#METABOLOMICS WORKBENCH engelmab_20241028_083820 DATATRACK_ID:5321 STUDY_ID:ST003554 ANALYSIS_ID:AN005843 PROJECT_ID:PR002188 VERSION 1 CREATED_ON November 6, 2024, 8:16 pm #PROJECT PR:PROJECT_TITLE Comprehension of the age-dependent gut and brain interaction of honeybee workers PR:PROJECT_TITLE by integration of multi omics approaches PR:PROJECT_SUMMARY In honeybees, division of labour is a key feature, with age-related behavioural PR:PROJECT_SUMMARY transitions being closely associated with molecular changes in the brain, gut, PR:PROJECT_SUMMARY and microbiota. In this study, to investigate these molecular changes and thus PR:PROJECT_SUMMARY better understand their contribution to behavioural responses and modulation, we PR:PROJECT_SUMMARY analysed the global metabolomic shifts in honeybee workers and their microbiota PR:PROJECT_SUMMARY throughout their lives. Overall, our findings provide new insights toward PR:PROJECT_SUMMARY developing potential biomarkers for evaluation of different functional changes PR:PROJECT_SUMMARY related to various environmental stressors. PR:INSTITUTE Helmholtz Centre for Environmental Research PR:DEPARTMENT Molecular Systems Biology PR:LAST_NAME Engelmann PR:FIRST_NAME Beatrice PR:ADDRESS Permoserstraße 15, Leipzipg, Saxony, 03418, Germany PR:EMAIL beatrice.engelmann@ufz.de PR:PHONE 004934160251099 #STUDY ST:STUDY_TITLE Investigation of age-dependent changes in the brain of honeybee workers using ST:STUDY_TITLE targeted metabolomics for amino acid and biogenic amine determination ST:STUDY_SUMMARY In the honeybee guts we measured the concentrations of 21 amino acids and 6 ST:STUDY_SUMMARY biogenic amines. Using a k-means clustering, the metabolites were grouped by ST:STUDY_SUMMARY their trends into six clusters. Tyramine and tyrosine are involved in the ST:STUDY_SUMMARY octopamine and dopamine biosynthesis. We found that all four metabolites were ST:STUDY_SUMMARY part of the same cluster, with their concentrations being significantly ST:STUDY_SUMMARY decreased in comparison to newly emerged workers from day 1 onwards for the ST:STUDY_SUMMARY entire duration of the study. ST:INSTITUTE Helmholtz Centre for Environmental Research ST:DEPARTMENT Molecular Systems Biology ST:LAST_NAME Engelmann ST:FIRST_NAME Beatrice ST:ADDRESS Permoserstraße 15, Leipzipg, Saxony, 03418, Germany ST:EMAIL beatrice.engelmann@ufz.de ST:PHONE 004934160251099 #SUBJECT SU:SUBJECT_TYPE Insect SU:SUBJECT_SPECIES Apis mellifera SU:TAXONOMY_ID 7460 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS H1_D0_1 H1_D0_1 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H1_D0_2 H1_D0_2 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H1_D0_3 H1_D0_3 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H1_D0_4 H1_D0_4 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=1; 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RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_D5_5 H1_D5_5_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_D7_1 H1_D7_1_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_D7_2 H1_D7_2_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_D7_3 H1_D7_3_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_D7_4 H1_D7_4_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_D7_5 H1_D7_5_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_NF_1 H1_NF_1_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H1_EF_1 H1_EF_1_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=1; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_1 H2_D0_1 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_2 H2_D0_2 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_3 H2_D0_3 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_4 H2_D0_4 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_5 H2_D0_5 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_1 H2_D1_1 life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_2 H2_D1_2 life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_3 H2_D1_3 life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_4 H2_D1_4 life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_5 H2_D1_5 life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_1 H2_D3_1 life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_2 H2_D3_2 life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_3 H2_D3_3 life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_4 H2_D3_4 life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_5 H2_D3_5 life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_1 H2_D5_1 life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_2 H2_D5_2 life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_3 H2_D5_3 life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_4 H2_D5_4 life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_5 H2_D5_5 life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_1 H2_D7_1 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_2 H2_D7_2 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_3 H2_D7_3 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_4 H2_D7_4 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_5 H2_D7_5 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_1 H2_NF_1 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_2 H2_NF_2 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_3 H2_NF_3 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_4 H2_NF_4 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_5 H2_NF_5 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_1 H2_EF_1 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_2 H2_EF_2 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_3 H2_EF_3 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_4 H2_EF_4 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_5 H2_EF_5 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_1 H2_D0_1_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_2 H2_D0_2_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_3 H2_D0_3_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_4 H2_D0_4_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D0_5 H2_D0_5_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_1 H2_D1_1_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_2 H2_D1_2_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_3 H2_D1_3_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_4 H2_D1_4_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D1_5 H2_D1_5_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_1 H2_D3_1_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_2 H2_D3_2_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_3 H2_D3_3_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_4 H2_D3_4_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D3_5 H2_D3_5_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_1 H2_D5_1_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_2 H2_D5_2_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_3 H2_D5_3_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_4 H2_D5_4_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D5_5 H2_D5_5_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_1 H2_D7_1_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_2 H2_D7_2_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_3 H2_D7_3_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_4 H2_D7_4_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_D7_5 H2_D7_5_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_1 H2_NF_1_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_2 H2_NF_2_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_3 H2_NF_3_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_4 H2_NF_4_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_NF_5 H2_NF_5_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_1 H2_EF_1_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_2 H2_EF_2_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_3 H2_EF_3_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_4 H2_EF_4_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H2_EF_5 H2_EF_5_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=2; RAW_FILE_NAME(Raw_data)=240405_CU_Eli_Batch2_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_1 H3_D0_1 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_2 H3_D0_2 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_3 H3_D0_3 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_4 H3_D0_4 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_5 H3_D0_5 life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_1 H3_D1_1 life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_2 H3_D1_2 life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_3 H3_D1_3 life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_4 H3_D1_4 life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_5 H3_D1_5 life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_1 H3_D3_1 life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_2 H3_D3_2 life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_3 H3_D3_3 life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_4 H3_D3_4 life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_5 H3_D3_5 life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_1 H3_D5_1 life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_2 H3_D5_2 life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_3 H3_D5_3 life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_4 H3_D5_4 life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_5 H3_D5_5 life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_1 H3_D7_1 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_2 H3_D7_2 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_3 H3_D7_3 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_4 H3_D7_4 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_5 H3_D7_5 life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_1 H3_NF_1 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_2 H3_NF_2 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_3 H3_NF_3 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_4 H3_NF_4 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_5 H3_NF_5 life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_1 H3_EF_1 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_2 H3_EF_2 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_3 H3_EF_3 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_4 H3_EF_4 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_5 H3_EF_5 life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240320_Eli_YY_Bee_Brain_undiluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_1 H3_D0_1_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_2 H3_D0_2_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_3 H3_D0_3_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_4 H3_D0_4_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D0_5 H3_D0_5_diluted life stage:newly_emerged | age of workers in days:0 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_1 H3_D1_1_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_2 H3_D1_2_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_3 H3_D1_3_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_4 H3_D1_4_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D1_5 H3_D1_5_diluted life stage:establishment | age of workers in days:1 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_1 H3_D3_1_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_2 H3_D3_2_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_3 H3_D3_3_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_4 H3_D3_4_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D3_5 H3_D3_5_diluted life stage:establishment | age of workers in days:3 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_1 H3_D5_1_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_2 H3_D5_2_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_3 H3_D5_3_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_4 H3_D5_4_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D5_5 H3_D5_5_diluted life stage:establishment | age of workers in days:5 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_1 H3_D7_1_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_2 H3_D7_2_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_3 H3_D7_3_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_4 H3_D7_4_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_D7_5 H3_D7_5_diluted life stage:nurse | age of workers in days:7-10 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_1 H3_NF_1_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_2 H3_NF_2_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_3 H3_NF_3_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_4 H3_NF_4_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_NF_5 H3_NF_5_diluted life stage:new_foragers | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_1 H3_EF_1_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_2 H3_EF_2_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_3 H3_EF_3_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_4 H3_EF_4_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff SUBJECT_SAMPLE_FACTORS H3_EF_5 H3_EF_5_diluted life stage:experienced_forager | age of workers in days:>12 | Sample source:brain hive=3; RAW_FILE_NAME(Raw_data)=240404_CU_Eli_Batch1_Brain_diluted.wiff #COLLECTION CO:COLLECTION_SUMMARY Workers were pooled to obtain a final sample number of n = 5. The brain(s) and CO:COLLECTION_SUMMARY gut(s) from the same workers were assigned the same sample names. In total, we CO:COLLECTION_SUMMARY had 15 samples from each age group, with the exception of the foraging stages, CO:COLLECTION_SUMMARY where owing to the lack of foragers from Hive 1, we only had 11. All sampled CO:COLLECTION_SUMMARY bees were placed directly in liquid nitrogen to prevent changes in the metabolic CO:COLLECTION_SUMMARY profile. They were then decapitated in the laboratory so that the head and body CO:COLLECTION_SUMMARY of each bee were kept in a separate Eppendorf tube (Eppendorf, Germany) for CO:COLLECTION_SUMMARY later dissection. Honeybee brains were dissected using a dissecting microscope CO:COLLECTION_SUMMARY (SZX7, Olympus). The head was placed on a petri dish lined with SYLGARD® 184 CO:COLLECTION_SUMMARY silicone. Before pinning the head for dissection, antennae and mouthpieces were CO:COLLECTION_SUMMARY removed. Subsequently, double-distilled water (ddH2O) was added until the head CO:COLLECTION_SUMMARY was fully submerged to facilitate dissection. The head cuticle was removed, and CO:COLLECTION_SUMMARY the hypopharyngeal glands and trachea were extracted from the inside. The brain CO:COLLECTION_SUMMARY was then separated from the cuticle and remaining tracheal remnants were removed CO:COLLECTION_SUMMARY before transferring the clean brain to a new Eppendorf tube. CO:SAMPLE_TYPE Bee brain #TREATMENT TR:TREATMENT_SUMMARY On the day of newly emerged workers (day 0), bees were samples as a baseline for TR:TREATMENT_SUMMARY no or low bacterial abundance in the gut. Moreover, bees from each hive were TR:TREATMENT_SUMMARY collected on Day 1, 3 and 5 to cover the establishment phase of the gut. Samples TR:TREATMENT_SUMMARY for the in-hive worker phase, defined as workers inside the hives that have a TR:TREATMENT_SUMMARY fully developed microbiome, were taken every day from Day 7 to Day 10. For the TR:TREATMENT_SUMMARY foraging phase, we differentiated between new and experienced foragers, as these TR:TREATMENT_SUMMARY show different behaviour due to different levels of experience.Based on TR:TREATMENT_SUMMARY experience, they were either collected one to two days after onset (new TR:TREATMENT_SUMMARY foragers) or at least four days after onset (experienced foragers). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY In brief, x mg pool brain was mixed with five times the volume (in µL) of SP:SAMPLEPREP_SUMMARY acetonitrile (ACN):Water (1:1, v/v) and homogenized using a TissueLyser II (30 SP:SAMPLEPREP_SUMMARY Hz, 10 min; Retsch Qiagen). After Centrifugation (2 min, 14000 rpm), 10 µL were SP:SAMPLEPREP_SUMMARY used for amino acid derivatization. First, the supernatant was evaporated to SP:SAMPLEPREP_SUMMARY dryness (SpeedVac, Eppendorf), resuspended in 50 µL of 5% phenyl isothiocyanate SP:SAMPLEPREP_SUMMARY (PITC) in ethanol: H2O: pyridine (1:1:1, v/v/v), and incubated for 25 min at RT. SP:SAMPLEPREP_SUMMARY Subsequently, the samples were dried to remove excess PITC and resuspended in 10 SP:SAMPLEPREP_SUMMARY µL 5 mM ammonium acetate in methanol. After incubation (10 min at 14,000 rpm) SP:SAMPLEPREP_SUMMARY 90 µL of H2O: ACN + 0.2% formic acid were added. Each derivative was measured SP:SAMPLEPREP_SUMMARY in an undiluted and diluted (1:25) manner. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Zorbax Eclipse XDB-C18 (100 x 3.0mm, 3.5um) CH:SOLVENT_A 100% water; 0.2% formic acid CH:SOLVENT_B 100% acetonitrile; 0.2% formic acid CH:FLOW_GRADIENT 0-0.5 min at 0% B, 0.5-4 min 0-70% B, 4-5.3 min 70% B, 5.3-5.4 min 70-0% B, CH:FLOW_GRADIENT 5.4-7.3 min 0% B CH:FLOW_RATE 0.5 mL/min CH:COLUMN_TEMPERATURE 50 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 6500+ Qtrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS For identification and quantitation, a scheduled MRM method was used, with MS:MS_COMMENTS specific transitions for every amino acid and biogenic amine. Data acquisition MS:MS_COMMENTS and peak integration were performed in SciexOS software (Version 3.0.0.). MS:MS_COMMENTS Calculation of concentration was done using external calibration curves. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS µMol MS_METABOLITE_DATA_START Samples H1_D0_1 H1_D0_2 H1_D0_3 H1_D0_4 H1_D0_5 H2_D0_1 H2_D0_2 H2_D0_3 H2_D0_4 H2_D0_5 H3_D0_1 H3_D0_2 H3_D0_3 H3_D0_4 H3_D0_5 H1_D1_1 H1_D1_2 H1_D1_3 H1_D1_4 H1_D1_5 H2_D1_1 H2_D1_2 H2_D1_3 H2_D1_4 H2_D1_5 H3_D1_1 H3_D1_2 H3_D1_3 H3_D1_4 H3_D1_5 H1_D3_1 H1_D3_2 H1_D3_3 H1_D3_4 H1_D3_5 H2_D3_1 H2_D3_2 H2_D3_3 H2_D3_4 H2_D3_5 H3_D3_1 H3_D3_2 H3_D3_3 H3_D3_4 H3_D3_5 H1_D5_1 H1_D5_2 H1_D5_3 H1_D5_4 H1_D5_5 H2_D5_1 H2_D5_2 H2_D5_3 H2_D5_4 H2_D5_5 H3_D5_1 H3_D5_2 H3_D5_3 H3_D5_4 H3_D5_5 H1_D7_1 H1_D7_2 H1_D7_3 H1_D7_4 H1_D7_5 H2_D7_1 H2_D7_2 H2_D7_3 H2_D7_4 H2_D7_5 H3_D7_1 H3_D7_2 H3_D7_3 H3_D7_4 H3_D7_5 H1_NF_1 H2_NF_1 H2_NF_2 H2_NF_3 H2_NF_4 H2_NF_5 H3_NF_1 H3_NF_2 H3_NF_3 H3_NF_4 H3_NF_5 H1_EF_1 H2_EF_1 H2_EF_2 H2_EF_3 H2_EF_4 H2_EF_5 H3_EF_1 H3_EF_2 H3_EF_3 H3_EF_4 H3_EF_5 Factors life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:newly_emerged | age of workers in days:0 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:1 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:3 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:establishment | age of workers in days:5 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:nurse | age of workers in days:7-10 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:new_foragers | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain life stage:experienced_forager | age of workers in days:>12 | Sample source:brain Tyramine 3.85E-02 3.68E-02 4.64E-02 3.51E-02 5.21E-02 3.26E-02 4.67E-02 7.88E-02 6.01E-02 6.81E-02 5.66E-02 3.19E-02 7.84E-02 1.01E-01 4.11E-02 2.18E-02 4.41E-02 3.41E-02 3.51E-02 5.05E-02 4.26E-02 2.34E-02 1.82E-02 2.82E-02 1.66E-02 2.49E-02 1.81E-02 2.22E-02 2.46E-02 1.75E-02 1.11E-02 1.17E-02 1.05E-02 1.11E-02 1.20E-02 1.10E-02 1.17E-02 1.22E-02 1.47E-02 1.20E-02 1.24E-02 1.11E-02 1.21E-02 1.23E-02 1.43E-02 1.15E-02 1.36E-02 1.24E-02 1.54E-02 1.24E-02 1.29E-02 1.19E-02 1.50E-02 1.23E-02 1.06E-02 1.30E-02 1.23E-02 1.22E-02 1.09E-02 1.17E-02 1.27E-02 1.33E-02 1.28E-02 1.22E-02 1.41E-02 1.25E-02 1.29E-02 1.19E-02 1.25E-02 1.35E-02 1.26E-02 1.25E-02 1.09E-02 1.11E-02 1.10E-02 1.45E-02 1.07E-02 1.15E-02 1.15E-02 1.38E-02 1.62E-02 1.22E-02 1.48E-02 1.08E-02 1.38E-02 1.47E-02 1.84E-02 1.60E-02 1.20E-02 1.46E-02 1.42E-02 1.42E-02 1.45E-02 1.45E-02 1.55E-02 1.29E-02 1.12E-02 Dopamine 5.52E-02 4.18E-02 7.68E-02 1.11E-01 9.92E-02 3.92E-02 6.33E-02 1.27E-01 4.78E-02 8.03E-02 4.27E-02 7.00E-02 1.36E-01 1.26E-01 5.68E-02 3.25E-02 1.26E-01 1.16E-01 8.54E-02 4.56E-02 4.20E-02 3.80E-02 2.96E-02 2.25E-02 3.90E-02 4.97E-02 7.58E-02 6.53E-02 6.90E-02 4.94E-02 2.23E-02 3.31E-02 3.35E-02 3.67E-02 3.23E-02 3.14E-02 2.88E-02 3.64E-02 4.24E-02 5.77E-02 2.19E-02 2.78E-02 4.62E-02 3.07E-02 7.18E-02 4.09E-02 3.78E-02 4.36E-02 6.26E-02 3.27E-02 3.46E-02 6.05E-02 9.96E-02 2.56E-02 3.66E-02 3.29E-02 3.62E-02 2.74E-02 3.51E-02 6.93E-02 4.08E-02 2.70E-02 3.56E-02 2.34E-02 4.74E-02 4.70E-02 3.65E-02 4.03E-02 4.27E-02 6.39E-02 2.52E-02 4.87E-02 2.44E-02 2.16E-02 2.91E-02 4.06E-02 2.83E-02 3.29E-02 5.70E-02 3.30E-02 3.10E-02 3.67E-02 3.65E-02 1.72E-02 2.35E-02 2.63E-02 2.71E-02 2.20E-02 2.04E-02 1.69E-02 2.01E-02 2.36E-02 2.46E-02 2.63E-02 2.68E-02 2.79E-02 1.81E-02 Kynurenine 2.21E-01 3.03E-01 2.37E-01 3.04E-01 1.88E-01 1.37E-01 2.08E-01 1.81E-01 2.50E-01 2.43E-01 1.56E-01 1.77E-01 1.68E-01 3.08E-01 2.87E-01 3.32E-01 2.81E-01 3.87E-01 2.95E-01 6.74E-01 1.95E-01 4.19E-01 1.73E-01 1.84E-01 8.97E-02 1.32E-01 2.54E-01 1.70E-01 2.09E-01 1.11E-01 2.00E-01 3.97E-01 1.37E-01 2.55E-01 1.62E-01 2.66E-01 3.28E-01 3.60E-01 2.65E-01 2.89E-01 2.90E-01 2.36E-01 2.91E-01 2.92E-01 2.57E-01 1.79E-01 1.33E-01 2.25E-01 1.49E-01 1.34E-01 1.78E-01 1.63E-01 3.63E-01 3.06E-01 2.55E-01 2.18E-01 2.27E-01 3.30E-01 2.62E-01 2.42E-01 1.70E-01 5.30E-02 8.94E-02 6.69E-02 6.67E-02 8.83E-02 1.23E-01 1.18E-01 1.02E-01 1.84E-01 1.75E-01 1.44E-01 1.90E-01 1.54E-01 8.65E-02 6.99E-02 3.18E-01 2.09E-01 2.96E-01 4.26E-02 2.06E-02 1.35E-01 1.26E-01 1.14E-01 6.23E-02 1.33E-01 7.31E-02 3.25E-02 7.30E-02 5.13E-02 4.24E-02 2.46E-02 3.34E-02 2.74E-02 4.15E-02 1.38E-01 5.33E-02 Citrulline 5.89E+00 3.52E+00 2.83E+00 3.08E+00 2.74E+00 1.55E+00 3.15E+00 2.38E+00 2.63E+00 2.29E+00 2.29E+00 2.99E+00 2.68E+00 2.67E+00 3.05E+00 2.22E+00 2.04E+00 2.52E+00 2.38E+00 3.53E+00 1.95E+00 3.03E+00 1.95E+00 2.57E+00 2.35E+00 2.69E+00 3.59E+00 2.52E+00 3.05E+00 2.82E+00 1.87E+00 2.41E+00 1.77E+00 1.96E+00 2.75E+00 3.09E+00 2.53E+00 1.99E+00 2.14E+00 2.24E+00 1.76E+00 2.08E+00 2.26E+00 2.34E+00 2.44E+00 3.34E+00 2.77E+00 2.70E+00 2.05E+00 3.19E+00 1.98E+00 2.34E+00 2.48E+00 2.09E+00 2.01E+00 2.91E+00 1.94E+00 2.25E+00 1.91E+00 1.80E+00 6.75E+00 2.19E+00 1.74E+00 1.77E+00 2.18E+00 1.71E+00 1.99E+00 1.58E+00 1.69E+00 1.32E+00 2.16E+00 1.99E+00 1.76E+00 1.51E+00 1.70E+00 2.02E+00 1.33E+00 2.09E+00 1.61E+00 2.08E+00 1.50E+00 2.17E+00 2.93E+00 2.26E+00 3.08E+00 2.41E+00 3.16E+00 3.81E+00 2.73E+00 2.49E+00 2.90E+00 2.85E+00 2.52E+00 2.31E+00 2.68E+00 2.09E+00 4.93E+00 Serotonin 9.46E-02 8.70E-02 9.01E-02 1.11E-01 1.16E-01 3.84E-02 9.65E-02 9.84E-02 9.38E-02 1.07E-01 9.61E-02 9.54E-02 1.09E-01 1.10E-01 8.12E-02 6.60E-02 1.21E-01 1.09E-01 9.25E-02 9.64E-02 7.58E-02 6.41E-02 9.08E-02 7.39E-02 8.71E-02 6.51E-02 8.15E-02 8.85E-02 8.28E-02 1.06E-01 2.96E-02 3.15E-02 3.70E-02 3.56E-02 4.87E-02 4.45E-02 4.55E-02 6.99E-02 8.60E-02 7.90E-02 5.75E-02 5.21E-02 7.21E-02 5.43E-02 9.27E-02 8.56E-02 8.15E-02 7.78E-02 1.09E-01 7.11E-02 1.16E-01 9.59E-02 1.19E-01 7.27E-02 6.90E-02 8.49E-02 8.38E-02 5.52E-02 8.14E-02 9.77E-02 9.49E-02 8.70E-02 8.81E-02 8.22E-02 1.01E-01 1.13E-01 1.13E-01 1.14E-01 1.07E-01 1.21E-01 9.22E-02 1.02E-01 7.57E-02 8.65E-02 8.93E-02 9.47E-02 6.84E-02 7.93E-02 1.07E-01 9.70E-02 8.25E-02 8.07E-02 8.23E-02 5.15E-02 9.05E-02 6.63E-02 4.07E-02 7.52E-02 4.86E-02 5.72E-02 6.20E-02 5.80E-02 5.23E-02 6.86E-02 6.92E-02 6.52E-02 5.76E-02 Octopamine 2.73E-02 3.78E-02 3.81E-02 4.72E-02 5.45E-02 3.04E-02 4.99E-02 5.64E-02 5.34E-02 6.76E-02 5.42E-02 3.87E-02 4.96E-02 7.18E-02 4.14E-02 3.50E-02 4.85E-02 3.91E-02 3.66E-02 2.84E-02 3.26E-02 2.42E-02 2.60E-02 2.78E-02 2.50E-02 3.19E-02 3.64E-02 2.99E-02 3.79E-02 3.00E-02 2.78E-02 2.59E-02 2.19E-02 2.85E-02 2.21E-02 2.29E-02 2.28E-02 1.79E-02 2.14E-02 2.09E-02 2.43E-02 1.90E-02 2.55E-02 2.98E-02 3.67E-02 3.25E-02 3.70E-02 3.27E-02 8.32E-02 3.04E-02 2.87E-02 2.71E-02 2.67E-02 2.48E-02 2.64E-02 2.30E-02 2.56E-02 2.06E-02 2.01E-02 2.54E-02 2.50E-02 3.07E-02 2.95E-02 4.06E-02 3.13E-02 2.28E-02 3.36E-02 2.84E-02 3.06E-02 3.57E-02 2.04E-02 2.42E-02 2.33E-02 2.02E-02 2.20E-02 3.35E-02 3.62E-02 2.90E-02 3.65E-02 2.74E-02 2.55E-02 2.20E-02 2.75E-02 2.10E-02 2.64E-02 2.03E-02 3.08E-02 2.24E-02 2.13E-02 2.22E-02 2.77E-02 2.41E-02 1.88E-02 2.23E-02 2.16E-02 2.55E-02 3.35E-02 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z KEGG Pubchem SID Pubchem CID Octopamine 289.95 C04227 6898 440266 Citrulline 311.2 C00327 3621 9750 Dopamine 289.2 C03758 6517 681 Kynurenine 344.2 C00328 3622 161166 Tyramine 273.21 C00483 3766 5610 Serotonin 312.2 C00780 4041 5202 METABOLITES_END #END