Fish assemblages from 54 oxbow lakes in the Mississippi Alluvial Valley were sampled to identify relationships among environmental variables and fish biodiversity in floodplain lake ecosystems. Environmental variables deterministic over fish biodiversity showed a hierarchical organization and were classified as primary or secondary depending on if one variable was considered to govern another. Primary variables included depth, surface area, surrounding land use, and degree of lake-river interconnectedness. Secondary variables included suites of water quality and primary productivity variables. Maximum depth and percentage of agricultural land surrounding lakes showed strongest relationships with other ecosystem components, significantly influencing water quality, primary productivity, and fish biodiversity. I found contrasting results regarding effect of lake-river interconnectedness on fish biodiversity, but maintain that connectivity is nonetheless an important floodplain lake ecosystem component and suggest that floodplain lake management efforts focus on depth, percentage of agricultural land, and restoration of connectivity.