Spatial and temporal variation of tropical insect communities has rarely been studied, although such variation influences estimates of global species richness. Therefore, we compared spatial and temporal variation of herbivorous insect communities on Neoboutonia macrocalyx trees among seven sites over 1 y in a primary tropical rain forest in Kibale National Park, Uganda. The distance between the study sites varied from 4.8 to 31.2 km and altitudinal differences ranged from 20 to 242 m. Permutational multivariate analysis of variance (PERMANOVA) revealed significant spatial changes in community composition of the herbivorous insects and study sites differed also in insect abundance (6.9–26.2 individuals m−2 of leaf area). This is likely to be caused by differences in vegetation, altitude and microclimate among the study sites. The similarity of insect species composition was negatively correlated with geographic and altitudinal distances among sites and positively correlated with the similarity of tree community composition. Species richness varied significantly between sampling dates, ranging from 33 to 41 species. Also community compositions changed between sampling dates, which likely follows from marked seasonal changes in climate and the phenology of other host plants used by the generalist insect species also living on Neoboutonia macrocalyx. In general our study supports the idea of high variability of herbivorous insect communities in primary rain forests even at a small spatial scale. This should be considered when estimations of insect biodiversity are made.