Development of a Method for the Genetic Identification of Mussel Species Belonging to Mytilus, Perna, Aulacomya, and Other Genera

Abstract

Legislation regarding the labeling of processed products is an important issue in the protection of consumer rights. This labeling is especially important in products that cannot be identified on the basis of their morphological characters, because these are removed from the animal in the transformation process. The goal of this study was the identification of mussel species using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Forensically Informative Nucleotide Sequencing (FINS) methodologies. The molecular marker selected was 18S rDNA (nuclear small-subunit rDNA gene), which allows identification at the genus level and at the species level in some cases. The genera included in this study were Mytilus, Perna, Aulacomya, Semimytilus, Brachidontes, Choromytilus, and Perumytilus. Different markers were used for genetic identification at the species level. To identify the species included in the genus Perna and Choromytilus, a fragment of ITS 1 (Internal Transcribed Spacer 1) was amplified by multiplex PCR and digested with restrictases. The species of Mytilus were identified by length polymorphism and RFLP of the polyphenolic adhesive protein gene. This methodology was validated with products manufactured in the authors' pilot plant and applied to commercial samples. Therefore, this sequential method can be completely or partially used to determine the mussel genus or species present in any food product.

Adaptive genetic variation distinguishes Chilean blue mussels (Mytilus chilensis) from different marine environments

“…In brief, primers Me15‐16 produce different sizes of amplicons by species: 180 pb for M. edulis , 168 pb M. trosuluss, and 126 pb for M. chilensis and M. galloprovincialis , posterior digestion with the enzyme Aci I cut the M. galloprovincialis amplicon while the M. chilensis amplicon remains uncut (Santaclara et al. 2006). Three specimens (1.6% of samples) were identified as hybrids and were removed from the study.…”

Development of a genetic method for the identification of salmon, trout, and bream in seafood products by means of PCR–RFLP and FINS methodologies

“…Moreover, it is especially useful for routine analysis when sequencing is not an aVordable option. The studies based on the identiWcation of marine species using PCR-RFLP analysis are numerous, both in invertebrate [12,13] and vertebrate species [11,33,34]. Therefore, the RFLP represents a suitable technique to determine the identity of the salmonid species included in this work.…”

“…The main disadvantage of all the previous studies is that these cannot be applied to highly processed products such as canned foods, because the ampliWed PCR product is longer than 200 bp. In this kind of products it is not possible to amplify DNA fragments of such a size [10][11][12][13]. Moreover, these methods do not include all the possible substitute species, and this fact can produce incorrect or not assignations.…”

Genetic composition of Mytilus species in mussel populations from southern Chile

“…Irrespective of the taxonomic status of Chilean mussels, regulatory and commercial interest to differentiate them from northern hemisphere M. galloprovincialis is a major concern to prevent improperly labeling [Regulations (CE)104/2000and 2065/2001] ENREF 20, to protect consumers rights, to achieve food traceability, and to fulfill other quality objectives, such as designation of origin. To differentiate both species, Santaclara et al (2006) developed a PCR-RFLP assay in polyphenolic adhesive protein cutting only M. galloprovincialis PCR product into two, allowing such discrimination.…”

“…To prevent in the samples the unintended presence of individuals from other genera inhabiting the same geographical region -Cholga (Aulacomya ater) and Choro zapato (Choromytilus chorus)-a genus assignment was made. PCR-RFLP technique was accomplished according to Santaclara et al (2006), using MusRFLP F and R primers to amplify a fragment of the gene encoding the small subunit rDNA (18S rDNA) and later digestion with endonuclease Bsa HI (New England Biolabs®).…”