Isolation of an equine coronavirus from adult horses with pyrogenic and enteric disease and its antigenic and genomic characterization in comparison with the NC99 strain.
Abstract: A new equine coronavirus was isolated from the feces of adult horses with pyrogenic and enteric disease. The disease outbreak was mainly observed among 2- to 4-year-old horses living in stables of a draft-horse racetrack in Japan. On comparing the isolated virus (isolate Tokachi09) with the equine coronavirus NC99 strain, no significant differences were observed in several biological properties such as hemagglutinating activity, antigenicity (in indirect immunofluorescence and neutralization tests), and one-step growth (in cell culture). The sequences of the nucleocapsid and spike genes of isolate Tokachi09 showed identical size (1341 and 4092 nucleotides, 446 and 1363 amino acids, respectively) and high similarity (98.0% and 99.0% at the nucleotides, 97.3% and 99.0% at the amino acids, respectively) to those of strain NC99. However, the isolate had a 185-nucleotide deletion from four bases after the 3'-terminal end of the spike gene, resulting in the absence of the open reading frame predicted to encode a 4.7-kDa nonstructural protein in strain NC99. These results suggest that the 4.7-kDa nonstructural protein is not essential for viral replication, at least in cell culture, and that the Japanese strain probably originated from a different lineage to the North American strain. This is the first equine coronavirus to be isolated from adult horses with pyrogenic and enteric disease.
Copyright © 2011 Elsevier B.V. All rights reserved.
Publication Date: 2011-01-11 PubMed ID: 21273011PubMed Central: PMC7117184DOI: 10.1016/j.vetmic.2011.01.004Google Scholar: Lookup
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Summary
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The research revolves around the isolation of a new strain of equine coronavirus, named Tokachi09, that was detected in adult horses exhibiting fever and digestive disease. The virus, with origins in a draft-horse racetrack in Japan, was analyzed and compared with an existing equine coronavirus (NC99 strain), showing no significant biological variations yet illustrating a unique genomic feature.
Isolation of the Equine Coronavirus
- The researchers isolated this new strain of the equine coronavirus from the feces of adult horses exhibiting symptoms of pyrogenic (fever-causing) and enteric (digestive) diseases.
- The disease outbreak mainly affected horses aged between 2 to 4 years at a draft-horse racetrack in Japan. The discovery of the virus in these adult horses marks the first time that an equine coronavirus has caused such symptoms in this demographic.
Comparative Analysis with the NC99 Strain
- The isolated virus, known as Tokachi09, was compared with the existing equine coronavirus NC99 strain to observe any significant differences.
- In the comparison, Tokachi09 did not exhibit significant differences from NC99 in several biological properties such as hemagglutinating activity, antigenicity (in indirect immunofluorescence and neutralization tests), and one-step growth (in cell culture).
Genomic Characterization of the Isolate
- Whilst the biological properties were similar, a unique genomic feature was discovered. The sequences of the nucleocapsid and spike genes of Tokachi09 displayed an identical size and high similarity to those of strain NC99, but the Tokachi09 isolate carried a 185-nucleotide deletion from four bases after the 3′-terminal end of the spike gene.
- This deletion resulted in the absence of the open reading frame that was predicted to encode a 4.7-kDa nonstructural protein in the NC99 strain.
- These results thereby propose that the 4.7-kDa nonstructural protein might not be crucial for viral replication, or at least in cell culture, and that the newly isolated Japanese strain possibly rooted from a different lineage than the North American strain.
Cite This Article
APA
Oue Y, Ishihara R, Edamatsu H, Morita Y, Yoshida M, Yoshima M, Hatama S, Murakami K, Kanno T.
(2011).
Isolation of an equine coronavirus from adult horses with pyrogenic and enteric disease and its antigenic and genomic characterization in comparison with the NC99 strain.
Vet Microbiol, 150(1-2), 41-48.
https://doi.org/10.1016/j.vetmic.2011.01.004 Publication
Researcher Affiliations
- Hokkaido Tokachi Livestock Hygiene Service Center, 59-6, Kisen, Kawanishicho, Obihiro, Hokkaido 089-1182, Japan.
MeSH Terms
- Animals
- Antigens, Viral / genetics
- Base Sequence
- Cell Line
- Coronavirus / genetics
- Coronavirus / growth & development
- Coronavirus / isolation & purification
- Coronavirus Infections / veterinary
- Coronavirus Infections / virology
- DNA, Viral / genetics
- Feces / virology
- Genomics
- Horse Diseases / epidemiology
- Horse Diseases / genetics
- Horse Diseases / virology
- Horses / virology
- Japan
- Molecular Sequence Data
- Neutralization Tests
- Open Reading Frames
- Phylogeny
- Sequence Analysis, DNA
References
This article includes 26 references
- Akashi H, Inaba Y, Miura Y, Tokuhisa S, Sato K, Satoda K. Properties of a coronavirus isolated from a cow with epizootic diarrhea. Vet. Microbiol. 1980;5:265–276.
- Bass EP, Sharpee RL. Coronavirus and gastroenteritis in foals.. Lancet 1975 Oct 25;2(7939):822.
- Brian DA, Baric RS. Coronavirus genome structure and replication.. Curr Top Microbiol Immunol 2005;287:1-30.
- Davis E, Rush BR, Cox J, DeBey B, Kapil S. Neonatal enterocolitis associated with coronavirus infection in a foal: a case report.. J Vet Diagn Invest 2000 Mar;12(2):153-6.
- de Haan CA, Masters PS, Shen X, Weiss S, Rottier PJ. The group-specific murine coronavirus genes are not essential, but their deletion, by reverse genetics, is attenuating in the natural host.. Virology 2002 Apr 25;296(1):177-89.
- Decaro N, Cirone F, Mari V, Nava D, Tinelli A, Elia G, Di Sarno A, Martella V, Colaianni ML, Aprea G, Tempesta M, Buonavoglia C. Characterisation of bubaline coronavirus strains associated with gastroenteritis in water buffalo (Bubalus bubalis) calves.. Vet Microbiol 2010 Oct 26;145(3-4):245-51.
- Durham PJ, Stevenson BJ, Farquharson BC. Rotavirus and coronavirus associated diarrhoea in domestic animals.. N Z Vet J 1979 Mar;27(3):30-2.
- Erles K, Shiu KB, Brownlie J. Isolation and sequence analysis of canine respiratory coronavirus.. Virus Res 2007 Mar;124(1-2):78-87.
- Gélinas AM, Boutin M, Sasseville AM, Dea S. Bovine coronaviruses associated with enteric and respiratory diseases in Canadian dairy cattle display different reactivities to anti-HE monoclonal antibodies and distinct amino acid changes in their HE, S and ns4.9 protein.. Virus Res 2001 Jul;76(1):43-57.
- Guy JS, Breslin JJ, Breuhaus B, Vivrette S, Smith LG. Characterization of a coronavirus isolated from a diarrheic foal.. J Clin Microbiol 2000 Dec;38(12):4523-6.
- Huang JC, Wright SL, Shipley WD. Isolation of coronavirus-like agent from horses suffering from acute equine diarrhoea syndrome.. Vet Rec 1983 Sep 17;113(12):262-3.
- Kanno T, Hatama S, Ishihara R, Uchida I. Molecular analysis of the S glycoprotein gene of bovine coronaviruses isolated in Japan from 1999 to 2006.. J Gen Virol 2007 Apr;88(Pt 4):1218-1224.
- Kanno T, Kamiyoshi T, Ishihara R, Hatama S, Uchida I. Phylogenetic studies of bovine coronaviruses isolated in Japan.. J Vet Med Sci 2009 Jan;71(1):83-6.
- Lai M M, Perlman S, Anderson L J. Coronaviridae. Fields Virology 2006;pp. 1305–1335.
- Mair TS, Taylor FG, Harbour DA, Pearson GR. Concurrent cryptosporidium and coronavirus infections in an Arabian foal with combined immunodeficiency syndrome.. Vet Rec 1990 Feb 10;126(6):127-30.
- Mounir S, Labonté P, Talbot PJ. Characterization of the nonstructural and spike proteins of the human respiratory coronavirus OC43: comparison with bovine enteric coronavirus.. Adv Exp Med Biol 1993;342:61-7.
- Page RD. TreeView: an application to display phylogenetic trees on personal computers.. Comput Appl Biosci 1996 Aug;12(4):357-8.
- Sasseville AM, Boutin M, Gélinas AM, Dea S. Sequence of the 3'-terminal end (8.1 kb) of the genome of porcine haemagglutinating encephalomyelitis virus: comparison with other haemagglutinating coronaviruses.. J Gen Virol 2002 Oct;83(Pt 10):2411-2416.
- Schwarz B, Routledge E, Siddell SG. Murine coronavirus nonstructural protein ns2 is not essential for virus replication in transformed cells.. J Virol 1990 Oct;64(10):4784-91.
- Sperry SM, Kazi L, Graham RL, Baric RS, Weiss SR, Denison MR. Single-amino-acid substitutions in open reading frame (ORF) 1b-nsp14 and ORF 2a proteins of the coronavirus mouse hepatitis virus are attenuating in mice.. J Virol 2005 Mar;79(6):3391-400.
- Storz J, Zhang XM, Rott R. Comparison of hemagglutinating, receptor-destroying, and acetylesterase activities of avirulent and virulent bovine coronavirus strains.. Arch Virol 1992;125(1-4):193-204.
- Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.. Nucleic Acids Res 1994 Nov 11;22(22):4673-80.
- Vieler E, Schlapp T, Anders C, Herbst W. Genomic relationship of porcine hemagglutinating encephalomyelitis virus to bovine coronavirus and human coronavirus OC43 as studied by the use of bovine coronavirus S gene-specific probes.. Arch Virol 1995;140(7):1215-23.
- Vijgen L, Keyaerts E, Moës E, Thoelen I, Wollants E, Lemey P, Vandamme AM, Van Ranst M. Complete genomic sequence of human coronavirus OC43: molecular clock analysis suggests a relatively recent zoonotic coronavirus transmission event.. J Virol 2005 Feb;79(3):1595-604.
- Woo PC, Lau SK, Huang Y, Yuen KY. Coronavirus diversity, phylogeny and interspecies jumping.. Exp Biol Med (Maywood) 2009 Oct;234(10):1117-27.
- Zhang J, Guy JS, Snijder EJ, Denniston DA, Timoney PJ, Balasuriya UB. Genomic characterization of equine coronavirus.. Virology 2007 Dec 5;369(1):92-104.
Citations
This article has been cited 36 times.- Kouadria W, Poder SL, van Maanen K, Seuberlich T, Dawson KLD, Zientara S, Laabassi F. First serological evidence of equine coronavirus and severe acute respiratory syndrome coronavirus 2 in horses in North Africa. Vet Res Commun 2025 Oct 4;49(6):347.
- Ricci I, Rosone F, Pacchiarotti G, Manna G, Cersini A, Carvelli A, La Rocca D, Cammalleri E, Giordani R, Tofani S, Conti R, Rombolà P, Nardini R, Minniti CA, Caforio R, Linardi B, Scicluna MT. Pegiviruses and Coronavirus: Biomolecular Prevalence and Phylogenetic Analysis of Strains Detected in Italian Horse Populations. Viruses 2025 Aug 2;17(8).
- Fukumoto N, Bannai H, Kawanishi N, Shibata M, Kishi D, Kambayashi Y, Tsujimura K, Nemoto M. The first outbreak of equine coronavirus infection in 13 years among draft horses at Obihiro Racecourse in Japan in 2025. J Vet Med Sci 2025 Oct 1;87(10):1158-1163.
- Kambayashi Y, Nemoto M, Ochi A, Kishi D, Ueno T, Tsujimura K, Bannai H, Kawanishi N, Ohta M, Suzuki T. Equine coronavirus infection and replication in equine intestinal enteroids. Vet Res 2024 Oct 10;55(1):135.
- Pusterla N, Lawton K, Barnum S. Investigation of the seroprevalence to equine coronavirus and SARS-CoV-2 in healthy adult horses recently imported to the United States. Vet Q 2024 Dec;44(1):1-6.
- Stummer M, Frisch V, Glitz F, Hinney B, Spergser J, Krücken J, Diekmann I, Dimmel K, Riedel C, Cavalleri JV, Rümenapf T, Joachim A, Lyrakis M, Auer A. Presence of Equine and Bovine Coronaviruses, Endoparasites, and Bacteria in Fecal Samples of Horses with Colic. Pathogens 2023 Aug 15;12(8).
- Bannai H, Kambayashi Y, Ohta M, Nemoto M, Tsujimura K. Prevalence of serum and salivary virus-neutralizing antibodies against equine coronavirus in four riding stables in Japan. J Equine Sci 2023 Mar;34(1):13-18.
- Xu Z, Kang X, Han P, Du P, Li L, Zheng A, Deng C, Qi J, Zhao X, Wang Q, Liu K, Gao GF. Binding and structural basis of equine ACE2 to RBDs from SARS-CoV, SARS-CoV-2 and related coronaviruses. Nat Commun 2022 Jun 21;13(1):3547.
- Mashin VV, Sergeev AN, Martynova NN, Oganov MD, Sergeev AA, Kataeva VV, Zagidullin NV. Ensuring Viral Safety of Equine Immunoglobulins during Production. Pharm Chem J 2022;56(2):283-288.
- McKendry R, Lemm NM, Papargyris L, Chiu C. Human Challenge Studies with Coronaviruses Old and New. Curr Top Microbiol Immunol 2024;445:69-108.
- Uzal FA, Arroyo LG, Navarro MA, Gomez DE, Asín J, Henderson E. Bacterial and viral enterocolitis in horses: a review. J Vet Diagn Invest 2022 May;34(3):354-375.
- Korath ADJ, Janda J, Untersmayr E, Sokolowska M, Feleszko W, Agache I, Adel Seida A, Hartmann K, Jensen-Jarolim E, Pali-Schöll I. One Health: EAACI Position Paper on coronaviruses at the human-animal interface, with a specific focus on comparative and zoonotic aspects of SARS-CoV-2. Allergy 2022 Jan;77(1):55-71.
- Schvartz G, Tirosh-Levy S, Barnum S, David D, Sol A, Pusterla N, Steinman A. Seroprevalence and Risk Factors for Exposure to Equine Coronavirus in Apparently Healthy Horses in Israel. Animals (Basel) 2021 Mar 21;11(3).
- Colina SE, Serena MS, Echeverría MG, Metz GE. Clinical and molecular aspects of veterinary coronaviruses. Virus Res 2021 May;297:198382.
- Zappulli V, Ferro S, Bonsembiante F, Brocca G, Calore A, Cavicchioli L, Centelleghe C, Corazzola G, De Vreese S, Gelain ME, Mazzariol S, Moccia V, Rensi N, Sammarco A, Torrigiani F, Verin R, Castagnaro M. Pathology of Coronavirus Infections: A Review of Lesions in Animals in the One-Health Perspective. Animals (Basel) 2020 Dec 11;10(12).
- Khamassi Khbou M, Daaloul Jedidi M, Bouaicha Zaafouri F, Benzarti M. Coronaviruses in farm animals: Epidemiology and public health implications. Vet Med Sci 2021 Mar;7(2):322-347.
- Goodrich EL, Mittel LD, Glaser A, Ness SL, Radcliffe RM, Divers TJ. Novel findings from a beta coronavirus outbreak on an American Miniature Horse breeding farm in upstate New York. Equine Vet Educ 2020 Mar;32(3):150-154.
- Pusterla N, Vin R, Leutenegger C, Mittel LD, Divers TJ. Equine coronavirus: An emerging enteric virus of adult horses. Equine Vet Educ 2016 Apr;28(4):216-223.
- Mattei DN, Kopper JJ, Sanz MG. Equine Coronavirus-Associated Colitis in Horses: A Retrospective Study. J Equine Vet Sci 2020 Apr;87:102906.
- Zhao S, Smits C, Schuurman N, Barnum S, Pusterla N, Kuppeveld FV, Bosch BJ, Maanen KV, Egberink H. Development and Validation of a S1 Protein-Based ELISA for the Specific Detection of Antibodies against Equine Coronavirus. Viruses 2019 Nov 30;11(12).
- Nemoto M, Schofield W, Cullinane A. The First Detection of Equine Coronavirus in Adult Horses and Foals in Ireland. Viruses 2019 Oct 14;11(10).
- Sanz MG, Kwon S, Pusterla N, Gold JR, Bain F, Evermann J. Evaluation of equine coronavirus fecal shedding among hospitalized horses. J Vet Intern Med 2019 Mar;33(2):918-922.
- Manship AJ, Blikslager AT, Elfenbein JR. Disease features of equine coronavirus and enteric salmonellosis are similar in horses. J Vet Intern Med 2019 Mar;33(2):912-917.
- Pusterla N, Vin R, Leutenegger CM, Mittel LD, Divers TJ. Enteric coronavirus infection in adult horses. Vet J 2018 Jan;231:13-18.
- Shaw SD, Stämpfli H. Diagnosis and Treatment of Undifferentiated and Infectious Acute Diarrhea in the Adult Horse. Vet Clin North Am Equine Pract 2018 Apr;34(1):39-53.
- Nemoto M, Kanno T, Bannai H, Tsujimura K, Yamanaka T, Kokado H. Antibody response to equine coronavirus in horses inoculated with a bovine coronavirus vaccine. J Vet Med Sci 2017 Nov 17;79(11):1889-1891.
- Kooijman LJ, James K, Mapes SM, Theelen MJ, Pusterla N. Seroprevalence and risk factors for infection with equine coronavirus in healthy horses in the USA. Vet J 2017 Feb;220:91-94.
- Nemoto M, Oue Y, Higuchi T, Kinoshita Y, Bannai H, Tsujimura K, Yamanaka T, Kondo T. Low prevalence of equine coronavirus in foals in the largest thoroughbred horse breeding region of Japan, 2012-2014. Acta Vet Scand 2015 Sep 22;57(1):53.
- Nemoto M, Oue Y, Murakami S, Kanno T, Bannai H, Tsujimura K, Yamanaka T, Kondo T. Complete genome analysis of equine coronavirus isolated in Japan. Arch Virol 2015 Nov;160(11):2903-6.
- Nemoto M, Morita Y, Niwa H, Bannai H, Tsujimura K, Yamanaka T, Kondo T. Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods 2015 Apr;215-216:13-6.
- Fielding CL, Higgins JK, Higgins JC, McIntosh S, Scott E, Giannitti F, Mete A, Pusterla N. Disease associated with equine coronavirus infection and high case fatality rate. J Vet Intern Med 2015 Jan;29(1):307-10.
- Nemoto M, Oue Y, Morita Y, Kanno T, Kinoshita Y, Niwa H, Ueno T, Katayama Y, Bannai H, Tsujimura K, Yamanaka T, Kondo T. Experimental inoculation of equine coronavirus into Japanese draft horses. Arch Virol 2014 Dec;159(12):3329-34.
- Miszczak F, Tesson V, Kin N, Dina J, Balasuriya UB, Pronost S, Vabret A. First detection of equine coronavirus (ECoV) in Europe. Vet Microbiol 2014 Jun 25;171(1-2):206-9.
- Barros IN, Silva SO, Nogueira Neto FS, Asano KM, Souza SP, Richtzenhain LJ, Brandao PE. A multigene approach for comparing genealogy of Betacoronavirus from cattle and horses. ScientificWorldJournal 2013;2013:349702.
- Kanno T, Ishihara R, Hatama S, Uchida I. Antigenic variation among recent Japanese isolates of bovine coronaviruses belonging to phylogenetically distinct genetic groups. Arch Virol 2013 May;158(5):1047-53.
- Pusterla N, Mapes S, Wademan C, White A, Ball R, Sapp K, Burns P, Ormond C, Butterworth K, Bartol J, Magdesian KG. Emerging outbreaks associated with equine coronavirus in adult horses. Vet Microbiol 2013 Feb 22;162(1):228-31.
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