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A multi-mesh trammel net has been developed and used for sampling freshwater fishes. However, little is known about the catch efficiency of the net. This research investigated the catch efficiency of a multi-mesh trammel net (nominal mesh size; 10.0, 30.0, 50.0, and 70.0 mm) for fish sampling in the Yangtze River of China. Catch composition and factors affecting catch per unit effort based on fish number (CPUE_N), weight (CPUE_W), and species number (CPUE_S) were analyzed. The results showed that the net was capable of capturing a wide range of fish sizes (total length, 5.5 to 121.0 cm) and species (<i>n</i> = 50). Increasing soak time from 9.4 to 24.0 h resulted in a decrease in CPUE_N and CPUE_W while a longer soak time increased CPUE_S. CPUE_N, CPUE_W, and CPUE_S varied significantly with fishing locations. The net provides a potential complement to the current fish sampling techniques used in freshwater ecosystems (e.g., large rivers, lakes, and reservoirs). The findings of this research help to improve our understanding of the catch efficiency of the multi-mesh trammel net and provides insight into better designs for gears and methods for sampling diverse fish sizes and species.