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Its eradication was achieved in several countries after sustained vaccination with the live attenuated <jats:italic>Brucella abortus<\/jats:italic> S19 vaccine, in combination with the slaughtering of serologically positive animals. S19 induces antibodies against the smooth lipopolysaccharide (S-LPS), making difficult the differentiation of infected from vaccinated bovines. We developed an S19 strain constitutively expressing the green fluorescent protein (S19-GFP) coded in chromosome II. The S19-GFP displays similar biological characteristics and immunogenic and protective efficacies in mice to the parental S19 strain. S19-GFP can be distinguished from S19 and <jats:italic>B<\/jats:italic>. <jats:italic>abortus<\/jats:italic> field strains by fluorescence and multiplex PCR. Twenty-five heifers were vaccinated withS19-GFP (5\u00d710<jats:sup>9<\/jats:sup> CFU) by the subcutaneous or conjunctival routes and some boosted with GFP seven weeks thereafter. Immunized animals were followed up for over three years and tested for anti-S-LPS antibodies by both the Rose Bengal test and a competitive ELISA. Anti-GFP antibodies were detected by an indirect ELISA and Western blotting. In most cases, anti-S-LPS antibodies preceded for several weeks those against GFP. The anti-GFP antibody response was higher in the GFP boosted than in the non-boosted animals. In all cases, the anti-GFP antibodies persisted longer, or at least as long, as those against S-LPS. The drawbacks and potential advantages of using the S19-GFP vaccine for identifying vaccinated animals in infected environments are discussed.<\/jats:p>","DOI":"10.1371\/journal.pone.0260288","type":"journal-article","created":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T18:27:49Z","timestamp":1637605669000},"page":"e0260288","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":17,"title":["Brucella abortus S19 GFP-tagged vaccine allows the serological identification of vaccinated cattle"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0865-0772","authenticated-orcid":true,"given":"Carlos","family":"Chac\u00f3n-D\u00edaz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Zabalza-Barangu\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beatriz","family":"San Rom\u00e1n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9-Mar\u00eda","family":"Blasco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maite","family":"Iriarte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dariana","family":"Salas-Alfaro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela","family":"Hern\u00e1ndez-Mora","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4428-3340","authenticated-orcid":true,"given":"El\u00edas","family":"Barquero-Calvo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1036-920X","authenticated-orcid":true,"given":"Caterina","family":"Guzm\u00e1n-Verri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Esteban","family":"Chaves-Olarte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1819-0186","authenticated-orcid":true,"given":"Mar\u00eda-Jes\u00fas","family":"Grill\u00f3","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edgardo","family":"Moreno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,11,22]]},"reference":[{"key":"pone.0260288.ref001","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1007\/0-387-30745-1_17","volume-title":"The Prokaryotes","author":"E Moreno","year":"2006"},{"issue":"1","key":"pone.0260288.ref002","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1051\/vetres:2003037","article-title":"Rough vaccines in animal brucellosis: structural and genetic basis and present status","volume":"35","author":"I Moriy\u00f3n","year":"2004","journal-title":"Vet Res"},{"key":"pone.0260288.ref003","first-page":"42","volume-title":"Brucellosis Clinical and Laboratory Aspects","author":"P Nicoletti","year":"1989"},{"key":"pone.0260288.ref004","doi-asserted-by":"crossref","first-page":"213","DOI":"10.3389\/fmicb.2014.00213","article-title":"Retrospective and prospective perspectives on zoonotic brucellosis","volume":"5","author":"E Moreno","year":"2014","journal-title":"Front Microbiol"},{"key":"pone.0260288.ref005","first-page":"1","volume-title":"Manual of diagnostic tests and vaccines for terrestrial animals.","author":"Office International des \u00c9pizooties","year":"2018"},{"key":"pone.0260288.ref006","first-page":"291","article-title":"Efficacy and safety of abortion vaccines prepared from Brucella abortus strains of different degrees of virulence","volume":"46","author":"WE Cotton","year":"1933","journal-title":"J Agric Res"},{"key":"pone.0260288.ref007","volume-title":"Bovine Brucellosis. 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