Characterization of Novel Reoviruses Wad Medani Virus (Orbivirus) and Kundal Virus (Coltivirus) Collected from Hyalomma anatolicum Ticks in India during Surveillance for Crimean Congo Hemorrhagic Fever

Abstract

In 2011, ticks were collected from livestock following an outbreak of Crimean Congo hemorrhagic fever (CCHF) in Gujarat state, India. CCHF-negative Hyalomma anatolicum tick pools were passaged for virus isolation, and two virus isolates were obtained, designated Karyana virus (KARYV) and Kundal virus (KUNDV), respectively. Traditional reverse transcription-PCR (RT-PCR) identification of known viruses was unsuccessful, but a next-generation sequencing (NGS) approach identified KARYV and KUNDV as viruses in the Reoviridae family, Orbivirus and Coltivirus genera, respectively. Viral genomes were de novo assembled, yielding 10 complete segments of KARYV and 12 nearly complete segments of KUNDV. The VP1 gene of KARYV shared a most recent common ancestor with Wad Medani virus (WMV), strain Ar495, and based on nucleotide identity we demonstrate that it is a novel WMV strain. The VP1 segment of KUNDV shares a common ancestor with Colorado tick fever virus, Eyach virus, Tai Forest reovirus, and Tarumizu tick virus from the Coltivirus genus. Based on VP1, VP6, VP7, and VP12 nucleotide and amino acid identities, KUNDV is proposed to be a new species of Coltivirus. Electron microscopy supported the classification of KARYV and KUNDV as reoviruses and identified replication morphology consistent with other orbi- and coltiviruses. The identification of novel tick-borne viruses carried by the CCHF vector is an important step in the characterization of their potential role in human and animal pathogenesis. IMPORTANCE Ticks and mosquitoes, as well Culicoides, can transmit viruses in the Reoviridae family. With the help of next-generation sequencing (NGS), previously unreported reoviruses such as equine encephalosis virus, Wad Medani virus (WMV), Kammavanpettai virus (KVPTV), and, with this report, KARYV and KUNDV have been discovered and characterized in India. The isolation of KUNDV and KARYV from Hyalomma anatolicum, which is a known vector for zoonotic pathogens, such as Crimean Congo hemorrhagic fever virus, Babesia, Theileria, and Anaplasma species, identifies arboviruses with the potential to transmit to humans. Characterization of KUNDV and KARYV isolated from Hyalomma ticks is critical for the development of specific serological and molecular assays that can be used to determine the association of these viruses with disease in humans and livestock.

Exploration of the Ixodes ricinus virosphere unveils an extensive virus diversity including novel coltiviruses and other reoviruses

“…Two of its members, Colorado tick fever virus and Eyach virus, are known human pathogens capable of causing severe disease ( Attoui et al, 2005 ). While the other members, Tarumizu tick virus, Tai Forest reovirus, and Kundal virus, have not been implicated in human disease, they have been shown to be capable of infecting mammalian and/or human cell lines ( Fujita et al, 2017 ; Weiss et al, 2017 ; Yadav et al, 2019 ). One of the two coltiviruses reported here appears to be a strain of Eyach virus, sharing >94 per cent nucleotide similarity for all but one segments.…”

Exploration of the Ixodes ricinus virosphere unveils an extensive virus diversity including novel Coltiviruses and other reoviruses

“…Two of its members, Colorado tick fever virus and Eyach virus, are known human pathogens capable of causing severe disease [36]. While the other members, Tarumizu tick virus, Tai Forest reovirus and Kundal virus, have not been implicated in human disease, they have been shown to be capable of infecting mammalian and/or human cell lines [37][38][39]. One of the two coltiviruses reported here appears to be a strain of Eyach virus, sharing >94% nucleotide similarity for all but one segments.…”

Genome Sequencing Reveals a Mixed Picture of SARS-CoV-2 Variant of Concern Circulation in Eastern Uttar Pradesh, India

“…Library preparation was done using the extracted quantified RNA using TruSeq Stranded mRNA Library Prep Kit. The detailed steps during the library preparation and sample loading are described elsewhere (15,16). The generated sequence reads were analyzed and mapped using the reference-based assembly method on the CLC Genomics Workbench version 20 (CLC, Qiagen) and used to call mutations in the SARS-CoV-2 genomes.…”