Objective To explore the applicability of the histological features of plexiform bone and osteon banding to species identification. Methods 35 ground cross sections of the tibial midshaft collected from 10 human cadavers, 10 pigs, 5 cattle, 5 dogs, and 5 sheep, were observed under light microscope at 100-fold magnification. Results Plexiform bone and osteon banding were mostly observed in nonhuman bones, whereas rarely in human bones. Additionally, histological structures also varied with different individuals within a species. Conclusion The plexiform bone and osteon banding are the two important characteristics for certifying nonhuman bone in species identification.