A Rapid Method for the Identification of Fresh and Processed Pagellus erythrinus Species against Frauds

Abstract

The commercialization of porgies or seabreams of the family Sparidae has greatly increased in the last decade, and some valuable species have become subject to seafood substitution. DNA regions currently used for fish species identification in fresh and processed products belong to the mitochondrial (mt) genes cytochrome b (Cytb), cytochrome c oxidase I (COI), 16S and 12S. However, these markers amplify for fragments with lower divergence within and between some species, failing to provide informative barcodes. We adopted comparative mitogenomics, through the analysis of complete mtDNA sequences, as a compatible approach toward studying new barcoding markers. The intent is to develop a specific and rapid assay for the identification of the common pandora Pagellus erythrinus, a sparid species frequently subject to fraudulent replacement. The genetic diversity analysis (Hamming distance, p-genetic distance, gene-by-gene sequence variability) between 16 sparid mtDNA genomes highlighted the discriminating potential of a 291 bp NAD2 gene fragment. A pair of species-specific primers were successfully designed and tested by end-point and real-time PCR, achieving amplification only in P. erythrinus among several fish species. The use of the NAD2 barcoding marker provides a rapid presence/absence method for the identification of P. erythrinus.

Dentex dentex Frauds: Establishment of a New DNA Barcoding Marker

“…Sixteen complete Sparidae mitogenome sequences were analyzed in this study, as reported in Ceruso et al [ 21 ]. Eleven specimens of D. dentex from Food and Agriculture Organization (FAO) area 37 were sampled ( Table 1 and Figure 1 ).…”

“…The fish species other than Dentex dentex were carefully chosen with the aim to include (i) those used to substitute Dentex dentex (e.g., Dentex gibbosus ), (ii) phylogenetically related sparid species (e.g., Pagellus erythrinus ) [ 18 ], and (iii) other species commonly found in Mediterranean fish markets. Ten specimens of Pagellus erythrinus , the most genetically close to Dentex dentex among sparids [ 21 ], were collected.…”

“…Our purpose was to identify a gene or gene fragment more specific for Dentex dentex and with a higher capacity of species differentiation. Therefore, we compared 16 sparid mitogenomes [ 17 , 18 ], 6 of which [ 19 , 20 , 21 , 22 , 23 , 24 ] were sequenced in our laboratories using a new and effective extraction protocol [ 25 ]. Our previous studies in this field led to the identification of two new barcode markers for the Sparidae family.…”

Geographic Pattern of Sushi Product Misdescription in Italy—A Crosstalk between Citizen Science and DNA Barcoding

“…Among the most used molecular markers, mitochondrial genes, such as Cytochrome b (Cytb), 16S rRNA (16S), Cytochrome Oxidase I (COI) and mtDNA Control Region (CR), have proven to be optimal tools for seafood species authentication. However, while the CR and Cytb have been successfully and widely used to study the genetic population structure [ 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ] rather than to authenticate fish species [ 27 , 28 , 29 ], COI has become the optimal DNA barcode for the identification of animal species [ 30 , 31 , 32 , 33 , 34 , 35 ] and particularly for fish species authentication in seafood products [ 15 , 36 , 37 , 38 , 39 , 40 , 41 ]. Furthermore, researchers have been also encouraged to look for rapid and low-cost molecular strategies to tackle substitution species frauds by large scale screening both using classic and new technologies [ 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 ].…”

Advancing traceability using DNA sequence on seafood product: fraudulence in shrimp crackers

“…Also, mtDNA undergoes faster evolution making it more effective for the identification of adjacent interspecies (Amaral et al 2017). Cytochrome oxidase subunit I (COI), 16S, and 12S ribosome subunits, as well as cytochrome b are the most widely used mtDNA for DNA barcoding (Ceruso et al 2020;Hanifaturahmah et al 2020). This study used COI gene from mitochondrial DNA as the target gene.…”