Detection of novel orthoreovirus genomes in shrew (<i>Crocidura hirta</i>) and fruit bat (<i>Rousettus aegyptiacus</i>)

Abstract

Orthoreoviruses have been indentified in several mammals, however, there is no information about orthoreoviruses in shrews. In this study, we screened wild animals in Zambia, including shrews, rodents, and bats for the detection of orthoreoviruses. Two orthoreovirus RNA genomes were detected from a shrew intestinal-contents (1/24) and a bat colon (1/96) sample by reverse-transcription (RT)-PCR targeting the RNA-dependent RNA polymerase gene of orthoreoviruses. Phylogenetic analyses revealed that each of the identified orthoreoviruses formed a distinct branch among members of the Orthoreovirus genus. This is the first report that shrews are susceptible to orthoreovirus infection. Our results suggest the existence of undiscovered orthoreoviruses in shrews and provide important information about the genetic diversity of orthoreoviruses.

Characterization of mammalian orthoreoviruses isolated from faeces of pigs in Zambia

“…Total RNAs were extracted from the supernatants using TRIzol-LS (Invitrogen, Waltham, MA, USA) according to the manufacturer’s instructions. RNA samples were examined to detect MRVs by RT-PCR using two specific primer sets based on the conserved sequence within the L1 and L3 segments as previously described [46–48] (Table S3). The protocol of RT-PCR for detecting the L3 segment was modified from that of the PrimeScript One Step RT-PCR Kit v2 (Takara, Shiga, Japan).…”

Attenuated infection by a Pteropine orthoreovirus isolated from an Egyptian fruit bat in Zambia

“…Total RNAs were extracted from colon homogenates using TRIzol-LS (Invitrogen, Waltham, MA, USA) or the QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany) according to the manufacturers' instructions. The PRV genome was examined by RT-PCR with pan-orthoreovirus primer sets targeting RNA-dependent RNA polymerase genes using total RNAs from colon tissues (S2 Table) [23,24]. After confirmation by gel electrophoresis, positive-PCR products were subjected to direct sequencing using the BigDye Terminator v3.1 Cycle Sequencing kit (Applied Biosystems, Foster City, CA, USA).…”

“…Therefore, there is the possibility that this high seroprevalence against PRV was due to cross-neutralization activity among orthoreoviruses, and that the straw-colored fruit bats were not infected with Nachunsulwe-57, but with other PRV-related viruses. In our previous study, a novel orthoreovirus species was identified in Egyptian fruit bats in Zambia [24], but the cross-reactivity of neutralizing antibodies between these bat-derived orthoreoviruses remains unevaluated. Considering the unavailability of the genetic sequence of PRV in straw-colored fruit bats, the bats might harbor PRV-related viruses that elicits cross-reactive antibodies.…”

Cryptic diversity of Crocidura shrews in the savannahs of Eastern and Southern Africa

“…Both C. flavescens and C. hirta are of interest for epidemiological studies because they are known to carry potentially zoonotic viruses (e.g. Harima et al, 2020;Sasaki et al, 2015) and haemoparasites (Katakweba et al, 2012). They often represent the most abundant shrews in open savannah-woodland habitats and cannot live in humid forests (Wilson and Mittermeier, 2018).…”