In Sweden, whole tree harvest is common practice, possibly leading to the depletion of mineral nutrients. Furthermore, the increased use of forestry residues for heat production has caused an increasingly growing amount of by-product consisting of wood ash. Therefore, the Swedish Forest Agency has recommended wood ash application (WAA) to replace the mineral nutrients removed by whole tree harvesting, as well as a means to mitigate the acidification of boreal forests and surface waters. In a multidisciplinary study during 2003–2006 in Bispgården (Sweden), we have investigated the limnological effects on a first order stream after WAA (conducted in 2004; 3000 kg ha−1) to a 50-ha forested catchment. In general, no significant effects on an annual basis were found for acidification parameters, such as pH, alkalinity and toxic forms of aluminum (Al). There was, however, evidence of an increased pH during the spring flood, accompanied by a simultaneous decrease in the frequency of low pH-values (<5.6). Moreover, alkalinity increased in the years 2005 and 2006 compared to that of 2003, although the increase in 2006 was not statistically different from that in 2005 or 2003. High concentrations of Al repeatedly occurred in the stream, and the WAA did not affect the frequencies of high concentrations of toxic Al forms (>50 μg l−1). The benthic diatom community did not change as a result of the wood ash treatment and the diatom-based index IPS (Indice de PulluoSensibilité) indicated no nutrient enrichment or organic pollution of the stream water. There were, however, indices of elevated concentrations of potassium (K) in the aquatic moss Fontinalis antipyretica and in leaves from Alder (Alnus incana). We conclude that wood ash treatment of a forested catchment with the dose and form of ash applied in this study did not modify the freshwater ecosystem of a first order stream.