Electrostatic properties of milt have a huge bearing on the reproductive success of externally fertilizing freshwater fish species. Considering the global significance of carp in aquaculture, we explored the charge profile of milt of ten commercially important carp species for the first time. Mean zeta potential of milt ranged from (−) 33.86 mV in Cirrhinus mrigala to (−) 56.34 mV in Systomus sarana at pH 7.0. The effect of pH on milt characteristics was studied with Labeo rohita as the model species. Zeta potential reduced significantly upon exposure to low pH (4.0) and increased at alkaline pH (10.0). Fertilization rate was significantly reduced at acidic pH. Our results indicate that changes in milt zeta potential due to acidic pH is a significant factor in reduced reproductive success of externally fertilizing freshwater fish. While low concentration of proteases were detected in milt, it remained in a stable colloidal form even after incubation for 18 h at room temperature with no significant difference in the mean zeta potential at the end of incubation. This seminal work on milt zeta potential of carp will open several avenues in aquaculture and fisheries for productivity enhancement and conservation. Study of the electrostatic properties of milt can also help explain puzzling questions of ecology, such as migratory behavior.