Abstract: To grasp the spatial-temporal distribution characteristics of fish resources in the Jianli and Hukou reaches of the middle Yangtze River, a synchronous fishery resources survey was conducted in June (wet season) and November (dry season) of 2023 using a combination of acoustic and fish catch methods. The results indicated that in terms of temporal distribution, the fish densities in the Jianli reach during the two periods were 47.66 and 25.27 individuals per 1 000m3, with average target strengths of -59.04 and -62.24 dB, respectively. Meanwhile, the fish densities in Hukou section were 43.50 and 23.08 individuals per 1 000 m3, with average target strengths of -55.01 and -59.28 dB, respectively. Significant differences (**P<0.01) were observed in fish density and target strength between the two periods in both Jianli and Hukou sections. In terms of spatial distribution, the fish density was unevenly distributed. The Jianli section demonstrated a higher concentration in the areas with winding river channels, while the Hukou section had a higher concentration in areas with dense sandbars. Both Jianli and Hukou sections exhibited the highest fish density in the bottom layer, followed by the middle layer, and the lowest in the surface layer during the high-water period. In contrast, during the dry season, the highest fish density was observed in the middle layer, followed by the bottom layer, and the lowest in the surface layer. However, differences in fish density and target strength among these layers during the dry and flood seasons were not statistically significant (P>0.05).Combined with the fishing catch survey data, it was found that the fish composition in both Jianli and Hukou sections was dominated by Cyprinidae, with a majority of small-sized fish species distributed mainly in the middle and lower water layers. The temporal and spatial distribution characteristics of fish in Jianli and Hukou sections were closely related to their ecological habits, such as reproduction, feeding, and overwintering behaviors. These findings provided data support for assessing the restoration and protection of fish resources in the middle Yangtze River after the implementation of the fishing ban policy.