Using a spatially extensive database from the Maryland Biological Stream Survey (MBSS), we describe nutrient relationships of small-order, non-tidal streams to Maryland watershed basins, Maryland Tributary Strategy basins, and stream order. In addition, we estimate the number of stream km affected by nutrient loading, using derived nutrient criteria. Based on the MBSS spring water quality sampling, we determined several important factors relating to nutrient levels in non-tidal streams. There are strong linear relationships of nutrients to the percentage of agriculture and forested land present within MBSS sampling strata. Both mean total nitrogen (TN) and mean total phosphorus (TP) levels for watershed basins by stream order show exceedances of derived nutrient reference criteria for Maryland. Four Maryland basins have over 85% of their stream kilometers exceeding the TN criterion, with three basins over 90% of the TP criterion. To protect small stream integrity in Maryland, we recommend an upper stream TN criterion between 1.34 and 1.68 mg/L and an upper stream TP criterion between 0.025 and 0.037 mg/L, based on quantile analyses. Elevated levels of both TN and TP are present in non-tidal streams, with subsequent nutrient inputs into the upper freshwater tidal reaches of the Chesapeake Bay.