The related topics of spatial variability of aquatic invertebrate community metrics, implications of spatial patterns of metric values to distributions of aquatic invertebrate communities, and ramifications of natural variability to the detection of human perturbations were investigated. Four metrics commonly used for stream assessment were computed for 9 stream reaches within a fairly homogeneous, minimally impaired stream segment of the San Pedro River, Arizona. Metric variability was assessed for differing sampling scenarios using simple permutation procedures. Spatial patterns of metric values suggest that aquatic invertebrate communities are patchily distributed on subsegment and segment scales, which causes metric variability. Wide ranges of metric values resulted in wide ranges of metric coefficients of variation (CVs) and minimum detectable differences (MDDs), and both CVs and MDDs often increased as sample size (number of reaches) increased, suggesting that any particular set of sampling reaches could yield misleading estimates of population parameters and effects that can be detected. Mean metric variabilities were substantial, with the result that only fairly large differences in metrics would be declared significant at α = 0.05 and β = 0.20. The number of reaches required to obtain MDDs of 10% and 20% varied with significance level and power, and differed for different metrics, but were generally large, ranging into tens and hundreds of reaches. Study results suggest that metric values from one or a small number of stream reach(es) may not be adequate to represent a stream segment, depending on effect sizes of interest, and that larger sample sizes are necessary to obtain reasonable estimates of metrics and sample statistics. For bioassessment to progress, spatial variability may need to be investigated in many systems and should be considered when designing studies and interpreting data.