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This study aimed to evaluate the effects of SC on splenic immunomodulation in broiler chickens by characterizing and quantifying immune cell populations\u2014including T lymphocytes (CD3\n                    <jats:sup>+<\/jats:sup>\n                    , CD4\n                    <jats:sup>+<\/jats:sup>\n                    , CD8\n                    <jats:sup>+<\/jats:sup>\n                    , and Treg CD25\n                    <jats:sup>+<\/jats:sup>\n                    ), macrophages (CD163\n                    <jats:sup>+<\/jats:sup>\n                    ), and B lymphocytes (CD20\n                    <jats:sup>+<\/jats:sup>\n                    )\u2014per mm\n                    <jats:sup>2<\/jats:sup>\n                    of spleen tissue using immunohistochemistry (IHC). Two treatment groups were compared: a control group (CG;\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u20098) without SC and an experimental group (PG;\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u20098) supplemented with SC at 10\n                    <jats:sup>7<\/jats:sup>\n                    \u00a0CFU\u00b7g\n                    <jats:sup>\u22121<\/jats:sup>\n                    . Splenic tissue sections were digitized at 20\u2009\u00d7\u2009magnification using a MoticEasyScan Infinity 60 slide scanner. Immune cell density and positivity percentages (CD3\n                    <jats:sup>+<\/jats:sup>\n                    , CD4\n                    <jats:sup>+<\/jats:sup>\n                    , CD8\n                    <jats:sup>+<\/jats:sup>\n                    , CD20\n                    <jats:sup>+<\/jats:sup>\n                    , CD25\n                    <jats:sup>+<\/jats:sup>\n                    , and CD163\n                    <jats:sup>+<\/jats:sup>\n                    ) were quantified using QuPath digital pathology software. SC supplementation significantly reduced cell density and positivity percentage of CD3\n                    <jats:sup>+<\/jats:sup>\n                    and CD4\n                    <jats:sup>+<\/jats:sup>\n                    T lymphocytes vs. CG and positivity percentage of CD25 cells in PG vs. CG. Supplementation with SC in broiler chickens significantly altered splenic immune cell morphometry, particularly in regions containing CD3\n                    <jats:sup>+<\/jats:sup>\n                    , CD4\n                    <jats:sup>+<\/jats:sup>\n                    , and positivity in CD25\n                    <jats:sup>+<\/jats:sup>\n                    , while no effects were observed on cell density or positivity in CD8\n                    <jats:sup>+<\/jats:sup>\n                    , CD20\n                    <jats:sup>+<\/jats:sup>\n                    , and CD163\n                    <jats:sup>+<\/jats:sup>\n                    macrophages. Additionally, digital pathology proved effective in enabling precise morphometric quantification of immunohistochemical expression in digitized whole slides.\n                  <\/jats:p>","DOI":"10.1007\/s12602-025-10736-7","type":"journal-article","created":{"date-parts":[[2025,9,27]],"date-time":"2025-09-27T09:41:08Z","timestamp":1758966068000},"page":"4467-4485","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Digital Pathology-Based Morphometric Analysis of Saccharomyces cerevisiae Effects on Splenic Cell Populations in Broiler Chickens"],"prefix":"10.1007","volume":"18","author":[{"given":"Elena S.","family":"Santana-Trujillo","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer D.","family":"Salazar-Rinc\u00f3n","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bryan S.","family":"S\u00e1nchez-Beltr\u00e1n","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gustavo","family":"Gonz\u00e1lez-Paya","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angel","family":"Cruz-Roa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3157-6777","authenticated-orcid":false,"given":"Julieta E.","family":"Ochoa-Amaya","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,9,27]]},"reference":[{"issue":"6","key":"10736_CR1","first-page":"211","volume":"15","author":"C Abraham","year":"2011","unstructured":"Abraham C, Medzhitov R (2011) Innate immunity to bacterial infections: the role of toll-like receptors. 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