Fish invasions can damage the ecological environment of invaded areas, causing negative effects such as monotony of ecological types in invaded waters and endangerment of native species. It is important to monitor their presence and spread in invaded areas. This study aims to update the available data on non-native fish species in the inland waters of Guangxi and to determine their seasonal and spatial distribution in this region. Taking the inland waters of Guangxi as the study area, 34 sampling sites were set up in the major river systems, and systematic sampling was conducted in four seasons in 2023. The data showed that a total of 7690 non-native fish were collected from 23 species, belonging to 7 orders, 13 families, and 20 genera. Of the non-native fish species, 19 species were introduced for aquaculture purposes, and three species were introduced for ornamental purposes. The most non-native fish species were found in summer with 21 species, which were followed by 20 species in winter, 18 species in spring, and only 15 species in autumn. However, the distribution composition was similar in each season, with Coptodon zillii being dominant in each season, which was followed by Oreochromis niloticus. The Hongshuihe River had the highest number of non-native fish species with 16 species, followed by the Xunjiang River with 14 species, the Qianjiang River with 13 species, and the Npanjiang River had the lowest number of non-native fish species with 7 species. In addition, the Xunjiang River and the Qianjiang River showed significant separation in the PCoA results, and the overall test showed significant differences in non-native fish composition among the river system. The main reasons for the differences in their spatial distribution are geographical location and temperature. O. niloticus and C. zillii were found in all river systems and have become the main invasive non-native fish species in the inland waters of Guangxi.