Skipped spawning, or variation in spawning periodicity, occurs in many annual spawning fish species and is an important consideration for population management. We assessed plasma sex steroid concentrations and measured gonad size and ovarian follicle diameter as metrics to nonlethally identify mass ovarian follicular atresia, which may contribute to skipped spawning in Burbot Lota lota. We maintained wild fish in captivity and exposed them to increasing water temperatures during a 3-wk period before the spawning season to induce mass ovarian follicular atresia. We collected ovarian follicles, blood plasma, and gonadal sonograms from fish weekly between January 28, 2018, and March 25, 2018. We histologically analyzed ovarian follicles to confirm stage of maturity. We measured concentrations of plasma sex steroids testosterone (T) and estradiol-17β (E2) by radioimmunoassay. We measured gonad diameter and circumference by ultrasonography and ovarian follicle diameter by image analysis. Mean plasma T concentration decreased from 8.94 ng/mL during late vitellogenesis to 1.83 ng/mL during atresia, suggesting that plasma T concentrations may be used to identify mass ovarian follicular atresia. We do not recommend using plasma E2 concentrations to identify mass ovarian follicular atresia because E2 concentrations rapidly decreased during the completion of vitellogenesis and the initiation of atresia in Burbot; therefore, plasma E2 may not accurately identify mass ovarian follicular atresia. Mean gonad diameter measured by ultrasonography decreased from 4.05 cm during late vitellogenesis to 3.65 cm during atresia. Mean diameter of ovarian follicles decreased during the final week of the study, suggesting that ovarian follicle diameter may be used to identify advanced mass ovarian follicular atresia. The nonlethal tools assessed—plasma sex steroid concentrations, ultrasonography, and ovarian follicle diameter—enable fisheries biologists to determine the occurrence and frequency of mass ovarian follicular atresia among Burbot in Lake Roosevelt and may be applied to other Burbot populations.