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. 2021 Jan;166(1):35-42.
doi: 10.1007/s00705-020-04826-w. Epub 2020 Oct 1.

Molecular characterization of canine coronaviruses: an enteric and pantropic approach

Mehmet Ozkan Timurkan  1 , Hakan Aydin  2 , Ender Dincer  3 , Nuvit Coskun  4
Affiliations

Molecular characterization of canine coronaviruses: an enteric and pantropic approach

Mehmet Ozkan Timurkan et al. Arch Virol. 2021 Jan.

Abstract

Canine coronavirus (CCoV) generally causes an infection with high morbidity and low mortality in dogs. In recent years, studies on coronaviruses have gained a momentum due to coronavirus outbreaks. Mutations in coronaviruses can result in deadly diseases in new hosts (such as SARS-CoV-2) or cause changes in organ-tissue affinity, as occurred with feline infectious peritonitis virus, exacerbating their pathogenesis. In recent studies on different types of CCoV, the pantropic strains characterized by hypervirulent and multi-systemic infections are believed to be emerging, in contrast to classical enteric coronavirus infections. In this study, we investigated emerging hypervirulent and multi-systemic CCoV strains using molecular and bioinformatic analysis, and examined differences between enteric and pantropic CCoV strains at the phylogenetic level. RT-PCR was performed with specific primers to identify the coronavirus M (membrane) and S (spike) genes, and samples were then subjected to DNA sequencing. In phylogenetic analysis, four out of 26 samples were classified as CCoV-1. The remaining 22 samples were all classified as CCoV-2a. In the CCoV-2a group, six samples were in branches close to enteric strains, and 16 samples were in the branches close to pantropic strains. Enteric and pantropic strains were compared by molecular genotyping of CCoV in dogs. Phylogenetic analysis of hypervirulent pantropic strains was carried out at the amino acid and nucleotide sequence levels. CCoV was found to be divergent from the original strain. This implies that some CCoV strains have become pantropic strains that cause multisystemic infections, and they should not be ruled out as the cause of severe diarrhea and multisystemic infections.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1
Phylogenetic tree based on canine coronavirus M gene sequences from Turkey and reference sequences from GenBank. The tree was constructed by the maximum-likelihood algorithm implemented in the MEGA 6 program using p-distance. Values at the nodes (based on 1,000 bootstrap replicates) represent the support for each node and the corresponding clade. The scale bar indicates substitutions per site. Sequences from Turkish pantropic CCoV2a strains are indicated by a square (∎), Turkish enteric CCoV2a strains are indicated by a black circle (●), Turkish CCoV1 strains are indicated by a triangle (▲), and reference sequences are indicated by their accession number
Fig. 2
Fig. 2
Phylogenetic tree based on canine coronavirus S (spike) gene sequences from Turkey and reference sequences from GenBank. The tree was constructed by the maximum-likelihood algorithm implemented in the MEGA 6 program using p-distance. Values at the nodes (based on 1,000 bootstrap replicates) represent the support for each node and the corresponding clade. The scale bar indicates substitutions per site. Sequences from Turkish pantropic CCoV2a strains are indicated by a square (∎), Turkish enteric CCoV2a strains are indicated by a black circle (●), Turkish CCoV1 strains are indicated by a triangle (▲), and reference sequences are indicated by their accession number
Fig. 3
Fig. 3
Analysis of the predicted amino acid sequences of the S (spike) proteins of enteric and pantropic CCoV strains
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