The objective of this study was to assess the effects of spatially and temporally varying abiotic conditions on the nekton residing in tidally influenced streams and determine the comparability of the assemblage response across the various gears needed to fully characterize the biota. Three tidal streams were sampled seasonally for 2 years for chemical (physiochemical profiles), physical (in-stream and riparian habitat classification), and biological (nekton sampled with bag seines, trawls, and gill nets) components of ecosystem integrity. Multiple sampling stations on each stream encompassed the transitional character of the tidally influenced ecosystem, from the freshwater of the river to the saltwater of the bay. Instead of characterizing the biota with traditional fish-based indexes of biotic integrity, analysis methodologies relying heavily on multivariate ordination techniques were used. This study showed that the temporal and spatial relationships among nekton assemblages and abiotic environmental conditions were quite gear dependent. The greatest degree of difference in indicators of ecosystem health all involved upstream-downstream gradients that appear to be driven by salinity structure. Based on the results of this study, dissolved oxygen concentration does not appear to be a major structuring factor in the physical, chemical, or biological component of ecosystem integrity.