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This study aimed to preliminary evaluate the immunogenic potential of an orally administered formulation comprising heat-inactivated ETEC strains expressing F4 and F18 fimbriae, combined with low-dose recombinant human interferon-alpha (IFN-\u03b1) as a mucosal adjuvant, in piglets. Piglets from two different litters were allocated into two experimental groups: a treated group (T) receiving the inactivated ETEC formulation with IFN-\u03b1 for 26 days, and a control group (C) receiving only IFN-\u03b1. According to the farmer, the sows had not been vaccinated. Immune responses were evaluated in the sows colostrum and piglet serum, saliva, and feces by ELISA. In mesenteric lymph nodes anti-F4\/F18 IgA and IgG antibodies were quantified by ELISPOT.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The inactivated product preserved fimbrial antigenicity and remained sterile. Colostrum from both sows displayed elevated levels of fimbriae-specific IgA and IgG despite neither sow being vaccinated against\n                      <jats:italic>E. coli.<\/jats:italic>\n                      Treated piglets showed a transient serum IgA increase against F4 whereas serum IgG levels were often comparable to the controls. Notably, elevated mucosal IgA responses were observed in saliva and feces against both F4 and F18 (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.01) in T group, accompanied by enhanced IgA-secreting B-cell activity in mesenteric lymph nodes. ELISA validation confirmed high assay reproducibility (R\u00b2 &gt; 0.98; CV\u2009&lt;\u200910%). ELISPOT analysis underscored the adjuvant role of F4 and IFN-\u03b1 in stimulating mucosal immunity.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Our preliminary findings may represent a promising strategy to control PWD and reduce antimicrobial use in pig production.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12917-026-05846-5","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:42:31Z","timestamp":1788309751000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of an inactivated, oral immunogenic product against post-weaning colibacillosis caused by enterotoxigenic Escherichia coli in piglets"],"prefix":"10.1186","volume":"22","author":[{"given":"Alessia","family":"Inglesi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joel","family":"Filipe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1270-6275","authenticated-orcid":false,"given":"Giulia","family":"Valli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Riva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Dall\u2019Ara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gianfilippo Alessio","family":"Clemente","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mario","family":"D\u2019Incau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivonne Laura","family":"Archetti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valentina","family":"Lorenzi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Battioni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara Rota","family":"Nodari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matteo","family":"Tonni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio Marco","family":"Maisano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Flavia","family":"Guarneri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Massimo","family":"Amadori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Fusi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,2]]},"reference":[{"key":"5846_CR1","doi-asserted-by":"publisher","unstructured":"Fairbrother JM, Nadeau \u00c9, Gyles CL. 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Immunopathol., vol. 120, no. 3\u20134, pp. 69\u201379, 2007, https:\/\/doi.org\/10.1016\/j.vetimm.2007.06.018","DOI":"10.1016\/j.vetimm.2007.06.018"}],"container-title":["BMC Veterinary Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12917-026-05846-5","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12917-026-05846-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12917-026-05846-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T02:55:39Z","timestamp":1789008939000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s12917-026-05846-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,2]]},"references-count":53,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["5846"],"URL":"https:\/\/doi.org\/10.1186\/s12917-026-05846-5","relation":{},"ISSN":["1746-6148"],"issn-type":[{"value":"1746-6148","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,2]]},"assertion":[{"value":"2 May 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"18 August 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 September 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"All animal procedures were conducted in full compliance with current animal welfare regulations. The experimental protocol was authorized by the Italian Ministry of Health under authorization No. 1230\/2020-PR (prot. 0028530-31\/12\/2020-DGSAF-MDS-P), in accordance with Article 31 of Legislative Decree 26\/2014. The study protocol was also reviewed and approved by the Animal Welfare Body (OPBA) and the Ethics Committee of the Istituto Zooprofilattico Sperimentale della Lombardia e dell\u2019Emilia-Romagna (Italy).Animals were euthanized using methods consistent with Annex IV of Legislative Decree 26\/2014, implementing European Directive 2010\/63\/EU for swine species.No pharmacological treatments were administered in order to avoid potential alterations of hematological parameters and\/or interactions with the immunological product.","order":1,"name":"Ethics","label":"Ethics approval and consent to participate","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","label":"Consent for publication","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"528"}}