{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000495","ANALYSIS_ID":"AN000761","VERSION":"1","CREATED_ON":"October 28, 2016, 12:11 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolomic profiles along the gastrointestinal tract of the healthy dog","PROJECT_SUMMARY":"Introduction: The fecal microbiome is relevant to the health and disease of many species. The importance of the fecal metabolome has more recently been appreciated, but our knowledge of the microbiome and metabolome at other sites along the gastrointestinal tract remains deficient. Objective: To analyze the gastrointestinal microbiome and metabolome of healthy domestic dogs at four anatomical sites. Methods: Samples of the duodenal, ileal, colonic, and rectal contents were collected from six adult dogs after humane euthanasia for an unrelated study. The microbiota were characterized using Illumina sequencing of 16S rRNA genes. The metabolome was characterized by mass spectrometry-based methods. Results: Prevalent phyla throughout the samples were Proteobacteria, Firmicutes, Fusobacteria, and Bacteroidetes, consistent with previous findings in dogs and other species. A total of 530 unique metabolites were detected; 199 of these were identified as previously named compounds, but 141 of them had at least one significantly different site-pair comparison. Noteworthy examples include amino acids, which decreased from the small to large intestine; pyruvate, which was at peak concentrations in the ileum; and several phenol-containing carboxylic acid compounds that increased in the large intestine. Conclusion: The microbiome and metabolome vary significantly at different sites along the canine gastrointestinal tract.","INSTITUTE":"Texas A&M University","DEPARTMENT":"Department of Small Animal Clinical Sciences","LABORATORY":"Gastrointestinal Laboratory","LAST_NAME":"Steiner","FIRST_NAME":"Jorg","ADDRESS":"College Station, TX 77843-4474","EMAIL":"JHonneffer@cvm.tamu.edu","PHONE":"(979)862-2861"},

"STUDY":{"STUDY_TITLE":"Metabolomic profiles along the gastrointestinal tract of the healthy dog","STUDY_SUMMARY":"Introduction: The fecal microbiome is relevant to the health and disease of many species. The importance of the fecal metabolome has more recently been appreciated, but our knowledge of the microbiome and metabolome at other sites along the gastrointestinal tract remains deficient. Objective: To analyze the gastrointestinal microbiome and metabolome of healthy domestic dogs at four anatomical sites. Methods: Samples of the duodenal, ileal, colonic, and rectal contents were collected from six adult dogs after humane euthanasia for an unrelated study. The microbiota were characterized using Illumina sequencing of 16S rRNA genes. The metabolome was characterized by mass spectrometry-based methods. Results: Prevalent phyla throughout the samples were Proteobacteria, Firmicutes, Fusobacteria, and Bacteroidetes, consistent with previous findings in dogs and other species. A total of 530 unique metabolites were detected; 199 of these were identified as previously named compounds, but 141 of them had at least one significantly different site-pair comparison. Noteworthy examples include amino acids, which decreased from the small to large intestine; pyruvate, which was at peak concentrations in the ileum; and several phenol-containing carboxylic acid compounds that increased in the large intestine. Conclusion: The microbiome and metabolome vary significantly at different sites along the canine gastrointestinal tract.","INSTITUTE":"University of California, Davis","DEPARTMENT":"Genome and Biomedical Sciences Facility","LABORATORY":"WCMC Metabolomics Core","LAST_NAME":"Fiehn","FIRST_NAME":"Oliver","ADDRESS":"1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616","EMAIL":"ofiehn@ucdavis.edu","PHONE":"(530) 754-8258","NUM_GROUPS":"4","TOTAL_SUBJECTS":"24","NUM_MALES":"4","NUM_FEMALES":"2"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Canis lupus familiaris","TAXONOMY_ID":"9615","GENOTYPE_STRAIN":"Hound-type","AGE_OR_AGE_RANGE":"Adult","WEIGHT_OR_WEIGHT_RANGE":"35-60 lbs","GENDER":"Male/Female","ANIMAL_INCLUSION_CRITERIA":"no evidence of gastrointestinal illness or other disease"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"Subject name (if any)",
"Sample ID":"Sample name",
"Factors":{"Organ":"Organ","Sex":"Sex"},
"Additional sample data":{"Species":"Species","Donor":"Donor"}
},
{
"Subject ID":"194106",
"Sample ID":"141217actsa19_1",
"Factors":{"Organ":"colon","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"A"}
},
{
"Subject ID":"194107",
"Sample ID":"141217actsa04_1",
"Factors":{"Organ":"colon","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"C"}
},
{
"Subject ID":"194108",
"Sample ID":"141217actsa05_1",
"Factors":{"Organ":"colon","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"D"}
},
{
"Subject ID":"194109",
"Sample ID":"141217actsa10_1",
"Factors":{"Organ":"colon","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"E"}
},
{
"Subject ID":"194110",
"Sample ID":"141217actsa23_1",
"Factors":{"Organ":"colon","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"F"}
},
{
"Subject ID":"194111",
"Sample ID":"141217actsa22_1",
"Factors":{"Organ":"colon","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"G"}
},
{
"Subject ID":"194099",
"Sample ID":"141217actsa06_1",
"Factors":{"Organ":"rectum","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"A"}
},
{
"Subject ID":"194100",
"Sample ID":"141217actsa12_1",
"Factors":{"Organ":"rectum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"C"}
},
{
"Subject ID":"194101",
"Sample ID":"141217actsa16_1",
"Factors":{"Organ":"rectum","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"D"}
},
{
"Subject ID":"194102",
"Sample ID":"141217actsa11_1",
"Factors":{"Organ":"rectum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"E"}
},
{
"Subject ID":"194103",
"Sample ID":"141217actsa08_1",
"Factors":{"Organ":"rectum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"F"}
},
{
"Subject ID":"194104",
"Sample ID":"141217actsa03_1",
"Factors":{"Organ":"rectum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"G"}
},
{
"Subject ID":"194092",
"Sample ID":"141217actsa17_1",
"Factors":{"Organ":"duod","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"A"}
},
{
"Subject ID":"194093",
"Sample ID":"141217actsa21_1",
"Factors":{"Organ":"duod","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"C"}
},
{
"Subject ID":"194094",
"Sample ID":"141217actsa13_1",
"Factors":{"Organ":"duod","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"D"}
},
{
"Subject ID":"194095",
"Sample ID":"141217actsa24_1",
"Factors":{"Organ":"duod","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"E"}
},
{
"Subject ID":"194096",
"Sample ID":"141217actsa20_1",
"Factors":{"Organ":"duod","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"F"}
},
{
"Subject ID":"194097",
"Sample ID":"141217actsa09_1",
"Factors":{"Organ":"duod","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"G"}
},
{
"Subject ID":"194085",
"Sample ID":"141217actsa07_1",
"Factors":{"Organ":"ileum","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"A"}
},
{
"Subject ID":"194086",
"Sample ID":"141217actsa02_1",
"Factors":{"Organ":"ileum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"C"}
},
{
"Subject ID":"194087",
"Sample ID":"141217actsa18_1",
"Factors":{"Organ":"ileum","Sex":"Female"},
"Additional sample data":{"Species":"Dog","Donor":"D"}
},
{
"Subject ID":"194088",
"Sample ID":"141217actsa14_1",
"Factors":{"Organ":"ileum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"E"}
},
{
"Subject ID":"194089",
"Sample ID":"141217actsa15_1",
"Factors":{"Organ":"ileum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"F"}
},
{
"Subject ID":"194090",
"Sample ID":"141217actsa01_1",
"Factors":{"Organ":"ileum","Sex":"Male"},
"Additional sample data":{"Species":"Dog","Donor":"G"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Animals were euthanized for an unrelated study. Within 1-3 hours after euthanasia, a ventral midline approach was used to access the gastrointestinal tract. A longitudinal incision was made through the serosal surface at each site sampled to expose a length of the lumen. Disposable plastic spatulae were used to collect intestinal contents from each site. Duodenal samples were collected aborally to the cranial flexure. The ileum was identified by the antimesenteric artery. The colon was identified by location in situ and samples were collected from the transverse or descending colon. Rectal contents were collected from as caudally as possible while still sampling through the abdominal incision.","SAMPLE_TYPE":"Other (digesta/intestinal contents)","COLLECTION_FREQUENCY":"Once","COLLECTION_DURATION":"Once","VOLUMEORAMOUNT_COLLECTED":"Five aliquots of 50-100mg","STORAGE_CONDITIONS":"-80°C","COLLECTION_VIALS":"2 mL screw cap polypropylene vials","STORAGE_VIALS":"2 mL screw cap polypropylene vials","COLLECTION_TUBE_TEMP":"room temp"},

"TREATMENT":{"TREATMENT_SUMMARY":"The variable tested across the samples was the site of the GI tract, so none of the animals received any treatment for the purpose of comparing the effect of it to a control. The individual dogs are indicated by the letter A, C, D, E, F, and G. For each dog, we collected samples from duodenum, ileum, colon, and rectum."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"1. Weigh 50 mg tissue sample in to a 25 ml conical polypropylene centrifuge tube. 2. Add 2.5mL extraction solvent to the tissue sample and homogenize for 45 seconds ensuring that sample resembles a powder. In between samples, clean the homogenizer in solutions of methanol, acetone, water, and the extraction solvent. 3. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500μl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 4. Evaporate one 500μl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 5. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) 6. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 7. Remove supernatant to a new Eppendorff tube. 8. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 9. Submit to derivatization."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Leco Pegasus III GC","COLUMN_NAME":"Restek Corporation Rtx-5Sil MS","FLOW_RATE":"1 ml/min","COLUMN_TEMPERATURE":"50-330C","METHODS_FILENAME":"Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf","COLUMN_PRESSURE":"7.7 PSI","INJECTION_TEMPERATURE":"50 C ramped to 250 C by 12 C/s","SAMPLE_INJECTION":"0.5 uL","OVEN_TEMPERATURE":"50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min","TRANSFERLINE_TEMPERATURE":"230C","WASHING_BUFFER":"Ethyl Acetate","SAMPLE_LOOP_SIZE":"30 m length x 0.25 mm internal diameter","RANDOMIZATION_ORDER":"Excel generated"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"WCMC Metabolomics Core","DETECTOR_TYPE":"TOF","SOFTWARE_VERSION":"ChromaTOF","DATA_FORMAT":".peg, .txt, .cdf"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Leco Pegasus III GC TOF","INSTRUMENT_TYPE":"GC-TOF","MS_TYPE":"EI","ION_MODE":"POSITIVE","ION_SOURCE_TEMPERATURE":"250 C","IONIZATION_ENERGY":"70 eV","MASS_ACCURACY":"Nominal","SOURCE_TEMPERATURE":"250 C","SCAN_RANGE_MOVERZ":"85-500 Da","SCANNING_CYCLE":"17 Hz","SCANNING_RANGE":"85-500 Da","SKIMMER_VOLTAGE":"1850 V"},

"MS_METABOLITE_DATA":{
"Units":"counts",

"Data":[{"Metabolite":"xylulose 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acid","141217actsa19_1":"25","141217actsa04_1":"1419","141217actsa05_1":"95","141217actsa10_1":"61","141217actsa23_1":"79","141217actsa22_1":"275","141217actsa06_1":"96","141217actsa12_1":"428","141217actsa16_1":"104","141217actsa11_1":"124","141217actsa08_1":"201","141217actsa03_1":"194","141217actsa17_1":"77","141217actsa21_1":"627","141217actsa13_1":"600","141217actsa24_1":"45","141217actsa20_1":"26","141217actsa09_1":"144","141217actsa07_1":"603","141217actsa02_1":"1193","141217actsa18_1":"705","141217actsa14_1":"226","141217actsa15_1":"222","141217actsa01_1":"784"},{"Metabolite":"xanthosine","141217actsa19_1":"137","141217actsa04_1":"77","141217actsa05_1":"346","141217actsa10_1":"64","141217actsa23_1":"343","141217actsa22_1":"240","141217actsa06_1":"264","141217actsa12_1":"50","141217actsa16_1":"214","141217actsa11_1":"37","141217actsa08_1":"147","141217actsa03_1":"482","141217actsa17_1":"288","141217actsa21_1":"4128","141217actsa13_1":"4095","141217actsa24_1":"704","141217actsa20_1":"321","141217actsa09_1":"3205","141217actsa07_1":"668","141217actsa02_1":"370","141217actsa18_1":"1708","141217actsa14_1":"967","141217actsa15_1":"741","141217actsa01_1":"437"},{"Metabolite":"xanthine","141217actsa19_1":"10189","141217actsa04_1":"7095","141217actsa05_1":"17674","141217actsa10_1":"2162","141217actsa23_1":"21217","141217actsa22_1":"6401","141217actsa06_1":"8582","141217actsa12_1":"2565","141217actsa16_1":"9422","141217actsa11_1":"6995","141217actsa08_1":"2247","141217actsa03_1":"14368","141217actsa17_1":"38441","141217actsa21_1":"142127","141217actsa13_1":"159664","141217actsa24_1":"62276","141217actsa20_1":"15556","141217actsa09_1":"116956","141217actsa07_1":"42108","141217actsa02_1":"50746","141217actsa18_1":"45848","141217actsa14_1":"36091","141217actsa15_1":"31998","141217actsa01_1":"47774"},{"Metabolite":"vanillic 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