The nervous systems of three Tylodelphys metacercariae (T. mashonense, Tylodelphys spp. 1 and 2) co-occurring in the cranial cavity of the catfish, Clarias gariepinus, were examined by the activity of acetylthiocholine iodide (AcThI), with the aim of better understanding the arrangement of sensillae on the body surface and the nerve trunks and commissures, for taxonomic purposes. Enzyme cytochemistry demonstrated a comparable orthogonal arrangement in the three metacercariae: the central nervous system (CNS) consisting of a pair of cerebral ganglia, from which anterior and posterior neuronal pathways arise and interlink by cross-connectives and commissures. However, the number of transverse nerves was significantly different in the three diplostomid metacercariae: Tylodelphys sp. 1 (30), Tylodelphys sp. 2 (21) and T. mashonense (15). The observed difference in the nervous system of the three metacercariae clearly separates them into three species. These findings suggest that consistent differences in the transverse nerves of digenean metacercariae could enable the differentiation of metacercariae to the species level in the absence of molecular techniques. This, however, might require further testing on a larger number of species of digenean metacercariae.