Bat Coronaviruses and Experimental Infection of Bats, the Philippines

Shumpei Watanabe,Naoya Ueda,Phillip Alviola,Joseph S Masangkay,Hiroomi Akashi,Kentaro Kato,Satoshi Taniguchi,Shumpei Tsuda,Tetsuya Mizutani,Yukinobu Tohya,Noriyo Nagata,Yasuhiro Yoshikawa,Shigeru Morikawa,Shigeru Kyuwa,Tsutomu Omatsu,Koichiro Iha,Maiko Endoh,Hikaru Fujii,Shuetsu Fukushi

doi:10.3201/eid1608.100208

Abstract

Fifty-two bats captured during July 2008 in the Philippines were tested by reverse transcription-PCR to detect bat coronavirus (CoV) RNA. The overall prevalence of virus RNA was 55.8%. We found 2 groups of sequences that belonged to group 1 (genus Alphacoronavirus) and group 2 (genus Betacoronavirus) CoVs. Phylogenetic analysis of the RNA-dependent RNA polymerase gene showed that groups 1 and 2 CoVs were similar to Bat-CoV/China/A515/2005 (95% nt sequence identity) and Bat-CoV/HKU9-1/China/2007 (83% identity), respectively. To propagate group 2 CoVs obtained from a lesser dog-faced fruit bat (Cynopterus brachyotis), we administered intestine samples orally to Leschenault rousette bats (Rousettus leschenaulti) maintained in our laboratory. After virus replication in the bats was confirmed, an additional passage of the virus was made in Leschenault rousette bats, and bat pathogenesis was investigated. Fruit bats infected with virus did not show clinical signs of infection.

Highlights

Fifty-two bats captured during July 2008 in the Philippines were tested by reverse transcription–PCR to detect bat coronavirus (CoV) RNA
Phylogenetic analysis of bat CoVs and other known CoVs suggested that the progenitor of Severe acute respiratory syndrome (SARS)-CoV and all other CoVs in other animal hosts originated in bats [5,7]
RT-PCRs for a 440-bp fragment in the RNA-dependent RNA polymerase (RdRp) gene of CoVs were performed for large intestine samples, including intestinal contents; 9 (17.3%) of 52 bats were positive

Summary

Introduction

Fifty-two bats captured during July 2008 in the Philippines were tested by reverse transcription–PCR to detect bat coronavirus (CoV) RNA. Epidemiologic studies have demonstrated that the first human cases of SARS were caused by CoVs closely related to those found in Himalayan palm civets and raccoon dogs in wildlife markets [1]. This finding accelerated surveys of CoVs specific for various animals in Southeast Asia to identify reser-. We attempted to isolate bat CoVs and virus RNA from cell cultures and from Leschenault rousette bats (Rousettus leschenaulti) orally infected with intestinal tissues and contents from a lesser dog-faced fruit bat (Cynopterus brachyotis). Clinical signs of infected bats were examined, and pathogenesis in bats was investigated

Methods

Results

Conclusion