Summary of Study ST002556

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001648. The data can be accessed directly via it's Project DOI: 10.21228/M8PF0K This work is supported by NIH grant, U2C- DK119886.

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Study IDST002556
Study TitleBlood metabolomics and impacted cellular mechanisms during transition into lactation in dairy cows that develop metritis
Study TypeCase-Control Study
Study SummaryThe objective of this study was to identify metabolites associated with metritis and use them for identification of cellular mechanisms affected during transition into lactation. Holstein cows (n = 104) had blood collected in the prepartum period (d-14 ± 6), at calving (d0), and at the day of metritis diagnosis (d7 ± 2). Cows with reddish or brownish, watery, and fetid discharge were diagnosed with metritis (n = 52). Cows with metritis were paired with herdmates without metritis (n = 52) based on DIM. The metabolome of plasma samples was evaluated using untargeted gas chromatography time-of-flight mass spectrometry. Univariate analyses included t-tests and fold change analyses. Metabolites with false discovery rate (FDR) adjusted P ≤ 0.10 on t-tests were used for partial least squares – discriminant analysis PLS-DA coupled with permutational analysis using 2,000 permutations. Metabolites with FDR adjusted P ≤ 0.10 on t-tests were also used for enriched pathway analyses and identification of cellular processes. Cows that developed metritis had affected cellular processes associated with lower amino acid metabolism in the prepartum period, greater lipolysis, cell death, and oxidative stress at calving and at metritis diagnosis, and greater leukocyte activation at calving, but lower immune cell activation at metritis diagnosis. In summary, cows that developed metritis had plasma metabolomic changes associated with greater lipolysis, oxidative stress, and a dysregulated immune response which may predispose cows to metritis development.
Institute
University of Florida
DepartmentCollege of Veterinary Medicine
LaboratoryLarge Animal Clinical Sciences
Last Namesegundocasaro
First NameSegundo
Address117 Deriso Hall, 2015 SW 16th Ave.
Emailsegundocasaro@ufl.edu
Phone3522844016
Submit Date2023-04-11
Num Groups2
Total Subjects104
Num Females104
Analysis Type DetailGC-MS
Release Date2023-04-28
Release Version1
Segundo segundocasaro Segundo segundocasaro
https://dx.doi.org/10.21228/M8PF0K
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001648
Project DOI:doi: 10.21228/M8PF0K
Project Title:Blood metabolomics and impacted cellular mechanisms during transition into lactation in dairy cows that develop metritis
Project Type:Case-Control Study
Project Summary:The objective of this study was to identify metabolites associated with metritis and use them for identification of cellular mechanisms affected during transition into lactation. Holstein cows (n = 104) had blood collected in the prepartum period (d-14 ± 6), at calving (d0), and at the day of metritis diagnosis (d7 ± 2). Cows with reddish or brownish, watery, and fetid discharge were diagnosed with metritis (n = 52). Cows with metritis were paired with herdmates without metritis (n = 52) based on DIM. The metabolome of plasma samples was evaluated using untargeted gas chromatography time-of-flight mass spectrometry. Univariate analyses included t-tests and fold change analyses. Metabolites with false discovery rate (FDR) adjusted P ≤ 0.10 on t-tests were used for partial least squares – discriminant analysis PLS-DA coupled with permutational analysis using 2,000 permutations. Metabolites with FDR adjusted P ≤ 0.10 on t-tests were also used for enriched pathway analyses and identification of cellular processes. Cows that developed metritis had affected cellular processes associated with lower amino acid metabolism in the prepartum period, greater lipolysis, cell death, and oxidative stress at calving and at metritis diagnosis, and greater leukocyte activation at calving, but lower immune cell activation at metritis diagnosis. In summary, cows that developed metritis had plasma metabolomic changes associated with greater lipolysis, oxidative stress, and a dysregulated immune response which may predispose cows to metritis development.
Institute:University of Florida
Department:College of Veterinary Medicine
Laboratory:Large Animal Clinical Sciences
Last Name:Casaro
First Name:Segundo
Address:117 Deriso Hall, 2015 SW 16th Ave.
Email:segundocasaro@ufl.edu
Phone:3522844016
Funding Source:USDA NIFA AFRI

Subject:

Subject ID:SU002657
Subject Type:Mammal
Subject Species:Bos taurus
Taxonomy ID:9913
Weight Or Weight Range:700 kg
Gender:Female
Animal Housing:Free-stalls
Animal Feed:TMR

Factors:

Subject type: Mammal; Subject species: Bos taurus (Factor headings shown in green)

mb_sample_id local_sample_id Group Time Parity
SA2568989794_Calving_ConCon Calving Mult
SA2568999778_Calving_ConCon Calving Mult
SA2569009979_Calving_ConCon Calving Mult
SA2569019971_Calving_ConCon Calving Mult
SA2569029802_Calving_ConCon Calving Mult
SA2569039966_Calving_ConCon Calving Mult
SA2569049804_Calving_ConCon Calving Mult
SA2569059536_Calving_ConCon Calving Mult
SA2569069985_Calving_ConCon Calving Mult
SA2569079280_Calving_ConCon Calving Mult
SA2569089268_Calving_ConCon Calving Mult
SA2569099945_Calving_ConCon Calving Mult
SA25691010001_Calving_ConCon Calving Mult
SA2569119992_Calving_ConCon Calving Mult
SA2569129823_Calving_ConCon Calving Mult
SA2569139987_Calving_ConCon Calving Mult
SA2569149509_Calving_ConCon Calving Mult
SA25691510038_Calving_ConCon Calving Mult
SA2569169924_Calving_ConCon Calving Mult
SA2569179918_Calving_ConCon Calving Mult
SA2569189917_Calving_ConCon Calving Mult
SA2569199925_Calving_ConCon Calving Mult
SA2569209927_Calving_ConCon Calving Mult
SA2569219957_Calving_ConCon Calving Mult
SA2569229944_Calving_ConCon Calving Mult
SA2569239950_Calving_ConCon Calving Mult
SA2569249942_Calving_ConCon Calving Mult
SA2569259954_Calving_ConCon Calving Mult
SA2569269956_Calving_ConCon Calving Mult
SA2569279875_Calving_ConCon Calving Mult
SA2569289872_Calving_ConCon Calving Mult
SA2569299904_Calving_ConCon Calving Mult
SA2569309876_Calving_ConCon Calving Mult
SA2569319907_Calving_ConCon Calving Mult
SA25693210303_Calving_ConCon Calving Prim
SA25693310264_Calving_ConCon Calving Prim
SA25693410254_Calving_ConCon Calving Prim
SA25693510212_Calving_ConCon Calving Prim
SA25693610246_Calving_ConCon Calving Prim
SA25693710262_Calving_ConCon Calving Prim
SA25693810268_Calving_ConCon Calving Prim
SA25693910274_Calving_ConCon Calving Prim
SA25694010311_Calving_ConCon Calving Prim
SA25694110261_Calving_ConCon Calving Prim
SA25694210229_Calving_ConCon Calving Prim
SA25694310290_Calving_ConCon Calving Prim
SA25694410245_Calving_ConCon Calving Prim
SA25694510194_Calving_ConCon Calving Prim
SA25694610171_Calving_ConCon Calving Prim
SA25694710165_Calving_ConCon Calving Prim
SA25694810199_Calving_ConCon Calving Prim
SA25694910189_Calving_ConCon Calving Prim
SA2569509985_Diagnosis_ConCon Diagnosis Mult
SA2569519904_Diagnosis_ConCon Diagnosis Mult
SA2569529907_Diagnosis_ConCon Diagnosis Mult
SA2569539918_Diagnosis_ConCon Diagnosis Mult
SA2569549954_Diagnosis_ConCon Diagnosis Mult
SA2569559917_Diagnosis_ConCon Diagnosis Mult
SA2569569872_Diagnosis_ConCon Diagnosis Mult
SA2569579876_Diagnosis_ConCon Diagnosis Mult
SA2569589804_Diagnosis_ConCon Diagnosis Mult
SA2569599794_Diagnosis_ConCon Diagnosis Mult
SA2569609778_Diagnosis_ConCon Diagnosis Mult
SA2569619536_Diagnosis_ConCon Diagnosis Mult
SA2569629802_Diagnosis_ConCon Diagnosis Mult
SA2569639268_Diagnosis_ConCon Diagnosis Mult
SA2569649509_Diagnosis_ConCon Diagnosis Mult
SA2569659823_Diagnosis_ConCon Diagnosis Mult
SA2569669280_Diagnosis_ConCon Diagnosis Mult
SA2569679875_Diagnosis_ConCon Diagnosis Mult
SA2569689925_Diagnosis_ConCon Diagnosis Mult
SA2569699979_Diagnosis_ConCon Diagnosis Mult
SA2569709971_Diagnosis_ConCon Diagnosis Mult
SA2569719987_Diagnosis_ConCon Diagnosis Mult
SA25697210038_Diagnosis_ConCon Diagnosis Mult
SA2569739992_Diagnosis_ConCon Diagnosis Mult
SA2569749924_Diagnosis_ConCon Diagnosis Mult
SA25697510001_Diagnosis_ConCon Diagnosis Mult
SA2569769966_Diagnosis_ConCon Diagnosis Mult
SA2569779944_Diagnosis_ConCon Diagnosis Mult
SA2569789942_Diagnosis_ConCon Diagnosis Mult
SA2569799927_Diagnosis_ConCon Diagnosis Mult
SA2569809957_Diagnosis_ConCon Diagnosis Mult
SA2569819945_Diagnosis_ConCon Diagnosis Mult
SA2569829956_Diagnosis_ConCon Diagnosis Mult
SA2569839950_Diagnosis_ConCon Diagnosis Mult
SA25698410254_Diagnosis_ConCon Diagnosis Prim
SA25698510194_Diagnosis_ConCon Diagnosis Prim
SA25698610171_Diagnosis_ConCon Diagnosis Prim
SA25698710189_Diagnosis_ConCon Diagnosis Prim
SA25698810261_Diagnosis_ConCon Diagnosis Prim
SA25698910165_Diagnosis_ConCon Diagnosis Prim
SA25699010199_Diagnosis_ConCon Diagnosis Prim
SA25699110212_Diagnosis_ConCon Diagnosis Prim
SA25699210245_Diagnosis_ConCon Diagnosis Prim
SA25699310262_Diagnosis_ConCon Diagnosis Prim
SA25699410229_Diagnosis_ConCon Diagnosis Prim
SA25699510274_Diagnosis_ConCon Diagnosis Prim
SA25699610303_Diagnosis_ConCon Diagnosis Prim
SA25699710311_Diagnosis_ConCon Diagnosis Prim
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Collection:

Collection ID:CO002650
Collection Summary:All cows had blood collected in the prepartum period (-14 DRP), calving (first 24h after calving), and at diagnosis (day of metritis diagnosis). Blood was sampled from the jugular vein using a 20-gauge x 2.54-cm needle and 10-mL evacuated tube containing lithium heparin (Vacutainer, Becton, Dickinson and Company, Franklin Lakes, NJ, USA). After collection, the blood tubes were placed on ice and transported to the laboratory within 2 hours. Once in the laboratory, the blood tubes were centrifuged at 4000 g, 4 °C, for 10 min, and the plasma was stored at -80 oC for further characterization of the plasma metabolome. The frozen plasma was submitted to the University of California’s West Coast Metabolomics Center in Davis, CA for metabolome analysis. Samples were analyzed by blinded technicians using untargeted gas chromatography with time-of-flight mass spectrometry (GC-TOF-MS) in a single batch.
Sample Type:Blood (plasma)

Treatment:

Treatment ID:TR002669
Treatment Summary:This was a case-control study, hence, there were not treatments applied. Cows were self assigned to the groups. Cows that developed metritis were paired with healthy cows by days in milk.

Sample Preparation:

Sampleprep ID:SP002663
Sampleprep Summary:After collection, the blood tubes were placed on ice and transported to the laboratory within 2 hours. Once in the laboratory, the blood tubes were centrifuged at 4000 g, 4 °C, for 10 min, and the plasma was stored at -80 oC for further characterization of the plasma metabolome. The frozen plasma was submitted to the University of California’s West Coast Metabolomics Center in Davis, CA for metabolome analysis

Combined analysis:

Analysis ID AN004211
Analysis type MS
Chromatography type GC
Chromatography system Leco Pegasus IV GC
Column Restek Rtx-5Sil MS (30m x 0.25mm, 0.25um)
MS Type EI
MS instrument type GC-TOF
MS instrument name Leco Pegasus IV TOF
Ion Mode POSITIVE
Units peak heights

Chromatography:

Chromatography ID:CH003122
Chromatography Summary:Data are acquired using the following chromatographic parameters, with more details to be found in Fiehn O. et al. Plant J. 53 (2008) 691–704. Column: Restek corporation Rtx-5Sil MS (30 m length x 0.25 mm internal diameter with 0.25 μm film made of 95% dimethyl/5%diphenylpolysiloxane) Mobile phase: Helium Column temperature: 50-330°C Flow- rate: 1 mL min-1 Injection volume: 0.5 μL Injection: 25 splitless time into a multi-baffled glass liner Injection temperature: 50°C ramped to 250°C by 12°C s-1 Oven temperature program: 50°C for 1 min, then ramped at 20°C min-1 to 330°C, held constant for 5 min. The analytical GC column is protected by a 10 m long empty guard column which is cut by 20 cm intervals whenever the reference mixture QC samples indicate problems caused by column contaminations. We have validated that at this sequence of column cuts, no detrimental effects are detected with respect to peak shapes, absolute or relative metabolite retention times or reproducibility of quantifications. This chromatography method yields excellent retention and separation of primary metabolite classes (amino acids, hydroxyl acids, carbohydrates, sugar acids, sterols, aromatics, nucleosides, amines and miscellaneous compounds) with narrow peak widths of 2–3 s and very good within-series retention time reproducibility of better than 0.2 s absolute deviation of retention times. We use automatic liner exchanges after each set of 10 injections which we could show to reduce sample carryover for highly lipophilic compounds such as free fatty acids. Mass spectrometry parameters are used as follows: a Leco Pegasus IV mass spectrometer is used with unit mass resolution at 17 spectra s-1 from 80-500 Da at - 70 eV ionization energy and 1800 V detector voltage with a 230°C transfer line and a 250°C ion source.
Instrument Name:Leco Pegasus IV GC
Column Name:Restek Rtx-5Sil MS (30m x 0.25mm, 0.25um)
Column Temperature:50-330
Flow Gradient:.
Flow Rate:1 mL min-1
Solvent A:.
Solvent B:.
Chromatography Type:GC

MS:

MS ID:MS003958
Analysis ID:AN004211
Instrument Name:Leco Pegasus IV TOF
Instrument Type:GC-TOF
MS Type:EI
MS Comments:See data acquisition, processing, and reporting in "Methods.pdf" file
Ion Mode:POSITIVE
Analysis Protocol File:Methods_SC.pdf
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