METABOLITE PROFILES IN STARVED DIPOREIA SPP. USING LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY (LC-MS) BASED METABOLOMICS

The holarctic amphipod Diporeia spp. was historically the most abundant benthic macroinvertebrate in the offshore region of the Laurentian Great Lakes basin. However, since the 1990's, the numbers of Diporeia have declined precipitously throughout the region. Competition for food with introduced dreissenid mussels may be partly to blame for this decline. Thus, a better understanding of how Diporeia responds and adjust to starvation is needed. For this purpose, we used liquid chromatography (LC) coupled with time-of-flight mass spectrometry (TOF-MS) to study the metabolite profiles of Diporeia during starvation. Diporeia were collected from Lake Michigan, brought to the laboratory and starved for up to 60 days. During the starvation period, metabolite levels were determined at 12-day intervals and compared to those of day 0. Principal component and cluster analyses revealed differential abundance of metabolite profiles across groups. Significantly down-regulated metabolites included polyunsaturated fatty acids, phospholipids, and amino acids and their derivatives. Overall, starved organisms relied predominantly on glycerophospolipid metabolism and protein based catabolism for energy production. This research demonstrates that LC-MS based metabolomics can be used to assess physiological status and has shown that unique metabolite profiles are distinguishable over several weeks of starvation in this freshwater amphipod. More importantly, these unique metabolites could be used to gain insights into the underlying cause(s) of Diporeia's decline in the Laurentian Great Lakes.