Benthic macroinvertebrate diversity influences stream food web dynamics, nutrient cycling and material exchange between the benthos and the water column. Stream bioassessment has moved to the forefront of water quality monitoring in terms of benthic macroinvertebrate diversity in the recent past. The objectives of this study were to determine optimum subsample size and level of taxonomic resolution necessary to accurately and precisely describe macroinvertebrate diversity in streams flowing in the Piedmont province of the James River watershed in Virginia. Forty-nine sampling sites were selected from streams within the Piedmont Physiographic Province of the James River watershed. Ten sites were randomly selected to have all macroinvertebrates in the sample identified to the genus level whenever possible. Optimum subsampling intensities and Virginia Stream Condition Index (VSCI) metrics and scores were determined. For samples with the total number of individuals at less than 500, the genus level of taxonomy provided lower overall optimum subsampling intensities. However, for samples with total individuals over 1000, optimum subsampling intensities at the genus level of taxonomy were higher than the family level for more than 50% of the metrics. For both family and genus levels of taxonomy, the majority of optimum subsampling intensities were well over 50% of the total individuals in the sample, with some as high as 100% of the individuals. While optimum subsampling intensities were valuable in comparing family and genus level taxonomy, they are not reasonable for stream bioassessment protocols; the cost:benefit ratio would be highly unbalanced. A minimum subsample size of 200 individuals is optimum for determining VSCI scores, while optimum taxonomic resolution is dependent on several factors. Thus, the level of taxonomic resolution for a particular study should be determined by the study objectives, level of site impairment and sample size.