Paleoecological analyses of biotic assemblages from cores collected throughout south Florida’s estuaries indicate gradually increasing salinities over approximately the last 2000 years, consistent with rising sea level. Around the beginning of the twentieth century these gradual patterns of change began to shift, corresponding to the beginning of human alteration of the environment via canal construction, railroad construction and other land use changes. Between 1950 and 1960, at a time of significant construction of water management structures another distinctive shift in the biological assemblages occurred. Analysis of the assemblages provides essential information on long-term patterns of change in the estuaries and provides a basis for predicting future trajectories of change. Paleosalinity estimates derived from the cores are providing input to linear regression models to determine related freshwater flow into the estuaries of south Florida. These analyses are being used to help establish performance measures and targets for the Comprehensive Everglades Restoration, established following an Act of Congress in 2000. Restoration of south Florida’s ecosystems is slated to be a 30–50 year effort that will require detailed knowledge of past decadal to centennial-scale changes in climate, freshwater flow and salinity. This historical perspective provides information that allows land managers to set realistic and sustainable goals for restoration, and provides insight into the potential response of south Florida’s ecosystem to various future scenarios of global change.