Skip to main content
SpringerLink
Log in

Toxicity of the Lessepsian pufferfish Lagocephalus sceleratus from eastern Mediterranean coasts of Turkey and species identification by rapid PCR amplification

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

Abstract

Pufferfish, Lagocephalus sceleratus, L. suezensis and L. spadiceus, are widely distributed Lessepsian species along the Turkish and other eastern Mediterranean coasts. L. sceleratus is considered one of the worst invasive species in the Mediterranean Sea and recently caused human intoxication by accidental consumption. Therefore, methods to assess pufferfish toxicity and accurately identify toxic pufferfish are important for food safety. Firstly, 20 specimens of L. sceleratus were collected from the eastern Mediterranean coasts of Turkey. Tetrodotoxin (TTX) content was determined among different tissues (muscle, liver, gonad, kidney, intestine and skin) by liquid chromatography-tandem mass spectrometry. All of the specimens contained TTX. The highest TTX concentration was detected in the ovary (80.0 µg TTX/g), followed by the intestine (48.8 µg TTX/g), kidney (34.0 µg TTX/g) and liver (25.4 µg TTX/g). The flesh and testis contained detectable amounts of TTX (3.4 µg TTX/g and 2.6 µg TTX/g, respectively). TTX analogs were also detected in the tissues, with 4,9-anhydro-5,6,11-trideoxyTTX and 5,6,11-trideoxyTTX being the major analog. A species-specific PCR method was developed to identify three species of genus Lagocephalus based on the nicotinamide adenine dinucleotide dehydrogenase subunit 2 or cytochrome oxidase subunit I regions of the mitochondrial DNA of L. sceleratus, L. suezensis, and L. spadiceus. The designed primers amplified only DNA of target species and not those of other fish and shellfish species. The species-specific PCR method successfully identified these pufferfish species and could be applied to thermally treated products of the pufferfish with only 1 ng of extracted DNA.

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. Bax N, Williamson A, Aguero M, Gonzalez E, Geeves W (2003) Marine invasive alien species: a threat to global biodiversity. Mar Policy 27:313–323

    Article  Google Scholar 

  2. Por FD (1978) Lessepsian migration: the influx of Red Sea biota into the Mediterranean by way of the Suez Canal. Springer, Berlin

    Book  Google Scholar 

  3. Turan C (2010) Status and trend of Lessepsian species in marine waters of Turkey. FAO EastMed 04:109–118

    Google Scholar 

  4. Akyol O, Ünal V, Ceyhan T, Bilecenoglu M (2005) First record of the silverside blaasop, Lagocephalus sceleratus (Gmelin, 1789), in the Mediterranean Sea. J Fish Biol 66:1183–1186

    Article  Google Scholar 

  5. Streftaris N, Zenetos A (2006) Alien marine species in the Mediterranean—the 100 ‘worst invasives’ and their impact. Mediterr Marine Sci 7:87–118

    Google Scholar 

  6. Sabrah M, El-Ganainy A, Zaky M (2006) Biology and toxicity of the puffer fish Lagocephalus sceleratus from the Gulf of Suez. Egypt J Aquat Res 32:283–297

    Google Scholar 

  7. Katikou P, Georgantelis D, Sinouris N, Petsi A, Fotaras T (2009) First report on toxicity assessment of the Lessepsian migrant pufferfish Lagocephalus sceleratus (Gmelin, 1789) from European waters (Aegean Sea, Greece). Toxicon 54:50–55

    Article  CAS  Google Scholar 

  8. Rodríguez P, Alfonso A, Otero P, Katikou P, Georgantelis D, Botana LM (2012) Liquid chromatography–mass spectrometry method to detect tetrodotoxin and its analogues in the puffer fish Lagocephalus sceleratus (Gmelin, 1789) from European waters. Food Chem 132:1103–1111

    Article  Google Scholar 

  9. Bentur Y, Ashkar J, Lurie Y, Levy Y, Azzam ZS, Litmanovich M, Golik M, Gurevych B, Golani D, Eisenman A (2008) Lessepsian migration and tetrodotoxin poisoning due to Lagocephalus sceleratus in the eastern Mediterranean. Toxicon 52:964–968

    Article  CAS  Google Scholar 

  10. Kheifets J, Rozhavsky B, Girsh Solomovovich Z, Marianna R, Soroksky A (2012) Severe tetrodotoxin poisoning after consumption of Lagicephalus sceleratus (pufferfish, fugu) fished in Mediterranean Sea, treated with cholinesterase inhibitor. Case Rep Crit Care 2012:782507

    Google Scholar 

  11. Bilecenoglu M, Taskavak E, Kunt KB (2002) Range extension of three lessepsian migrant fish (Fistularia commersoni, Sphyraena flavicauda, Lagocephalus suezensis) in the Mediterranean Sea. J Mar Biol Ass UK 82:525–526

    Article  Google Scholar 

  12. Armani A, Guardone L, La Castellana R, Gianfaldoni D, Guidi A, Castigliego L (2015) DNA barcoding reveals commercial and health issues in ethnic seafood sold in the Italian market. Food Control 55:206–214

    Article  CAS  Google Scholar 

  13. Ortea I, Canas B, Calo-Malta P, Barros-Velazquez J, Gallardo JM (2010) Identification of commercial prawn and shrimp species of food interest by native isoelectric focusing. Food Chem 121:569–574

    Article  CAS  Google Scholar 

  14. Rasmussen RS, Morrissey MT (2008) DNA-based methods for the identification of commercial fish and seafood species. Comp Rev Food Sci F 7:280–295

    Article  CAS  Google Scholar 

  15. Cohen NJ, Deeds JR, Wong ES, Hanner RH, Yancy HF, White KD, Thompson TM, Wahl M, Pham TD, Guichard FM, Huh I, Austin C, Dizikes G, Gerber SI (2009) Public health response to puffer fish (tetrodotoxin) poisoning from mislabeled product. J Food Protect 4:810–817

    Article  Google Scholar 

  16. Teletcha F (2009) Molecular identification methods of fish species: reassessment and possible applications. Rev Fish Biol Fisher 19:265–293

    Article  Google Scholar 

  17. Hsieh HS, Hwang DF (2004) Molecular phylogenetic relationships of puffer fish inferred from partial sequences of cytochrome b gene and restriction fragment length polymorphism analysis. J Agric Food Chem 52:4159–4165

    Article  CAS  Google Scholar 

  18. Ishizaki S, Yokoyama Y, Oshiro N, Teruya N, Nagashima Y, Shiomi K, Watabe S (2006) Molecular identification of pufferfish species using PCR amplification and restriction analysis of a segment of the 16S rRNA gene. Comp Biochem Phys D 1:134–144

    Google Scholar 

  19. Hsieh CH, Chang WT, Chang HC, Hsieh HS, Chung YH, Hwang DF (2010) Puffer fish-based commercial fraud identification in a segment of cytochrome b region by PCR-RFLP analysis. Food Chem 121:1305–1311

    Article  CAS  Google Scholar 

  20. Luekasemsuk T, Panvisavas N, Chaturongakul S (2015) TaqMan qPCR for detection and quantification of mitochondrial DNA from toxic pufferfish species. Toxicon 102:43–47

    Article  CAS  Google Scholar 

  21. Tuncer S, Cihangir HA, Bilecenoglu M (2008) First record of the Lessepsian migrant Lagocephalus spadiceus (Tetraodontidae) in the Sea of Marmara. Cybium 32:347–348

    Google Scholar 

  22. Kodama M, Sato S (2005) Standard methods of analysis in food safety regulation. Chemistry, Ministry of Health, Labour and Welfare Ed., Japan Food Hygiene Association, Tokyo

    Google Scholar 

  23. Kiriake A, Ohta A, Suga E, Matsumoto T, Ishizaki S, Nagashima Y (2016) Comparison of tetrodotoxin uptake and gene expression in the liver between juvenile and adult tiger pufferfish, Takifugu rubripes. Toxicon 111:6–12

    Article  CAS  Google Scholar 

  24. Nakagawa T, Jang J, Yotsu-Yamashita M (2006) Hydrophilic interaction liquid chromatography–electrospray ionization mass spectrometry of tetrodotoxin and its analogs. Anal Biochem 352:142–144

    Article  CAS  Google Scholar 

  25. Nagashima Y, Matsumoto T, Kadoyama K, Ishizaki S, Terayama M (2011) Toxicity and molecular identification of green toadfish Lagocephalus lunaris collected from Kyushu Coast. J Toxicol, Japan. doi:10.1155/2011/801285

    Google Scholar 

  26. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  Google Scholar 

  27. Noguchi T, Ebesu JSM (2001) Puffer poisoning: epidemiology and treatment. J Toxicol-Toxin Rev 20:1–10

    Article  CAS  Google Scholar 

  28. Noguchi T, Arakawa O (2008) Tetrodotoxin—distribution and accumulation in aquatic organisms, and cases of human intoxication. Mar Drug 6:220–242

    Article  CAS  Google Scholar 

  29. Walker JA, Hughes DA, Anders BA, Shewale J, Sinha SK, Batzer MA (2003) Quantitative intra-short interspersed element PCR for species-specific DNA identification. Anal Biochem 316:259–269

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo, 108-8477, Japan

    Caner Acar, Shoichiro Ishizaki & Yuji Nagashima

Authors
  1. Caner Acar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shoichiro Ishizaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuji Nagashima
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Caner Acar.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Compliance with ethics requirements

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Acar, C., Ishizaki, S. & Nagashima, Y. Toxicity of the Lessepsian pufferfish Lagocephalus sceleratus from eastern Mediterranean coasts of Turkey and species identification by rapid PCR amplification. Eur Food Res Technol 243, 49–57 (2017). https://doi.org/10.1007/s00217-016-2721-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00217-016-2721-1

Keywords

Search

Navigation