Abstract The litter layer of forest soils harbors high amounts of labile organic matter, and anaerobic decomposition processes can be initiated when oxygen is consumed more rapidly than it is supplied by diffusion. In this study, two adjacent moderately acidic forest sites, a spruce and a beech-oak forest, were selected to compare the anaerobic bacterial and fungal activities and populations of conifer and deciduous leaf litter. Most probable number (MPN) estimates of general heterotrophic aerobes and anaerobes from conifer litter equaled those from deciduous leaf litter. H(2), ethanol, formate, and lactate were initially produced with similar rates in both anoxic conifer and deciduous leaf litter microcosms. These products were rapidly consumed in deciduous leaf but not in conifer litter microcosms. Supplemental ethanol and H(2) were consumed only by deciduous leaf litter and yielded additional amounts of acetate in stoichiometries indicative of ethanol- or H(2)-dependent acetogenesis. The negligible turnover of primary fermentation products in conifer litter might be due to the low numbers of acetogens and secondary fermenters present in conifer litter compared to deciduous leaf litter. Fungi capable of anaerobic growth made up only 0.01-0.1% of the total anaerobic microorganisms cultured from conifer and deciduous leaf litter, respectively. Metabolic product profiles obtained from the highest anoxic, growth-positive MPN dilutions supplemented with antibacterial agents indicated that the dominant population of fungi, apparently mainly yeast-like cells, produced H(2), ethanol, acetate, and lactate both in conifer and deciduous leaf litter. Thus, despite acidic conditions, bacteria appear to dominate in the decomposition of carbon in anoxic microsites of both conifer and deciduous leaf litter.