Summary of Study ST003675

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 PR002280. The data can be accessed directly via it's Project DOI: 10.21228/M8X534 This work is supported by NIH grant, U2C- DK119886.

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Study IDST003675
Study TitlePolar Metabolite Profiles Distinguish Between Early and Severe Sub-maintenance Nutritional States of Wild Bighorn Sheep
Study SummaryUnderstanding the metabolic adaptations of wild bighorn sheep (Ovis c. canadensis) to nutritional stress is crucial for their conservation. This study employed 1H nuclear magnetic resonance (NMR) metabolomics to investigate the biochemical responses of these animals to varying sub-maintenance nutritional states. Serum samples from 388 wild bighorn sheep collected between 2014 and 2017 during December through March across Wyoming and Montana were analyzed. Multivariate statistics and machine learning analyses were employed to identify characteristic metabolic patterns and metabolic interactions between early and severe sub-maintenance nutritional states. Significant differences were observed in the levels of 15 of the 49 quantified metabolites, including formate, thymine, glucose, choline, and others, pointing to disruptions in one-carbon, amino acid, and central carbon metabolic pathways. These metabolites may serve as indicators of critical physiological processes such as nutritional intake, immune function, energy metabolism, and protein catabolism, essential for understanding how wild bighorn sheep adapt to nutritional stress. The study has generated valuable insights into molecular networks underlying the metabolic resilience of wild bighorn sheep, highlighting the potential for using specific biochemical markers to evaluate nutritional and energetic states in free-ranging ungulates. These insights may help wildlife man-agers and ecologists compare populations across different times in seasonal cycles, providing information to assess the adequacy of seasonal ranges and support conservation efforts. This research strengthens our understanding of metabolic adaptations to envi-ronmental stressors in wild ruminants, offering a foundation for improving management practices to maintain healthy bighorn sheep populations.
Institute
Montana State University
DepartmentChemistry and Biochemistry
LaboratoryPI: Dr. Valerie Copie
Last NameO'Shea-Stone
First NameGalen
Address103 Chemistry and Biochemistry Building
Emailgalenoshea@gmail.com
Phone3035237132
Submit Date2025-01-15
Num Groups3
Total Subjects398
Num Females398
Analysis Type DetailNMR
Release Date2025-02-11
Release Version1
Galen O'Shea-Stone Galen O'Shea-Stone
https://dx.doi.org/10.21228/M8X534
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002280
Project DOI:doi: 10.21228/M8X534
Project Title:Polar Metabolite Profiles Distinguish Between Early and Severe Sub-maintenance Nutritional States of Wild Bighorn Sheep
Project Type:Untargeted NMR Metabolomics
Project Summary:Understanding the metabolic adaptations of wild bighorn sheep (Ovis c. canadensis) to nutritional stress is crucial for their conservation. This study employed 1H nuclear magnetic resonance (NMR) metabolomics to investigate the biochemical responses of these animals to varying sub-maintenance nutritional states. Serum samples from 388 wild bighorn sheep collected between 2014 and 2017 during December through March across Wyoming and Montana were analyzed. Multivariate statistics and machine learning analyses were employed to identify characteristic metabolic patterns and metabolic interactions between early and severe sub-maintenance nutritional states. Significant differences were observed in the levels of 15 of the 49 quantified metabolites, including formate, thymine, glucose, choline, and others, pointing to disruptions in one-carbon, amino acid, and central carbon metabolic pathways. These metabolites may serve as indicators of critical physiological processes such as nutritional intake, immune function, energy metabolism, and protein catabolism, essential for understanding how wild bighorn sheep adapt to nutritional stress. The study has generated valuable insights into molecular networks underlying the metabolic resilience of wild bighorn sheep, highlighting the potential for using specific biochemical markers to evaluate nutritional and energetic states in free-ranging ungulates. These insights may help wildlife man-agers and ecologists compare populations across different times in seasonal cycles, providing information to assess the adequacy of seasonal ranges and support conservation efforts. This research strengthens our understanding of metabolic adaptations to envi-ronmental stressors in wild ruminants, offering a foundation for improving management practices to maintain healthy bighorn sheep populations.
Institute:Montana State University
Department:Chemistry and Biochemistry
Laboratory:PI: Dr. Valerie Copie
Last Name:O'Shea-Stone
First Name:Galen
Address:103 Chemistry and Biochemistry Building
Email:galenoshea@gmail.com
Phone:3035237132

Subject:

Subject ID:SU003807
Subject Type:Mammal
Subject Species:Ovis canadensis canadensis
Taxonomy ID:112262

Factors:

Subject type: Mammal; Subject species: Ovis canadensis canadensis (Factor headings shown in green)

mb_sample_id local_sample_id Sheep type Nurtitional State
SA4018021372Mtn Early-submaint
SA4018031629Mtn Early-submaint
SA4018041630Mtn Early-submaint
SA4018051358Mtn Early-submaint
SA4018061346Mtn Early-submaint
SA4018071347Mtn Early-submaint
SA4018081359Mtn Early-submaint
SA4018091348Mtn Early-submaint
SA4018101360Mtn Early-submaint
SA4018111361Mtn Early-submaint
SA4018121627Mtn Early-submaint
SA4018131349Mtn Early-submaint
SA4018141362Mtn Early-submaint
SA4018151363Mtn Early-submaint
SA4018161364Mtn Early-submaint
SA4018171350Mtn Early-submaint
SA4018181351Mtn Early-submaint
SA4018191352Mtn Early-submaint
SA4018201365Mtn Early-submaint
SA4018211373Mtn Early-submaint
SA4018221628Mtn Early-submaint
SA4018231626Mtn Early-submaint
SA4018241366Mtn Early-submaint
SA4018251614Mtn Early-submaint
SA4018261743Mtn Early-submaint
SA4018271729Mtn Early-submaint
SA4018281727Mtn Early-submaint
SA4018291738Mtn Early-submaint
SA4018301733Mtn Early-submaint
SA4018311611Mtn Early-submaint
SA4018321612Mtn Early-submaint
SA4018331613Mtn Early-submaint
SA4018341615Mtn Early-submaint
SA4018351625Mtn Early-submaint
SA4018361616Mtn Early-submaint
SA4018371617Mtn Early-submaint
SA4018381618Mtn Early-submaint
SA4018391619Mtn Early-submaint
SA4018401620Mtn Early-submaint
SA4018411621Mtn Early-submaint
SA4018421622Mtn Early-submaint
SA4018431623Mtn Early-submaint
SA4018441624Mtn Early-submaint
SA4018451353Mtn Early-submaint
SA4018461367Mtn Early-submaint
SA4018471725Mtn Early-submaint
SA4018481675Mtn Early-submaint
SA4018491667Mtn Early-submaint
SA4018501668Mtn Early-submaint
SA4018511669Mtn Early-submaint
SA4018521670Mtn Early-submaint
SA4018531671Mtn Early-submaint
SA4018541672Mtn Early-submaint
SA4018551673Mtn Early-submaint
SA4018561674Mtn Early-submaint
SA4018571676Mtn Early-submaint
SA4018581664Mtn Early-submaint
SA4018591677Mtn Early-submaint
SA4018601567Mtn Early-submaint
SA4018611568Mtn Early-submaint
SA4018621569Mtn Early-submaint
SA4018631570Mtn Early-submaint
SA4018641574Mtn Early-submaint
SA4018651575Mtn Early-submaint
SA4018661579Mtn Early-submaint
SA4018671580Mtn Early-submaint
SA4018681666Mtn Early-submaint
SA4018691663Mtn Early-submaint
SA4018701354Mtn Early-submaint
SA4018711651Mtn Early-submaint
SA4018721355Mtn Early-submaint
SA4018731356Mtn Early-submaint
SA4018741357Mtn Early-submaint
SA4018751374Mtn Early-submaint
SA4018761368Mtn Early-submaint
SA4018771369Mtn Early-submaint
SA4018781370Mtn Early-submaint
SA4018791375Mtn Early-submaint
SA4018801371Mtn Early-submaint
SA4018811652Mtn Early-submaint
SA4018821662Mtn Early-submaint
SA4018831653Mtn Early-submaint
SA4018841654Mtn Early-submaint
SA4018851655Mtn Early-submaint
SA4018861656Mtn Early-submaint
SA4018871657Mtn Early-submaint
SA4018881658Mtn Early-submaint
SA4018891659Mtn Early-submaint
SA4018901660Mtn Early-submaint
SA4018911661Mtn Early-submaint
SA4018921726Mtn Early-submaint
SA4018931739Mtn Early-submaint
SA4018941714Mtn Early-submaint
SA4018951736Mtn Early-submaint
SA4018961735Mtn Early-submaint
SA4018971741Mtn Early-submaint
SA4018981716Mtn Early-submaint
SA4018991731Mtn Early-submaint
SA4019001730Mtn Early-submaint
SA4019011715Mtn Early-submaint
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Collection:

Collection ID:CO003800
Collection Summary:A total of 388 serum samples were obtained from wild bighorn sheep populations in Wyoming (7 locations) and Montana (4 locations) via helicopter netgun capture techniques as described below: Animal captures occurred from December through March during the winters of 2014–15, 2015–16, and 2016–17, when all animals were on senescent native forages resulting in sub-maintenance diets. The majority of these animals, 385, were captured using net guns fired from a helicopter. This method required close pursuit by the helicopter, normally for 2–5 min, until a small net was deployed from a shoulder mounted gun that entangled the animal. A handler was place on the ground to physically restrain the captured animal via a blindfold and hobbles. Animals captured in remote wilderness were processed and released at the capture site, generally within 15–35 min of capture. In other situations with good ground access, the blindfolded and hobbled animals were placed in transport bags and slung under the helicopter on a long cable to a central processing site where they were processed, with blood samples normally collected between 20 and 60 min after capture. Large nets suspended over baited sites (dropnet) were dropped on 104 animals. Once the net was dropped, a large crew of handlers physically restrained the animals with blindfolds and hobbles, extracted each animal from the net, and carried it to a central processing area within 100 m of the dropnet site. Because the large nets captured 10–30 animals each, captured animals were queued for processing with blood samples generally drawn from 20 to 90 min following deployment of the nets. Ground-based delivery of immobilizing drugs, i.e. a cocktail of butorphanol, azaperone, and medetomidine; via dart rifles was used to capture 73 animals12. Animals were approached to within 5–15 m for effective dart delivery. Once stuck by the dart, the animals normally ran 5–20 m, and resumed pre-darting behaviors with their social group until the drugs began to take effect, causing the darted animal to bed down until sedated, normally 20–35 min following drug delivery. Sedated animals were blindfolded and hobbled, sampled, and drug antagonists administered, with processing time normally requiring 10–20 min. For all capture techniques, a blood sample was drawn from the jugular vein of each animal and immediately placed under refrigeration until serum was harvested 2–6 h after capture. Serum was frozen at − 20 °C for transport to research facilities where all samples were stored at − 80 °C until further processed. The majority of the samples originated from unique animals, but small numbers of marked animals were repeatedly captured and sampled in consecutive years in three of the Wyoming herds.
Sample Type:Blood (serum)

Treatment:

Treatment ID:TR003816
Treatment Summary:Due to seasonal changes, these wild animals were sustained on senescent native forages, resulting in sub-maintenance diets from late summer/early fall until the onset of the growing season the following spring. Animals captured in December months (n = 170) were classified as “early submaintenance” (Early-SM), animals captured in January (n = 35) were classified as “moderate submaintenance” (Mod-SM), and animals captured from February through March (n=183), which subsisted approximately 6 months on senescent forages, were classified as “severe submaintenance” (Se-vere-SM).

Sample Preparation:

Sampleprep ID:SP003814
Sampleprep Summary:Serum samples were prepared for small molecule polar metabolite extraction and 1H NMR metabolomics as follows: Samples were thawed at room temperature following storage at − 80 °C with reagents kept at − 20 °C until used. A 1:3 500 μL serum: 1500 μL acetone solution was added to 2 mL plastic, flat-cap conical vials13. The resulting solution was mixed thoroughly by inverting the sample tubes 10 times, and incubated at − 20 °C for 60 min to allow for protein precipitation, followed by sample centrifugation at 13,000×g for 30 min at room temperature. Clarified supernatants containing the polar metabolite mixtures were subsequently transferred to new 2.0 mL tubes and dried overnight using a Speedvac vacuum centrifuge with no heat, and stored at − 80 °C until further use. For NMR, dried metabolite extracts were resuspended in 600 μL of NMR buffer consisting of 25 mM of NaH2PO4/Na2HPO4, 0.4 mM of imidazole, 0.25 mM of 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS) in 90% H2O/10% D2O, pH 7.0.

Analysis:

Analysis ID:AN006036
Analysis Type:NMR
Num Factors:5
Num Metabolites:47
Units:Micromolar

NMR:

NMR ID:NM000304
Analysis ID:AN006036
Instrument Name:Bruker 600 MHz Avance III
Instrument Type:FT-NMR
NMR Experiment Type:1D-1H
NMR Comments:Bruker 600 MHz Avance III NMR Spectrometer (actively shielded) with 600 MHz TCI (H-C/N-D05Z) LT (liquid helium cooled) Helium CryoProbeTM. Available working temperature range: -40oC+80oC. Three channels: 1H, X1=13C, X2=15N, lock. Auto-tune/Auto-shim. SampleJetTM automated sample loading system.
Spectrometer Frequency:600 MHz
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