Bacterial communities of urban soils have not been thoroughly investigated up to now. Therefore, soil samples from the urban park Tiergarten in the centre of Berlin were taken from a profile in 15, 30 and 90 cm depth. The total number of bacteria (4′,6-diamidino-2-phenylindole (DAPI) counts) as well as biomass declined one order of magnitude from topsoil to subsoil. Soil texture changed comparably and water content and amount of organic matter dropped 3–10-fold. The number of culturable bacteria (colony forming units = CFU) also decreased with increasing soil depth. Amplified ribosomal DNA restriction analysis (ARDRA) revealed similar bacterial communities in the two upper soil layers in contrast to the deepest layer. The number of bacterial cells which were detected with probe EUB338 in relation to total cell counts differed between 43 and 35% in the three soil layers. With the probe active count method (PAC) this number could be increased up to 72% of total cell counts in topsoil whereas activation of cells declined with increasing depth. In relation to total cell counts (DAPI) α- Proteobacteria and β- Proteobacteria are equally distributed in all three depths, whereas γ- Proteobacteria declined within the soil profile. With the BIOLOG system we observed the general trend that the capability of utilizing diverse substrates decreased with soil depth whereas a few substrates, such as Tween 40 and Tween 80 could be utilised by the bacteria of all soil depths.