{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001187","ANALYSIS_ID":"AN001979","VERSION":"1","CREATED_ON":"June 2, 2019, 12:30 pm"},

"PROJECT":{"PROJECT_TITLE":"Untargeted metabolomics of honey bees exposed to selenate or cadmium","PROJECT_SUMMARY":"Effects of selenate and cadmium exposure on honey bees.","INSTITUTE":"UC Riverside","LAST_NAME":"Rothman","FIRST_NAME":"Jason","ADDRESS":"900 University Ave., Riverside, CA, 91766, USA","EMAIL":"jroth002@ucr.edu","PHONE":"9518275817"},

"STUDY":{"STUDY_TITLE":"Effects of selenate and cadmium exposure on the honey bee metabolome","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":"UC Riverside","LAST_NAME":"Rothman","FIRST_NAME":"Jason","ADDRESS":"900 University Ave.","EMAIL":"jroth002@ucr.edu","PHONE":"9518275817"},

"SUBJECT":{"SUBJECT_TYPE":"Insect","SUBJECT_SPECIES":"Apis mellifera","TAXONOMY_ID":"7460"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"QC-1",
"Factors":{"Treatment":"Pool"}
},
{
"Subject ID":"-",
"Sample ID":"HBCT-4",
"Factors":{"Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"HBSE-2",
"Factors":{"Treatment":"Selenate"}
},
{
"Subject ID":"-",
"Sample ID":"HBSE-4",
"Factors":{"Treatment":"Selenate"}
},
{
"Subject ID":"-",
"Sample ID":"QC-2",
"Factors":{"Treatment":"Pool"}
},
{
"Subject ID":"-",
"Sample ID":"HBSE-5",
"Factors":{"Treatment":"Selenate"}
},
{
"Subject ID":"-",
"Sample ID":"HBCT-3",
"Factors":{"Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"HBCD-3",
"Factors":{"Treatment":"Cadmium"}
},
{
"Subject ID":"-",
"Sample ID":"QC-3",
"Factors":{"Treatment":"Pool"}
},
{
"Subject ID":"-",
"Sample ID":"HBSE-1",
"Factors":{"Treatment":"Selenate"}
},
{
"Subject ID":"-",
"Sample ID":"HBCT-1",
"Factors":{"Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"HBCD-4",
"Factors":{"Treatment":"Cadmium"}
},
{
"Subject ID":"-",
"Sample ID":"HBCT-2",
"Factors":{"Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"QC-4",
"Factors":{"Treatment":"Pool"}
},
{
"Subject ID":"-",
"Sample ID":"HBCD-2",
"Factors":{"Treatment":"Cadmium"}
},
{
"Subject ID":"-",
"Sample ID":"HBSE-3",
"Factors":{"Treatment":"Selenate"}
},
{
"Subject ID":"-",
"Sample ID":"HBCD-1",
"Factors":{"Treatment":"Cadmium"}
},
{
"Subject ID":"-",
"Sample ID":"QC-5",
"Factors":{"Treatment":"Pool"}
}
],
"COLLECTION":{"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_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."},

"SAMPLEPREP":{"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."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Normal phase","INSTRUMENT_NAME":"Waters Acquity I-Class","COLUMN_NAME":"Waters Acquity CSH C18 (100 x 2.1mm, 1.7um)","FLOW_GRADIENT":"0 min, 1% B; 1 min, 1% B; 8 min, 40% B; 24 min, 100% B; 26.5 min, 100% B; 27 min, 1% B","FLOW_RATE":"250 ul/min","COLUMN_TEMPERATURE":"40","SOLVENT_A":"0.1% formic acid","SOLVENT_B":"Acetonitrile with 0.1% formic acid"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"UC Riverside Metabolomics Core Facility"},

"MS":{"INSTRUMENT_NAME":"Waters Synapt G2 Si QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"We processed the metabolite data (peak picking, alignment, deconvolution, integration, normalization, and spectral matching) with Progenesis Qi software (Nonlinear Dynamics, Durham, NC).","MS_RESULTS_FILE":"ST001187_AN001979_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}