The mountain pine beetle, Dendroctonus ponderosae (Coleoptera: Scolytinae), is an economically important bark beetle species with a wide geographic range spanning from the southwestern United States into northern Canada. This beetle causes extensive tree mortality to 13 pine species. Mites (Acari) are common and abundant symbionts of mountain beetles that may influence their fitness through positive and negative interactions. We present a unique assessment of the mite associates of mountain pine beetles using measures of alpha and beta diversity. We sampled phoretic mites from five beetle populations: Arizona, Colorado, South Dakota, Utah (USA), and Alberta (Canada) that varied in host tree species, local climate, and beetle population level. We collected 4848 mites from 8 genera and 12 species. Fifty to seventy percent of beetles carried mites in flight with the highest mite loads occurring in middle and southern populations; decreasing in northern populations. Mite assemblages (i.e., both richness and composition) varied along a south to north latitudinal gradient and were driven by species turnover (i.e., species replacement). Differences in mite composition increased with distance between populations. We discuss climatic variation, environmental filtering, and host tree differences as factors that could affect differences in mite composition between beetle populations and discuss implications for functional shifts. Our results could represent a model for estimating diversity patterns of mite symbionts associated with other major insect pests in coniferous forest systems.