Mysis diluviana is a major component of prey fish diets in the Great Lakes, so annual production of M. diluviana is important for understanding and modeling energy flow through Great Lakes food webs. However, only three lake-wide measurements of M. diluviana annual production in Lake Ontario are currently available (1971, 1990, 1995). During 2013, lake-wide coverage of Lake Ontario was achieved during four periods from April to November. Annual mean density and biomass of M. diluviana in 2013 were 99 #/m² (SE: 8) and 318 mg dw/m² (SE: 28) – approximately half of values observed in 1990s. M. diluviana comprised 13–30% of offshore zooplankton biomass in each period. Reproduction peaked in fall, with mean brood size of 32 embryos (range: 11–49), at least 10% larger than in 1990s. Generation time was two years from embryo to initial reproduction. Growth rates were 0.052 mm/d for the age-0 cohort and 0.027 mm/d for the age-1 cohort. Age-0 growth rate was significantly higher than in 1980s–90s (0.035 mm/d). Annual production in 2013 was 0.85 g dw/m²/yr (SE: 0.03) which was 30–40% of values observed in 1990 and 1995 (2.23 and 2.53 g/m²/yr). Annual production to biomass ratio (P/B) in 2013 was 2.65 /yr which was 80–85% of values observed in 1990 and 1995 (3.24 and 3.11 /yr), but this difference was not statistically significant. Our results suggest that changes in annual production over time can be estimated using changes in biomass over time and a mean P/B ratio.