{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:01:52Z","timestamp":1760058112999,"version":"build-2065373602"},"reference-count":54,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,3,11]],"date-time":"2025-03-11T00:00:00Z","timestamp":1741651200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"ANIBOTNET project","award":["98"],"award-info":[{"award-number":["98"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background\/Objectives. Bovine botulism, although relatively rare, presents significant economic losses due to high mortality rates and restrictions on livestock product trade. Vaccination remains the most effective strategy for preventing botulism-related mortality. This study evaluated the efficacy of a bivalent recombinant vaccine targeting the C-terminal portion of the heavy chain (Hc) of botulinum neurotoxin serotype C (BoNT\/C) (Hc BoNT\/C) and botulinum neurotoxin serotype D (BoNT\/D) (Hc BoNT\/D) in inducing neutralizing antibodies against these toxins and their mosaic variants BoNT\/CD and BoNT\/DC in cattle. This comparison aims to improve the design of an optimal recombinant vaccine for preventing bovine botulism caused by the most common serotypes. Methods. Twenty, four-month-old Holstein Friesian calves were randomly assigned to two groups of ten animals: vaccinated group and control group. Sera were collected at various time points to assess antibody titers using ELISA and neutralizing antibody titers using a mouse protection assay. Neutralizing antibody titers were compared to those obtained with a commercially available toxoid vaccine. Results. The recombinant vaccine elicited significant increases in anti-HcBoNT\/C and anti-HcBoNT\/D IgG antibody levels in vaccinated animals compared to controls animals with no adverse effects. Specifically, post-vaccination, the calves showed no local reactions (swelling, warmth) or behavioral changes suggestive of systemic illness. Neutralizing antibody titers against BoNT\/C and BoNT\/D were significantly higher in the recombinant vaccine group compared to the toxoid vaccine group. However, the recombinant vaccine showed lower neutralizing activity against BoNT\/DC compared to the toxoid vaccine. Conclusions. The bivalent recombinant vaccine demonstrated promising immunogenicity in cattle, inducing high neutralizing antibody titers against BoNT\/C and BoNT\/D. While effective against these toxins, the lower efficacy against BoNT\/DC highlights the need for further research to optimize the vaccine formulation, potentially by incorporating a BoNT\/DC Hc component, to provide broader protection against bovine botulism.<\/jats:p>","DOI":"10.3390\/vaccines13030299","type":"journal-article","created":{"date-parts":[[2025,3,11]],"date-time":"2025-03-11T06:52:30Z","timestamp":1741675950000},"page":"299","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Effectiveness of a Bivalent Recombinant Vaccine on the Production of Neutralizing Antibodies Against BoNT\/C, BoNT\/D, BoNT\/CD e BoNT\/DC in Bovines"],"prefix":"10.3390","volume":"13","author":[{"given":"Ilenia","family":"Drigo","sequence":"first","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale delle Venezie, SCT2 Sez. Territoriale di Treviso, 31020 Villorba, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luca","family":"Zandon\u00e0","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale delle Venezie, SCT2 Sez. Territoriale di Treviso, 31020 Villorba, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2595-8488","authenticated-orcid":false,"given":"Elena","family":"Tonon","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale delle Venezie, SCT2 Sez. Territoriale di Treviso, 31020 Villorba, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4351-7934","authenticated-orcid":false,"given":"Katia","family":"Capello","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale delle Venezie, SCS4 Epidemiologia Analitica e Biostatistica, 35020 Legnaro, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5034-3704","authenticated-orcid":false,"given":"Luca","family":"Bano","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale delle Venezie, SCT2 Sez. Territoriale di Treviso, 31020 Villorba, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1365","DOI":"10.1590\/1678-5150-pvb-5643","article-title":"Economic impact of an outbreak of botulism in a cattle feedlot","volume":"38","author":"Soares","year":"2018","journal-title":"Pesqui. Veterin\u00e1ria Bras."},{"key":"ref_2","unstructured":"European Parliament, Council of the European Union (2025, February 26). 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