Abstract

Bats are recognized reservoirs for many emerging zoonotic viruses of public health importance. Identifying and cataloguing the viruses of bats is a logical approach to evaluate the range of potential zoonoses of bat origin. We characterized the fecal pathogen microbiome of both insectivorous and frugivorous bats, incorporating 281 individual bats comprising 20 common species, which were sampled in three locations of Yunnan province, by combining reverse transcription polymerase chain reaction (RT-PCR) assays and next-generation sequencing. Seven individual bats were paramyxovirus-positive by RT-PCR using degenerate primers, and these paramyxoviruses were mainly classified into three genera (Rubulavirus, Henipavirus and Jeilongvirus). Various additional novel pathogens were detected in the paramyxovirus-positive bats using Illumina sequencing. A total of 7066 assembled contigs (≥200 bp) were constructed, and 105 contigs matched eukaryotic viruses (of them 103 belong to 2 vertebrate virus families, 1 insect virus, and 1 mycovirus), 17 were parasites, and 4913 were homologous to prokaryotic microorganisms. Among the 103 vertebrate viral contigs, 79 displayed low identity (<70%) to known viruses including human viruses at the amino acid level, suggesting that these belong to novel and genetically divergent viruses. Overall, the most frequently identified viruses, particularly in bats from the family Hipposideridae, were retroviruses. The present study expands our understanding of the bat virome in species commonly found in Yunnan, China, and provides insight into the overall diversity of viruses that may be capable of directly or indirectly crossing over into humans.

Highlights

  • IntroductionBats (order Chiroptera) comprise the second largest mammalian group in the world and are widely distributed across six continents [1]

  • Bats comprise the second largest mammalian group in the world and are widely distributed across six continents [1]

  • Seven fecal samples of seven individual bats were identified as paramyxovirus-positive, and in these bats, two individuals belong to Hipposideros cineraceus, one of Rousettus leschenaultii, one of Eonycteris spelaea, one of Hipposideros armiger, and three

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Summary

Introduction

Bats (order Chiroptera) comprise the second largest mammalian group in the world and are widely distributed across six continents [1]. The outbreak of SARS coronavirus in 2002, which originated in horseshoe bats in China and infected more than 8000 people globally, has spurred increased efforts from the international research community to characterize the diversity and distribution of bat viruses around the world [4,14]. Over 120 bat species have been identified in China, and many are widely distributed throughout the southern provinces of Yunnan, Guangdong, Guangxi, and Fujian [15]. These more ubiquitous bat species, which are in the genera of Rousettus, Myotis, Miniopterus and Hipposideros, naturally reside in close proximity to humans, increasing the potential of transmission of zoonotic pathogens to humans.

Sample Collection and Viral Nucleic Acids Preparation
Screening of Paramyxoviruses and Phylogenetic Analysis
High-throughput Sequencing and Pathogen Analysis
Identification of Viral Homologous Sequences
Phylogenetic Analysis of Viral Sequences
Nucleotide Sequence Accession Numbers
Detection of Paramyxoviruses
In-Depth Analysis of Pathogens by Illumina High-throughput Sequencing
Assembled Contigs and BLASTx Analysis
Identification of Bacteriophage Sequences
Analysis of Vertebrate Viral Sequences by Family
Retroviridae
Polyomaviridae
Discussion
Conclusions
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