{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T03:06:28Z","timestamp":1790651188681,"version":"4.1.0"},"reference-count":48,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T00:00:00Z","timestamp":1765843200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research & Development Program of China","award":["2022YFC2304100"],"award-info":[{"award-number":["2022YFC2304100"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>The ongoing global monkeypox outbreak since 2022 has highlighted the urgent need for vaccine development. Current vaccination strategies rely on cross-protective immunity provided by orthopoxvirus-based live-attenuated vaccines. However, these vaccines not only exhibit suboptimal efficacy against monkeypox virus (MPXV) but also raise safety concerns, particularly given the significant global overlap between MPXV infections and HIV. Owing to their superior safety profile and accessibility, recombinant subunit vaccines represent a highly promising platform for monkeypox vaccine development. In this study, we developed a subunit vaccine comprising A29L, B6R, and M1R antigens formulated with a proprietary nanoemulsion adjuvant and evaluated its immunogenicity and protective efficacy. In mice immunized with a prime-boost regimen of the three individual antigens combined with the nanoemulsion adjuvant, comparable serum IgG levels against each antigen were elicited. Both A29 and M1 formulations induced serum antibodies with potent neutralizing activity against MPXV and Vaccinia virus Western Reserve strain (VACV-WR). Notably, M1 antiserum exhibited stronger neutralization than A29 antiserum, whereas B6R immune serum showed no significant neutralizing activity. Splenocytes from B6R-immunized mice mounted a robust IFN-\u03b3 response, which was markedly lower in those immunized with A29 or M1. All three monovalent vaccines conferred complete survival following an intranasal lethal MPXV challenge, with M1 providing the strongest protection. In a lethal VACV-WR challenge model, only M1 immunization conferred significant protection. Histopathological analysis of lung tissues on day 5 post-infection revealed more pronounced inflammatory features in B6R-immunized mice compared to the nanoemulsion adjuvant control group. Furthermore, the nanoemulsion-adjuvanted bivalent A29L + B6R formulation induced significantly higher IgG and neutralizing antibody titers and demonstrated superior protective efficacy compared to the aluminum hydroxide-adjuvanted formulation. This comparative preclinical evaluation provides important evidence to support the development of a safe and effective subunit vaccine against monkeypox.<\/jats:p>","DOI":"10.3390\/pathogens14121293","type":"journal-article","created":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T12:47:26Z","timestamp":1765889246000},"page":"1293","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Immune Responses and Protective Efficacy of Nanoemulsion-Adjuvanted Monkeypox Virus Recombinant Vaccines Against Lethal Challenge in Mice"],"prefix":"10.3390","volume":"14","author":[{"given":"Congcong","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nuo","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanqi","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhendong","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dawei","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanda","family":"Tang","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanhua","family":"He","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-9859-1909","authenticated-orcid":false,"given":"Xu","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongtian","family":"Qi","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinxin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China"},{"name":"Key Laboratory of Biosafety Defense (Naval Medical University), Ministry of Education of the People\u2019s Republic of China, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1038\/s41577-022-00775-4","article-title":"Monkeypox: Disease Epidemiology, Host Immunity and Clinical Interventions","volume":"22","author":"Lum","year":"2022","journal-title":"Nat. Rev. Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1822","DOI":"10.1016\/S0140-6736(23)00588-3","article-title":"Mpox Neglect and the Smallpox Niche: A Problem for Africa, a Problem for the World","volume":"401","author":"Adetifa","year":"2023","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1056\/NEJMoa2207323","article-title":"Monkeypox Virus Infection in Humans across 16 Countries\u2014April\u2013June 2022","volume":"387","author":"Thornhill","year":"2022","journal-title":"N. Engl. J. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1016\/S1473-3099(22)00354-1","article-title":"Monkeypox Outbreaks Outside Endemic Regions: Scientific and Social Priorities","volume":"22","author":"Zumla","year":"2022","journal-title":"Lancet Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Sukhdeo, S., Mishra, S., and Walmsley, S. (2022). Human Monkeypox: A Comparison of the Characteristics of the New Epidemic to the Endemic Disease. BMC Infect. Dis., 22.","DOI":"10.1186\/s12879-022-07900-7"},{"key":"ref_6","unstructured":"(2025, November 15). Epidemiological Update\u2014Week 35\/2024: Mpox Due to Monkeypox Virus Clade I. Available online: https:\/\/www.ecdc.europa.eu\/en\/news-events\/mpox-epidemiological-update-monkeypox-2-september-2024."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"106794","DOI":"10.1016\/j.ijsu.2022.106794","article-title":"The Global Human Monkeypox Outbreak in 2022: An Overview","volume":"104","author":"Ahmed","year":"2022","journal-title":"Int. J. Surg."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"734","DOI":"10.15585\/mmwr.mm7122e1","article-title":"Use of JYNNEOS (Smallpox and Monkeypox Vaccine, Live, Nonreplicating) for Preexposure Vaccination of Persons at Risk for Occupational Exposure to Orthopoxviruses: Recommendations of the Advisory Committee on Immunization Practices\u2014United States, 2022","volume":"71","author":"Rao","year":"2022","journal-title":"MMWR Morb. Mortal. Wkly. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.tips.2022.10.005","article-title":"Monkeypox: Potential Vaccine Development Strategies","volume":"44","author":"Lozano","year":"2023","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1007\/s44197-025-00432-8","article-title":"Safety and Efficacy of Repurposed Smallpox Vaccines Against Mpox: A Critical Review of ACAM2000, JYNNEOS, and LC16","volume":"15","author":"Wahid","year":"2025","journal-title":"J. Epidemiol. Glob. Health"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.1001\/jamaneurol.2022.3491","article-title":"Neurologic Complications of Smallpox and Monkeypox: A Review","volume":"79","author":"Billioux","year":"2022","journal-title":"JAMA Neurol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"106926","DOI":"10.1016\/j.ijsu.2022.106926","article-title":"Monkeypox Vaccination: Does It Cause Neurologic and Psychiatric Manifestations?\u2014Correspondence","volume":"106","author":"Farahat","year":"2022","journal-title":"Int. J. Surg."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1093\/infdis\/jis753","article-title":"Safety, Immunogenicity, and Surrogate Markers of Clinical Efficacy for Modified Vaccinia Ankara as a Smallpox Vaccine in HIV-Infected Subjects","volume":"207","author":"Greenberg","year":"2013","journal-title":"J. Infect. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.bsheal.2024.04.005","article-title":"Human Mpox Co-Infection with Advanced HIV-1 and XDR-TB in a MSM Patient Previously Vaccinated against Smallpox: A Case Report","volume":"6","author":"Fang","year":"2024","journal-title":"Biosaf. Health"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e349","DOI":"10.1016\/S1473-3099(22)00574-6","article-title":"Prevention of Monkeypox with Vaccines: A Rapid Review","volume":"22","author":"Poland","year":"2022","journal-title":"Lancet Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1111\/hiv.13661","article-title":"Mpox in People with HIV: A Narrative Review","volume":"25","author":"Nakamura","year":"2024","journal-title":"HIV Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1038\/s41422-023-00792-5","article-title":"Polyvalent mRNA Vaccination Elicited Potent Immune Response to Monkeypox Virus Surface Antigens","volume":"33","author":"Fang","year":"2023","journal-title":"Cell Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"105392","DOI":"10.1016\/j.ebiom.2024.105392","article-title":"Single-Chain A35R-M1R-B6R Trivalent mRNA Vaccines Protect Mice against Both Mpox Virus and Vaccinia Virus","volume":"109","author":"Kong","year":"2024","journal-title":"eBioMedicine"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Garcia-Atutxa, I., Mondragon-Teran, P., Huerta-Saquero, A., and Villanueva-Flores, F. (2024). Advancements in Monkeypox Vaccines Development: A Critical Review of Emerging Technologies. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1456060"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4726","DOI":"10.1038\/s41467-025-59826-8","article-title":"Mpox Multiprotein Virus-like Nanoparticle Vaccine Induces Neutralizing and Protective Antibodies in Mice and Non-Human Primates","volume":"16","author":"Belghith","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"106156","DOI":"10.1016\/j.micpath.2023.106156","article-title":"Progress and Prospects on Vaccine Development against Monkeypox Infection","volume":"180","author":"Saadh","year":"2023","journal-title":"Microb. Pathog."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1597","DOI":"10.1038\/nm.3409","article-title":"Key Roles of Adjuvants in Modern Vaccines","volume":"19","author":"Reed","year":"2013","journal-title":"Nat. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3321","DOI":"10.1016\/j.apsb.2023.01.006","article-title":"Advanced Subunit Vaccine Delivery Technologies: From Vaccine Cascade Obstacles to Design Strategies","volume":"13","author":"Hou","year":"2023","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.addr.2021.01.001","article-title":"Recombinant Protein Vaccines, a Proven Approach against Coronavirus Pandemics","volume":"170","author":"Pollet","year":"2021","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1038\/nri2510","article-title":"Towards an Understanding of the Adjuvant Action of Aluminium","volume":"9","author":"Marrack","year":"2009","journal-title":"Nat. Rev. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Awate, S., Babiuk, L.A., and Mutwiri, G. (2013). Mechanisms of Action of Adjuvants. Front. Immunol., 4.","DOI":"10.3389\/fimmu.2013.00114"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Cao, H., Yang, S., Wang, Y., Luan, N., Yin, X., Lin, K., and Liu, C. (2021). An Established Th2-Oriented Response to an Alum-Adjuvanted SARS-CoV-2 Subunit Vaccine Is Not Reversible by Sequential Immunization with Nucleic Acid-Adjuvanted Th1-Oriented Subunit Vaccines. Vaccines, 9.","DOI":"10.3390\/vaccines9111261"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"102229","DOI":"10.1016\/j.coi.2022.102229","article-title":"Resolving Adjuvant Mode of Action to Enhance Vaccine Efficacy","volume":"77","author":"Turley","year":"2022","journal-title":"Curr. Opin. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/j.coi.2009.05.003","article-title":"Immunology of TLR-Independent Vaccine Adjuvants","volume":"21","author":"Wack","year":"2009","journal-title":"Curr. Opin. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1038\/s41392-023-01557-7","article-title":"Vaccine Adjuvants: Mechanisms and Platforms","volume":"8","author":"Zhao","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"454","DOI":"10.1038\/s41573-021-00163-y","article-title":"Emerging Concepts in the Science of Vaccine Adjuvants","volume":"20","author":"Pulendran","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"9563","DOI":"10.1016\/j.vaccine.2011.08.089","article-title":"Immunomodulatory and Physical Effects of Oil Composition in Vaccine Adjuvant Emulsions","volume":"29","author":"Fox","year":"2011","journal-title":"Vaccine"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1038\/s41423-025-01306-6","article-title":"Multiscale Dynamic Immunomodulation by a Nanoemulsified Trojan-TLR7\/8 Adjuvant for Robust Protection against Heterologous Pandemic and Endemic Viruses","volume":"22","author":"Yoo","year":"2025","journal-title":"Cell. Mol. Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2461","DOI":"10.1016\/j.vaccine.2011.01.011","article-title":"Adjuvant System AS03 Containing \u03b1-Tocopherol Modulates Innate Immune Response and Leads to Improved Adaptive Immunity","volume":"29","author":"Morel","year":"2011","journal-title":"Vaccine"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"11608","DOI":"10.1021\/acsomega.3c06552","article-title":"Immune Potentiation of PLGA Controlled-Release Vaccines for Improved Immunological Outcomes","volume":"9","author":"Cassaidy","year":"2024","journal-title":"ACS Omega"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Yoshioka, Y., Kobiyama, K., Hayashi, T., Onishi, M., Yanagida, Y., Nakagawa, T., Hashimoto, M., Nishinaka, A., Hirose, J., and Asaoka, Y. (2023). A-910823, a Squalene-Based Emulsion Adjuvant, Induces T Follicular Helper Cells and Humoral Immune Responses via \u03b1-Tocopherol Component. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1116238"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2402059","DOI":"10.1002\/smtd.202402059","article-title":"Opportunities and Challenges for Nanomaterials as Vaccine Adjuvants","volume":"9","author":"Li","year":"2025","journal-title":"Small Methods"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/s41421-021-00370-2","article-title":"CoVac501, a Self-Adjuvanting Peptide Vaccine Conjugated with TLR7 Agonists, against SARS-CoV-2 Induces Protective Immunity","volume":"8","author":"Long","year":"2022","journal-title":"Cell Discov."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2482702","DOI":"10.1080\/22221751.2025.2482702","article-title":"Rational Mpox Vaccine Design: Immunogenicity and Protective Effect of Individual and Multicomponent Proteins in Mice","volume":"14","author":"Cheng","year":"2025","journal-title":"Emerg. Microbes Infect."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3100","DOI":"10.1038\/s41467-025-58180-z","article-title":"Generation and Characterization of Neutralizing Antibodies against M1R and B6R Proteins of Monkeypox Virus","volume":"16","author":"Qu","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_41","unstructured":"(2025, November 15). Interim Recommendations for the Use of Protein Subunit COVID-19 Vaccines. Available online: https:\/\/www.who.int\/publications\/i\/item\/WHO-2019-nCoV-vaccines-SAGE_recommendation-protein_subunit-2023.1."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1186\/s12934-022-01929-8","article-title":"Recombinant Vaccines in 2022: A Perspective from the Cell Factory","volume":"21","author":"Corchero","year":"2022","journal-title":"Microb. Cell. Factories"},{"key":"ref_43","unstructured":"(2025, November 15). CDC Storage and Handling of Immunobiologics, Available online: https:\/\/www.cdc.gov\/vaccines\/hcp\/imz-best-practices\/storage-handling-immunobiologics.html."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Protopapas, K., Dimopoulou, D., Kalesis, N., Akinosoglou, K., and Moschopoulos, C.D. (2024). Mpox and Lessons Learned in the Light of the Recent Outbreak: A Narrative Review. Viruses, 16.","DOI":"10.3390\/v16101620"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2204151","DOI":"10.1080\/22221751.2023.2204151","article-title":"Mpox Multi-Antigen mRNA Vaccine Candidates by a Simplified Manufacturing Strategy Afford Efficient Protection against Lethal Orthopoxvirus Challenge","volume":"12","author":"Zeng","year":"2023","journal-title":"Emerg. Microbes Infect."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"126384","DOI":"10.1016\/j.vaccine.2024.126384","article-title":"Systematic Evaluation of the Induction of Efficient Neutralizing Antibodies by Recombinant Multicomponent Subunit Vaccines against Monkeypox Virus","volume":"42","author":"Tan","year":"2024","journal-title":"Vaccine"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.virol.2005.08.008","article-title":"Identification and Preliminary Characterization of Vaccinia Virus (Dryvax) Antigens Recognized by Vaccinia Immune Globulin","volume":"343","author":"Garcia","year":"2005","journal-title":"Virology"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Li, J., Li, X., Dong, J., Wei, J., Guo, X., Wang, G., Xu, M., and Zhao, A. (2024). Enhanced Immune Responses in Mice by Combining the Mpox Virus B6R-Protein and Aluminum Hydroxide-CpG Vaccine Adjuvants. Vaccines, 12.","DOI":"10.3390\/vaccines12070776"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/12\/1293\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T13:01:13Z","timestamp":1765890073000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/12\/1293"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,16]]},"references-count":48,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["pathogens14121293"],"URL":"https:\/\/doi.org\/10.3390\/pathogens14121293","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,16]]}}}