First Isolation and Genetic Characterization of Avian Nephritis Virus 4 from Commercial Poultry in India.

Riyesh Thachamvally, Yogesh Chander, Ram Kumar, Garvit Kumar, Nitin Khandelwal, Anagha G, Anju Manuja, Rajesh Kumar Vaid, Naveen Kumar, Sanjay Barua, Yash Pal, B N Tripathi, T K Bhattacharya
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Abstract

Avian nephritis virus (ANV), which belongs to the family Astroviridae, is associated with different clinical manifestations (enteric and kidney disorders) in poultry. Despite being a significant pathogen of the avian industry worldwide, information regarding genetic features of these viruses in India is scarce. In this study, 386 intestinal samples collected from 37 slaughterhouses in two north Indian states (Rajasthan and Haryana) were screened for ANV with reverse transcription PCR (RT-PCR) targeting the conserved ORF1b gene, followed by nucleotide sequencing of the amplified product. RT-PCR and sequencing confirmed the presence of ANV in 32 clinical samples (8.29%), with concurrent infections of infectious bronchitis virus, chicken astrovirus, and fowl adenoviruses observed in some clinical samples (n = 4). Virus isolations were successful from four out of 12 ANV-positive clinical samples passaged via the yolk-sac route in specific-pathogen-free embryonated chicken eggs. Additionally, the near-complete genomes of two viruses were determined through sequencing. Phylogenetic analysis based on full-length capsid protein sequences classified the viruses into ANV genotype 4 (ANV4), and to the best of our knowledge this is the first report of ANV4 from India. This study revealed the presence and circulation of new strains of ANV in Indian poultry. Genetic profiling and isolation of the viruses in this study will not only aid in the development of diagnostic tools and vaccines for ANV but also offer valuable insights into its epidemiology.

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首次从印度商用家禽中分离出禽肾炎病毒 4 并确定其遗传特征。
禽肾炎病毒(ANV)属于 Astroviridae 科,与家禽的不同临床表现(肠道和肾脏疾病)有关。尽管禽肾炎病毒是全球养禽业的一种重要病原体,但有关这些病毒在印度遗传特征的信息却很少。在这项研究中,通过针对保守的 ORF1b 基因进行反转录 PCR(RT-PCR),对从印度北部两个邦(拉贾斯坦邦和哈里亚纳邦)的 37 个屠宰场采集的 386 份肠道样本进行了 ANV 筛查,随后对扩增产物进行了核苷酸测序。RT-PCR和测序证实了32份临床样本(8.29%)中存在ANV,在一些临床样本(n = 4)中观察到同时感染了传染性支气管炎病毒、鸡星状病毒和鸡腺病毒。在 12 份 ANV 阳性的临床样本中,有 4 份样本通过无特定病原体的胚胎鸡卵黄-卵巢途径成功分离出病毒。此外,还通过测序确定了两种病毒近乎完整的基因组。据我们所知,这是印度首次报告ANV4病毒。据我们所知,这是印度首次报告 ANV4。这项研究揭示了印度家禽中 ANV 新毒株的存在和流通情况。这项研究中对病毒的基因分析和分离不仅有助于开发 ANV 的诊断工具和疫苗,还能对其流行病学提供有价值的见解。
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