Traits provide an informative approach to examine species-environment interactions. Often, species-by-species approaches are inefficient to generate generalizable ecological relationships and do not predict species responses to environmental changes based on specific traits species possess. Multiple lines of inquiry and multi-scale approaches are best for assessing environment-trait responses. This thesis examines important questions not specifically addressed before in traits-based research. Chapter one explores biological and ecological trait associations incorporating ontogenetic diet shifts for New River fishes. Niche shift analysis as a chapter one sub-objective quantitatively support where species-specific diet shifts likely occur. Strong biological-ecological trait associations, some intuitive and others not so intuitive, were found that relate biological structure to ecological function. Improved understanding of trait associations, including what factors influence others, supports inference of ecology of fishes. Chapters two and three examine spatial and intraspecific trait variability. Chapter two specifically examines large-scale life history trait variability along latitudinal gradients for twelve widely distributed fish species, including directionality of trait variation, and hypothesizing how optimal traits change with large-scale environmental factors. Strong positive and negative patterns found include average total length of newly hatched larvae, average total length at maturation, average spawning temperature, average egg diameter, and maximum length. These five traits are correlated with other adaptive attributes (i.e. growth rate, reproductive output, and longevity/population turnover rate). In contrast to latitudinal scale, Chapter three examines trait variability of white sucker (Catostomus commersonii) and fantail darter (Etheostoma flabellare) as a function of small-watershed scale spatial factors and anthropogenic disturbance. Toms Creek and Chestnut Creek white sucker and fantail darter displayed positive response to disturbance, contrary to past studies. Lower resource competition, and / or competitive exclusion of fishes with similar niche requirements are possible mechanisms. All three objectives support understanding of trait association and variability as a useful foundation in ecological applications and for formulating plans for conservation and management of species. / Master of Science