Temperature–oxygen profiles, collected biweekly to monthly for ~40 years, were used to calculate end-of-summer volume-weighted hypolimnetic oxygen (VWHO) concentrations in six small lakes located in south-central Ontario, Canada. Coherent decreases in thermocline depth and increases in hypolimnetic volume, mean hypolimnetic dissolved oxygen (DO) concentration, and VWHO were observed in five of the six study lakes. All lakes underwent an abrupt increase in VWHO and mean hypolimnetic DO after 2010. In four of the six study lakes, the highest hypolimnetic DO concentrations were observed in years where chlorophyll a concentrations were low, whereas at five of the six study lakes the highest hypolimnetic volumes were observed when dissolved organic carbon concentrations were relatively high. Warmer spring or winter air temperatures were associated with higher hypolimnetic DO concentrations at two sites, and longer ice-free periods were associated with smaller hypolimnetic volumes at two sites. These results suggest that the recent VWHO increases in the studied south-central Ontario lakes may be a function of multiple drivers that include changes in primary production, lake water transparency, and regional climatic factors.