{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T12:12:43Z","timestamp":1788351163914,"version":"build-2803163510"},"reference-count":39,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:00:00Z","timestamp":1788307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"DairyBio"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>The current clinical test for ketosis in dairy cows, which is based on measuring blood \u03b2-hydroxybutyrate (BHB) concentrations, is an invasive procedure that impacts herd welfare. Alternative biomarkers, especially non-invasive ones, are urgently needed for subclinical ketosis (SCK) diagnosis. To address this, we examined the milk lipidomic profiles of healthy and SCK cows from a research dairy farm over two consecutive years. A total of 125 polar lipid species were quantified in milk samples using a targeted liquid chromatography\u2013mass spectrometry (LC-MS) approach. Chemometric analysis identified four potential biomarkers (PC 28:0, PC 29:0, PC 30:1 and PC 31:0), which met the thresholds for p value (&lt;0.0004) and fold change (FC &gt; 2). Global fatty acid (FA) profiling by gas chromatography\u2013flame ionization detection (GC-FID) after transesterification of all lipids revealed that the levels of C8:0, C10:0, C12:0, C14:0, C15:0 and C18:1t11 were significantly suppressed in the SCK group (15\u201352% lower compared to the control group, p &lt; 0.01), and C15:0 and C18:1t11 were the most promising markers for SCK in dairy cows at the FA level (43\u201350% reduction compared to the control group, p &lt; 0.0001). Tandem LC-MS and FA data generated by GC demonstrated that SCK also caused significant changes in regio-isomer and double bond isomer composition of a model phospholipid molecule (PC 34:1) and a model triglyceride molecule TAG 52:2; these fine-structural alterations could also be explored for discovering novel SCK biomarkers in dairy cows. The PLS-DA predictive model built from lipidomic data showed excellent accuracy (&gt;95%) in predicting cows with SCK. All of the promising biomarkers identified and the predictive model built in this study need further validation using larger SCK cohorts from different herds.<\/jats:p>","DOI":"10.3390\/foods15173117","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T10:57:23Z","timestamp":1788346643000},"page":"3117","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Lipidomic Profiling of Milk Enables Identification of Novel Biomarkers for Subclinical Ketosis in Dairy Cows"],"prefix":"10.3390","volume":"15","author":[{"given":"Zhiqian","family":"Liu","sequence":"first","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vilnis","family":"Ezernieks","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-6905-9887","authenticated-orcid":false,"given":"Joanne E.","family":"Hemsworth","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3902-1314","authenticated-orcid":false,"given":"Coralie M.","family":"Reich","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1157-6844","authenticated-orcid":false,"given":"Carolyn R.","family":"Bath","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-1807-4524","authenticated-orcid":false,"given":"Monique J.","family":"Berkhout","sequence":"additional","affiliation":[{"name":"School of Applied Systems Biology, La Trobe University, Bundoora, VIC 3083, Australia"},{"name":"Ellinbank Centre, Agriculture Victoria, Ellinbank, VIC 3821, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad S.","family":"Tahir","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9698-8401","authenticated-orcid":false,"given":"Leah C.","family":"Marett","sequence":"additional","affiliation":[{"name":"School of Applied Systems Biology, La Trobe University, Bundoora, VIC 3083, Australia"},{"name":"Ellinbank Centre, Agriculture Victoria, Ellinbank, VIC 3821, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9395-1299","authenticated-orcid":false,"given":"Amanda J.","family":"Chamberlain","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"},{"name":"School of Applied Systems Biology, La Trobe University, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mike E.","family":"Goddard","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"},{"name":"School of Agriculture and Food, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Melbourne, VIC 3010, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruidong","family":"Xiang","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"},{"name":"School of Applied Systems Biology, La Trobe University, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8442-6081","authenticated-orcid":false,"given":"Simone J.","family":"Rochfort","sequence":"additional","affiliation":[{"name":"Centre for AgriBiosciences, Agriculture Victoria, 5 Ring Road, Bundoora, VIC 3083, Australia"},{"name":"School of Applied Systems Biology, La Trobe University, Bundoora, VIC 3083, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Tufarelli, V., Puva\u010da, N., Glamo\u010di\u0107, D., Pugliese, G., and Colonna, M.A. (2024). 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