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Zearalenone (ZEN) is a frequently detected mycotoxin with well-known estrogenic activity. This study aimed to evaluate the effects of ingesting ZEN-contaminated feed on the methylation and hydroxymethylation of CpG islands located in the candidate genes\n                    <jats:italic>CYP1A1<\/jats:italic>\n                    ,\n                    <jats:italic>CYP21A2<\/jats:italic>\n                    , and\n                    <jats:italic>CYP1B1<\/jats:italic>\n                    in Nellore heifers. Twenty heifers were confined for 12 weeks during the finishing phase and assigned to two groups: a control group (non-contaminated feed) and a ZEN-treated group (approximately 300 ppb). After slaughter, liver samples were collected, and genomic DNA was extracted. DNA was treated with T4-\u03b2-glucosyltransferase and digested with the restriction enzymes\n                    <jats:italic>MspI<\/jats:italic>\n                    and\n                    <jats:italic>HpaII<\/jats:italic>\n                    . CpG island methylation and hydroxymethylation were quantified by real-time PCR. Statistical analyses were performed using analysis of variance followed by\n                    <jats:italic>t<\/jats:italic>\n                    -tests, and\n                    <jats:italic>p<\/jats:italic>\n                    -values were adjusted for multiple comparisons using the false discovery rate (FDR). No significant differences were observed in methylation or hydroxymethylation levels of\n                    <jats:italic>CYP1A1<\/jats:italic>\n                    between groups after FDR adjustment. In contrast, animals exposed to ZEN-contaminated feed exhibited increased DNA methylation in the CpG island of\n                    <jats:italic>CYP1B1<\/jats:italic>\n                    , and this effect remained statistically significant after FDR correction. For\n                    <jats:italic>CYP21A2<\/jats:italic>\n                    , differences in hydroxymethylation observed in the unadjusted analysis were attenuated after FDR adjustment and are therefore interpreted as a trend. These findings indicate that ZEN exposure induces gene-specific epigenetic modulation in the liver of Nellore heifers, with a robust effect observed for\n                    <jats:italic>CYP1B1<\/jats:italic>\n                    . The results highlight the importance of monitoring ZEN contamination in cattle feed and support the need for further studies integrating epigenetic, transcriptomic, and endocrine analyses to clarify the biological implications of mycotoxin exposure in beef cattle.\n                  <\/jats:p>","DOI":"10.1007\/s11250-026-04978-y","type":"journal-article","created":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T12:02:14Z","timestamp":1775908934000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The role of Zearalenone in epigenetic modifications of candidate genes in Nellore heifers"],"prefix":"10.1007","volume":"58","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-8219-5523","authenticated-orcid":false,"given":"Lethissia Amorim da Silva","family":"Coelho","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1577-395X","authenticated-orcid":false,"given":"Rondineli Pavezzi","family":"Barbero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro Ruiz Martins Tapajos","family":"Pereira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina Emiliano Bastos","family":"Polido","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Clara Souza Resende","family":"de Aguiar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8218-3626","authenticated-orcid":false,"given":"Huarrisson Azevedo","family":"Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9790-4764","authenticated-orcid":false,"given":"Marco Roberto Bourg","family":"de Mello","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5323-2353","authenticated-orcid":false,"given":"Elisandra Lurdes","family":"Kern","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0049-0201","authenticated-orcid":false,"given":"Denise Carleto","family":"Andia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7033-5995","authenticated-orcid":false,"given":"Marina Mortati Dias","family":"Barbero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,4,11]]},"reference":[{"key":"4978_CR1","doi-asserted-by":"publisher","unstructured":"Assis RIF, Wiench M, Silv\u00e9rio KG, da Silva RA, da Silva Feltran G, Sallum EA, Casati MZ, Nociti FH, Andia DC (2018) RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation. 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