The third-stage larvae of several genera of anisakid nematodes are important etiological agents for zoonotic human anisakiasis. The present study investigated the prevalence of potentially zoonotic anisakid larvae in fish collected on the coastal shelves off Madang and Rabaul in Papua New Guinea (PNG) where fish represents a major component of the diet. Nematodes were found in seven fish species including Decapterus macarellus, Gerres oblongus, Pinjalo lewisi, Pinjalo pinjalo, Selar crumenophthalmus, Scomberomorus maculatus and Thunnus albacares. They were identified by both light and scanning electron microscopy as Anisakis Type I larvae. Sequencing and phylogenetic analysis of the ribosomal internal transcribed spacer (ITS) and the mitochondrial cytochrome C oxidase subunit II (cox2) gene identified all nematodes as Anisakis typica. This study represents the first in-depth characterisation of Anisakis larvae from seven new fish hosts in PNG. The overall prevalence of larvae was low (7.6%) and no recognised zoonotic Anisakis species were identified, suggesting a very low threat of anisakiasis in PNG. © 2012 Elsevier B.V.