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Journal of Virology, May 2007, p. 4981-4990, Vol. 81, No. 10
0022-538X/07/$08.00+0     doi:10.1128/JVI.02361-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Biologic, Antigenic, and Full-Length Genomic Characterization of a Bovine-Like Coronavirus Isolated from a Giraffe{triangledown}

Mustafa Hasoksuz ,1,{dagger},{ddagger} Konstantin Alekseev,1,{ddagger} Anastasia Vlasova,1,{ddagger} Xinsheng Zhang,1 David Spiro,2 Rebecca Halpin,2 Shiliang Wang,2 Elodie Ghedin,2 and Linda J. Saif1*

Food Animal Health Research Program, Department of Veterinary Preventive Medicine, Ohio Agricultural Research and Development Center, The Ohio State University, 1680 Madison Avenue, Wooster, Ohio 44691-4096,1 The Institute for Genomic Research, Rockville, Maryland2

Received 27 October 2006/ Accepted 1 March 2007

Coronaviruses (CoVs) possess large RNA genomes and exist as quasispecies, which increases the possibility of adaptive mutations and interspecies transmission. Recently, CoVs were recognized as important pathogens in captive wild ruminants. This is the first report of the isolation and detailed genetic, biologic, and antigenic characterization of a bovine-like CoV from a giraffe (Giraffa camelopardalis) in a wild-animal park in the United States. CoV particles were detected by immune electron microscopy in fecal samples from three giraffes with mild-to-severe diarrhea. From one of the three giraffe samples, a CoV (GiCoV-OH3) was isolated and successfully adapted to serial passage in human rectal tumor 18 cell cultures. Hemagglutination assays, receptor-destroying enzyme activity, hemagglutination inhibition, and fluorescence focus neutralization tests revealed close biological and antigenic relationships between the GiCoV-OH3 isolate and selected respiratory and enteric bovine CoV (BCoV) strains. When orally inoculated into a BCoV-seronegative gnotobiotic calf, GiCoV-OH3 caused severe diarrhea and virus shedding within 2 to 3 days. Sequence comparisons and phylogenetic analyses were performed to assess its genetic relatedness to other CoVs. Molecular characterization confirmed that the new isolate belongs to group 2a of the mammalian CoVs and revealed closer genetic relatedness between GiCoV-OH3 and the enteric BCoVs BCoV-ENT and BCoV-DB2, whereas BCoV-Mebus was more distantly related. Detailed sequence analysis of the GiCoV-OH3 spike gene demonstrated the presence of a deletion in the variable region of the S1 subunit (from amino acid 543 to amino acid 547), which is a region associated with pathogenicity and tissue tropism for other CoVs. The point mutations identified in the structural proteins (by comparing GiCoV-OH3, BCoV-ENT, BCoV-DB2, and BCoV-Mebus) were most conserved among GiCoV-OH3, BCoV-ENT, and BCoV-DB2, whereas most of the point mutations in the nonstructural proteins were unique to GiCoV-OH3. Our results confirm the existence of a bovine-like CoV transmissible to cattle from wild ruminants, namely, giraffes, but with certain genetic properties different from those of BCoVs.

* Corresponding author. Mailing address: Food Animal Health Research Program, Department of Veterinary Preventive Medicine, Ohio Agricultural Research and Development Center, The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691. Phone: (330) 263-3744. Fax: (330) 263-3677. E-mail: saif.2{at}osu.edu

{triangledown} Published ahead of print on 7 March 2007.

{dagger} Present address: Department of Virology, Faculty of Veterinary Medicine, Istanbul University, Avcilar 34320, Istanbul, Turkey.

{ddagger} M.H., K.A., and A.V. contributed equally to this paper.

Journal of Virology, May 2007, p. 4981-4990, Vol. 81, No. 10
0022-538X/07/$08.00+0     doi:10.1128/JVI.02361-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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