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The suitability of the crayfish metabolome as a bioindicator of stream conditions was investigated because there is a need for fast responding, sensitive, diagnostic, and ecologically relevant bioindicators. A laboratory study assessed Procambarus clarkii metabolome sensitivity to six different treatments of food or DO stress. A field study in subcatchments of the Red River Valley, Manitoba, Canada assessed the effects of land-use type (reference, agriculture, wastewater effluent) on the Orconectes virilis metabolome. Crayfish were sacrificed, and hepatopancreas, gill, and tail tissues were dissected. Analyses found the metabolome was sensitive enough to potentially diagnose food and DO stress in P. clarkii tail muscle. O. virilis gill metabolomes were the most sensitive to land-use treatments associated with substantive differences in the stressor environments. Although this study shows promise in using the crayfish metabolome in bioassay monitoring, further metabolomics studies are needed to assess the predictability and ecological relevance of crayfish metabolic changes.