The spectral absorbance of photoreceptor visual pigments and the opsin gene class of the visual pigments was investigated in Sardinops melanostictus. Microspectrophotometric (MSP) measurements showed that the rod photoreceptors had peak absorbance spectra (λmax) at 502 nm. The spectral sensitivity of single cones was centered at 393 nm. Double cones had a λmax of 493/522 nm, but a few displayed a red-shifted absorbance of the long-wave member at 542 nm. The mRNAs of six different opsins were isolated from the retina, retrotranscribed, cloned, and sequenced. Three genes encoded opsins in the green-sensitive class (RH2), and three genes encoded opsins in the red-sensitive class (LWS), the ultraviolet (UV)-sensitive (SWS1) class, and the rod class (RH1). A Southern blot analysis showed that the blue-sensitive (SWS2) opsin gene is absent from this species, hence it was concluded that the λmax of 393 nm was generated from the SWS1 opsin. Phylogenetic analyses of S. melanostictus RH1, LWS, and SWS1 sequences placed them with orthologs from other species (e.g., the cyprinids Danio rerio and Carrasius auratus) in Otomorpha. However, unexpectedly, the RH2 sequences were more similar to orthologs in members of the Euteleosteomorpha (e.g., Oryzias latipes and Takifugu rubripes) than to cyprinid RH2 opsins.