The abundance and activity of methane-oxidizing bacteria (MOB) in the water column were investigated in three lakes with different contents of nutrients and humic substances. The abundance of MOB was determined by analysis of group-specific phospholipid fatty acids from type I and type II MOB, and in situ activity was measured with a 14 CH 4 transformation method. The fatty acid analyses indicated that type I MOB most similar to species of Methylomonas, Methylomicrobium, and Methylosarcina made a substantial contribution (up to 41%) to the total bacterial biomass, whereas fatty acids from type II MOB generally had very low concentrations. The MOB biomass and oxidation activity were positively correlated and were highest in the hypo-and metalimnion during summer stratification, whereas under ice during winter, maxima occurred close to the sediments. The methanotroph biomass-specific oxidation rate (V) ranged from 0.001 to 2.77 mg CH 4 -C mg ؊1 C day ؊1 and was positively correlated with methane concentration, suggesting that methane supply largely determined the activity and biomass distribution of MOB. Our results demonstrate that type I MOB often are a large component of pelagic bacterial communities in temperate lakes. They represent a potentially important pathway for reentry of carbon and energy into pelagic food webs that would otherwise be lost as evasion of CH 4 .Microbiological oxidation of methane takes place in many kinds of aquatic systems and soils (28,32,33,58) and plays an important role in the global budget of this greenhouse gas (15,43). It can proceed both under aerobic and anaerobic conditions, but the latter seems to be extensive primarily in waters with high ionic strength, e.g., saline lakes and marine systems (29,31,55). Many strains capable of aerobic methane oxidation have been isolated and characterized, and phylogenetically all belong to either the ␥-proteobacteria (commonly referred to as "type I" methanotrophs) or the ␣-proteobacteria ("type II" methanotrophs) (10,12,24).Aerobic methane-oxidizing bacteria (MOB) in the water columns of lakes consume a significant part of the methane produced in the sediment or in anaerobic layers of water (46,54). Besides metabolizing methane that would otherwise be emitted to the atmosphere, they represent a route for reentry of carbon back into the food-webs (2,26,30,46). In spite of this, direct studies of MOB populations in the pelagic systems of lakes are rare and, with the exception of a small oxbow lake (45) and a tropical hydropower reservoir (20, 21), very little is known about the abundance and population structure of pelagic methanotrophs in lakes.The aerobic methanotrophic bacteria contain some phospholipid fatty acids (PLFAs) that seem to be unique for this group of organisms (16:18c in type I MOB and 18:18c in type II MOB (10, 40). Analysis of these PLFAs has been shown to be a very sensitive tool for the direct study of populations of type I and type II MOB in samples from several different kinds of ecosystems (7,9,16,39,51).In the prese...