Title: investigating how multiple stressors drive thiamine deficiency linked to declines of pacific salmonid populations biological samples:O EGG samples from chinook salmon and steelhead trout…

Title: investigating how multiple stressors drive thiamine deficiency linked to declines of pacific salmonid populations biological samples:O EGG samples from chinook salmon and steelhead trout (collected by collaborators at 60 hatcheries along the pacific coast).O prey samples (collected by NOAA research vessels in the pacific ocean).O samples will be stored at -80C until biochemical analyses.O each sample will be assigned a unique identifier and logged into a database. Analytical data and supporting materials:O thiamine concentrations (free thiamine, thiamine monophosphate, thiamine pyrophosphate) in EGG and prey samples (nmol G). Protocols described in sops available in DR. Rinchards lab (SOP for thiamine extraction and measurement are based on brown ET AL. 1998 futia ET AL. 2017, and zajicek ET AL. 2005)O lipid contents (%) and fatty acid signatures (% of total fatty acids) in EGG and prey samples. Protocols described in sops available in DR. Rinchards lab (SOP for lipid extraction, fatty acid transmethylation, and measurement are based on folch ET AL. 1957, metcalfe and schmitz 1961, futia ET AL. 2019)O quality control and instrument performance data (e.g., hplc and GC MS calibration curves, replicate measurements):use of auto-injector to lessen amount of human error. Blank samples run regularly with the samples to account for any background noise. Data are reviewed and compared to notebook and computer files by PI. Student and PI check for outliers and consistency. Data entered by student who analyze the samples. For thiamine analysis, use of standard to establish standard curve, peaks of thiamine (free, mono and pyrophosphate) are checked and integrated using chemstation software for each sample. Samples are run in duplicate.