Shallow marine ecosystems are experiencing significant environmental alterations as a result of changing climate and increasing human activities along coasts. Intensive urbanization of the southeast Florida coast and intensification of climate change over the last few centuries changed the character of coastal ecosystems in the semi-enclosed Biscayne Bay, Florida. In order to develop management policies for the Bay, it is vital to obtain reliable scientific evidence of past ecological conditions. The long-term records of subfossil diatoms obtained from No Name Bank and Featherbed Bank in the Central Biscayne Bay, and from the Card Sound Bank in the neighboring Card Sound, were used to study the magnitude of the environmental change caused by climate variability and water management over the last ~ 600 yr. Analyses of these records revealed that the major shifts in the diatom assemblage structures at No Name Bank occurred in 1956, at Featherbed Bank in 1966, and at Card Sound Bank in 1957. Smaller magnitude shifts were also recorded at Featherbed Bank in 1893, 1942, 1974 and 1983. Most of these changes coincided with severe drought periods that developed during the cold phases of El Niño Southern Oscillation (ENSO), Atlantic Multidecadal Oscillation (AMO) and Pacific Decadal Oscillation (PDO), or when AMO was in warm phase and PDO was in the cold phase. Only the 1983 change coincided with an unusually wet period that developed during the warm phases of ENSO and PDO. Quantitative reconstructions of salinity using the weighted averaging partial least squares (WA-PLS) diatom-based salinity model revealed a gradual increase in salinity at the three coring locations over the last ~ 600 yr, which was primarily caused by continuously rising sea level and in the last several decades also by the reduction of the amount of freshwater inflow from the mainland. Concentration of sediment total nitrogen (TN), total phosphorus (TP) and total organic carbon (TOC) increased in the second half of the 20th century, which coincided with the construction of canals, landfills, marinas and water treatment plants along the western margin of Biscayne Bay. Increased magnitude and rate of the diatom assemblage restructuring in the mid- and late-1900s, suggest that large environmental changes are occurring more rapidly now than in the past.