Molecular prey identification in wild Octopus vulgaris paralarvae

The trophic ecology of Octopus vulgaris paralarvae collected in 2008 off the Ría de Vigo, NW Spain (42º12.80’N–9º00.00’W) was approached by both morphological and molecular methods. External digestion of prey and posterior suction of the liquefied contents by wild O. vulgaris paralarvae made the morphological identification of gut contents impossible. Thus, a PCR-based method using group specific primers was selected to identify prey consumed by O. vulgaris paralarvae in the pelagic realm. The mitochondrial ribosomal 16S gene region was chosen for designing group specific primers, which targeted a broad range of crustaceans and fishes but avoided the amplification of predator DNA. These primers successfully amplified DNA of prey by using a semi-nested PCR-based approach and posterior cloning. Homology search and phylogenetic analysis were then conducted with the 20 different operational taxonomic units (OTUs) obtained to identify the putative organisms ingested. The phylogenetic analysis clustered ingested prey into 12 families of crustaceans (11 belonging to the order Decapoda and 1 to the order Euphausiacea) and two families of fishes (Gobiidae and Carangidae). According to the Czekanowski's Index (CI), the trophic niche breadth of O. vulgaris paralarvae is low (CI = 0.13), which means that these paralarvae are specialist predators at least during the first weeks of their life cycle. It is the first time that natural prey has been identified in O. vulgaris paralarvae collected from the wild and such knowledge may be critical to increasing the survival of O. vulgaris hatchlings in captivity, a goal that has been actively pursued since the 1960s by aquaculture researchers.