In order to clarify and enhance awareness of freshwater venomous fishes, the fauna composition, spatial distribution and feeding characteristics of venomous fishes in the Yellow River Basin were studied based on historical and watershed fishes survey data. The results showed that 67 species from 41 genera, 16 families and 10 orders of venomous fishes were found in the river basin, which accounted for 33.33 percent of the total species of fishes. In taxonomic level, the proportion of venomous fish in taxa decreased gradually from higher taxa to lower taxa. In spatial distribution level, the number of poisonous fish species increases from the source to the downstream estuary segment. Schizopygopsis pylzovi and Platypharodon extremus are only present in the source and upstream reach while other venomous fish species such as Takifugu ocellatus, Takifugu obscurus, Takifugu rubripes and Takifugu niphobles were only distributed in the estuary of the Yellow River and other venomous fishes were distributed in whole reach. The similarity analysis showed that the distribution of venomous fish fauna form was similar in all river segments and chi-square test (χ2) showed no significant difference in the distribution of venomous fish fauna in all river segments (P > 0.05). There were twenty-five species of gall-bladder posinous fishes, twenty-four species of acanthotoxic fishes, seven species of ichthyootoxic fishes, five species of ichthyohepatoxic fishes, four species of puffer-like poisonous fishes and two species of ichthyohemotoxic fishes among the sixty-seven venomous fish species. The dominant toxic types in the Yellow river were gall-bladder posinous fishes and acanthotoxic fishes, which were consistent with the main toxin types of freshwater poisonous fish, and mainly concentrated on cypriniformes, siluriformes and perciformes. There were seven kinds of trophic guilds in venomous fishes in the river basin, which mainly concentrated on carnivorous and invertebrate feeding guild, accounting for 32.83 percent and 28.36 percent of the poisonous fish species, respectively. The comprehensive statistical analysis showed that the venomous fishes in the Yellow River basin are widely distributed and the poisonous types are diverse. The results can provide scientific support for the identification, development and utilization of venomous fish resources in the Yellow River Basin.