Background: Madhya Pradesh is rich in biodiversity having diversified flora and fauna. But now a days, apart from various threat like loss of habitat poisoning, accident, disease facing wildlife today and poaching remain one of the important causes for fauna declination. poaching remains one of important cause for fauna declination. Poaching of wild animals particularly chital and sambar deer is more for the meat, ornamental and trophy purposes. In forensic laboratory where there is lack of molecular facilities, comparative gross morphological study plays an important role in solving wildlife crime cases. skull is a best material to compare the closely relative species. hence, the study was conducted on comparative gross morphology of 4 male chital and sambar deer skull. Methods: The study was conducted 4 male adult skulls each of chital and sambar deer. The skulls were de-skinned and kept under maceration for a period of one month. The gross morphological differentiating features of dorsal surface, lateral surface, basal surface and nuchal surface were observed and recorded with the help of digital camera. Result: In this study, unique differentiating features were sufficient to distinguish among the skull of male chital and sambar. The facial tuberosity was present caudal to infraorbital foramen, dorsal to the superior third premolar tooth in chital deer, whereas; it was present dorsally the superior first molar tooth in sambar deer. Rostral extremities of nasal bones in skull of male chital deer were notched; however, these were blunt and without notch in the skull of male sambar deer. Incisive (Premaxilla) bone was directed upward in chital deer; whereas, it was directed caudally in case of sambar deer. The present comparative study will provide reliable information for identification of these closely related species specially needed for wildlife forensic cases.