The largest Florida manatee (Trichechus manatus latirostris) aggregation at a natural warm-water refuge occurs in Kings Bay, Crystal River, FL. Over the last 32 years, the U.S. Fish and Wildlife Service and the State of Florida have created a network of manatee protection areas within Kings Bay including a year-round refuge designation and seven Federal manatee sanctuaries during the winter manatee season (November 15 – March 31). Aerial survey data collected between 1983 and 2012 was used to examine the seasonal change in manatee distribution within Kings Bay in order to assess the effectiveness of current sanctuary sizes and locations. Regression analysis indicated a significant change in manatee abundance among the winter seasons (p < 0.05). The average winter manatee counts increased by 4.81 animals per year over the 30 year period. In contrast, no significant changes in average or peak manatee abundance was detected among the summer seasons (p = 0.71 and p = 0.45 respectively). The average manatee counts increased by only 0.109 animals per year over the summer periods. Spatially explicit models using Geographic Information System (GIS) analysis revealed a strong correlation between high manatee density and artesian springs during the winter seasons. Highest abundances were identified at three locations: King’s Spring, Three Sisters Springs, and Magnolia Springs. These three locations coincide well with pre-existing sanctuary designations, but additional coverage is needed to support the overflow of manatees outside of sanctuary boundaries. Manatees continued to use Kings Bay in the summer seasons but in lower numbers and densities. Because density patterns were not uniform across summer periods, a heavier reliance on boat speed regulation is recommended to provide adequate protection to the endangered Florida manatee. Within a habitat type, the Magnolia Springs, South Banana Island, and Three Sisters Springs sanctuaries exhibited a significant influence on manatee density, suggesting differences in quality among sanctuaries. Years coinciding with extreme cold weather events also had a significant influence on manatee density. Using GIS to investigate seasonal shifts in manatees can be very informative regarding many issues including habitat selection and may improve the design and management of protected areas.