The role and identity of host and non-host chemical cues (semiochemicals) in host location and non-host avoidance for copepodid larvae of sea lice, Lepeophtheirus salmonis, was investigated using Y-tube behavioural bioassays, solid-phase extraction (SPE), and coupled gas chromatography mass spectrometry (GCMS). Using artificial seawater conditioned with the preferred salmonid host, Salmo salar, L. salmonis displayed high activation and directional responses in Y-tube assays to salmon-conditioned water (SCW), to an extract of SCW prepared by SPE, and to a vacuum distillate of the SPE extract. Similar responses were observed to two chemicals identified from SCW by coupled GCMS: isophorone and 6-methyl-5-hepten-2-one. Dose-response studies with isophorone showed that copepodid responses across the range tested were maximised at 0.01 and 0.1 mg·mL1. A mixture of isophorone and 6-methyl-5-hepten-2-one also induced high activation and directional responses. Semiochemicals were also isolated from the non-host fish, turbot (Scophthalmus maximus (Rafinesque)), by SPE and analysed by GCMS. Two non-host-specific chemicals were identified as 2-aminoacetophenone and 4-methylquinazoline. When SCW was mixed with either of the non-host chemicals, activation and directional responses to SCW were eliminated in the Y tube.