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While soil extracellular enzyme assays (EEAs) are frequently used to infer soil microbial function, the data typically reflect a small number of sampling points across a season, and it is unclear to what extent soil EEA may vary on the time scale of days to weeks. Rain events, in particular, may cause rapid shifts in EEA, and fine scale temporal data are needed to properly assess the generality of EEA data collected at coarser time scales. We examined soil EEA 2-3 times per week in the field from June to November in the context of natural rain events and temperature fluctuations, and explored how long-term water addition altered EEA responses. We also tested the short-term effects of water addition on the distribution of EEA in intact soil mesocoms and leachate. There was little temporal variation in EEA for the hydrolases phosphatase. N-acetyl-glucosaminidase and beta-glucosidase, despite the occurrence of multiple large rain events and large soil temperature fluctuations. Phenol oxidase activity correlated significantly with seasonal trends in temperature and soil moisture, but was highly variable at short time scales, and the latter did not correlate significantly with short-term changes in soil microclimate. EEA generally increased in response to long-term water addition, and in soil mesocosms water addition did not significantly redistribute EEA among the upper and lower soil layers, and leachate EEA was three orders of magnitude lower than soil EEA. Overall, our results reveal relatively minor short-term variation in EEA for hydrolase enzymes, and no discernable response to temperature fluctuations or precipitation over the short term. However, high short-term variation in phenol oxidase activity suggests that it may be difficult to infer temporal trends in EEA for this enzyme from a limited number of sampling points. (C) 2011 Elsevier GmbH. All rights reserved.