Hysterothylacium zhoushanensis sp. nov. collected from the intestine of the flatfish Pseudorhombus oligodon (Bleeker) (Pleuronectiformes: Paralichthyidae) in the East China Sea is described and illustrated by light and scanning electron microscopy. The new species can be easily distinguished from its congeners by the presence of remarkable lateral alae, the very short intestinal caecum, the unusually long ventricular appendix (ratio of intestinal caecum to ventricular appendix 1:8.74–23.8), the short spicules (0.58–0.81 mm long, representing 1.70–2.08 % of body length) and the number and arrangement of male caudal papillae (35–42 pairs in total, arranged as 26–32 pairs of precloacal, two pairs of paracloacal and six to eight pairs of postcloacal). In addition, the adults and the putative third-stage larvae identified morphologically of the new species are characterised by sequencing and analysing the internal transcribed spacer (ITS) of the ribosomal DNA. The result reveals that they are homogeneous genetically, and all belong to the same species. Molecular analysis by comparing the ITS gene of H. zhoushanensis sp. nov. with these species of Hysterothylacium available in GenBank also seem to support the validity of the new species based on the morphological observation.