{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T06:59:04Z","timestamp":1762325944828,"version":"build-2065373602"},"reference-count":33,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2024,6,21]],"date-time":"2024-06-21T00:00:00Z","timestamp":1718928000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Bovine coronavirus (BCoV) is distributed globally and mainly causes different clinical manifestations: enteric diarrhea in calves, winter dysentery in adults, and respiratory symptoms in cattle of all ages. Low mortality and high morbidity are the hallmarks of BCoV infection, usually associated with substantial economic losses for the livestock industry. Vaccination, combined with the implementation of biosecurity measures, is the key strategy for the prevention of infections. This pilot study evaluates the immunogenicity of a recombinant vaccine containing two BCoV antigens (S and M) in sheep, compared to vaccines containing only the M or S protein. Three groups of sheep were inoculated intramuscularly at day 0 and day 21 with recombinant adenoviruses expressing BCoV S protein (AdV-BCoV-S), BCoV M protein (AdV-BCoV-M), or both proteins (AdV-BCoV-S + M). Serum antibodies were evaluated using immunofluorescence (IF) and serum neutralization (SN) tests. Moderate seroconversion was observed by day 21, but serum antibodies detected via SN increased from 1:27.5 (day 21) to 1:90 (day 28) in sheep inoculated with the recombinant AdV expressing both the S- and M-BCoV proteins. Based on the SN results, a repeated-measures ANOVA test indicated a more significant difference in immune response between the three groups (F = 20.47; p &lt; 0.001). The experimental investigation produced satisfactory results, highlighting that the S + M recombinant vaccine was immunogenic, stimulating a valid immune response. Despite some inherent limitations, including a small sample size and the absence of challenge tests, the study demonstrated the efficacy of the immune response induced via the recombinant vaccine containing both S and M proteins compared to that induced via the individual proteins S or M.<\/jats:p>","DOI":"10.3390\/pathogens13070523","type":"journal-article","created":{"date-parts":[[2024,6,21]],"date-time":"2024-06-21T05:33:28Z","timestamp":1718948008000},"page":"523","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Humoral Immune Response in Immunized Sheep with Bovine Coronavirus Glycoproteins Delivered via an Adenoviral Vector"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6862-1792","authenticated-orcid":false,"given":"Annamaria","family":"Pratelli","sequence":"first","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5563-1131","authenticated-orcid":false,"given":"Paolo","family":"Capozza","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-1569-9500","authenticated-orcid":false,"given":"Sergio","family":"Minesso","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science, University of Parma, 43126 Parma, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Stella","family":"Lucente","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0026-8472","authenticated-orcid":false,"given":"Francesco","family":"Pellegrini","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4300-8822","authenticated-orcid":false,"given":"Maria","family":"Tempesta","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3945-2285","authenticated-orcid":false,"given":"Valentina","family":"Franceschi","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science, University of Parma, 43126 Parma, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Canio","family":"Buonavoglia","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gaetano","family":"Donofrio","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science, University of Parma, 43126 Parma, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1016\/j.rvsc.2020.11.002","article-title":"Bovine respiratory disease in beef calves supported long transport stress: An epidemiological study and strategies for control and prevention","volume":"135","author":"Pratelli","year":"2021","journal-title":"Res. Vet. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e646","DOI":"10.1016\/S2666-5247(21)00282-2","article-title":"One world, one health, one virology of the mysterious labyrinth of coronaviruses: The canine coronavirus affair","volume":"2","author":"Pratelli","year":"2021","journal-title":"Lancet Microbe"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2809","DOI":"10.1007\/s11250-020-02354-y","article-title":"Coronaviruses in cattle","volume":"52","author":"Hodnik","year":"2020","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"643220","DOI":"10.3389\/fvets.2021.643220","article-title":"Bovine Coronavirus and the Associated Diseases","volume":"8","author":"Vlasova","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"854844","DOI":"10.3389\/fvets.2022.854844","article-title":"Editorial: Evolving Prospects of Bovine Respiratory Diseases and Management in Feedlot Cattle","volume":"9","author":"Pratelli","year":"2022","journal-title":"Front. Vet. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Buonavoglia, A., Pellegrini, F., Decaro, N., Galgano, M., and Pratelli, A. (2023). A One Health Perspective on Canine Coronavirus: A Wolf in Sheep\u2019s Clothing?. Microorganisms, 11.","DOI":"10.3390\/microorganisms11040921"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"213","DOI":"10.5326\/JAAHA-MS-Canine-Vaccination-Guidelines","article-title":"2022 AAHA Canine Vaccination Guidelines","volume":"58","author":"Ellis","year":"2022","journal-title":"J. Am. Anim. Hosp. Assoc."},{"key":"ref_8","first-page":"147","article-title":"What is the evidence that bovine coronavirus is a biologically significant respiratory pathogen in cattle?","volume":"60","author":"Ellis","year":"2019","journal-title":"Can. Vet. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1128\/JVI.75.3.1312-1324.2001","article-title":"The membrane M protein carboxy terminus binds to transmissible gastroenteritis coronavirus core and contributes to core stability","volume":"75","author":"Escors","year":"2001","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2020","DOI":"10.1002\/j.1460-2075.1996.tb00553.x","article-title":"Nucleocapsid-independent assembly of coronavirus-like particles by co-expression of viral envelope protein genes","volume":"15","author":"Vennema","year":"1996","journal-title":"EMBO J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/j.cvfa.2010.04.005","article-title":"Bovine respiratory coronavirus","volume":"26","author":"Saif","year":"2010","journal-title":"Vet. Clin. N. Am. Food Anim. Pract."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Zhu, Q., Li, B., and Sun, D. (2022). Advances in Bovine Coronavirus Epidemiology. Viruses, 14.","DOI":"10.3390\/v14051109"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Cirone, F., Padalino, B., Tullio, D., Capozza, P., Losurdo, M., Lanave, G., and Pratelli, A. (2019). Prevalence of Pathogens Related to Bovine Respiratory Disease Before and After Transportation in Beef Steers: Preliminary Results. Animals, 9.","DOI":"10.3390\/ani9121093"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1367128","DOI":"10.3389\/fvets.2024.1367128","article-title":"Editorial: Respiratory diseases and management in livestock","volume":"11","author":"Pratelli","year":"2024","journal-title":"Front. Vet. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"627894","DOI":"10.3389\/fvets.2021.627894","article-title":"Factors Affecting the Development of Bovine Respiratory Disease: A Cross-Sectional Study in Beef Steers Shipped From France to Italy","volume":"8","author":"Padalino","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0378-1119(85)90017-4","article-title":"The quasispecies (extremely heterogeneous) nature of viral RNA genome populations: Biological relevance\u2014A review","volume":"40","author":"Domingo","year":"1985","journal-title":"Gene"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3264","DOI":"10.1128\/jcm.33.12.3264-3269.1995","article-title":"Isolation of coronaviruses antigenically indistinguishable from bovine coronavirus from wild ruminants with diarrhea","volume":"33","author":"Tsunemitsu","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Franzo, G., Drigo, M., Legnardi, M., Grassi, L., Pasotto, D., Menandro, M.L., Cecchinato, M., and Tucciarone, C.M. (2020). Bovine Coronavirus: Variability, Evolution, and Dispersal Patterns of a No Longer Neglected Betacoronavirus. Viruses, 12.","DOI":"10.3390\/v12111285"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1017\/S1466252318000117","article-title":"Bovine-like coronaviruses in domestic and wild ruminants","volume":"19","author":"Amer","year":"2018","journal-title":"Anim. Health Res. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"41","DOI":"10.3201\/eid1601.090726","article-title":"Recombinant canine coronaviruses in dogs, Europe","volume":"16","author":"Decaro","year":"2010","journal-title":"Emerg. Infect. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7270","DOI":"10.1128\/JVI.02675-05","article-title":"Evolutionary history of the closely related group 2 coronaviruses: Porcine hemagglutinating encephalomyelitis virus, bovine coronavirus, and human coronavirus OC43","volume":"80","author":"Vijgen","year":"2006","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/j.virol.2007.06.035","article-title":"Genomic characterization of equine coronavirus","volume":"369","author":"Zhang","year":"2007","journal-title":"Virology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2401","DOI":"10.1007\/s007050170011","article-title":"Cross-protection studies between respiratory and calf diarrhea and winter dysentery coronavirus strains in calves and RT-PCR and nested PCR for their detection","volume":"146","author":"Cho","year":"2001","journal-title":"Arch. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.virusres.2016.06.017","article-title":"Bovine herpesvirus-1: Genetic diversity of field strains from cattle with respiratory disease, genital, fetal disease and systemic neonatal disease and their relationship to vaccine strains","volume":"223","author":"Fulton","year":"2016","journal-title":"Virus Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s44149-024-00118-x","article-title":"Decoding bovine coronavirus immune targets: An epitope informatics approach","volume":"4","author":"Rani","year":"2024","journal-title":"Anim. Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.1002\/jmv.27487","article-title":"COVID-19 vaccinations: The unknowns, challenges, and hopes","volume":"94","author":"Mohamed","year":"2022","journal-title":"J. Med. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2509","DOI":"10.1073\/pnas.95.5.2509","article-title":"A simplified system for generating recombinant adenoviruses","volume":"95","author":"He","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8802","DOI":"10.1073\/pnas.91.19.8802","article-title":"An efficient and flexible system for construction of adenovirus vectors with insertions or deletions in early regions 1 and 3","volume":"91","author":"Bett","year":"1994","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"5437","DOI":"10.1128\/JVI.79.9.5437-5444.2005","article-title":"Replication-competent adenovirus formation in 293 cells: The recombination-based rate is influenced by structure and location of the transgene cassette and not increased by overproduction of HsRad51, Rad51-interacting, or E2F family proteins","volume":"79","author":"Duigou","year":"2005","journal-title":"J. Virol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"637152","DOI":"10.3389\/fimmu.2021.637152","article-title":"Bovine Coronavirus Immune Milk Against COVID-19","volume":"12","author":"Arenas","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Ning, S., and Firinu, D. (2024). Current Understanding of the Immune Response after COVID-19 Vaccination. Vaccines, 12.","DOI":"10.3390\/vaccines12030250"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Bonifacio, M.A., Laterza, R., Vinella, A., Schirinzi, A., Defilippis, M., Di Serio, F., Ostuni, A., Fasanella, A., and Mariggio, M.A. (2022). Correlation between In Vitro Neutralization Assay and Serological Tests for Protective Antibodies Detection. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23179566"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"618685","DOI":"10.3389\/fimmu.2020.618685","article-title":"Distinct Features and Functions of Systemic and Mucosal Humoral Immunity Among SARS-CoV-2 Convalescent Individuals","volume":"11","author":"Butler","year":"2020","journal-title":"Front. Immunol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/13\/7\/523\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:02:17Z","timestamp":1760108537000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/13\/7\/523"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,21]]},"references-count":33,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2024,7]]}},"alternative-id":["pathogens13070523"],"URL":"https:\/\/doi.org\/10.3390\/pathogens13070523","relation":{},"ISSN":["2076-0817"],"issn-type":[{"type":"electronic","value":"2076-0817"}],"subject":[],"published":{"date-parts":[[2024,6,21]]}}}