Establishment of Intestinal Organoid from Rousettus leschenaultii and the Susceptibility to Bat-Associated Viruses, SARS-CoV-2 and Pteropine Orthoreovirus.

Mohamed Elbadawy,Masayuki Saijo,Mio Kobayashi,Amira Abugomaa,Kazuaki Sasaki,Masahiro Kaneda,Toshinori Yoshida,Chang-Kweng Lim,Hideyuki Yamawaki,Tetsuya Mizutani,Makoto Shibutani,Tsutomu Omatsu,Nagisa Saito,Yuki Kato,Tatsuya Usui,Kimika Hayashi

doi:10.3390/ijms221910763

Abstract

Various pathogens, such as Ebola virus, Marburg virus, Nipah virus, Hendra virus, Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and SARS-CoV-2, are threatening human health worldwide. The natural hosts of these pathogens are thought to be bats. The rousette bat, a megabat, is thought to be a natural reservoir of filoviruses, including Ebola and Marburg viruses. Additionally, the rousette bat showed a transient infection in the experimental inoculation of SARS-CoV-2. In the current study, we established and characterized intestinal organoids from Leschenault’s rousette, Rousettus leschenaultii. The established organoids successfully recapitulated the characteristics of intestinal epithelial structure and morphology, and the appropriate supplements necessary for long-term stable culture were identified. The organoid showed susceptibility to Pteropine orthoreovirus (PRV) but not to SARS-CoV-2 in experimental inoculation. This is the first report of the establishment of an expandable organoid culture system of the rousette bat intestinal organoid and its sensitivity to bat-associated viruses, PRV and SARS-CoV-2. This organoid is a useful tool for the elucidation of tolerance mechanisms of the emerging rousette bat-associated viruses such as Ebola and Marburg virus.

Highlights

Bats are one of the most important natural reservoirs for a variety of emerging viruses that induce severe illness, including severe acute respiratory syndrome coronavirus (SARSCoV), Middle East respiratory syndrome coronavirus (MERS-CoV), Hendra virus, Ebola virus, Marburg virus, and SARS-CoV-2 [1,2,3,4]
Rousette bat intestinal organoids were successfully generated and analysed from R. leschenaultii that died of natural causes at the zoo and were collected Figure 1A
The structure of the organoid mimicked the multicellular structure of the bat small intestinal epithelium (Figure 1B), and the organoids recapitulated the histology of the original tissue (Figure 1C)

Summary

Introduction

Bats are one of the most important natural reservoirs for a variety of emerging viruses that induce severe illness, including severe acute respiratory syndrome coronavirus (SARSCoV), Middle East respiratory syndrome coronavirus (MERS-CoV), Hendra virus, Ebola virus, Marburg virus, and SARS-CoV-2 [1,2,3,4]. Among the Rousettus spp., the rousette bat is thought to be a natural reservoir of Ebola and Marburg viruses (family Filoviridae) and is a source of virus spillover into human populations; these viruses have frequently caused outbreaks in African countries [5,6]. These days, betacoronavirus genome [7,8] and CoV RNA-dependent RNA polymerase gene [9] were detected in Rousettus spp. In an experiment involving inoculation of R. aegyptiacs with Marburg virus, viremia was detected, and a systemic infection developed without any symptoms [14]

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