To quantify species-specific relationships between bivalve carbonate isotope geochemistry (δ18Oc) and water conditions (temperature and salinity, related to water isotopic composition <δ18Ow>), an aquaculture-based methodology was developed and applied to Mytilus edulis (blue mussel). The four-by-three factorial design consisted of four circulating temperature baths (7, 11, 15, and 19¿C) and three salinity ranges (23, 28, and 32 parts per thousand (ppt); monitored for δ18Ow weekly). In mid-July of 2003, 4800 juvenile mussels were collected in Salt Bay, Damariscotta, Maine, and were placed in each configuration. The size distribution of harvested mussels, based on 105 specimens, ranged from 10.9 mm to 29.5 mm with a mean size of 19.8 mm. The mussels were grown in controlled conditions for up to 8.5 months, and a paleotemperature relationship based on juvenile M. edulis from Maine was developed from animals harvested at months 4, 5, and 8.5. This relationship 18Oc VPBD - δ18Ow VSMOW} + 0.17 (¿0.13) {δ18Oc VPBD - δ18Ow VSMOW}2; r2 = 0.99; N = 105; P < 0.0001> is nearly identical to the Kim and O'Neil (1997) abiogenic calcite equation over the entire temperature range (7--19¿C), and it closely resembles the commonly used paleotemperature equations of Epstein et al. (1953) and Horibe and Oba (1972). Further, the comparison of the M. edulis paleotemperature equation with the Kim and O'Neil (1997) equilibrium-based equation indicates that M. edulis specimens used in this study precipitated their shell in isotopic equilibrium with ambient water within the experimental uncertainties of both studies. The aquaculture-based methodology described here allows similar species-specific isotope paleothermometer calibrations to be performed with other bivalve species and thus provides improved quantitative paleoenvironmental reconstructions.