Summary of Study ST000049
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 PR000024. The data can be accessed directly via it's Project DOI: 10.21228/M85P4V This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000049 |
| Study Title | Metabolomics Analysis of Thermally Challenged Mayfly Larvae (NMR analysis) |
| Study Type | Metabolomic analysis of mayflies |
| Study Summary | The purpose of this study was to examine the metabolic profiles of mayfly (Centroptilum triangulifer) larvae subjected to thermal challenge. This species is unusual in terms of its ease of culture, and its suitability as a laboratory test organism. Our purpose here was to examine how an environmentally realistic thermal challenge affects the physiology of this organism. In this study, we obtained several types of insect species and we were able to show that NMR Metabolomics could be used to distinguish among the different types of larvae. |
| Institute | University of North Carolina |
| Department | Discovery Science Technology |
| Laboratory | Sumner Lab |
| Last Name | Sumner |
| First Name | Susan |
| Address | Eastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081 |
| susan_sumner @unc.edu | |
| Phone | 704-250-5066 |
| Submit Date | 2014-02-28 |
| Num Groups | 7 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | fid |
| Uploaded File Size | 7.6 M |
| Analysis Type Detail | NMR |
| Release Date | 2015-05-01 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR000024 |
| Project DOI: | doi: 10.21228/M85P4V |
| Project Title: | Metabolomics Analysis of Thermally Challenged Mayfly Larvae |
| Project Type: | MS/NMR |
| Project Summary: | The purpose of this study was to examine the metabolic profiles of mayfly (Centroptilum triangulifer) larvae subjected to a thermal challenge. This species is unusual in terms of its ease of culture, and its suitability as a laboratory test organism. Studies complementary to this metabolic study have focused on thermal effects on metabolic rates (respiration) and gene expression patterns in this species and do not support Prtners oxygen limitation paradigm. Our purpose here was to examine how an environmentally realistic thermal challenge affects the physiology of this organism. |
| Institute: | North Carolina State University |
| Department: | Environmental and Molecular Toxicology |
| Laboratory: | Buchwalter Laboratory |
| Last Name: | Buchwalter |
| First Name: | David |
| Address: | North Carolina State University Raleigh, NC 27695 |
| Email: | dbbuchwa@ncsu.edu |
| Phone: | 919-513-1129 |
Subject:
| Subject ID: | SU000067 |
| Subject Type: | Insect |
| Subject Species: | Centroptilum triangulifer;Atherix;Megaloptera |
| Taxonomy ID: | 2078957;124299;50553 |
| Species Group: | Insects |
Factors:
Subject type: Insect; Subject species: Centroptilum triangulifer;Atherix;Megaloptera (Factor headings shown in green)
| mb_sample_id | local_sample_id | species | Treatment | wing pads |
|---|---|---|---|---|
| SA002143 | AD_01 | Athorix Diptoa | Athorix Diptoa control | no wing pads |
| SA002144 | Pooled_QC_01 | Mayfly, Megalopterans, Athorix Diptoa | Pooled QC sample | no wing pads |
| SA002145 | Pooled_QC_02 | Mayfly, Megalopterans, Athorix Diptoa | Pooled QC sample | no wing pads |
| SA002146 | Pooled_QC_03 | Mayfly, Megalopterans, Athorix Diptoa | Pooled QC sample | no wing pads |
| SA002150 | MF_01 | Mayfly | Mayfly control from different study | no wing pads |
| SA002152 | Control_02 | Mayfly | Mayfly control | no wing pads |
| SA002153 | Control_03 | Mayfly | Mayfly control | no wing pads |
| SA002154 | Control_01 | Mayfly | Mayfly control | no wing pads |
| SA002151 | Control_X | Mayfly | Mayfly controlX | wing pads |
| SA002147 | Heat_Treated_03 | Mayfly | Mayfly Heat Treated | no wing pads |
| SA002148 | Heat_Treated_02 | Mayfly | Mayfly Heat Treated | no wing pads |
| SA002149 | Heat_Treated_01 | Mayfly | Mayfly Heat Treated | no wing pads |
| SA002155 | MG_01 | Megalopterans | Megalopterans control | no wing pads |
| Showing results 1 to 13 of 13 |
Collection:
| Collection ID: | CO000050 |
| Collection Summary: | whole insects were collected |
| Collection Protocol Comments: | Original culture material was obtained from the Stroud Water Research Center (Avondale, PA). Larvae were reared at 22°C and fed a periphytic diet. When larvae reached a suitable size, they were separated from their food source for 12 hours to evacuate gut contents. Larvae were then divided into two treatment groups a control group maintained at the culturing temperature of 22°C, and a thermal challenge group. The thermal challenge group was subjected to a temperature increase at a rate of 1°C per hour. This rate of temperature change is commonly observed in temperate streams. When the treatment temperature reached 30°C, larvae from both control and treatment groups were flash frozen in liquid nitrogen in groups of 12-13 larvae. Each grouping of 12-13 larvae comprised a composite replicate for the metabolic profiling analysis. A total of 3 replicates each for the control and thermal treatments were used in this study. Some larvae had developed dark wing pads during the experiment. This is a signal that the larvae were very close to emerging from the aquatic larval phase to a winged sub-adult phase. Enough of these larvae were present in the control treatment to be removed from the control cohort and considered as an independent sample. |
| Sample Type: | whole insect |
| Volumeoramount Collected: | 50 mg |
| Storage Conditions: | -80C |
Treatment:
| Treatment ID: | TR000068 |
| Treatment Summary: | Metabolomics profile of mayfly larvae exposed to heat |
| Treatment Protocol Comments: | Heat Treated | Control | ControlX |
| Treatment: | Abiotic |
| Animal Acclimation Duration: | exposed to heat (22C to 30C ramp); fasted for 8 hours | control (fasted for 8 hours) | control (fasted for 6 hours) |
Sample Preparation:
| Sampleprep ID: | SP000063 |
| Sampleprep Summary: | Control and heat-treated mayfly samples were provided in triplicate and were processed individually. One sample was provided for AD, ControlX, MF control (from a previous study) and MG larvae. Three replicates of the larvae listed above having only a single sample were created. Aliquots of 50-300mg of insect larvae were mixed with degassed 1:1 Acetonitrile:Water solution in a 2 mL snap cap tube at a concentration of 50 mg/mL. Megalopterans were substantially larger than the other larvae and were prepared at 200 mg/mL. Samples were homogenized and then centrifuged at 4?C for 5 minutes at 14000rcf. The supernatant was removed and placed into a new tube. Samples were centrifuged again at 4?C for 5 minutes at 14000rcf and a volume of the homogenate corresponding to 40 mg insect larvae were taken. After homogenization, the three larval replicates from AD, ControlX, MG and MF control (from a previous study) were pooled and a single sample created for each larval type. There was insufficient Mayfly (heat and control) sample material to create an internal pool, so an external pool was made by combining equal concentration amounts of AD, MG and MF control (from a previous study) pool quality check (QC) samples. Samples were completely dried by vacuum centrifuge and were reconstituted by adding 630 µL of D2O (Aldrich) and 70 µL of Chenomx ISTD solution (Chenomx, Edmonton, Alberta, Canada). The samples were vortexed and centrifuged at 14000rcf for 2 minutes. 650 µL of sample was transferred into 5mm NMR tubes and analyzed by NMR. |
| Sampleprep Protocol Filename: | RTI_Metabolomic_Analysis_of_Thermally_Challenged_Larvae_Procedure.docx |
| Processing Method: | Homogenization and Solvent Removal with SpeedVac |
| Processing Storage Conditions: | 4°C |
| Extraction Method: | 50% Aqueous Acetonitrile |
| Extract Concentration Dilution: | dried on SpeedVac |
| Extract Storage: | -80C |
| Sample Resuspension: | resuspended in D2O + Chenomx ISTD solution |
Analysis:
| Analysis ID: | AN000085 |
| Laboratory Name: | DHMRI |
| Analysis Type: | NMR |
| Acquisition Date: | 41624 |
| Software Version: | TopSpin 3.0 |
| Operator Name: | Kevin Knagge, Wimal Pathmasiri |
| Randomization Order: | Yes |
| Detector Type: | NMR |
| Data Format: | Bruker |
| Chromatography ID: | CH000054 |
| Num Factors: | 7 |
NMR:
| NMR ID: | NM000022 |
| Analysis ID: | AN000085 |
| Instrument Name: | Bruker Avance III |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D 1H |
| Field Frequency Lock: | D20 |
| Standard Concentration: | 0.5 mM |
| Spectrometer Frequency: | 950 MHz |
| NMR Probe: | 5mm Cryogenically Cooled ATMA |
| NMR Solvent: | D20 |
| NMR Tube Size: | 5mm |
| Shimming Method: | Topshim (Gradient) |
| Pulse Sequence: | noesypr1d |
| Water Suppression: | yes |
| Pulse Width: | 8.68 |
| Power Level: | 12.589W |
| Receiver Gain: | 4 |
| Offset Frequency: | 4468.30 Hz |
| Chemical Shift Ref Cpd: | DSS |
| Temperature: | 298 K |
| Number Of Scans: | 256 |
| Dummy Scans: | 4 |
| Acquisition Time: | 1.73 sec |
| Relaxation Delay: | 2s |
| Spectral Width: | 18939.395 Hz |
| Num Data Points Acquired: | 65536 |
| Real Data Points: | 65536 |
| Line Broadening: | 0.5 |
| Zero Filling: | Yes |
| Apodization: | Lorentzian |
| Baseline Correction Method: | Polynomial |
| Chemical Shift Ref Std: | DSS |
