We used field-derived data from streams in Nevada, USA, to quantify relationships between stream biological condition, in-stream stressors, and potential sources of stress (land use). We used 2 freshwater macroinvertebrate-based indices to measure biological condition: a multimetric index (MMI) and an observed to expected (O/E) index of taxonomic completeness. We considered 4 categories of potential stressors: dissolved metals, total dissolved solids, nutrients, and flow alteration. For physicochemical factors that varied predictably across natural environmental gradients, we quantified potential stress as the site-specific difference between observed (O) and expected (E) levels of each factor (O–Estress). We then used 2 sets of Random Forest models to quantify relationships between: 1) biological condition and potential stressors, and 2) stressor values and land uses. The 2 indices of biological condition were differentially responsive to stressors, indicating that no single measure of biological condition could fully characterize assemblage response to stress. Total dissolved solids (as measured by electrical conductivity [EC]) and metal contamination were the stressors most strongly associated with biological degradation. The most likely sources of these stressors were agriculture, urban development, and mining. Our findings highlight the need to develop EC criteria for streams. Measures of biological condition and stress that account for natural variability should reduce errors of inference and increase confidence in causal analyses. This approach will require development of robust models capable of predicting physical and chemical reference conditions. Causal analyses for individual sites require appropriate hypotheses about which stressors and what levels of stress can cause biological degradation. Our study demonstrates the usefulness of field data collected from multiple sites within a region for developing these hypotheses.