This study was designed to test the hypothesis that desert ant species that build nests that remain viable at a particular point in space for more than a decade produce soil conditions that enhance microbial biomass and functional diversity. We studied the effects of a seed-harvester ant, Pogonomyrmex rugosus, and two generalist ant species, Aphaeonogaster cockerelli and Myrmecocystus depilis, on soil microbial communities. Microbial biomass was higher in P. rugosus-modified soils than in reference soils when soil water content was higher than 3%. Microbial biomass was either higher in reference soils or exhibited no difference in reference soils and nest-modified soils of A. cockerelli and M. depilis. There were differences in microbial functional diversity and microbial community level physiological profiles (MicroResp method) between ant-nest-modified and reference soils of the three ant species on some sampling dates. Temporal pattenrs of soil microbial communities associated with the ant species resulted from differences in soil moisture, density, and species composition of the annual plant c0ommunities associated with the ant nests and in reference areas. Differences in annual plant communities associated with ant nests and surrounding areas resulted in different chemical inputs into the soil organic-matter pools. This study shows that generalizations about the effects of long-lived ant nests on soil biota in arid regions must consider feeding behaviors of the ant species and temporal patterns of rainfall.