Persistent metal contamination limits lotic ecosystem heterotrophic metabolism after more than 100 years of exposure: A novel application of the resazurin resorufin smart tracer

Persistent stress from anthropogenic metal deposition in lotic ecosystems is a global concern. This long-term selective pressure shapes hyporheic microbial assemblages and influences ecosystem functional integrity. We hypothesized that, even after 100 years of adaptation opportunity, ecosystem function remains inhibited by sediment-associated metal stress and that the Resazurin Resorufin Smart Tracer can be used to quantify this impact. The Resazurin Resorufin Smart Tracer system is applied here in a novel capacity as an indicator of ecosystem function by quantifying ecosystem respiration of microbial communities. Hyporheic microbial communities exposed to differing magnitudes of chronic metal stress were compared to pristine reference sites in controlled column experiments. A Markov chain Monte Carlo technique was developed to solve the inverse smart tracer transport equation to derive community respiration data. Results suggest metals inhibit respiration by 13-30% relative to reference sites and this inhibition is directly related to the level of in situ metal stress. We demonstrate the first application of a hydrologic smart tracer as a functional indicator of ecological integrity within anthropogenically influenced flowing water systems and provide data suggesting resilience is limited in hyporheic ecosystems even after more than a century of microbial adaption to chronic pollutants.