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In the mantled karst terrane of northern Florida, the water quality of the Upper Floridan aquifer is influenced by the degree of connectivity between the aquifer and the surface. Chemical and isotopic analyses [18O/16O (δ18O), 2H/1H (δD), 13C/12C (δ13C), tritium (3H), and strontium‐87/strontium‐86 (87Sr/86Sr)] along with geochemical mass‐balance modeling were used to identify the dominant hydrochemical processes that control the composition of ground water as it evolves downgradient in two systems. In one system, surface water enters the Upper Floridan aquifer through a sinkhole located in the Northern Highlands physiographic unit. In the other system, surface water enters the aquifer through a sinkhole lake (Lake Bradford) in the Woodville Karst Plain. Differences in the composition of water isotopes (δ18O and <δD) in rainfall, ground water, and surface water were used to develop mixing models of surface water (leakage of water to the Upper Floridan aquifer from a sinkhole lake and a sinkhole) and ground water. Using mass‐balance calculations, based on differences in δ18O and δD, the proportion of lake water that mixed with meteoric water ranged from 7 to 86% in water from wells located in close proximity to Lake Bradford. In deeper parts of the Upper Floridan aquifer, water enriched in 18O and D from five of 12 sampled municipal wells indicated that recharge from a sinkhole (1 to 24%) and surface water with an evaporated isotopic signature (2 to 32%) was mixing with ground water. The solute isotopes, δ13C and 87Sr/86Sr, were used to test the sensitivity of binary and ternary mixing models, and to estimate the amount of mass transfer of carbon and other dissolved species in geochemical reactions. In ground water downgradient from Lake Bradford, the dominant processes controlling carbon cycling in ground water were dissolution of carbonate minerals, aerobic degradation of organic matter, and hydrolysis of silicate minerals. In the deeper parts of the Upper Floridan aquifer, the major processes controlling the concentrations of major dissolved species included dissolution of calcite and dolomite, and degradation of organic matter under oxic conditions. The Upper Floridan aquifer is highly susceptible to contamination from activities at the land surface in the Tallahassee area. The presence of post‐ 1950s concentrations of 3H in ground water from depths greater than 100 m below land surface indicates that water throughout much of the Upper Floridan aquifer has been recharged during the last 40 years. Even though mixing is likely between ground water and surface water in many parts of the study area, the Upper Floridan aquifer produces good quality water, which due to dilution effects shows little if any impact from trace elements or nutrients that are present in surface waters.

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