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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14061/m_MTBLS14061_LC-MS_alternating_reverse-phase_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14061/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14061/a_MTBLS14061_LC-MS_alternating_reverse-phase_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14061/s_MTBLS14061.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14061Targeted Bile Acid Analysis A volume of 100 μL of sample was accurately transferred and mixed with 50 μL of internal standard working solution 2 (200 ng/mL), followed by the addition of 350 μL of extraction solvent (methanol). The mixture was vortexed for 30 s, subjected to low-temperature ultrasonication for 30 min (5 °C, 40 kHz), and then left at −20℃ for 30 min. After centrifugation at 13,000 rpm for 15 min at 4℃, the supernatant was collected and evaporated to dryness under nitroge  arrow_drop_downmoreMetaboLightsPublicLiquid Chromatography MS - alternating - reverse-phaseLC-ESI-MS/MS (UHPLC-Qtrap),ExionLC AD system, Waters BEH C18 (150*2.1 mm,1.7 μm) liquid chromatography column, column temperature 50 ° C, injection volume 5 μL. Mobile phase A (0.1% formic acid aqueous solution), mobile phase B (0.1% formic acid acetonitrile)Multi-omics and intervention analyses reveal a hindgut microbiota–secondary bile acid–mammary axis linked to milk yield in dairy cows.jiaxin wangjilin universityserummass spectrometry assayPrecisely remove 100 μL sample, add 50 μL internal standard working solution 2 (200 ng/mL), then add 350 μL extraction solution (methanol), The mixture was vortexed and mixed for 30 s, then sonicated at low temperature (5℃, 40 KHz) for 30 min, left at -20℃ for 30 min, centrifuged at 13000 rcf at 4℃ for 15 min, and the supernatant was dried under nitrogen gas. Then 100 μl of 50% acetonitrile water was added to redissolve, and the mixture was vortexed and mixed for 30 s, and sonicated at  arrow_drop_downmoreBos taurushttps://www.ebi.ac.uk/metabolights/MTBLS14061jiaxin wang. Jilin university. 1252978953@qq.com.shoupeng fu. jilin university. fushoupeng@jlu.edu.cn.Each ion fragment was automatically identified and integrated using default parameters in the AB Sciex quantification software OS, assisted by manual inspection. The ratio of the peak area of the analyte to the peak area of the internal standard was plotted as the ordinate, and the concentration of the analyte was plotted as the abscissa. sample This concentration calculation: the ratio of the peak area of the sample analyte to the peak area of the internal standard was substitute  arrow_drop_downmoreTreatment1252978953@qq.comHolstein dairy cows were recruited from a commercial dairy farm in Changchun, China, housing approximately 3,000 cows. Among them, second-parity cows in mid-lactation (120 ± 10 days in milk) were categorized into an H group (milk yield: 49.04 ± 1.28 kg/d) and an N group (milk yield: 34.55 ± 2.06 kg/d), with 12 cows in each group. None of the cows had received antibiotics or other medications in the six months prior to sampling. All cows were maintained under identical environmental and arrow_drop_downmoreMetabolomicsTargetbile acidSerumCowTargetbile acidSerumCowMass spectrum conditions: AB SCIEX QTRAP 6500+, detected in negative mode, Curtain Gas (CUR) 35, Collision Gas (CAD) was Medium, IonSpray Voltage (IS) was -4500, Temperature (TEM) was 550, Ion Source Gas1 (GS1) was 50, Ion Source Gas2 (GS2) was 50.falseMulti-omics and intervention analyses reveal a hindgut microbiota–secondary bile acid–mammary axis linked to milk yield in dairy cows_serumShotgun metagenomics, untargeted metabolomics, and targeted metabolomics of high versus normal-yield cows revealed that high-yield animals harbored distinct hindgut microbial communities enriched in bile salt hydrolase–positive taxa, and elevated levels of deoxycholic acid (DCA). Fecal microbiota transplantation from high-yield donors into antibiotic-treated recipient mice increased their milk production, accompanied by a shift in circulating bile acid profiles towards higher DCA. Oral administration of DCA to antibiotic-treated mice partially recapitulated these mammary and lactation phenotypes, supporting a causal contribution of bile acids downstream of the microbiota. In lactating cows, supplementation with a rumen-protected DCA formulation enhanced cumulative energy-corrected milk yield and tended to increase daily milk yield. DCA can directly upregulate the expression of genes related to the cell cycle progression in dairy cow mammary glands, thereby promoting and maintaining the activity of mammary epithelial cells in mid-lactation cows to support milk production.2026-04-152026-03-17MTBLS14061