The reproductive cycles of turtles are linked to environmental factors, such as photoperiod and temperature. Currently, the reproductive physiology of diamondback terrapins (Malaclemys terrapin) is poorly understood, especially in Gulf of Mexico. The reproductive cycles of terrapins are thought to follow typical seasonal patterns. However, latitudinal variations in temperature regimens lead to longer-lasting warm periods, which can facilitate extended reproductive periods in some turtle species. This suggests that terrapins may show a similar change in the southern parts of their range. To elucidate aspects of the terrapin reproductive cycle, we sampled during the known reproductive season of a southern population of terrapins (May–July), as well as during the winter in late December and early January. We used enzyme-linked immunosorbent assays to quantify concentrations of the plasma sex hormones estradiol and testosterone, and the egg yolk protein precursor vitellogenin. Additionally, we used radiography and ultrasonography to monitor the ovarian status and egg development in females. Follicles showed no significant difference in average diameter across sampling periods with preovulatory class follicles existing in both summer and winter. Eggs were only detected from May to July, with radiographic data showing second clutches in 4 individuals. Testosterone and estradiol showed elevated concentrations throughout the nesting season, coinciding with multiple clutches of eggs, before both showed a significant decrease in winter. Vitellogenin showed peak concentrations in June with other months showing lower but detectable concentrations. Our results suggest that in southwestern Florida, terrapins may have extended reproductive potential and continuous vitellogenic cycles given the presence of preovulatory follicles and high quantities of vitellogenin found in summer and winter. However, true continuous reproduction was not detected in this study.