Investigating the long-term fluctuations of the feeding ecology of megaherbivores such as sirenians is important, as any changes could be indicative of shifts in resource availability. The Indian River Lagoon (IRL), eastern Florida, USA, is a critical habitat for the Florida manatee (Trichechus manatus latirostris). However, the IRL has experienced a substantial decline in seagrass due to the persistence of several harmful algal blooms. Using microhistological analysis, we examined the diet of manatees over a discontinuous sampling period spanning over 38 years using stomach contents collected from carcasses recovered in the IRL. Samples collected between 2013–2015 (post-seagrass die-off, n = 90) were compared to archived stomach samples collected between 1977–1989 (pre-seagrass die-off, n = 103). Samples analyzed from 1977–1989 contained primarily seagrasses (61.7%), followed by algae (28.4%) and vascular plants (1.7%). In contrast, stomach samples from the post-seagrass die-off primarily contained algae (49.5%), followed by seagrasses (34%) and vascular plants (2.7%). Between 1977–1989 and 2013–2015, manatees in the IRL experienced a 44.9% decline in seagrass consumption, and a 74.3% increase in algal consumption. This dietary shift was not influenced by body length, a proxy of age, or sex. Our results indicate that the dietary shift experienced by manatees is due to the decline of available seagrass forage in the IRL, and highlight the dietary plasticity of manatees in the face of changes in resource availability. However, the individual health and population-level consequences of this dietary shift are unknown. An increase in mortality due to undetermined causes in this region since 2012 can be associated with deteriorating body conditions of manatees in the IRL, possibly resulting from a lack of seagrass diet. Future research should further investigate behavioral changes affecting manatees in relation to seagrass decline in the IRL, including the energetic costs of this dietary change.