{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T05:02:27Z","timestamp":1790744547834,"version":"4.1.0"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:00:00Z","timestamp":1790726400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Bovine mastitis is generally viewed as a localized intramammary infection, but the extent to which gastrointestinal, circulating, and mammary tissue features covary across clinical phenotypes remains unclear. We aimed to characterize concurrent alterations across the rumen, hindgut, serum, milk, and mammary gland in healthy control (HC), subclinical mastitis (SCM), and clinical mastitis (CM) cows and to evaluate circulating 5-androstene-3\u03b2,17\u03b2-diol (5-AED) as a correlate of mastitis phenotype.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>In a discovery cohort, fecal and rumen microbiota were characterized by 16S rRNA gene sequencing, and untargeted metabolomics was performed across four biological matrices. Mammary tissue was analyzed using tandem mass tag (TMT)-based proteomics, parallel reaction monitoring (PRM), and histology, while circulating inflammatory and metabolic mediators were measured in serum. A later cohort from the same herd was analyzed independently to replicate the serum 5-AED findings using targeted analysis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Systemic inflammatory and lipid mediators differed in SCM cows despite the absence of overt clinical signs. Fecal and rumen microbial community structures differed among HC, SCM, and CM groups, with lower rumen microbial richness in mastitis cows. 16S rRNA-based functional predictions indicated differences in carbohydrate- and amino acid-related functional potential. Serum 5-AED showed the highest distribution in HC, an intermediate distribution in SCM, and the lowest distribution in CM, and this group ordering was reproduced in the later cohort. Mammary tissue exhibited lower abundance of claudin-8, fatty acid synthase, acetyl-CoA carboxylase, and ABCG2, together with enrichment of complement and coagulation pathways and annotation of a neutrophil extracellular trap-associated pathway. Targeted proteomics supported the direction of selected barrier-, lipid-, complement-, and fibrinogen-related differences. Hydroxysteroid 17\u03b2-dehydrogenase abundance also differed among groups; however, enzyme activity and a direct biochemical relationship with 5-AED were not established.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>These findings reveal coordinated differences across gastrointestinal, circulating, and mammary compartments associated with distinct mastitis phenotypes, including subclinical disease. However, the cross-sectional design does not establish temporal progression, mastitis specificity, or a causal gut\u2013mammary pathway. The association between circulating 5-AED and mastitis phenotype warrants prospective evaluation in independent herds using an independently validated analytical assay before its potential clinical utility can be assessed.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1955642","type":"journal-article","created":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T04:53:38Z","timestamp":1790744018000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Multi-compartment omics identifies a reproducible association between circulating 5-androstene-3\u03b2,17\u03b2-diol and mastitis clinical status in dairy cows"],"prefix":"10.3389","volume":"17","author":[{"given":"Yuqiong","family":"Li","sequence":"first","affiliation":[{"name":"Institute of Animal Science, Ningxia Academy of Agricultural and Forestry Sciences","place":["Yinchuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haihui","family":"Gao","sequence":"additional","affiliation":[{"name":"Institute of Animal Science, Ningxia Academy of Agricultural and Forestry Sciences","place":["Yinchuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhen","family":"Dong","sequence":"additional","affiliation":[{"name":"Hunan Province Key Laboratory of Traditional Chinese Veterinary Medicine, Hunan Agricultural University","place":["Changsha, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Gao","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang","family":"Huang","sequence":"additional","affiliation":[{"name":"Ningxia Agricultural Land Reclamation Changshan Tou Farm Company Limited","place":["Yinchuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Song","family":"Hua","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University","place":["Xianyang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuming","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Ningxia University","place":["Yinchuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongbing","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Animal Science, Ningxia Academy of Agricultural and Forestry Sciences","place":["Yinchuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenglian","family":"Zhang","sequence":"additional","affiliation":[{"name":"Ningxia Hui Autonomous Region Animal Disease Prevention and Control Center","place":["Yinchuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meizhou","family":"Huang","sequence":"additional","affiliation":[{"name":"Metabolic Hepatobiliary and Pancreatic Diseases Key Laboratory of Luzhou City, Academician (Expert) Workstation of Sichuan Province, Department of General Surgery (Hepatopancreatobiliary Surgery), The Affiliated Hospital of Southwest Medical University","place":["Luzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,30]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"108801","DOI":"10.1016\/j.intimp.2022.108801","article-title":"The protective and therapeutic effects of 5-androstene-3\u03b2,17\u03b2-diol (ADIOL) in abdominal post-operative adhesions in rat: suppressing TLR4\/NF-\u03baB\/HMGB1\/TGF-\u03b21\/\u03b1-SMA pathway","volume":"109","author":"Abbas","year":"2022","journal-title":"Int. Immunopharmacol."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"53.04","DOI":"10.4049\/jimmunol.206.Supp.53.04","article-title":"Emerging role of the mitochondrial branched-chain aminotransferase (BCATm) as an immunosuppressive enzyme during CD4+ T cell activation","volume":"206","author":"Ananieva-Stoyanova","year":"2021","journal-title":"J. Immunol."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1111\/j.2517-6161.1995.tb02031.x","article-title":"Controlling the false discovery rate: a practical and powerful approach to multiple testing","volume":"57","author":"Benjamini","year":"1995","journal-title":"J. R. Stat. Soc. Ser. B: Stat. Methodol."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"2626","DOI":"10.1182\/blood.2021014966","article-title":"S100A8\/A9 drives the formation of procoagulant platelets through GPIb\u03b1","volume":"140","author":"Colicchia","year":"2022","journal-title":"Blood"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1186\/s40781-015-0079-8","article-title":"Innate immunity and carbohydrate metabolism alterations precede occurrence of subclinical mastitis in transition dairy cows","volume":"57","author":"Dervishi","year":"2015","journal-title":"J. Anim. Sci. Technol."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"15880","DOI":"10.1073\/pnas.1005743107","article-title":"Extracellular DNA traps promote thrombosis","volume":"107","author":"Fuchs","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1111\/imr.12453","article-title":"Clearance of apoptotic neutrophils and resolution of inflammation","volume":"273","author":"Greenlee-Wacker","year":"2016","journal-title":"Immunol. Rev."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"4089","DOI":"10.3390\/ijms24044089","article-title":"The gut\u2013organ-axis concept: advances the application of gut-on-chip technology","volume":"24","author":"Guo","year":"2023","journal-title":"Int. J. Mol. Sci."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1016\/j.mce.2018.11.012","article-title":"17\u03b2-hydroxysteroid dehydrogenases as acyl thioester metabolizing enzymes","volume":"489","author":"Hiltunen","year":"2019","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"1897","DOI":"10.1038\/s41396-020-0651-1","article-title":"The gut microbiota contributes to the development of Staphylococcus aureus -induced mastitis in mice","volume":"14","author":"Hu","year":"2020","journal-title":"ISME J."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1016\/j.jare.2023.02.009","article-title":"Gut\/rumen-mammary gland axis in mastitis: gut\/rumen microbiota\u2013mediated \u201cgastroenterogenic mastitis\u201d","volume":"55","author":"Hu","year":"2024","journal-title":"J. Adv. Res."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"9004","DOI":"10.3168\/jds.2024-26210","article-title":"Invited review: multiomics insights into the molecular and regulatory mechanisms underlying bovine mastitis","volume":"108","author":"Ibeagha-Awemu","year":"2025","journal-title":"J. Dairy Sci."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"1332","DOI":"10.1016\/j.cell.2016.05.041","article-title":"From dietary fiber to host physiology: short-chain fatty acids as key bacterial metabolites","volume":"165","author":"Koh","year":"2016","journal-title":"Cell"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"4423","DOI":"10.3168\/jds.2025-27842","article-title":"Milk metabolomic alterations correlated with intramammary infection in dairy cows: from healthy status to clinical mastitis","volume":"109","author":"Lisuzzo","year":"2026","journal-title":"J. Dairy Sci."},{"key":"ref19","doi-asserted-by":"publisher","DOI":"10.1681\/ASN.0000001102","article-title":"Neutrophil extracellular traps aggravate injury in complement-mediated thrombotic microangiopathy","author":"Liu","year":"2026","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"949","DOI":"10.1038\/s41590-026-02456-z","article-title":"Lipid asymmetry disruption by XKR8 orchestrates neutrophil extracellular trap formation and inhibits fungal infection","volume":"27","author":"Liu","year":"2026","journal-title":"Nat. Immunol."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1093\/advances\/nmx009","article-title":"Butyrate: a double-edged sword for health?","volume":"9","author":"Liu","year":"2018","journal-title":"Adv. Nutr."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1111\/j.1574-6968.2011.02424.x","article-title":"Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures","volume":"325","author":"Minas","year":"2011","journal-title":"FEMS Microbiol. Lett."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1007\/s13668-023-00461-4","article-title":"Butyrate: more than a short chain fatty acid","volume":"12","author":"Mohamed Elfadil","year":"2023","journal-title":"Curr. Nutr. Rep."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"846","DOI":"10.1038\/nature01320","article-title":"Points of control in inflammation","volume":"420","author":"Nathan","year":"2002","journal-title":"Nature"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"998","DOI":"10.1055\/s-0044-1782660","article-title":"The role of myeloid cells in thromboinflammatory disease","volume":"50","author":"Noone","year":"2024","journal-title":"Semin. Thromb. Hemost."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1038\/nri.2017.105","article-title":"Neutrophil extracellular traps in immunity and disease","volume":"18","author":"Papayannopoulos","year":"2018","journal-title":"Nat. Rev. Immunol."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"277","DOI":"10.3389\/fimmu.2019.00277","article-title":"Short chain fatty acids (SCFAs)-mediated gut epithelial and immune regulation and its relevance for inflammatory bowel diseases","volume":"10","author":"Parada Venegas","year":"2019","journal-title":"Front. Immunol."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"557","DOI":"10.1007\/s00018-014-1762-5","article-title":"Co-operation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling","volume":"72","author":"P\u0142\u00f3ciennikowska","year":"2015","journal-title":"Cell. Mol. Life Sci."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"e1005752","DOI":"10.1371\/journal.pcbi.1005752","article-title":"mixOmics: an R package for 'omics feature selection and multiple data integration","volume":"13","author":"Rohart","year":"2017","journal-title":"PLoS Comput. Biol."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"10381","DOI":"10.3168\/jds.2017-13023","article-title":"A 100-year review: mastitis detection, management, and prevention","volume":"100","author":"Ruegg","year":"2017","journal-title":"J. Dairy Sci."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"1191","DOI":"10.1038\/ni1276","article-title":"Resolution of inflammation: the beginning programs the end","volume":"6","author":"Serhan","year":"2005","journal-title":"Nat. Immunol."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"2498","DOI":"10.1101\/gr.1239303","article-title":"Cytoscape: a software environment for integrated models of biomolecular interaction networks","volume":"13","author":"Shannon","year":"2003","journal-title":"Genome Res."},{"key":"ref38","doi-asserted-by":"publisher","first-page":"1103836","DOI":"10.3389\/fmicb.2022.1103836","article-title":"Butyrate producers, \u201cthe sentinel of gut\u201d: their intestinal significance with and beyond butyrate, and prospective use as microbial therapeutics","volume":"13","author":"Singh","year":"2023","journal-title":"Front. Microbiol."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1038\/s41569-021-00552-1","article-title":"Interplay between inflammation and thrombosis in cardiovascular pathology","volume":"18","author":"Stark","year":"2021","journal-title":"Nat. Rev. Cardiol."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1111\/imm.12617","article-title":"Lipid rafts in immune signalling: current progress and future perspective","volume":"149","author":"Varshney","year":"2016","journal-title":"Immunology"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"100141","DOI":"10.1016\/j.xinn.2021.100141","article-title":"clusterProfiler 4.0: a universal enrichment tool for interpreting omics data","volume":"2","author":"Wu","year":"2021","journal-title":"Innovation (Cambridge (Mass.))"},{"key":"ref42","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1016\/j.rvsc.2021.06.019","article-title":"Neutrophil extracellular traps in cattle health and disease","volume":"139","author":"Xie","year":"2021","journal-title":"Res. Vet. Sci."},{"key":"ref43","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1186\/s13045-023-01453-1","article-title":"Amino acid metabolism in immune cells: essential regulators of the effector functions, and promising opportunities to enhance cancer immunotherapy","volume":"16","author":"Yang","year":"2023","journal-title":"J. Hematol. Oncol."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"112186","DOI":"10.1016\/j.celrep.2023.112186","article-title":"Accumulation of branched-chain amino acids reprograms glucose metabolism in CD8+ T cells with enhanced effector function and anti-tumor response","volume":"42","author":"Yao","year":"2023","journal-title":"Cell Rep."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"eadj1397","DOI":"10.1126\/sciadv.adj1397","article-title":"NET formation is a default epigenetic program controlled by PAD4 in apoptotic neutrophils","volume":"9","author":"Zhu","year":"2023","journal-title":"Sci. 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