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A comprehensive molecular analysis of bovine coronavirus strains isolated from Brazil and comparison of a wild-type and cell culture-adapted strain associated with respiratory disease

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Abstract

Bovine coronavirus (BCoV) has dual tropisms that can trigger enteric and respiratory diseases in cattle. Despite its global distribution, BCoV field strains from Brazil remain underexplored in studies investigating the virus's worldwide circulation. Another research gap involves the comparative analysis of S protein sequences in BCoV isolates from passages in cell lines versus direct sequencing from clinical samples. Therefore, one of the objectives of our study was to conduct a comprehensive phylogenetic analysis of BCoV strains identified from Brazil, including a respiratory strain obtained during this study, comparing them with global and ancestral BCoV strains. Additionally, we performed a comparative analysis between wild-type BCoV directly sequenced from the clinical sample (nasal secretion) and the cell culture-adapted strain, utilizing the Sanger method. The field strain and multiple cell passage in cell culture (HRT-18) adapted BCoV strain (BOV19 NS) detected in this study were characterized through molecular and phylogenetic analyses based on partial fragments of 1,448 nt covering the hypervariable region of the S gene. The analyses have demonstrated that different BCoV strains circulating in Brazil, and possibly Brazilian variants, constitute a new genotype (putative G15 genotype). Compared with the ancestral prototype (Mebus strain) of BCoV, 33 nt substitutions were identified of which 15 resulted in non-synonymous mutations (nine transitions and six transversions). Now, compared with the wild-type strain was identified only one nt substitution in nt 2,428 from the seventh passage onwards, which resulted in transversion, neutral-neutral charge, and one substitution of asparagine for tyrosine at aa residue 810 (N810Y).

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Acknowledgements

The authors would like to thank the Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES/MEC) for granting a scholarship to the first author.

Funding

This work was supported by Universidade Federal do Paraná-UFPR, Universidade Estadual de Londrina-UEL, CAPES/MEC-Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, and National Institute of Science and Technology of Dairy Production Chain (INCT-Leite / CNPq, CAPES, and Araucaria Foundation-FAP/PR).

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Authors and Affiliations

  1. Departament of Veterinary Sciences, Universidade Federal do Paraná-UFPR, Rua Pioneiro, 2153, Palotina, Paraná, 85950-000, Brazil

    Janaina Lustosa de Mello, Daniela Lorencena, Ruana Renostro Delai, Andressa Fernanda Kunz & Elisabete Takiuchi

  2. Department of Preventive Veterinary Medicine, Universidade Estadual de Londrina-UEL, PO Box 6001, Londrina, Paraná, 86051-990, Brazil

    Flávia Possatti & Amauri Alcindo Alfieri

  3. Multi-User Animal Health Laboratory, Molecular Biology Unit, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Londrina, Paraná, 86057-970, Brazil

    Amauri Alcindo Alfieri

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  1. Janaina Lustosa de Mello
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  2. Daniela Lorencena
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Contributions

Janaina Lustosa de Mello: Conceptualization, Methodology, Investigation, Data curation, Formal analysis, Writing-original draft. Daniela Lorencena: Investigation. Ruana Renostro Delai: Investigation, Data curation. Andressa Fernanda Kunz: Investigation, Validation. Flávia Possatti: Investigation, Data curation. Amauri Alcindo Alfieri: Visualization, Writing-review & editing. Elisabete Takiuchi: Conceptualization, Formal analysis, Supervision, Visualization, Writing-review & editing.

Corresponding author

Correspondence to Elisabete Takiuchi.

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This study was approved by the Ethics Committee for the Use of Animals of the Federal University of Paraná, under protocol nº. 34/2016.

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de Mello, J.L., Lorencena, D., Delai, R.R. et al. A comprehensive molecular analysis of bovine coronavirus strains isolated from Brazil and comparison of a wild-type and cell culture-adapted strain associated with respiratory disease. Braz J Microbiol (2024). https://doi.org/10.1007/s42770-024-01287-0

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