Skip to main content
SpringerLink
Log in

A rapid PCR–RFLP method for the identification of Lophius species

  • Original Paper
  • Published:
European Food Research and Technology Aims and scope Submit manuscript

Abstract

The seven Anglerfish species, which belong to the genus Lophius, have a different value on the market, worldwide. If whole fishes can be identified by their morphological characteristics, they become indistinguishable when prepared or processed. In this study, a rapid method based on polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) was developed for the authentication of the seven Lophius species, using a cytochrome b gene fragment of 566 bp. After a genus-specific PCR, a fast digestion with the restriction enzyme BfaI, followed by agarose gel electrophoresis, allowed a clear species identification by producing specific restriction patterns. The total time required was as low as 6 h, DNA extraction included. The method was then used to analyse 48 commercial samples, whose phylogenetic analysis confirmed the PCR–RFLP response at 100 %. Results showed that mislabelling occurs on the market regardless the kind of processing.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Caruso JH (1985) The systematics and distribution of the lophiid anglerfishes. III. Intergeneric relationships. Copeia 4:870–875

    Article  Google Scholar 

  2. Fishbase. http://www.fishbase.org/

  3. ZipcodeZoo. http://zipcodezoo.com/

  4. Farina AC, Azevedo M, Landa J, Duarte R, Sampedro P, Costas G, Torres MA, Canas L (2008) Lophius in the world: a synthesis on the common features and life strategies. ICES J Mar Sc 65:1272–1280

    Article  Google Scholar 

  5. Food and Agriculture Organization (2007) FAO yearbook fishery statistics, capture production, captures 2005. FAO, Rome

    Google Scholar 

  6. Leslie RW, Grant WS (1994) Meristic and morphometric variation among anglerfish of the genus Lophius (Lophiiformes). J Zool Lond 232:565–584

    Google Scholar 

  7. Council Regulation (EC) No. 104/2000 of 17 December 1999. On the common organisation of the markets in fishery and aquaculture products. OJ Eur Comm L17:22–52

  8. Commission Regulation (EC) No. 2065/2001 of 22 October 2001. Laying down detailed rules for the application of Council Regulation (EC) No. 104/2000 as regards informing consumers about fishery and aquaculture products. OJ Eur Comm L287:6–8

    Google Scholar 

  9. Teletchea F (2009) Molecular identification methods of fish species: reassessment and possible applications. Rev Fish Biol Fish 19(3):265–293

    Article  Google Scholar 

  10. Kochzius M, Seidel C, Antoniou A, Botla SK, Campo D, Cariani A, Vazquez EG, Hauschild J, Hervet C, Hjörleifsdottir S, Hreggvidsson G, Kappel K, Landi M, Magoulas A, Marteinsson V, Nölte M, Planes S, Tinti F, Turan C, Venugopal MN, Weber H, Blohm D (2010) Identifying. Fishes through DNA barcodes and microarrays. PLoS ONE 5(9):e12620. doi:10.1371/journal.pone.0012620

    Article  Google Scholar 

  11. Armani A, Castigliego L, Tinacci L, Gianfaldoni D, Guidi A (2011) Molecular characterization of icefish (Salangidae family), using direct sequencing of mitochondrial cytochrome b gene. Food Contr 22:888–895

    Article  CAS  Google Scholar 

  12. Vicari N, Maniero S, Arcangeli G, Tisato E (2001) Identificazione di rana pescatrice (Lophius budegassa e Lophius piscatorius) gallinella (Trigla lucerna), pesce prete (Uranoscopus scaber) e pesce palla (Arothron nigropunctatus) mediante analisi di restrizione di prodotti di PCR. Ing Al 5:24–27

    Google Scholar 

  13. Sanjuan J, Raposo-Guillán J, Comesaña AS (2002) Genetic Identification of Lophius budegassa and L. piscatorius by PCR–RFLP analysis of a mitochondrial tRNAGlu/cytochrome b segment. J Food Sci 67(7):2644–2648

    Article  Google Scholar 

  14. Espiñeira M, González-Lavin N, Vieites JM, Santaclara FJ (2008) Authentication of Anglerfish Species (Lophius spp.) by means of polymerase chain reaction-restriction fragment length polymorphism (PCR–RFLP) and forensically informative nucleotide sequencing (FINS) methodologies. J Agric Food Chem 56(22):10594–10599

    Article  Google Scholar 

  15. Sevilla RG, Diez A, Norén M, Mouchel O, Jérôme M, Verrez-Bagnis V, Van Pelt H, Favre-Krey L, Krey G, Bautista JM (2007) Primers and polymerase chain reaction conditions for DNA barcoding teleost fish based on the mitochondrial cytochrome b and nuclear rhodopsin genes. Mol Ecol Notes 7(5):730–734

    Article  CAS  Google Scholar 

  16. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp 41:95–98

    CAS  Google Scholar 

  17. Palumbi SR (1996) Nucleic acids II: the polymerase chain reaction. In: Hillis DM, Moritz C, Mable BK (eds). Molecular systematics. Second edition. Sinauer Associates, Inc., Sunderland, Massachusetts, pp 205–247

  18. Van de Peer Y, DeWatcher R (1994) TREECON for windows. A software package for the construction and drawing of evolutionary trees for Microsoft windows environment. Computer Appl Biosci 10:569–570

    Google Scholar 

  19. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  Google Scholar 

  20. Maartens L, Booth AJ (2001) Quantifying commercial catch and effort of monkfish Lophius vomerinus and L. vaillanti off Namibia. S Afr J Mar Sci 23:291–306

    Article  Google Scholar 

  21. Resolución de 22 de marzo de 2011, de la Secretaría General del Mar, por la que se establece y se publica el listado de denominaciones comerciales de especies pesqueras y de acuicultura admitidas en España. Ministerio De Medio Ambiente, Y Medio Rural Y Marino, Boletín officia del estado, Núm. 82, Sec.III., Pág.35515. http://www.boe.es/boe/dias/2011/04/06/pdfs/BOE-A-2011-6222.pdf. Accessed 05.07.2011

  22. talian Ministry of Agricultural, Food and Forestry Policies (MIPAAF). Decree N_. 27.03.02. Etichettatura dei prodotti ittici e sistema di controllo. Official Gazette of the Italian Republic 84:33–59

  23. Italian Ministry of Agricultural, Food and Forestry Policies (MIPAAF). Decree N_ 31 January 2008. Denominazione in lingua italiana delle specie ittiche di interesse commerciale e Modifiche e integrazioni dell’elenco di cui al decreto 25 luglio 2005. Official Gazette of the Italian Republic 45:11–32

  24. Italian Ministry of Agricultural, Food and Forestry Policies (MIPAAF). Decree N_ 5 March 2010. Denominazione in lingua italiana delle specie ittiche di interesse commerciale. Official Gazette of the Italian Republic 124

  25. Ward RD, Hanner R, Hebert PDN (2009) The campaign to DNA barcode all fishes, FISH-BOL. J Fish Biol 74:329–356

    Article  CAS  Google Scholar 

  26. Aranishi F (2005) Rapid PCR–RFLP method for discrimination of imported and domestic mackerel. Mar Biotech 7(6):571–575

    Article  CAS  Google Scholar 

  27. Cespedes A, Garcia T, Carrera E, Gonzalez I, Fernandez A, Asensio L, Hernandez PE, Martin R (2000) Genetic differentiation between sole (Solea solea) and Greenland halibut (Reinhardtius hippoglossoides) by PCR–RFLP analysis of a 12S rRNA gene fragment. J Sci Food Agric 80:29–32

    Article  CAS  Google Scholar 

  28. Dooley JJ, Sage HD, Clarke MA, Brown HM, Garrett SD (2005) Fish species identification using PCR–RFLP analysis and lab-on-a-chip capillary electrophoresis: application to detect white fish species in food products and an interlaboratory study. J Agric Food Chem 53(9):3348–3357

    Article  CAS  Google Scholar 

  29. Cocolin L, D’Agaro E, Manzano M, Lanari D, Comi G (2000) Rapid PCR–RFLP Method for the Identification of Marine Fish Fillets (Seabass, Seabream, Umbrine, and Dentex). J Food Sci 65(8):1315–1317

    Article  CAS  Google Scholar 

  30. Food Standard Agency (2011) Fish commercial designation list. http://www.food.gov.uk/multimedia/spreadsheets/fishcomdes.xls. Accessed 05.07.2011

  31. Yoneda M, Tokimura M, Fujita H, Takeshita N, Takeshita K, Matsuyama M, Matsuura S (1998) Ovarian structure and batch fecundity in Lophiomus setigerus. J Fish Biol 52:94–106

    Article  Google Scholar 

  32. Choi SH, Kim J, Jo JO, Cho MK, Yu HS, Cha HJ, Ock MS (2011) Anisakis simplex larvae: infection status in marine fish and cephalopods purchased from the Cooperative Fish Market in Busan, Korea. Korean J Parasitol 49(1):39–44

    Article  Google Scholar 

  33. Guidi A, Armani A, Castigliego L, Li XN, Fanzone F, Fusco S, Facibeni E, Gianfaldoni D (2010) Labeling of ethnic food in the Prato Chinese community. Vet Res Commun 34(1):163–166

    Article  Google Scholar 

Download references

Acknowledgments

The authors wish to thank Dr. Paolo Manzoni for his help in species identifications.

Author information

Authors and Affiliations

  1. Department of Animal Pathology, Prophylaxis and Food Hygiene, University of Pisa, Via delle Piagge 2, 56124, Pisa, Italy

    Andrea Armani, Lorenzo Castigliego, Lara Tinacci, Daniela Gianfaldoni & Alessandra Guidi

  2. Ministry of Health DGSAN UVAC Veneto-PIF, Veneto, Italy

    Gabriele Gandini

Authors
  1. Andrea Armani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lorenzo Castigliego
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lara Tinacci
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gabriele Gandini
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Daniela Gianfaldoni
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Alessandra Guidi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andrea Armani.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Armani, A., Castigliego, L., Tinacci, L. et al. A rapid PCR–RFLP method for the identification of Lophius species. Eur Food Res Technol 235, 253–263 (2012). https://doi.org/10.1007/s00217-012-1754-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00217-012-1754-3

Keywords

Search

Navigation