There has been much debate as to whether H. placei is a separate species to H. contortus. The aim of this study is to provide molecular information to assess the species status of H. placei. Using the polymerase chain reaction, the second internal transcribed spacer (ITS-2) of ribosomal DNA was amplified and sequenced. Comparison of the 231 base pair ITS-2 sequences showed no intraspecific variation in H. contortus, among one isolate from each of the United Kingdom. Switzerland and China and 5 isolates from within Australia, or among 3 Australian isolates of H. placei. Three (1.3%) nucleotide differences were detected between the ITS-2 sequences of H. contortus and H. placei. In addition, 2 diagnostic sites for the endonucleases BfaI and FokI allowed the delineation of H. placei from H. contortus. The data presented herein support previous morphological and genetic evidence that H. placei is a separate species to H. contortus.