Comparative studies, such as the analysis of physicochemical properties and the microbiological composition of soil, are burdened with many problems resulting from the various locations of soils-often, different weather conditions among the experimental fields and varying time between the sample collection and analysis. The aim of this study was to assess the differences in the physiological profiles of bacterial communities from eight different types of soils from Poland, used in the microplot experiment that was established in 1881. The same plant species were continuously grown at all plots, at the same time, and the soil received the same type of fertilization. Moreover, the soils were always under the same weather conditions. The community-level physiological profiles of microorganisms were evaluated by using the Biolog EcoPlate™ method. The analysis demonstrated that good quality soils, especially the Gleyic Chernozem, Cambic Leptosol, and the Fluvic Cambisol exhibit a significantly higher enzyme activity, compared with the dystric soils. The dehydrogenases activity in the different time-points indicates a wide soil microbiome buffering capacity, which allows the persistence of a relatively permanent physiological profile, over many years.