Coelobothrium gambusiense n. sp. (Bothriocephalidae) was collected and described from the intestine of the freshwater fish Gambusia affinis (Baird and Girard) (Poeciliidae) in Fujian Province, Peoples' Republic of China. It is the first record of Coelobothrium in China. The parasite closely resembles Coelobothrium monodi Dollfus, 1970, from Capoeta damascina (Valenciennes, Cyprinidae) in Iran and Coelobothrium oitense Kugi and Matsuo, 1990, from Tribolodon hakonensis (Günther, Cyprinidae) in Japan in general morphological characters, the scolex, and the incomplete proglottids. The third species of Coelobothrium is distinguished from its congeners by its much shorter strobila, presence of a neck, a bilobed ovary instead of a transversely elongated ovary, larger eggs, different final host and locality, and other morphological characters.