Identification of barramundi (Lates calcarifer) and tilapia (Oreochromis spp.) fillets by DNA- and protein-analytical methods

Abstract

Fish and fillet of barramundi (Lates calcarifer) and tilapia (Oreochromis species) obtained from wholesale and retail trade were assigned to species by sequencing of PCR products. Two segments (358 and 464 bp) of the cytochrome b gene (cytb) were amplified using universal primers. The amplicons gave characteristic patterns in SSCP-analysis (single strand conformation polymorphism) suitable for differentiation of Lates calcarifer from Lates niloticus and Lateolabrax japonicus. Intra-specific variation of sequences and SSCP patterns were observed for barramundi. In case of tilapia species, it was found to be difficult to identify samples by BLAST due to the high similarity of cytb sequences of O. niloticus, O. mossambicus, O. aureus and Sarotherodon galileus. Four different patterns of single strand DNA (ssDNA) were obtained by SSCP analysis of the 464 bp amplicon of tilapia. Different patterns of ssDNA matched to variations in sequences. Protein profiles obtained by IEF (isoelectric focusing) of water-soluble proteins from raw fillet were found to be suitable for rapid differentiation of Lates calcarifer from Lateolabrax japonicus, but the three different Oreochromis species expressed only minor differences in protein patterns. The patterns of the tilapia and barramundi species showed a number of acidic, heat-stable proteins, presumably representing parvalbumin.