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Devils Hole, a groundwater-filled fracture in the carbonate aquifer of the southern Nevada Mojave Desert, represents a unique ecohydrological setting, as home to the only extant population of Cyprinodon diabolis, the endangered Devils Hole pupfish. Using water column temperatures collected with a fiber-optic distributed temperature sensor (DTS) during four field campaigns in 2009, evidence of deep circulation and nutrient export are, for the first time, documented. The DTS was deployed to measure vertical temperature profiles in the system, and the raw data returned were postprocessed to refine the calibration beyond the precision of the instrument’s native calibration routines. Calibrated temperature data serve as a tracer for water movement and reveal a seasonal pattern of convective mixing that is supported by numerical simulations of the system. The periodic presence of divers in the water is considered, and their impacts on the temperature profiles are examined and found to be minimal. The seasonal mixing cycle may deplete the pupfish’s food supplies when nutrients are at their scarcest. The spatial and temporal scales of the DTS observations make it possible to observe temperature gradients on the order of 0.001 degrees C m(-1), revealing phenomena that would have been lost in instrument noise and uncertainty.