Heterosentis holospinus Amin, Heckmann & Ha, 2011 (Arhythmacanthidae) was first described from the striped eel catfish, Plotosus lineatus (Plotosidae) in Halong Bay, Vietnam. New morphological information, scanning electron microscope images, molecular analysis, and Energy Dispersive X-ray analysis (EDXA) of hooks of specimens of H. holospinus from a new collection from the common ponyfish, Leiognathus equulus (Leiognathidae), in Quang Binh, Gulf of Tonkin, Vietnam are reported here for the first time. Additional details of the anterior trunk cone, proboscis hooks, wholly spined trunk, duck-bill-like spines with micropores, and micropore distribution, are described. The unique metal composition of hooks (EDXA) demonstrated a considerably higher level of calcium and phosphorus but lower level of sulfur at the hook basal arch than at the hook tip and edge. An analysis of our new sequences of cytochrome oxidase 1 (COI) showed that H. holospinus had low genetic variation and two haplotypes.