Temperature fluctuations and climate change impacts may substantially affect spawning success of fish, especially migratory species with a limited spawning window. Factors affecting American shad (Alosa sapidissima) spawning success and survival were investigated at different temperatures and periods (peak- and late-spawning periods) during the Connecticut River, USA, spawning migration in 2017. Wild caught American shad were exposed to constant temperatures regimes of 15, 18, 21, 24 and 27 °C for 2 weeks. During the peak-spawning period, an increase in temperature (15–24 °C) was shown to increase spawning success factors, including spawning probability, number of eggs, and fertilization success, but decreased egg size. Temperatures between 18 and 27 °C did not affect these factors during the late-spawning period. Glochidia infection by the alewife floater (Anodonta implicata) was much higher in the late-spawning period and significantly decreased the survival of American shad. Further research should investigate the parasite-host relationship between the alewife floater and American shad to determine annual variability of glochidia infections and how they affect American shad from physiological and passage perspectives. Higher temperatures were shown to increase spawning success of American shad during the peak-spawning period, but temperature had no effect during the late-spawning period. However, any effect during the late-spawning period may have been masked by a high level of glochidia infection.