Northeastern North America has been experiencing warmer winters with reduced snow accumulation, with more frequent winter freeze-thaw cycles.We conducted a laboratory experiment to investigate how increased frequency of freeze-thaw cycles (FTC) would alter soil C and N availability.Organic (O) and mineral (B) horizon soils were collected from a coniferous forest in Maine, processed to exclude roots, and then frozen in the laboratory (-10 °C) with one (FTC-1), two (FTC-2), or six (FTC-6) thaw periods (+5 °C).Soils were analyzed for extractable ammonium (NH4-N), water extractable organic carbon (WEOC), carbon dioxide flux (respiration), and total C and N.Extractable NH4-N increased following FTC (all levels), for both horizons.While WEOC concentrations did not change for FTC vs. control, the WEOC in O horizons had a lower SUVA254 in FTC soils compared to control, indicating a stronger microbial influence (i.e., microbial cell lysis) in these soils after FTC.Respiration in O horizon soils decreased post-incubation and did not differ between FTC and Control soils.In the B horizon, however, FTC soils showed greater respiration than Control soils, suggesting that the newly available nutrients may have stimulated microbial activity.In contrast to these results, total C and N remained unaltered by FTC, presumably because the FTC disturbances represented mostly a translocation of C and N from one pool into another, and losses due to respiration were too small to significantly influence the large TC and TN pools.The effect of FTC on NH4-N did not change with FTC frequency, suggesting that a single FTC is sufficient to alter both C and N availability and/or quality, and that addnl. FTC may not have a significant further effect.This study provides fresh insights on how organic and mineral horizon soils might respond to increased freeze-thaw frequency in winter.