In the present study we evaluated how management practices, structural habitat parameters and arthropod availability affect bat activity, species richness and species diversity (estimated by Shannon's diversity index H') in a cultural landscape composed by a mosaic of different habitats in Brandenburg (Germany). Over a study period of two years (2012–2013), a standardised acoustic monitoring was conducted on 12 sampling sites comprising different land use types in forests (pine, mixed and deciduous forest) and agricultural areas (arable land and grassland). The focus was on the effect of small-scale changes in structural habitat parameters resulting from management practices within one-and-the-same land use type. We applied a paired sampling design and compared two (forest) and three (agricultural areas) sampling sites (complexity categories) per land use type. An effect of structural complexity was tested using parametric and non-parametric analyses. Sampling sites differ in a complexity index derived from vegetation measurements related to: a) vertical vegetation structure (pine forests), b) tree species composition (mixed forests), c) tree age (deciduous forests), d) crop type (arable land), and e) management intensity (grassland). Within the pine and mixed forest, management practices leading to an increase in structural habitat complexity were associated with a significant increase in bat activity, species richness, and species diversity (H'). This effect is only partially explained by increased prey abundance. On grassland, increased bat abundance is associated with low-intensity management practices. This effect is explainable by an increase in arthropod abundance associated with structural complexity but no additional effect of structural complexity beyond that. On arable land, the difference in structural complexity among different crops (and not prey abundance) significantly affects bat communities. Our approach employed proxy measures of habitat quality to estimate which management practices within the study area theoretically result in suitable and available habitats to meet the requirements of local bat species. Thus, our findings have implications for forest and agricultural management. Even minor changes in silvicultural management (understory development in pine monocultures and increased percentage of deciduous trees in mixed forests) may lead to a significant increase in forest habitat quality for bats. The findings also stress the importance of extensive management practices in grassland, as well as structure-rich crops on arable land to achieve a more environmentally sustainable farmland management.