An Authentication Survey on Retail Seafood Products Sold on the Bulgarian Market Underlines the Need for Upgrading the Traceability System

Economically motivated or accidental species substitutions lead to economic and potential health damage to consumers with a loss of confidence in the fishery supply chain. In the present study, a three–year survey on 199 retail seafood products sold on the Bulgarian market was addressed to assess: (1) product authenticity by molecular identification; (2) trade name compliance to the list of official trade names accepted in the territory; (3) adherence of the list in force to the market supply. DNA barcoding on mitochondrial and nuclear genes was applied for the identification of whitefish (WF), crustaceans (C) and mollusks (cephalopods—MC; gastropods—MG; bivalves—MB) except for Mytilus sp. products for which the analysis was conducted with a previously validated RFLP PCR protocol. Identification at the species level was obtained for 94.5% of the products. Failures in species allocation were reconducted due to low resolution and reliability or the absence of reference sequences. The study highlighted an overall mislabeling rate of 11%. WF showed the highest mislabeling rate (14%), followed by MB (12.5%), MC (10%) and C (7.9%). This evidence emphasized the use of DNA–based methods as tools for seafood authentication. The presence of non–compliant trade names and the ineffectiveness of the list to describe the market species varieties attested to the need to improve seafood labeling and traceability at the national level.

[1]  C. Malloggi,et al.  Mislabeling in seafood products sold on the Italian market: A systematic review and meta-analysis , 2022, Food Control.

[2]  L. Tinacci,et al.  New official Bulgarian list of seafood trade names: coping with EU labelling requirements and market trends to enhance consumers’ informed choice , 2022, Journal of Consumer Protection and Food Safety.

[3]  M. Dean,et al.  The 11 sins of seafood: Assessing a decade of food fraud reports in the global supply chain. , 2022, Comprehensive reviews in food science and food safety.

[4]  U. R. Sumaila,et al.  Drivers of Seafood Consumption at Different Geographical Scales , 2022, Journal of Sustainability Research.

[5]  C. Malloggi,et al.  Mislabeling assessment and species identification by PCR-RFLP of mussel-based products (Mytilus spp.) sold on the Italian market , 2021, Food Control.

[6]  P. M. Galetti,et al.  DNA Barcoding of Penaeidae (Decapoda; Crustacea): Non-Distance-Based Species Delimitation of the Most Economically Important Shrimp Family , 2021, Diversity.

[7]  Lidiya Wilwet,et al.  In-house and on-field validation of the multiplex PCR assay developed for authentication of three commercially important shrimp species , 2021 .

[8]  A. Pulvirenti,et al.  Bioinformatics Approach to Mitigate Mislabeling in EU Seafood Market and Protect Consumer Health , 2021, International journal of environmental research and public health.

[9]  E. Garcia-Vazquez,et al.  DNA Analysis Detects Different Mislabeling Trend by Country in European Cod Fillets , 2021, Foods.

[10]  Sudhir Kumar,et al.  MEGA11: Molecular Evolutionary Genetics Analysis Version 11 , 2021, Molecular biology and evolution.

[11]  C. Román,et al.  Determinants of fishery and aquaculture products consumption at home in the EU28 , 2021 .

[12]  M. Dean,et al.  A comprehensive review of food fraud terminologies and food fraud mitigation guides , 2021 .

[13]  Rosalee S. Hellberg,et al.  Seafood fraud , 2021, Food Fraud.

[14]  Rosalee S. Hellberg,et al.  DNA-based techniques for seafood species authentication. , 2021, Advances in food and nutrition research.

[15]  Prashant Singh,et al.  High resolution real-time PCR melting curve assay for identification of top five Penaeidae shrimp species , 2020 .

[16]  E. Garcia-Vazquez,et al.  Seventeen years analysing mislabelling from DNA barcodes: Towards hake sustainability , 2020 .

[17]  Isabel Mafra,et al.  DNA barcode markers applied to seafood authentication: an updated review , 2020, Critical reviews in food science and nutrition.

[18]  C. Haldar,et al.  DNA barcoding for fish species identification: current status and future prospective , 2020 .

[19]  Athanassios C. Tsikliras,et al.  Seafood mislabeling in Greek market using DNA barcoding , 2020 .

[20]  L. Tinacci,et al.  Nationwide survey of the Bulgarian market highlights the need to update the official seafood list based on trade inputs , 2020, Food Control.

[21]  Wei‐Jen Chen,et al.  Molecular phylogeny and diversity of penaeid shrimps (Crustacea: Decapoda) from South‐East Asian waters , 2020 .

[22]  L. Rodríguez-Muñiz,et al.  Lab experience with seafood control at the undergraduate level: Cephalopods as a case study , 2020, Biochemistry and molecular biology education : a bimonthly publication of the International Union of Biochemistry and Molecular Biology.

[23]  L. Tinacci,et al.  Labelling compliance and species identification of herring products sold at large scale retail level within the Italian market , 2019 .

[24]  N. Knowlton,et al.  GenBank is a reliable resource for 21st century biodiversity research , 2019, Proceedings of the National Academy of Sciences.

[25]  L. Manning,et al.  Food fraud vulnerability assessment: Reliable data sources and effective assessment approaches , 2019, Trends in Food Science & Technology.

[26]  Gloria M. Luque,et al.  The characterization of seafood mislabeling: A global meta-analysis , 2019, Biological Conservation.

[27]  R. Hanner,et al.  Survey of mislabelling across finfish supply chain reveals mislabelling both outside and within Canada. , 2019, Food research international.

[28]  Kelly A Meiklejohn,et al.  Assessment of BOLD and GenBank – Their accuracy and reliability for the identification of biological materials , 2019, PloS one.

[29]  L. Webster,et al.  DNA barcoding validates species labelling of certified seafood , 2019, Current Biology.

[30]  Alexander M. Weigand,et al.  DNA barcode reference libraries for the monitoring of aquatic biota in Europe: Gap-analysis and recommendations for future work , 2019, bioRxiv.

[31]  P. Acutis,et al.  Authentication of ready-to-eat anchovy products sold on the Italian market by BLAST analysis of a highly informative cytochrome b gene fragment , 2019, Food Control.

[32]  A. Giusti,et al.  The new Italian official list of seafood trade names (annex I of ministerial decree n. 19105 of September the 22nd, 2017): Strengths and weaknesses in the framework of the current complex seafood scenario , 2019, Food Control.

[33]  Amanda M. Naaum,et al.  DNA barcoding as a regulatory tool for seafood authentication in Canada , 2018, Food Control.

[34]  Jónas R. Viðarsson,et al.  DNA barcoding revealing mislabeling of seafood in European mass caterings , 2018, Food Control.

[35]  L. Tinacci,et al.  Seafood labelling compliance with European legislation and species identification by DNA barcoding: A first survey on the Bulgarian market , 2018, Food Control.

[36]  A. Giusti,et al.  DNA barcoding for the verification of supplier’s compliance in the seafood chain: How the lab can support companies in ensuring traceability , 2018, Italian journal of food safety.

[37]  F. Volckaert,et al.  Seafood substitution and mislabeling in Brussels' restaurants and canteens , 2018 .

[38]  M. Dean,et al.  The seafood supply chain from a fraudulent perspective , 2016, Food Security.

[39]  I. Mafra,et al.  High resolution melting analysis of a COI mini-barcode as a new approach for Penaeidae shrimp species discrimination , 2017 .

[40]  P. Acutis,et al.  An insight into the Chinese traditional seafood market: Species characterization of cephalopod products by DNA barcoding and phylogenetic analysis using COI and 16SrRNA genes , 2017 .

[41]  A. Benvenuti,et al.  Is raw better? A multiple DNA barcoding approach (full and mini) based on mitochondrial and nuclear markers reveals low rates of misdescription in sushi products sold on the Italian market , 2017 .

[42]  I. Mafra,et al.  DNA barcoding coupled to HRM analysis as a new and simple tool for the authentication of Gadidae fish species. , 2017, Food chemistry.

[43]  Karen Everstine Supply Chain Complexity and Economically Motivated Adulteration , 2017 .

[44]  EU CONSUMER HABITS REGARDING FISHERY AND AQUACULTURE PRODUCTS , 2017 .

[45]  D. Pauly,et al.  The Marine Fisheries in Bulgaria's Exclusive Economic Zone, 1950–2013 , 2017, Frontiers in Marine Science.

[46]  R. Hanner,et al.  A systematic analysis across North Atlantic countries unveils subtleties in cod product labelling , 2016 .

[47]  Shunping He,et al.  DNA barcoding for the identification of common economic aquatic products in Central China and its application for the supervision of the market trade , 2016 .

[48]  D. Cawthorn,et al.  Fish species substitution and misnaming in South Africa: An economic, safety and sustainability conundrum revisited. , 2015, Food chemistry.

[49]  L. Tinacci,et al.  Fish species identification in canned pet food by BLAST and Forensically Informative Nucleotide Sequencing (FINS) analysis of short fragments of the mitochondrial 16s ribosomal RNA gene (16S rRNA) , 2015 .

[50]  L. Guardone,et al.  DNA and Mini-DNA barcoding for the identification of Porgies species (family Sparidae) of commercial interest on the international market , 2015 .

[51]  G. Rajkumar,et al.  Molecular identification of shrimp species, Penaeus semisulcatus, Metapenaeus dobsoni, Metapenaeus brevicornis, Fenneropenaeus indicus, Parapenaeopsis stylifera and Solenocera crassicornis inhabiting in the coromandel coast (Tamil Nadu, India) using MT-COI gene , 2015 .

[52]  G. Carvalho,et al.  Fish Product Mislabelling: Failings of Traceability in the Production Chain and Implications for Illegal, Unreported and Unregulated (IUU) Fishing , 2014, PloS one.

[53]  Xiong Xiong,et al.  Development of a Simple and Cost-Effective Bead-Milling Method for DNA Extraction from Fish Muscles , 2014, Food Analytical Methods.

[54]  Ayşe Gündüz Hoşgör,et al.  Managing Rapana in the Black Sea: Stakeholder workshops on both sides , 2014 .

[55]  G. Vermeij,et al.  Global phylogeny and new classification of the Rapaninae (Gastropoda: Muricidae), dominant molluscan predators on tropical rocky seashores. , 2013, Molecular phylogenetics and evolution.

[56]  J. Vieites,et al.  Rapid method for controlling the correct labeling of products containing common octopus (Octopus vulgaris) and main substitute species (Eledone cirrhosa and Dosidicus gigas) by fast real-time PCR. , 2012, Food chemistry.

[57]  S. Mariani,et al.  Seafood mislabelling: comparisons of two western European case studies assist in defining influencing factors, mechanisms and motives , 2012 .

[58]  H. Min,et al.  DNA Barcoding of Fish, Insects, and Shellfish in Korea , 2012, Genomics & informatics.

[59]  Sara M. Handy,et al.  A single-laboratory validated method for the generation of DNA barcodes for the identification of fish for regulatory compliance. , 2011, Journal of AOAC International.

[60]  K. Chu,et al.  Phylogeny of Decapoda using two nuclear protein-coding genes: origin and evolution of the Reptantia. , 2008, Molecular phylogenetics and evolution.

[61]  O. Mouchel,et al.  Primers and polymerase chain reaction conditions for DNA barcoding teleost fish based on the mitochondrial cytochrome b and nuclear rhodopsin genes , 2007 .

[62]  R. Bieler,et al.  Phylogeny of Veneroidea (Mollusca: Bivalvia) based on morphology and molecules , 2006 .

[63]  M. Vecchione,et al.  A molecular systematic evaluation of the squid genus Illex (Cephalopoda: Ommastrephidae) in the North Atlantic Ocean and Mediterranean Sea. , 2006, Molecular phylogenetics and evolution.

[64]  M. Kimura A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences , 1980, Journal of Molecular Evolution.

[65]  T. A. Hall,et al.  BIOEDIT: A USER-FRIENDLY BIOLOGICAL SEQUENCE ALIGNMENT EDITOR AND ANALYSIS PROGRAM FOR WINDOWS 95/98/ NT , 1999 .

[66]  S. Palumbi,et al.  Nucleic acids II: the polymerase chain reaction , 1996 .

[67]  R. Vrijenhoek,et al.  DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. , 1994, Molecular marine biology and biotechnology.

[68]  N. Saitou,et al.  The neighbor-joining method: a new method for reconstructing phylogenetic trees. , 1987, Molecular biology and evolution.