Providing freshwater to human populations while protecting or rehabilitating ecosystem health is a significant challenge to water resource managers and requires accurate knowledge of aquatic resources. Previous studies of fish assemblages in the San Francisco Estuary and watershed have focused on specific habitat types, water bodies, or geographic subregions. In this study, we use seining data from two monitoring programs to provide an integrated view of spring nearshore resident fish species composition and life history characteristics in five regions: the San Joaquin River, the upper Sacramento River, the lower Sacramento River, the northern Sacramento-San Joaquin Delta (North Delta), and the Interior Delta. Data for the period March-May from 1994 to 2002, showed that spring species composition of the San Joaquin River was very different from the other four regions. Total catch in the San Joaquin River was dominated by small, short-lived batch spawning alien species (93%), particularly red shiner Cyprinella lutrensis (>75% of total catch). The upper and lower Sacramento River were very similar in species composition and life history characteristics and less dominated by alien fish (<45% of total catch). Ordination of species percentage abundances by non–metric multidimensional scaling confirmed that the major gradient was from assemblages dominated by native species to assemblages dominated by alien species. Two-way analysis of variance of ordination scores indicated that spatial variability was more important than annual variability in explaining patterns in species composition. The potential benefits of San Joaquin River native fish restoration appear high because there is so much potential for improvement; however, it is unclear how to best manipulate the system to achieve such restoration. Addressing such uncertainties is necessary if society desires the reservation and restoration of native biodiversity as human demands on water resources increase.