Summary of Study ST001222
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 PR000799. The data can be accessed directly via it's Project DOI: 10.21228/M8D98R This work is supported by NIH grant, U2C- DK119886.
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| Study ID | ST001222 |
| Study Title | Effects of selenate and cadmium exposure on the honey bee metabolome (part-II) |
| Study Summary | We moved one frame of brood each from five healthy honey bee colonies with marked Italian queens and housed them in a hive body at 35°C and 50% humidity under constant darkness. We then allowed the bees to emerge, mixed the newly emerged workers (NEWs) to randomize their colony of origin and placed NEWs into 13 cm x 10.5 cm x 6.5 cm wire cages equipped with feeders containing 35mL of deionized water and 35mL 50% sucrose. We also provided a pollen patty to each cage of bees consisting of 269g corn syrup, 113g sucrose and 113g of Bee Pro (Mann Lake, Hackensack, MN). To inoculate the newly emerged workers with their “core” microbiome, we collected 50 mL of foragers from the source hives of the NEWs, immobilized the bees at 4°C, aseptically dissected out the abdomens and macerated the whole abdomens in 50% sucrose. We added 1 mL of the resulting slurry to 34 mL of 50% sucrose solution and fed it to the NEWs. We allowed the bees to feed ad libitium on the mixture for two days before replacing the feeders with 50% sucrose. We allowed the bees to feed for three more days to fully establish a microbiome. Once the bees had an established microbiome, we prepared treatment feeding solutions of 50% sucrose (as a no metal/metalloid control), 50% sucrose spiked with 0.6 mg/L sodium selenate or 50% sucrose with 0.24 mg/L cadmium chloride (Alfa Aesar, Ward Hill, MA) and pollen patties spiked with either 6.0 mg/L selenium or 0.46 mg/L cadmium as in Hladun et al 2015. We again allowed the bees to feed ad libitium. We sampled three bees from 13 cages after four days of continuous exposure to the above-mentioned treatments and immediately placed the samples on dry ice, followed by long-term storage at -80 °C. |
| Institute | University of California, Riverside |
| Last Name | Rothman |
| First Name | Jason |
| Address | 900 University Ave. |
| jroth002@ucr.edu | |
| Phone | 9518275817 |
| Submit Date | 2019-07-17 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Waters) |
| Analysis Type Detail | LC-MS |
| Release Date | 2019-09-23 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR000799 |
| Project DOI: | doi: 10.21228/M8D98R |
| Project Title: | Untargeted metabolomics of honey bees exposed to selenate or cadmium |
| Project Summary: | Effects of selenate and cadmium exposure on honey bees. |
| Institute: | University of California, Riverside |
| Last Name: | Rothman |
| First Name: | Jason |
| Address: | 900 University Ave., Riverside, CA, 91766, USA |
| Email: | jroth002@ucr.edu |
| Phone: | 9518275817 |
Subject:
| Subject ID: | SU001289 |
| Subject Type: | Invertebrate |
| Subject Species: | Apis mellifera |
| Taxonomy ID: | 7460 |
Factors:
Subject type: Invertebrate; Subject species: Apis mellifera (Factor headings shown in green)
| mb_sample_id | local_sample_id | Factor |
|---|---|---|
| SA086278 | cadmium-3 | Cadmium |
| SA086279 | cadmium-4 | Cadmium |
| SA086280 | cadmium-1 | Cadmium |
| SA086281 | cadmium-2 | Cadmium |
| SA086282 | control-2 | Control |
| SA086283 | control-3 | Control |
| SA086284 | control-1 | Control |
| SA086285 | control-4 | Control |
| SA086286 | QC-2 | QCPool |
| SA086287 | QC-3 | QCPool |
| SA086288 | QC-1 | QCPool |
| SA086289 | selenium-1 | Selenate |
| SA086290 | selenium-2 | Selenate |
| SA086291 | selenium-3 | Selenate |
| SA086292 | selenium-4 | Selenate |
| SA086293 | selenium-5 | Selenate |
| Showing results 1 to 16 of 16 |
Collection:
| Collection ID: | CO001283 |
| Collection Summary: | We sampled three bees from 13 cages after four days of continuous exposure to the treatments and immediately placed the samples on dry ice, followed by long-term storage at -80 °C. |
| Sample Type: | Insect tissue |
| Storage Conditions: | Described in summary |
Treatment:
| Treatment ID: | TR001304 |
| Treatment Summary: | Once the bees had an established microbiome, we prepared treatment feeding solutions of 50% sucrose (as a no metal/metalloid control), 50% sucrose spiked with 0.6 mg/L sodium selenate or 50% sucrose with 0.24 mg/L cadmium chloride (Alfa Aesar, Ward Hill, MA) and pollen patties spiked with either 6.0 mg/L selenium or 0.46 mg/L cadmium and allowed the bees to feed ad libitium. |
Sample Preparation:
| Sampleprep ID: | SP001297 |
| Sampleprep Summary: | Samples were freeze-dried, homogenized, and extracted with 30:30:20:20 acetonitrile:methanol:water:isopropanol. Samples were then sonicated, vortexed, and centrifuged before analysis. |
Combined analysis:
| Analysis ID | AN002035 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Waters Acquity I-Class |
| Column | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Waters Xevo TQ-S |
| Ion Mode | POSITIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH001475 |
| Chromatography Summary: | We analyzed targeted, polar primary metabolites on a Xevo TQ-XS triple quadrupole mass spectrometer (Waters) coupled to an I-class UPLC system (Waters) in the UC Riverside Metabolomics Core Facility. We used a ZIC-pHILIC column (2.1 x 150 mm, 5 µM) (EMD Millipore) for separations with the following mobile phases:(A) water with 15 mM ammonium bicarbonate adjusted to pH 9.6 with ammonium hydroxide and (B) acetonitrile. We set the flow rate to 200 µL/min and held the column 50° C with an injection volume of 1 µL. We used the following gradient: 0 min, 90% B; 1.5 min, 90% B; 16 min, 20% B; 18 min, 20% B; 20 min, 90% B; 28 min, 90% B. |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0 min, 90% B; 1.5 min, 90% B; 16 min, 20% B; 18 min, 20% B; 20 min, 90% B; 28 min, 90% B |
| Flow Rate: | 200 ul/min |
| Solvent A: | 100% water; 15 mM ammonium bicarbonate, pH 9.6 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS001887 |
| Analysis ID: | AN002035 |
| Instrument Name: | Waters Xevo TQ-S |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | We operated the MS in selected reaction monitoring mode with source and desolvation temperatures at 150° C and 500° C, respectively, with the desolvation gas set to 1000 L/hr, cone gas set to 150 L/hr and collision gas set to 0.15 mL/min. We used nitrogen gas for each step, except for the collision gas, which was argon, and we set capillary voltage to 1 kV in positive ion mode and 2 kV in negative ion mode. Similar to the untargeted methods, we generated a quality control sample by pooling equal aliquots of each sample and analyzed the QC sample every 3-4 injections to monitor system stability and performance. We processed the metabolite data (peak picking, alignment, deconvolution, integration, normalization, and spectral matching) with Progenesis Qi software (Nonlinear Dynamics, Durham, NC). We used Skyline software (MacLean et al. 2010) for data processing our targeted metabolites. |
| Ion Mode: | POSITIVE |
