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Full text release has been delayed at the author's request until May 05, 2027

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Isolation, Characterization, and Reverse Genetics Platform Construction for Bovine Coronaviruses

Li, Yu
http://rave.ohiolink.edu/etdc/view?acc_num=osu1713308997086987

Abstract Details

2024, Master of Science, Ohio State University, Animal Sciences.
Bovine coronaviruses (BCoVs) are important members of the Betacoronavirus genus, Embecovirus subgenus in Orthocoronavirinae subfamily within the Coronaviridae family. They can cause enteric or/and respiratory diseases in cattle. However, the mechanisms for BCoV tissue tropism and pathogenesis are still unknown and could be due to interactions of viral, host, and environmental factors. There are limited studies to investigate whether co-infection with other bovine pathogens, environmental factors (such as season of the year) and host factors (such as beef vs dairy; age, such as pre- vs post-weaning; and the level of pre-existing BCoV antibodies) contribute to enteric or respiratory tropism. In our study, we isolated recent BCoV strains and compared them with historical strains to study the evolution of BCoVs. Bovine fecal samples were collected from dairy calves, dairy cows, and beef cattle in Georgia by Dr. Palomares Lab at the University of Georgia. Bovine fecal and nasal samples from asymptomatic veal calves from an Ohio farm were provided by Dr. Habing’s Lab at The Ohio State University. We detected BCoVs from both nasal (n=10) and fecal samples (n=156) and isolated BCoV strains from enteric samples. The BCoV genomic sequences of five strains BC7, BC8, BC9, BC39, and BC47 were characterized, and they all belong to genogroup II by phylogenetic analyses. Sequence analyses were performed to compare the mutations between the historical and current viruses and between the viruses isolated from respiratory and enteric samples. We found that one pair of samples BC8 and BC18 from the same calf, but different collection sites (fecal and nasal sites), showed two amino acid differences in the spike (S) protein; We predicted the locations of these two amino acid residues in the 3D protein structures. Reverse genetics is a state-of-the-art technology to study viral factors, but until now no infectious clones have been generated for BCoVs, and this has hampered in-depth research on disease mechanisms and vaccine development. In our study, we are constructing an infectious clone for BCoV. We will introduce the viral genomic RNA (gRNA) into HRT-18 cells to generate the infectious clone-derived BCoV (icBCoV). The construction of this platform will enable future studies of which viral genes are critical for its virulence, tissue tropism, and antagonism to host innate immunity. Our long-term goal is to develop approaches to prevent and control BCoV infections to improve cattle health.
Qiuhong Wang (Advisor)
Linda Saif (Committee Member)
Roberto Palomares (Committee Member)
111 p.
Animal Sciences; Virology
Bovine coronavirus, virus isolation, genomic sequence analysis, reverse genetics, infectious clone

Recommended Citations

Citations

  • Li, Y. (2024). Isolation, Characterization, and Reverse Genetics Platform Construction for Bovine Coronaviruses [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1713308997086987

    APA Style (7th edition)

  • Li, Yu. Isolation, Characterization, and Reverse Genetics Platform Construction for Bovine Coronaviruses. 2024. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1713308997086987.

    MLA Style (8th edition)

  • Li, Yu. "Isolation, Characterization, and Reverse Genetics Platform Construction for Bovine Coronaviruses." Master's thesis, Ohio State University, 2024. http://rave.ohiolink.edu/etdc/view?acc_num=osu1713308997086987

    Chicago Manual of Style (17th edition)

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© 2024, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.
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