In many species, including the dog (Canis familiaris), semen analysis protocols has improved in recent years. In vitro assessment of semen quality provides important information; however, the ultimate parameter for determining semen quality is pregnancy outcome. The objective of the present study was to investigate the efficiency of in vitro sperm tests in predicting seminal quality in dogs by correlating those results with pregnancy success after artificial insemination (AI). Forty-eight semen samples refrigerated for 24, 48 or 72 hours were examined for sperm motility and progressive motility, membrane integrity, and zona pellucida-binding capacity before being used to artificially inseminate bitches. Results indicated positive correlations between pregnancy success and sperm motility (0.77), vigor (0.72), membrane integrity (0.72) and zona pellucida-binding (0.81). Both pregnancy success and the sperm characteristics declined with increased semen storage time. In conclusion, the semen characteristics evaluated in this study can be used to predict the conception potential of semen samples in the dog.