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Fecal biomarkers represent a promising alternative, as they reflect gut inflammation without invasive sampling. This study evaluated four biomarkers\u2014calprotectin (fCal), lipocalin-2 (LCN-2), myeloperoxidase (MPO), and adenosine deaminase (ADA)\u2014using a zinc oxide (ZnO) model to assess their value as gut-health indicators in piglets. A total of 384 weaned piglets (23\u2009d) were assigned to two diets: Control and ZnO-supplemented (1\u2009g\/kg feed). Piglets were weighed on d 0, 3, 8, and 14. On d 3 and 14, nine average-weight piglets per group were sampled for feces and intestinal tissue. Calprotectin was measured by immunoturbidimetry, MPO byss o-dianisidine oxidation, ADA by enzymatic colorimetry, and LCN-2 by ELISA. Microbiota was analyzed by 16S sequencing and tissues by qPCR and histology. On d 3, ZnO piglets showed fewer intraepithelial lymphocytes (P\u2009=\u20090.062), higher IL-10 expression (P\u2009=\u20090.029), and lower Escherichia\u2013Shigella abundance (P\u2009=\u20090.045). By d 14, they were \u223c300\u2009g heavier (P\u2009=\u20090.071), had higher villus-to-crypt ratios (P\u2009=\u20090.026), maintained IL-10 upregulation (P\u2009=\u20090.083), and increased OCLN expression (P\u2009&amp;lt;\u20090.001). Their microbiota shifted toward Odoribacter and Akkermansia (P\u2009\u2264\u20090.002), with reductions in Clostridium sensu stricto 1 (P\u2009&amp;lt;\u20090.001). fCal and LCN-2 were reduced in the ZnO group on d 3 (P\u2009\u2264\u20090.002), but only fCal remained reduced by d 14 (P\u2009=\u20090.012). No diet effects were observed for MPO or ADA (P\u2009&amp;gt;\u20090.05). On d 3, fCal correlated with CRHR1 (R\u2009=\u20090.59, P\u2009=\u20090.025) and negatively with barrier-related genes MUC13 and OCLN (R \u2264 \u22120.50, P\u2009\u2264\u20090.050). MPO tended to correlate with CRHR1, crypt depth (R\u2009\u2265\u20090.44, P\u2009\u2264\u20090.069), and Streptococcus abundance (R\u2009=\u20090.45, P\u2009=\u20090.073). LCN-2 was negatively associated with inflammatory genes, such as TNF-\u03b1 and NF-\u03baB (R \u2264 \u22120.55, P\u2009\u2264\u20090.041). ADA showed positive trends with potentially pathogenic phyla, such as Campilobacterota (R\u2009\u2265\u20090.43, P\u2009\u2264\u20090.084). By d 14, fCal, LCN-2, and MPO were correlated with immune-related genes (eg TNF-\u03b1, TGF-\u03b2; R\u2009\u2265\u20090.52, P\u2009\u2264\u20090.071), while ADA was associated with the stress-related gene HSD11B1 (R\u2009=\u20090.74, P\u2009=\u20090.002). Overall, fecal biomarkers were valuable indicators of post-weaning gut inflammation and barrier function. Specifically, fCal was the most consistent, LCN-2 and MPO may serve complementary roles, whereas ADA primarily reflected cellular stress rather than inflammation.<\/jats:p>","DOI":"10.1093\/jas\/skag132","type":"journal-article","created":{"date-parts":[[2026,4,17]],"date-time":"2026-04-17T12:51:20Z","timestamp":1776430280000},"source":"Crossref","is-referenced-by-count":0,"title":["Evaluating fecal inflammation biomarkers for non-invasive detection of weaning-related intestinal inflammation in piglets"],"prefix":"10.1093","volume":"104","author":[{"given":"J\u00falia","family":"Suppi","sequence":"first","affiliation":[{"name":"Animal Nutrition and Welfare Service, Department of Animal and Food Sciences, Universitat Aut\u00f2noma de Barcelona , Bellaterra, 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08193,","place":["Spain"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yolanda","family":"Saco","sequence":"additional","affiliation":[{"name":"Veterinary Clinical Biochemistry Service (SBCV), Department of Biochemistry and Molecular Biology, School of Veterinary, Universitat Aut\u00f2noma de Barcelona , Bellaterra, 08193,","place":["Spain"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4213-2274","authenticated-orcid":false,"given":"Anna","family":"Bassols","sequence":"additional","affiliation":[{"name":"Veterinary Clinical Biochemistry Service (SBCV), Department of Biochemistry and Molecular Biology, School of Veterinary, Universitat Aut\u00f2noma de Barcelona , Bellaterra, 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