The dwindling number of the tri-spine horseshoe crab, Tachypleus tridentatus has been reported globally and its status in Malaysia is not much known. Study on dimorphism in adult body sizes and population size estimation were conducted using capture–mark–recapture method of adult T. tridentatus in Tawau, Sabah. Camry the estimated population sizes of T. tridentatus ranged from 182 to 1095 with 95% confident limits of 56–42,942 individuals (Schnabel formula). The multivariate discriminant Hotelling’s T2 test verifies the sexual size dimorphism among the adult T. tridentatus with 97.7% separation among sexes (Hotelling’s T2 = 778.49, F = 152.85, p < 0.001) with females being larger and heavier than the male individuals. The number estimated from the study is the first reported for T. tridentatus in Malaysia, particularly in Sabah. Even though this number may slightly overestimate the actual population size in the area owing to the low number of individuals recaptured, for now it could serve as baseline data for horseshoe crab management purpose.