Summary1. Habitat fragmentation is a major threat to biodiversity and can lead to the loss of both species and ecosystem services, but fragmentation effects vary greatly between studies and studied organisms. Understanding the distinct effects of habitat amount and isolation at the patch and landscape scale may account for some of this variation. 2. We studied biodiversity in 30 traditional orchards that were selected for independent variation in habitat amount and habitat isolation at the patch and landscape scale. We analysed species richness and abundance of snails, beetles, true bugs, spiders and breeding birds that avoid open farmland but occur in woody vegetation types. Additionally, the abundances of nine single species were analysed using specific habitat definitions. 3. Surprisingly, the effects of habitat isolation were more important than the effects of habitat amount. Effects at the patch scale were more frequent than landscape-scale effects. 4. Spider species richness decreased with increasing patch-scale habitat amount. Abundance of the weevil Phyllobius oblongus increased with landscape-scale habitat amount. Negative effects of patch isolation were greater for predatory birds and spiders, while the predominately herbivorous beetles, true bugs and snails were less affected. Species richness of birds, spiders and beetles, and abundance of birds, Cyanistes caeruleus, Parus major and Fringilla coelebs, decreased with increasing patchscale habitat isolation. In contrast, species richness of spiders and beetles increased with increasing landscape-scale habitat isolation. 5. Synthesis and applications. The effects of habitat fragmentation differed between taxonomic groups, with stronger and more consistent responses in birds than invertebrates. Our understanding of fragmentation effects may be biased due to the dominance of bird studies in the literature, and further invertebrate studies are encouraged. Landscape management to improve biodiversity or ecosystem services requires a group-specific approach and coordinated priority setting. High habitat connectivity benefited wood-preferring birds, spiders and beetles, lending support to national initiatives for increased habitat connectedness. The negative effects of patch isolation were greater for natural pest regulators, birds and spiders than for herbivorous beetles and bugs.