The objective of this study was to develop tools for distinguishing between sexes of the two Black-crowned Night-Heron (Nycticorax nycticorax) subspecies (N. n. obscurus and N. n. hoactli) using discriminant function analysis. Significant differences were found in length of the culmen, length of the bill from the gape, and length of the wing chord between the sexes of each subspecies, with males being larger than females. The resulting discriminant functions were able to differentiate between the sexes of each studied subspecies and between subspecies after determining the sex of the individuals (with a correct classification of 77.8% for females and 97.8% for males). In females, all morphometrics were greater for N. n. obscurus than N. n. hoactli; this was also the case for males, except for bill width, which was greater in N. n. hoactli. Wing chord length was the most useful measurement for constructing the discriminant functions. External morphometrics are a valuable tool not only for discriminating between N. n. hoactli and N. n. obscurus but also for sexing these subspecies.