema in the embryos, whereas the 14-day-old did not show any alteration. Viral RNA was detected in the embryos of both ages of inoculation, and the same viral particles were visualized. The embryos from the mock group showed no alteration and were negative for all the tests. The viral cDNA was sequenced, and the molecular and phylogenetic analyses showed that the Brazilian isolates are more related with the ANV-1 serotype group; the sequences of these isolates showed a high percentage of nucleotide (86.4 to 94.9%) and amino acid (92.3 to 98.7%) similarity with other sequences from China, Japan, Australia, and the United States that belong to this serotype previously classified group. In this study, we isolated 8 samples of ANV in SPF-CEE from enteric content samples from chickens with RSS. In doing so, we showed the pathological injuries to the embryo caused by the virus and the molecular characterization of a part of the ORF 1b gene of the virus.

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Isolation of chicken astrovirus from specific pathogen-free chicken embryonated eggs

Cited by 23 publications

References 31 publications

Detection of enteric viruses in pancreas and spleen of broilers with runting-stunting syndrome (RSS)

Detection of enteric viruses in pancreas and spleen of broilers with runting-stunting syndrome (RSS)

Complete genome sequence analysis of chicken astrovirus isolate from India

Isolation of avian nephritis virus from chickens showing enteric disorders

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