Objective To investigate the infection in marine fishes from the local markets in Liaoning Province and identify the third larvae of Anisakis by molecular biological method. Methods The microscopically morphologies were performed after the larvae of Anisakis were detected by direct dissection. The rDNA internal transcribed space (ITS) sequences of Anisakis were extracted and amplified, and the sequences of ITS1 and ITS2 in the ribosome gene spacer were compared and analyzed. Results The larvae of Anisakis was detected from 70 fishes, the infection rate was 31.53%(70/222). Larimichthys polyactis and Trichiurus haumel had higher infection rates, and the highest degree of infection was 233 larvae per fish. The larvae were divided into different genus by sequence alignment and evolutionary analysis. Five Anisdkis nematodes including Anisakis simplex, Anisakis pegreffii, Anisakis typical, Hysterothylcaium aduncum and Hysterothylcaium amoyense were identified. Conclusion The infection situation of marine fish was quite serious in Liaoning Province, and the dominant species of infection was Anisakis pegreffii.