{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001885","ANALYSIS_ID":"AN003049","VERSION":"1","CREATED_ON":"July 22, 2021, 1:20 pm"},

"PROJECT":{"PROJECT_TITLE":"Differentiating toxic and nontoxic cogeneric harmful algae using the non-polar metabolome.","PROJECT_SUMMARY":"Recognition and rejection of chemically defended prey is critical to maximizing fitness for predators. Paralytic shellfish toxins (PSTs) which strongly inhibit voltage-gated sodium channels in diverse animal taxa are produced by several species of the bloom-forming algal genus Alexandrium where they appear to function as chemical defenses against grazing copepods. Despite PSTs being produced and localized within phytoplankton cells, some copepods distinguish toxic from non-toxic prey, selectively ingesting less toxic cells, in ways that suggest cell surface recognition perhaps associated with non-polar metabolites. In this study LC/MS and NMR-based metabolomics revealed that the non-polar metabolomes of two toxic species (Alexandrium catenella and Alexandrium pacificum) vary considerably from their non-toxic congener Alexandrium tamarense despite all three being very closely related. Toxic and non-toxic Alexandrium spp. were distinguished from each other by metabolites belonging to seven lipid classes. Of these, 17 specific metabolites were significantly more abundant in both toxic A. catenella and A. pacificum compared to non-toxic A. tamarense suggesting that just a small portion of the observed metabolic variability is associated with toxicity. Future experiments aimed at deciphering chemoreception mechanisms of copepod perception of Alexandrium toxicity should consider these metabolites, and the broader lipid classes phosphatidylcholines and sterols, as potential candidate cues.","INSTITUTE":"Georgia Institute of Technology","LAST_NAME":"Brown","FIRST_NAME":"Emily","ADDRESS":"950 Atlantic Dr Atlanta GA 30332, USA","EMAIL":"julia.kubanek@biosci.gatech.edu","PHONE":"404-894-8424"},

"STUDY":{"STUDY_TITLE":"MS Differentiating toxic and nontoxic cogeneric harmful algae using the non-polar metabolome.","STUDY_SUMMARY":"Recognition and rejection of chemically defended prey is critical to maximizing fitness for predators. Paralytic shellfish toxins (PSTs) which strongly inhibit voltage-gated sodium channels in diverse animal taxa are produced by several species of the bloom-forming algal genus Alexandrium where they appear to function as chemical defenses against grazing copepods. Despite PSTs being produced and localized within phytoplankton cells, some copepods distinguish toxic from non-toxic prey, selectively ingesting less toxic cells, in ways that suggest cell surface recognition perhaps associated with non-polar metabolites. In this study LC/MS and NMR-based metabolomics revealed that the non-polar metabolomes of two toxic species (Alexandrium catenella and Alexandrium pacificum) vary considerably from their non-toxic congener Alexandrium tamarense despite all three being very closely related. Toxic and non-toxic Alexandrium spp. were distinguished from each other by metabolites belonging to seven lipid classes. Of these, 17 specific metabolites were significantly more abundant in both toxic A. catenella and A. pacificum compared to non-toxic A. tamarense suggesting that just a small portion of the observed metabolic variability is associated with toxicity. Future experiments aimed at deciphering chemoreception mechanisms of copepod perception of Alexandrium toxicity should consider these metabolites, and the broader lipid classes phosphatidylcholines and sterols, as potential candidate cues.","INSTITUTE":"Georgia Institute of Technology","LAST_NAME":"Brown","FIRST_NAME":"Emily","ADDRESS":"950 Atlantic Dr Atlanta GA 30332, USA","EMAIL":"julia.kubanek@biosci.gatech.edu","PHONE":"404-894-8424"},

"SUBJECT":{"SUBJECT_TYPE":"Other organism","SUBJECT_SPECIES":"Alexandrium catenella; Alexandrium tamarense; Alexandrium pacificum","SPECIES_GROUP":"Not Human -- Alexandrium catenella; Alexandrium tamarense; Alexandrium pacificum"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_1",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_1"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_2",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_2"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_3",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_3"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_4",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_4"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_5",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_5"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_6",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_6"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_7",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_7"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_8",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_8"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_9",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_9"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_10",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_10"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_11",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_11"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_12",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_12"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_13",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_13"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_14",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_14"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AC_15",
"Factors":{"factor":"Treatment;Alexandrium_catenella"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AC_15"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_1",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_1"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_2",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_2"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_3",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_3"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_4",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_4"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_5",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_5"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_6",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_6"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_7",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_7"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_8",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_8"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_9",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_9"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_10",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_10"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_11",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_11"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_12",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_12"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_13",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_13"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_14",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_14"}
},
{
"Subject ID":"-",
"Sample ID":"74_3ERB74_AT_15",
"Factors":{"factor":"Treatment;Alexandrium_tamarense"},
"Additional sample data":{"RAW_FILE_NAME":"74_3ERB74_AT_15"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_1",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_1"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_2",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_2"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_3",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_3"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_4",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_4"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_5",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_5"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_6",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_6"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_7",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_7"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_9",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_9"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_10",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_10"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_11",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_11"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_13",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_13"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_14",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_14"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AP_15",
"Factors":{"factor":"Treatment;Alexandrium_pacificum"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AP_15"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_1",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_1"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_2",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_2"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_3",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_3"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_4",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_4."}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_5",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_5"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_6",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_6"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_7",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_7"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_8",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_8"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_9",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_9"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_10",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_10"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_11",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_11"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_12",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_12"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_13",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_13"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_14",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_14"}
},
{
"Subject ID":"-",
"Sample ID":"75_3ERB75_AT_15",
"Factors":{"factor":"Treatment;Alexandrium_tamarense2"},
"Additional sample data":{"RAW_FILE_NAME":"75_3ERB75_AT_15"}
},
{
"Subject ID":"-",
"Sample ID":"Blank01",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"Blank01"}
},
{
"Subject ID":"-",
"Sample ID":"Blank02",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"Blank02"}
},
{
"Subject ID":"-",
"Sample ID":"QC01",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC01"}
},
{
"Subject ID":"-",
"Sample ID":"QC02",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC02"}
},
{
"Subject ID":"-",
"Sample ID":"QC03",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC03"}
},
{
"Subject ID":"-",
"Sample ID":"QC04",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC04"}
},
{
"Subject ID":"-",
"Sample ID":"QC05",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC05"}
},
{
"Subject ID":"-",
"Sample ID":"QC06",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC06"}
},
{
"Subject ID":"-",
"Sample ID":"QC07",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC07"}
},
{
"Subject ID":"-",
"Sample ID":"QC08",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC08"}
},
{
"Subject ID":"-",
"Sample ID":"QC09",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC09"}
},
{
"Subject ID":"-",
"Sample ID":"QC10",
"Factors":{"factor":"QC"},
"Additional sample data":{"RAW_FILE_NAME":"QC10"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Alexandrium cells were collected by vacuum filtration onto GF/F filters and quenched with liquid nitrogen. Frozen cells with filters were stored in foil(previously muffled for 3 h at 450 °C) at -80 °C until extraction.","SAMPLE_TYPE":"Algae"},

"TREATMENT":{"TREATMENT_SUMMARY":"Metabolomes of toxic versus non-toxic species were compared using the following experimental pairings: A. tamarense (n=15) with A. catenella (n=15) (Experiment 1) and A. tamarense (n=15) with A. pacificum (n=15) (Experiment 2). The same non-toxic strain of A. tamarense was used in both experiments but the two experiments were conducted separately, in different months, to make the experiment manageable based on availability of batches grown from stock cultures. For both experiments, Alexandrium spp. cultures at a cell density of 12,000 to 13,000 cells mL-1 were split into fifteen 300 mL subcultures of each species which grew for two days. At the end of each experiment, during harvesting, a 1.0 mL aliquot from each culture was preserved with Lugol’s solution to measure cell concentrations"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"IPA: ACN (2:1) extract 4.54e6 cells per 1mL"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Thermo Accucore C30 (150 x 2.1 mm, 2.6um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Orbitrap ID-X tribrid","INSTRUMENT_TYPE":"orbitrap and ion trap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"compound discoverer","MS_RESULTS_FILE":"ST001885_AN003049_Results.txt UNITS:peak area Has m/z:Neutral masses Has RT:Yes RT units:Minutes"}

}