{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T01:08:45Z","timestamp":1770772125399,"version":"3.50.0"},"reference-count":41,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,12,30]],"date-time":"2024-12-30T00:00:00Z","timestamp":1735516800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"EUGLOHRIA","award":["2023"],"award-info":[{"award-number":["2023"]}]},{"name":"EUGLOHRIA","award":["44104"],"award-info":[{"award-number":["44104"]}]},{"name":"EUGLOHRIA","award":["2021.09461.BD"],"award-info":[{"award-number":["2021.09461.BD"]}]},{"name":"Fund of the Hedda and John Forssman Foundation","award":["2023"],"award-info":[{"award-number":["2023"]}]},{"name":"Fund of the Hedda and John Forssman Foundation","award":["44104"],"award-info":[{"award-number":["44104"]}]},{"name":"Fund of the Hedda and John Forssman Foundation","award":["2021.09461.BD"],"award-info":[{"award-number":["2021.09461.BD"]}]},{"name":"FCT","award":["2023"],"award-info":[{"award-number":["2023"]}]},{"name":"FCT","award":["44104"],"award-info":[{"award-number":["44104"]}]},{"name":"FCT","award":["2021.09461.BD"],"award-info":[{"award-number":["2021.09461.BD"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>SARS-CoV-2 can cause clinical and inapparent disease and mortality in several animals cohabitating with humans, and sheep are susceptible to SARS-CoV-2 due to virus\u2013receptor interactions similar to those in humans. Hence, sheep have the potential to be infected, spread, and develop neutralising antibodies (NAbs) against SARS-CoV-2. The aim of this study was to investigate the prevalence of SARS-CoV-2 NAbs in farm animals after natural exposure to the virus. Serum samples were collected from sheep in the Serra da Estrela region in Portugal, both prior to and during the COVID-19 pandemic. The sera were tested by established SARS-CoV-2 pseudovirus systems for multiple SARS-CoV-2 variants (early\u2014Wuhan, mid\u2014Delta, Omicron\u2014BA.1, and late\u2014Omicron XBB, BQ.1.1). Partial neutralisation activity in Pre-pandemic and Mid-pandemic samples was observed, while no NAb activity was observed in Late-pandemic samples tested. Different levels of NAbs were observed between Pre-pandemic samples and those collected during the Mid-pandemic and Late-pandemic periods (p \u2264 0.01). Our results indicate that SARS-CoV-2 cross-species transmission may have occurred through human\u2013sheep contacts on sheep farms during the pandemic, and that farm animals could contribute to the One Health Approach in zoonotic virus surveillance and pandemic preparedness.<\/jats:p>","DOI":"10.3390\/microorganisms13010049","type":"journal-article","created":{"date-parts":[[2024,12,31]],"date-time":"2024-12-31T14:21:12Z","timestamp":1735654872000},"page":"49","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Investigating SARS-CoV-2 Neutralising Antibody Response in Sheep"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5685-4027","authenticated-orcid":false,"given":"Milena","family":"Samojlovi\u0107","sequence":"first","affiliation":[{"name":"Systems Virology, Faculty of Medicine, Lund University, 223 62 Lund, Sweden"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8769-8103","authenticated-orcid":false,"given":"Jo\u00e3o R.","family":"Mesquita","sequence":"additional","affiliation":[{"name":"Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar, University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9742-5643","authenticated-orcid":false,"given":"S\u00e9rgio","family":"Santos-Silva","sequence":"additional","affiliation":[{"name":"Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar, University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6930-1720","authenticated-orcid":false,"given":"Malin","family":"Neptin","sequence":"additional","affiliation":[{"name":"Systems Virology, Faculty of Medicine, Lund University, 223 62 Lund, Sweden"}]},{"given":"Joakim","family":"Esbj\u00f6rnsson","sequence":"additional","affiliation":[{"name":"Systems Virology, Faculty of Medicine, Lund University, 223 62 Lund, Sweden"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"549","DOI":"10.3906\/sag-2004-127","article-title":"Coronaviruses and SARS-CoV-2","volume":"50","year":"2020","journal-title":"Turk. J. Med. Sci."},{"key":"ref_2","unstructured":"World Health Organization (WHO) (2024, May 17). WHO-Convened Global Study of Origins of SARS-CoV-2. Infectious Diseases & Immunity. Available online: https:\/\/www.who.int\/publications\/i\/item\/who-convened-global-study-of-origins-of-sars-cov-2-china-part."},{"key":"ref_3","first-page":"162","article-title":"SARS-CoV-2 variant biology: Immune escape, transmission and fitness","volume":"21","author":"Carabelli","year":"2023","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1177\/03009858231225500","article-title":"Mortality associated with SARS-CoV-2 in nondomestic felids","volume":"61","author":"Drozd","year":"2024","journal-title":"Vet. Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Giraldo-Ramirez, S., Rendon-Marin, S., Jaimes, J.A., Martinez-Gutierrez, M., and Ruiz-Saenz, J. (2021). SARS-CoV-2 Clinical Outcome in Domestic and Wild Cats: A Systematic Review. Animals, 11.","DOI":"10.3390\/ani11072056"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"100653","DOI":"10.1016\/j.onehlt.2023.100653","article-title":"Data on SARS-CoV-2 events in animals: Mind the gap!","volume":"17","author":"Nerpel","year":"2023","journal-title":"One Health"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e28147","DOI":"10.1002\/jmv.28147","article-title":"SARS-CoV-2 and natural infection in animals","volume":"95","author":"Qiu","year":"2023","journal-title":"J. Med. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1289\/ehp.9399351","article-title":"The use of mammals as sentinels for human exposure to toxic contaminants in the environment","volume":"99","author":"Kaneene","year":"1993","journal-title":"Environ. Health Perspect."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"10-1128","DOI":"10.1128\/JVI.00083-21","article-title":"Susceptibility of White-Tailed Deer (Odocoileus virginianus) to SARS-CoV-2","volume":"95","author":"Palmer","year":"2021","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Martins, M., Boggiatto, P.M., Buckley, A., Cassmann, E.D., Falkenberg, S., Caserta, L.C., Fernandes, M.H.V., Kanipe, C., Lager, K., and Palmer, M.V. (2022). From Deer-to-Deer: SARS-CoV-2 is efficiently transmitted and presents broad tissue tropism and replication sites in white-tailed deer. PLoS Pathog., 18.","DOI":"10.1371\/journal.ppat.1010197"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2011","DOI":"10.1038\/s41564-022-01268-9","article-title":"Divergent SARS-CoV-2 variant emerges in white-tailed deer with deer-to-human transmission","volume":"7","author":"Pickering","year":"2022","journal-title":"Nat. Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Di Teodoro, G., Valleriani, F., Puglia, I., Monaco, F., Di Pancrazio, C., Luciani, M., Krasteva, I., Petrini, A., Marcacci, M., and D\u2019Alterio, N. (2021). SARS-CoV-2 replicates in respiratory ex vivo organ cultures of domestic ruminant species. Vet. Microbiol., 252.","DOI":"10.1016\/j.vetmic.2020.108933"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1080\/22221751.2022.2037397","article-title":"Susceptibility of sheep to experimental co-infection with the ancestral lineage of SARS-CoV-2 and its alpha variant","volume":"11","author":"Gaudreault","year":"2022","journal-title":"Emerg. Microbes Infect."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1741","DOI":"10.1007\/s11259-022-10044-3","article-title":"First serological evidence of SARS-CoV-2 natural infection in small ruminants","volume":"47","author":"Fusco","year":"2023","journal-title":"Vet. Res. Commun."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e01283-20","DOI":"10.1128\/JVI.01283-20","article-title":"SARS-CoV-2 and Three Related Coronaviruses Utilize Multiple ACE2 Orthologs and Are Potently Blocked by an Improved ACE2-Ig","volume":"94","author":"Li","year":"2020","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Villanueva-Saz, S., Giner, J., Fern\u00e1ndez, A., Lacasta, D., Ort\u00edn, A., Ramos, J.J., Ferrer, L.M., Ruiz de Arcaute, M., Tobajas, A.P., and P\u00e9rez, M.D. (2021). Absence of SARS-CoV-2 Antibodies in Natural Environment Exposure in Sheep in Close Contact with Humans. Animals, 11.","DOI":"10.3390\/ani11071984"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Gaspar, D., Usi\u00e9, A., Le\u00e3o, C., Guimar\u00e3es, S., Pires, A.E., Matos, C., Ramos, A.M., and Ginja, C. (2023). Genome-wide assessment of the population structure and genetic diversity of four Portuguese native sheep breeds. Front. Genet., 14.","DOI":"10.3389\/fgene.2023.1109490"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1515\/opag-2021-0024","article-title":"Characterization of dairy sheep farms in the Serra da Estrela PDO region","volume":"6","author":"Monteiro","year":"2021","journal-title":"Open Agric."},{"key":"ref_19","unstructured":"(2024, August 29). Di\u00e1rio da Republica, Decreto-Lei n.\u00ba 39 209, of 14 May 1953 and Decreto-Lei n.\u00ba 244\/2000, of 27 September 2000. Available online: https:\/\/diariodarepublica.pt\/dr\/home."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1261\/rna.2192803","article-title":"Lentivirus-delivered stable gene silencing by RNAi in primary cells","volume":"9","author":"Stewart","year":"2003","journal-title":"RNA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"956","DOI":"10.1126\/science.abc7520","article-title":"Isolation of potent SARS-CoV-2 neutralizing antibodies and protection from disease in a small animal model","volume":"69","author":"Rogers","year":"2020","journal-title":"Science"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"eabj5413","DOI":"10.1126\/scitranslmed.abj5413","article-title":"Bispecific antibodies targeting distinct regions of the spike protein potently neutralize SARS-CoV-2 variants of concern","volume":"13","author":"Cho","year":"2021","journal-title":"Sci. Transl. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1126\/science.abq3773","article-title":"Broadly neutralizing antibodies target the coronavirus fusion peptide","volume":"377","author":"Dacon","year":"2022","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1016\/j.immuni.2023.02.005","article-title":"Broadly neutralizing anti-S2 antibodies protect against all three human betacoronaviruses that cause deadly disease","volume":"56","author":"Zhou","year":"2023","journal-title":"Immunity"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1016\/S1473-3099(22)00129-3","article-title":"Neutralisation sensitivity of the SARS-CoV-2 omicron (B.1.1.529) variant: A cross-sectional study","volume":"22","author":"Sheward","year":"2022","journal-title":"Lancet Infect. Dis."},{"key":"ref_26","unstructured":"(2023, December 28). Portuguese National Institute of Health Official Webpage. Available online: https:\/\/insaflu.insa.pt\/covid19\/."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1080\/22221751.2020.1743767","article-title":"Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2","volume":"9","author":"Nie","year":"2020","journal-title":"Emerg. Microbes Infect."},{"key":"ref_28","unstructured":"World Organisation for Animal Health (WOAH) (2024, August 27). Considerations on Monitoring SARS-CoV-2 in Animals. Available online: https:\/\/www.woah.org\/app\/uploads\/2022\/08\/en-sars-cov-2-surveillance.pdf."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Greaney, A.J., Eguia, R.T., Starr, T.N., Khan, K., Franko, N., Logue, J.K., Lord, S.M., Speake, C., Chu, H.Y., and Sigal, A. (2022). The SARS-CoV-2 Delta variant induces an antibody response largely focused on class 1 and 2 antibody epitopes. PLoS Pathog., 18.","DOI":"10.1101\/2022.03.12.484088"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1038\/s41591-022-01705-6","article-title":"Neutralizing antibodies against the SARS-CoV-2 Delta and Omicron variants following heterologous CoronaVac plus BNT162b2 booster vaccination","volume":"28","author":"Lucas","year":"2022","journal-title":"Nat. Med."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Govindaraj, S., Cheedarla, N., Cheedarla, S., Irby, L.S.S., Neish, A.S., Roback, J.D., Smith, A.K., and Velu, V. (2023). COVID-19 vaccine induced poor neutralization titers for SARS-CoV-2 omicron variants in maternal and cord blood. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1211558"},{"key":"ref_32","first-page":"337","article-title":"SARS coronavirus 2-reactive antibodies in bovine colostrum","volume":"64","author":"Ellis","year":"2023","journal-title":"Can. Vet. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1038\/s41586-021-04385-3","article-title":"Omicron escapes the majority of existing SARS-CoV-2 neutralizing antibodies","volume":"602","author":"Cao","year":"2022","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"eabi8452","DOI":"10.1126\/scitranslmed.abi8452","article-title":"A high-throughput cell- and virus-free assay shows reduced neutralization of SARS-CoV-2 variants by COVID-19 convalescent plasma","volume":"13","author":"Fenwick","year":"2021","journal-title":"Sci. Transl. Med."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Hulst, M., Kant, A., Harders-Westerveen, J., Hoffmann, M., Xie, Y., Laheij, C., Murk, J.L., and Van der Poel, W.H.M. (2023). Cross-Reactivity of Human, Wild Boar, and Farm Animal Sera from Pre- and Post-Pandemic Periods with Alpha- and \u0392eta-Coronaviruses (CoV), Including SARS-CoV-2. Viruses, 16.","DOI":"10.3390\/v16010034"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Wang, R., Zhang, Q., Zhang, R., Aw, Z.Q., Chen, P., Wong, Y.H., Hong, J., Ju, B., Shi, X., and Ding, Q. (2022). SARS-CoV-2 Omicron Variants Reduce Antibody Neutralization and Acquire Usage of Mouse ACE2. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.854952"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Mazet, J.A., Clifford, D.L., Coppolillo, P.B., Deolalikar, A.B., Erickson, J.D., and Kazwala, R.R. (2009). A \u201cOne Health\u201d approach to address emerging zoonoses: The HALI project in Tanzania. PLoS Med., 6.","DOI":"10.1371\/journal.pmed.1000190"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s42522-019-0007-9","article-title":"Implementing One Health approaches to confront emerging and re-emerging zoonotic disease threats: Lessons from PREDICT","volume":"2","author":"Kelly","year":"2020","journal-title":"One Health Outlook"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1186\/s42522-023-00082-5","article-title":"Strengthening coordination and collaboration of one health approach for zoonotic diseases in Africa","volume":"5","author":"Alimi","year":"2023","journal-title":"One Health Outlook"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"100487","DOI":"10.1016\/j.onehlt.2023.100487","article-title":"A new One Health Framework in Qatar for future emerging and re-emerging zoonotic diseases preparedness and response","volume":"16","author":"Bansal","year":"2023","journal-title":"One Health"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Horefti, E. (2023). The Importance of the One Health Concept in Combating Zoonoses. Pathogens, 12.","DOI":"10.3390\/pathogens12080977"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/1\/49\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:57:08Z","timestamp":1760115428000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/1\/49"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,30]]},"references-count":41,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["microorganisms13010049"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms13010049","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,30]]}}}