{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002456","ANALYSIS_ID":"AN004006","VERSION":"1","CREATED_ON":"January 27, 2023, 1:12 am"},

"PROJECT":{"PROJECT_TITLE":"1H NMR metabolomics applied to assess the metabolic response of Ruditapes philippinarum clams to sea warming and 17-α-ethinylestradiol exposure","PROJECT_TYPE":"1H NMR metabolomics to study the effects of warming conditions and exposure to 17-α-ethinylestradiol (EE2) on the polar metabolome of Ruditapes philippinarum clams","PROJECT_SUMMARY":"Hormones correspond to one of the most important classes of PhACs and are usually classified as endocrine-disrupting chemicals (EDCs), due to their ability to alter endocrine system functions, resulting in known adverse health effects on non-target organisms. Among the known EDCs, 17-α-ethinylestradiol (EE2) is characterized by high estrogenic potency, chemical stability, and a tendency to accumulate in aquatic biota. This hormone reaches the environment mainly through human and animal excretion, with conventional WWTP processes not fully managing its removal from treated effluents. Bivalves are excellent model organisms to assess the toxicity of several environmental stressors. Alongside the impacts of pollutants, organisms in coastal ecosystems are also subjected to climate change-related factors, such as temperature rise. Water temperature may increase up to 4 °C by 2100 and this is expected to severely impact several aspects of marine organisms’ biology. In addition, these environmental changes are also likely to affect the sensitivity of organisms to pollutants and, hence, pollutants toxicity. In this study, a comprehensive untargeted 1H NMR metabolomics strategy was applied to measure the metabolic impact of sea warming, in tandem with exposure to EE2, on Ruditapes philippinarum clams. The clams were exposed to different EE2 concentrations, either at 17 °C as control temperature or at 21 °C (representing a 4 °C increase, which corresponds to the worst-case warming scenario). The obtained data added important knowledge, unveiling individual metabolic effects of temperature rise and synergetic effects upon EE2 exposure, and paving the way for the definition of new metabolic markers for the monitoring of environmental stressors.","INSTITUTE":"University of Aveiro","DEPARTMENT":"CICECO – Aveiro Institute of Materials, Department of Chemistry","LABORATORY":"Metabolomics group","LAST_NAME":"Gil","FIRST_NAME":"Ana M.","ADDRESS":"University of Aveiro, Campus Universitario de Santiago, 3810-193 Aveiro, Portugal","EMAIL":"agil@ua.pt","PHONE":"+351234370707","FUNDING_SOURCE":"This work was developed within the CICECO-Aveiro Institute of Materials project (UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020) financed by national funds through the FCT/MEC (PIDDAC). We are also grateful to the Portuguese National NMR Network (PTNMR), supported by FCT funds as the NMR spectrometer used is part of PTNMR and partially supported by Infrastructure Project No. 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL, and the FCT through PIDDAC). This work was also financially supported by the project BISPECIAl: BIvalveS under Polluted Environment and ClImate chAnge (POCI-01-0145-FEDER- 028425) funded by FEDER, through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI), and by national funds (OE), through FCT/MCTES. Mónica G. Silva benefited from Research Grant (MSc) (BI/CESAM/0043_2019/POCI-01-0145-FEDER-028425) under the project BISPECIAl: BIvalveS under Polluted Environment and ClImate change PTDC/CTA-AMB/28425/2017 (POCI-01-0145-FEDER-028425)."},

"STUDY":{"STUDY_TITLE":"1H NMR metabolomics applied to assess the metabolic response of Ruditapes philippinarum clams to sea warming and 17-α-ethinylestradiol exposure","STUDY_TYPE":"1H NMR metabolomics to study the effects of warming conditions and exposure to 17-α-ethinylestradiol (EE2) on the polar metabolome of Ruditapes philippinarum clams","STUDY_SUMMARY":"In this study, a comprehensive untargeted 1H NMR metabolomics strategy was applied to measure the metabolic impact of sea warming, in tandem with exposure to EE2, on Ruditapes philippinarum clams. The clams were exposed to five different EE2 concentrations: 0 (control group), 5, 25, 125 and 625 ng/L; either at 17 °C as control temperature or at 21 °C (representing a 4 °C increase, which corresponds to the worst-case warming scenario). The obtained data added important knowledge, unveiling individual metabolic effects of temperature rise and synergetic effects upon EE2 exposure, and paving the way for the definition of new metabolic markers for the monitoring of environmental stressors.","INSTITUTE":"University of Aveiro","DEPARTMENT":"CICECO – Aveiro Institute of Materials, Department of Chemistry","LABORATORY":"Metabolomics Group- CICECO","LAST_NAME":"Rodrigues","FIRST_NAME":"Joao A.","ADDRESS":"University of Aveiro, Campus Universitario de Santiago, 3810-193 Aveiro, Portugal","EMAIL":"joao.e.a.rodrigues@gmail.com","PHONE":"00351963481841","NUM_GROUPS":"10","TOTAL_SUBJECTS":"103","STUDY_COMMENTS":"This work was developed within the CICECO-Aveiro Institute of Materials project (UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020) financed by national funds through the FCT/MEC (PIDDAC). We are also grateful to the Portuguese National NMR Network (PTNMR), supported by FCT funds as the NMR spectrometer used is part of PTNMR and partially supported by Infrastructure Project No. 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL, and the FCT through PIDDAC). This work was also financially supported by the project BISPECIAl: BIvalveS under Polluted Environment and ClImate chAnge (POCI-01-0145-FEDER- 028425) funded by FEDER, through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI), and by national funds (OE), through FCT/MCTES. Mónica G. Silva benefited from Research Grant (MSc) (BI/CESAM/0043_2019/POCI-01-0145-FEDER-028425) under the project BISPECIAl: BIvalveS under Polluted Environment and ClImate change PTDC/CTA-AMB/28425/2017 (POCI-01-0145-FEDER-028425)."},

"SUBJECT":{"SUBJECT_TYPE":"Other organism","SUBJECT_SPECIES":"Ruditapes philippinarum","TAXONOMY_ID":"129788","HEIGHT_OR_HEIGHT_RANGE":"length: 3.81 ± 0.42 cm; and width: 3.06 ± 0.51 cm","GENDER":"Male and female"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_01",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_001.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_02",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_002.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_03",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_003.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_04",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_004.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_05",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_005.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_06",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_006.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_07",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_007.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_08",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_008.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_09",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_010.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_10",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_011.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C0_11",
"Factors":{"Factor":"bivalves_17C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_012.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_01",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_013.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_02",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_014.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_03",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_015.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_04",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_016.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_05",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_017.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_06",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_018.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_07",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_019.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_08",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_020.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_09",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_021.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_10",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_022.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C5_11",
"Factors":{"Factor":"bivalves_17C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_024.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_01",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_025.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_02",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_026.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_03",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_027.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_04",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_028.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_05",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_029.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_06",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_030.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_07",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_031.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_08",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_032.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_09",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_033.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_10",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_034.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_11",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_035.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C25_12",
"Factors":{"Factor":"bivalves_17C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_036.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_01",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_037.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_02",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_038.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_03",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_039.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_04",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_040.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_05",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_041.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_06",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_042.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_07",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_043.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_08",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_044.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_09",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_045.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_10",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_046.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C125_11",
"Factors":{"Factor":"bivalves_17C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_047.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_01",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_049.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_02",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_050.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_03",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_051.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_04",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_052.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_05",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_053.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_06",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_054.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_07",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_055.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_08",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_056.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_09",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_057.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_10",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_058.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_17C_C625_11",
"Factors":{"Factor":"bivalves_17C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_059.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_01",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_061.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_02",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_062.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_03",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_063.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_04",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_064.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_05",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_065.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_06",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_066.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_07",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_067.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_08",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_069.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_09",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_070.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C0_10",
"Factors":{"Factor":"bivalves_21C_C0"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_071.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_01",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_073.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_02",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_074.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_03",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_075.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_04",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_077.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_05",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_078.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_06",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_079.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_07",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_080.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_08",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_081.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_09",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_083.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C5_10",
"Factors":{"Factor":"bivalves_21C_C5"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_084.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_01",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_085.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_02",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_086.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_03",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_087.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_04",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_088.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_05",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_089.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_06",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_093.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_07",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_094.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_08",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_095.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C25_09",
"Factors":{"Factor":"bivalves_21C_C25"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_096.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_01",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_097.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_02",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_098.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_03",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_099.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_04",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_100.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_05",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_101.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_06",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_102.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_07",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_105.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_08",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_107.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C125_09",
"Factors":{"Factor":"bivalves_21C_C125"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_108.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_01",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_109.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_02",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_110.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_03",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_111.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_04",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_112.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_05",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_113.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_06",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_114.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_07",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_115.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_08",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_117.raw"}
},
{
"Subject ID":"-",
"Sample ID":"bivalves_21C_C625_09",
"Factors":{"Factor":"bivalves_21C_C625"},
"Additional sample data":{"RAW_FILE_NAME":"bivalves_119.raw"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Ruditapes philippinarum clams were collected at the Ria de Aveiro, a shallow coastal system located on the Northwest Atlantic coast of Portugal. Individuals of similar size (length: 3.81 ± 0.42 cm; and width: 3.06 ± 0.51 cm) were selected. For depuration and acclimation to laboratory conditions, all clams were maintained in artificial seawater for 10 days (salinity: 30 ± 1, Tropic Marin® SEA SALT, from Tropic Marine Center), under continuous aeration, constant temperature (17 ± 1 ºC) and a natural photoperiod. Artificial seawater was renewed every 2-3 days and clams were fed every 2-3 days with Algamac Protein Plus (150,000 cells/animal/day) after the 3rd day. After depuration, the organisms were subjected to a chronic toxicity test for 28 days, consisting of exposure to five different EE2 concentrations (Sigma-Aldrich, purity ≥ 98%, MW = 296.40 g/mol, 1 mg/L stock solution in ultrapure water): 0 (control group), 5, 25, 125 and 625 ng/L. To assess the effects of a warming scenario on the impacts of EE2, the experiments were carried out at 17 ± 1 °C (control; mean temperature of sampling area during September: 16 - 19 °C) and at 21 ± 1 °C (worst-case climate change scenario, IPCC, 2021). The aquaria were placed in distinct climatic rooms for each temperature. To reach 21 °C, the temperature was raised by 2 °C, every 2–3 days. For each concentration level and temperature, 12 samples were considered: 4 individuals per aquarium and 3 aquaria per treatment. In each aquarium, a total of 3 L of artificial seawater (salinity: 30 ± 1), continuous aeration, and a natural photoperiod were used. The exposure medium for each condition was renewed weekly, after which EE2 concentration levels were re-established. Mortality was checked daily and found to be null. At the end of the 28-day exposure period, the clams were frozen in liquid nitrogen and stored at – 80 ºC.","COLLECTION_PROTOCOL_FILENAME":"Bivalves Experimental Procedure","SAMPLE_TYPE":"Tissue","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Sea warming and 17-α-ethinylestradiol exposure. After depuration, the organisms were subjected to a chronic toxicity test for 28 days, consisting of exposure to five different EE2 concentrations: 0 (control group), 5, 25, 125 and 625 ng/L. To assess the effects of a warming scenario on the impacts of EE2, the experiments were carried out at 17 ± 1 °C (control; mean temperature of sampling area during September) and at 21 ± 1 °C (worst-case climate change scenario). The aquaria were placed in distinct climatic rooms for each temperature. To reach 21 °C, the temperature was raised by 2 °C, every 2–3 days. For each concentration level and temperature, 12 samples were considered: 4 individuals per aquarium and 3 aquaria per treatment. In each aquarium, a total of 3 L of artificial seawater (salinity: 30 ± 1), continuous aeration, and a natural photoperiod were used. The exposure medium for each condition was renewed weekly, after which EE2 concentration levels were re-established. Mortality was checked daily and found to be null. At the end of the 28-day exposure period, the clams were frozen in liquid nitrogen and stored at – 80 ºC.","TREATMENT_PROTOCOL_FILENAME":"Bivalves Experimental Procedure","TREATMENT_COMPOUND":"Sea warming and 17-α-ethinylestradiol exposure","TREATMENT_DOSE":"Clams were exposed to five different EE2 concentrations: 0 (control group), 5, 25, 125 and 625 ng/L. To assess the effects of a warming scenario on the impacts of EE2, the experiments were carried out at 17 ± 1 °C (control) and at 21 ± 1 °C (worst-case climate change scenario)."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Metabolite extraction was performed using a water/methanol/chloroform method, as described in (Hines, Oladiran, Bignell et al., 2007). Briefly, the clams´ soft tissue (0.15 g per sample) was ground with a pestle and mortar, in liquid nitrogen, and then transferred to a microtube, followed by the addition of cold methanol (600 µL), ultrapure water (128 µL) and chloroform (300 µL). The mixture was vortexed, left in ice for 10 min and centrifuged (2,500 g, 4 °C, 10 min). The top layer was transferred into a microtube to which chloroform (300 µL) and water (300 µL) were added. The mixture was vortexed and centrifuged (2,500 g, 4 °C, 10 min). The upper layer (aqueous) was transferred into vials, dried in a centrifugal vacuum concentrator (UNIVAP 100H) and stored at −80 °C until NMR analysis.","SAMPLEPREP_PROTOCOL_FILENAME":"Bivalves Experimental Procedure","PROCESSING_STORAGE_CONDITIONS":"-80℃","EXTRACTION_METHOD":"Water/methanol/chloroform method, as described in (Hines, Oladiran, Bignell et al., 2007)","EXTRACT_STORAGE":"-80℃","SAMPLE_RESUSPENSION":"The dried polar extracts of clam samples were resuspended in 600 μL of sodium phosphate buffer (0.1 M in D2O, 99.96% D, pH 7.4, containing 0.5 mM sodium salt of 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid, TSP-d4, chemical shift referencing). The mixture was vortexed and centrifuged (16,000 g, 10 min, room temperature) and 550 μL were transferred into 5 mm NMR tubes.","SAMPLE_SPIKING":"0.5 mM sodium salt of 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid (TSP-d4), as a chemical shift reference."},

"ANALYSIS":{"ANALYSIS_TYPE":"NMR","LABORATORY_NAME":"Metabolomics group","OPERATOR_NAME":"Joao A. Rodrigues","DETECTOR_TYPE":"Bruker Avance III 500 MHz spectrometer","SOFTWARE_VERSION":"TopSpin 3.2 and Amix 3.9.14","ANALYSIS_PROTOCOL_FILE":"Bivalves Experimental Procedure","DATA_FORMAT":"fid, 1r"},

"NM":{"INSTRUMENT_NAME":"Bruker AVANCE III 500 spectrometer","INSTRUMENT_TYPE":"FT-NMR","NMR_EXPERIMENT_TYPE":"1D-1H","SPECTROMETER_FREQUENCY":"500 MHz","NMR_PROBE":"TXI probe","NMR_SOLVENT":"D2O","NMR_TUBE_SIZE":"5 mm NMR tubes","SHIMMING_METHOD":"Topshim","PULSE_SEQUENCE":"noesypr1d","WATER_SUPPRESSION":"presat","PULSE_WIDTH":"90-degree","RECEIVER_GAIN":"203","TEMPERATURE":"298 K","NUMBER_OF_SCANS":"256","DUMMY_SCANS":"8","ACQUISITION_TIME":"2.34 s","RELAXATION_DELAY":"3 s","SPECTRAL_WIDTH":"7,002.8 Hz","NUM_DATA_POINTS_ACQUIRED":"32 k","LINE_BROADENING":"0.3 Hz","ZERO_FILLING":"64 k","BASELINE_CORRECTION_METHOD":"manual","CHEMICAL_SHIFT_REF_STD":"0 ppm for TSP-d4","NMR_RESULTS_FILE":"4._Bivalves_results_data.txt UNITS:ppm"}

}