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1、S H O R T C O M M U N I C A T I O N Coronaviruses in guano from Pteropus medius bats in Peradeniya, Sri Lanka H. D. W. S. Kudagammana1,2|V. Thevanesam2|D. K. W. Chu3| N. B. Eriyagama2|J. S. M. Peiris3|F. Noordeen2 1Faculty of Allied Health Sciences, University of Peradeniya, Peradeniya, Sri Lanka 2D
2、epartment of Microbiology, Faculty of Medicine, University of Peradeniya, Peradeniya, Sri Lanka 3School of Public Health, University of Hong Kong, Hong Kong, Hong Kong Correspondence H. D. W. S. Kudagammana, Faculty of Allied Health Sciences, University of Peradeniya, Peradeniya, Sri Lanka. Email: w
3、asanakudagammanayahoo.co.uk Funding information University of Peradeniya, Sri Lanka, Grant/ Award Number: RG/AF/13/38/M; US National Institutes of Health, Grant/Award Number: Contract No. HHSN272201500006C Summary Bats are a unique group of mammals well suited to be hosts for emerging viruses. With
4、current rates of deforestation and urbanization, redistribution of bat habitats to urban and suburban areas may bring bats into closer contact with livestock and humans. Common flying fox, Pteropus medius (previously known as Pteropus gigan- teus), forms large communal roosts on treetops, often in c
5、lose proximity to human habitation in Sri Lanka. This report describes the detection of coronavirus RNA in P. medius bat guano collected in Peradeniya, Sri Lanka. These viruses had 97% nucleotide identity with coronaviruses detected in Cynopterus sphinx, Scotophilus heathii and S. kuhlii bats in Tha
6、iland. Pteropus medius is widespread in Asia and appears to excrete group D coronaviruses, which are hitherto confined to bats; however, these findings may have public health implications in the future. K E Y W O R DS coronavirus, flying fox, Sri Lanka 1|INTRODUCTION Bats contribute to maintain a ba
7、lanced ecosystem through dissemi- nating plant seeds or eating some types of insects. Some bat species are at risk and endangered. The displacement and redistribution of bats from their natural habitats expose susceptible human and ani- mal populations to micro-organisms (Han et al., 2015). Bats are
8、 a unique group of mammals that can fly long distances for daily forag- ing and aggregate in large colonies. Daily movement patterns, com- pact roosting structure, hibernation, periodic migration and long lifespan may make bats suitable hosts for emerging viruses (Han et al., 2015). Coronaviruses (C
9、oV) are a group of enveloped, single-stranded (positive-sense) RNA viruses that belong to the order Nidovirales, family Coronaviridae. They are divided into four major groups: alpha, beta, gamma and delta coronaviruses, only the alpha and beta coron- aviruses being known to have infected humans (de
10、Groot et al., 2013). The earliest known human coronaviruses (HCoV), HCoV-229E and HCoV-OC43 are causes of mild upper respiratory infections. The identification of the precursor of Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) in Rhinolophid bats led to more intensive investigations of cor
11、onaviruses in humans and in bats (Han et al., 2015). NL63 and HKU1 are newly discovered coronaviruses endemic in humans (Woo et al., 2005; Pyrc, Berkhout, 2018 Blackwell Verlag GmbH | 1 selected trees were covered with clean polythene sheets. The accu- mulations of droppings were observed and identi
12、fied immediately prior to collection to confirm the origin of the guano. About 12 g of each bat guano sample was collected into a vial with viral trans- port medium (VTM). The guano samples were transported to the Virology Laboratory, Faculty of Medicine, University of Peradeniya for screening with
13、RT-PCR work. The ethical clearance (EC No: 2012/EC/52) for the study was obtained from the Ethical Review Committee of the Faculty of Medicine, University of Peradeniya, Sri Lanka. Viral RNA was extracted using QIAGEN QIAamp Viral RNA kit (QIAGEN, Hilden, Germany) as recommended by the manufacturer.
14、 The cDNA was generated from RNA using Superscript III reverse transcriptase (Invitrogen) with random hexamers. The cDNA was screened for the presence of coronavirus using a broad range hemi- nested RT-PCR targeting conserved regions of the RdRp gene for detecting known and unknown coronaviruses as
15、previously described (Hemida et al., 2017). Positive and negative controls were included for each run. RT-PCR products were analysed using agarose gel elec- trophoresis to identify a product size of 440 base pairs indicating the presence of known or unknown coronaviruses. DNA sequencing was carried
16、out on four strongly positive RT-PCR products at School of Public Health, University of Hong Kong, Hong Kong. 3|RESULTS AND DISCUSSION The bats from which the guano samples taken were morphologically identified as Pteropus medius (previously known as P. giganteus) belonging to the family Pteropodedae. Eight (16%) of the 50 bat guano samples were positive for coronavirus; three of 10 collected on 1 February, four of 10 collected on 8 February, zero of 16 FIGURE 1Phylogenetic analysis of coronav
