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 IDST001222
Study TitleEffects of selenate and cadmium exposure on the honey bee metabolome (part-II)
Study SummaryWe 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 NameRothman
First NameJason
Address900 University Ave.
Emailjroth002@ucr.edu
Phone9518275817
Submit Date2019-07-17
Raw Data AvailableYes
Raw Data File Type(s).d, .xsd, .ini, .bin, .xml, .m
Analysis Type DetailLC-MS
Release Date2019-09-23
Release Version1
Jason Rothman Jason Rothman
https://dx.doi.org/10.21228/M8D98R
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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
SA086278cadmium-3Cadmium
SA086279cadmium-4Cadmium
SA086280cadmium-1Cadmium
SA086281cadmium-2Cadmium
SA086282control-2Control
SA086283control-3Control
SA086284control-1Control
SA086285control-4Control
SA086286QC-2QCPool
SA086287QC-3QCPool
SA086288QC-1QCPool
SA086289selenium-1Selenate
SA086290selenium-2Selenate
SA086291selenium-3Selenate
SA086292selenium-4Selenate
SA086293selenium-5Selenate
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 Normal phase
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:water with 15 mM ammonium bicarbonate adjusted to pH 9.6 with ammonium hydroxide
Solvent B:acetonitrile
Chromatography Type:Normal phase

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
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