The Gambia Estuary is one of the last estuaries in West Africa not to have been affected by strong environmental changes and adverse human disturbances. In order to describe the spatio-temporal change in fish biomass in this estuary, and the seasonal changes in fish size classes, five research surveys have been conducted at characteristic periods in the river's water regime and meteorological seasons. The fish assemblages were sampled by two vertical echosounding protocols: mobile surveys (zigzag transects) in the estuary and moored stations. The two sampling methods produced a similar image of the fish assemblages in terms of the distribution of total biomass and target strength distributions. Large fish aggregations were detected in the most marine area at the end of the wet season, and fish biomass increased through the dry season in the upstream zone. Fish were smaller at the beginning of the wet season and they were always larger in downstream areas.