Skip to main content
SpringerLink
Log in

Tetrodotoxin Detection in Japanese Bivalves: Toxification Status of Scallops

  • Research
  • Published:
Marine Biotechnology Aims and scope Submit manuscript

Abstract

Tetrodotoxin (TTX), or pufferfish toxin, has been frequently detected in edible bivalves around the world during the last decade and is problematic in food hygiene and safety. It was reported recently that highly concentrated TTX was detected in the midgut gland of the akazara scallop Chlamys (Azumapecten) farreri subsp. akazara collected in coastal areas of the northern Japanese archipelago. The toxification of the bivalve was likely to involve the larvae of the flatworm, Planocera multitentaculata. However, the overall status of bivalve TTX toxification has not been elucidated. In this study, 14 species/subspecies of bivalves from various Japanese waters were subjected to LC–MS/MS analysis to reveal TTX toxification state, demonstrating that the Pectinidae, including C. farreri akazara, Chlamys farreri nipponensis, Chlamys (Mimachlamys) nobilis, and Mizuhopecten yessoensis, accumulated TTX in their midgut gland. Many individuals of C. farreri akazara and C. farreri nipponensis were found with high concentrations of TTX, while C. nobilis and M. yessoensis exhibited low concentrations. The extent of TTX accumulation in C. farreri akazara and C. farreri nipponensis varied widely by region and season. Curiously, no other bivalve species investigated in this study showed evidence of TTX. These results suggest that monitoring for TTX, like other shellfish toxins, is necessary to ensure that pectinid bivalves are a safe food resource.

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
Fig. 4

Similar content being viewed by others

Data Availability

All data generated or analyzed during this study are included in this article and supplementary information.

References

  • Antonelli P, Salerno B, Bordin P, Peruzzo A, Orsini M, Arcangeli G, Barco L, Losasso C (2022) Tetrodotoxin in live bivalve mollusks from Europe: is it to be considered an emerging concern for food safety? Compr Rev Food Sci Food Saf 21:719–737

    Article  CAS  PubMed  Google Scholar 

  • Asano Y, Yoshida A, Isozaki N, Ishida S (2001) Production of intestine-specific monoclonal antibody and interspecific cross-reaction in triclads and polyclads. Belg J Zool 131:137–141

    Google Scholar 

  • Asia-Pacific Economic Cooperation (APEC) (2009) Commercial importance of Pectinidae characterization within Asia-Pacific region. In Harmonizing Quality and Traceability Standards for Pecten Trade in Asia-Pacific Region; APEC Secretariat: Singapore

  • Biessy L, Boundy MJ, Smith KS, Harwood DT, Hawes I, Wood SA (2019) Tetrodotoxin in marine bivalves and edible gastropods: a mini-review. Chemosphere 236:124404

  • Blanco L, Lago J, González V, Paz B, Rambla-Alegre M, Cabado AG (2019) Occurrence of tetrodotoxin in bivalves and gastropods from harvesting areas and other natural spaces in Spain. Toxins 11:1–9

    Article  Google Scholar 

  • Bordin P, Dall'Ara S, Tartaglione L, Antonelli P, Calfapietra A, Varriale F, Guiatti D, Milandri A, Dell'Aversano C, Arcangeli G, Barco L (2021) First occurrence of tetrodotoxins in bivalve mollusks from Northern Adriatic Sea (Italy). Food Control 120:107510

  • Cao R, Wang D, Wei Q, Wang Q, Yang D, Liu H, Dong Z, Zhang X, Zhang Q, Zhao J (2018) Integrative biomarker assessment of the influence of saxitoxin on marine bivalves: a comparative study of the two bivalve species oysters, Crassostrea gigas, and scallops, Chlamys farreri. Front Physiol 9:1173

    Article  PubMed  PubMed Central  Google Scholar 

  • Colquhon D, Henderson R, Ritchie JM (1972) The binding of labeled tetrodotoxin to non-myelinated nerve fibres. J Physiol 227:95–126

    Article  Google Scholar 

  • Daguer H, Hoff RB, Molognoni L, Kleemann CR, Felizardo LV (2018) Outbreaks, toxicology, and analytical methods of marine toxins in seafood. Curr Opin Food Sci 24:43–55

    Article  Google Scholar 

  • EFSA CONTAM Panel, Knutsen HK, Alexander J, Barregård L, Bignami M, Brüschweiler B, Ceccatelli S, Cottrill B, Dinovi M, Edler L, Grasl-Kraupp B, Hogstrand C, Hoogenboom L, Nebbia CS, Oswald IP, Rose M, Roudot A-C, Schwerdtle T, Vleminckx C, Vollmer G, Wallace H, Arnich N, Benford D, Botana L, Viviani B, Arcella D, Binaglia M, Horvath Z, Steinkellner H, van Manen M, Petersen A (2017) Risks for public health related to the presence of tetrodotoxin (TTX) and TTX analogues in marine bivalves and gastropods. EFSA J 15:e04752

  • Food and Agriculture Organization of the United Nations (FAO) (2022) In brief to the state of world fisheries and aquaculture 2022. Towards Blue Transformation. Rome, FAO. https://doi.org/10.4060/cc0463en

  • Gerssen A, Bovee T, Klijnstra M, Poelman M, Portier L, Hoogenboom R (2018) First report on the occurrence of tetrodotoxins in bivalve mollusks in the Netherlands. Toxins 10:1–12

    Article  Google Scholar 

  • Goto T, Kishi Y, Takahashi S, Hirata Y (1965) Tetrodotoxin. Tetrahedron 21:2059–2088

  • Hallegraeff GM (2010) Ocean climate change, phytoplankton community responses, and harmful algal blooms: a formidable predictive challenge. J Phycol 46:220–235

    Article  CAS  Google Scholar 

  • Han C, Zhang X, Li L, Chen S, Yan Z, Gao X, Chang J (2018) Analysis and evaluation of tetrodotoxin in coastal aquatic products of Zhejiang Province. J Coast Res 83:380–385

    Article  CAS  Google Scholar 

  • Hort V, Arnich N, Guérin T, Lavison-Bompard G, Nicolas M (2020) First detection of tetrodotoxin in bivalves and gastropods from the French mainland coasts. Toxins 12:599

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ito M, Furukawa R, Yasukawa S, Sato M, Oyama H, Okabe T, Suo R, Sugita H, Takatani T, Arakawa O, Adachi M, Nishikawa T, Itoi S (2022) Local differences in the toxin amount and composition of tetrodotoxin and related compounds in pufferfish (Chelonodon patoca) and toxic goby (Yongeichthys criniger) juveniles. Toxins 14:150

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Itoi S, Sato T, Takei M, Yamada R, Ogata R, Oyama H, Teranishi S, Kishiki A, Wada T, Noguchi K, Abe M, Okabe T, Akagi H, Kashitani M, Suo R, Koito T, Takatani T, Arakawa O, Sugita H (2020) The planocerid flatworm is a main supplier of toxin to tetrodotoxin-bearing fish juveniles. Chemosphere 249:126217

  • Itoi S, Ueda H, Yamada R, Takei M, Sato T, Oshikiri S, Wajima Y, Ogata R, Oyama H, Shitto T, Okuhara K, Tsunashima T, Sawayama E, Sugita H (2018) Including planocerid flatworms in the diet effectively toxifies the pufferfish, Takifugu niphobles. Sci Rep 8:12302

    Article  PubMed  PubMed Central  Google Scholar 

  • Kato K (1944) Polycladida of Japan. J Sigenkagaku Kenkyusyo 1:257–318

  • Kashitani M, Okabe T, Oyama H, Noguchi K, Yamazaki H, Suo R, Mori T, Sugita H, Itoi S (2020) Taxonomic distribution of tetrodotoxin in acotylean flatworms (Polycladida: Platyhelminthes). Mar Biotechnol 22:805–811

    Article  CAS  Google Scholar 

  • Kawashiro I, Tanabe H, Ishii A, Kondo T (1962) Studies on the poison in AKASARA shellfish (Chlamys nipponensis Kuroda). J Food Hyg Soc Jpn 3:273–277

    Article  CAS  Google Scholar 

  • Kodama M, Sato S, Ogata T (1993) Alexandrium tamarense as a source of tetrodotoxin in the scallop Patinopecten yessoensis. In Toxic Phytoplankton Blooms in the Sea; Smayda TJ, Shimizu Y, Eds; Elsevier: New York, NY, USA. pp 401–406

  • McNabb PS, Taylor DI, Ogilvie SC, Wilkinson L, Anderson A, Hamon D, Wood SA, Peake BM (2014) First detection of tetrodotoxin in the bivalve Paphies australis by liquid chromatography coupled to triple quadrupole mass spectrometry with and without precolumn reaction. J AOAC Int 97:325–333

    Article  CAS  PubMed  Google Scholar 

  • Makarova M, Rycek L, Hajicek J, Baidilov D, Hudlicky T (2019) Tetrodotoxin: history, biology, and synthesis. Angew Chem Int Ed 58:18338–18387

    Article  CAS  Google Scholar 

  • Miyazawa K, Jeon JK, Maruyama J, Noguchi T, Ito K, Hashimoto K (1986) Occurrence of tetrodotoxin in the flatworm Planocera multitentaculata (Platyhelminthes). Toxicon 24:645–650

    Article  CAS  PubMed  Google Scholar 

  • Miyazawa K, Jeon JK, Noguchi T, Ito K, Hashimoto K (1987) Distribution of tetrodotoxin in the tissues of the flatworm Planocera multitentaculata (Platyhelminthes). Toxicon 25:975–980

    Article  CAS  PubMed  Google Scholar 

  • Miyazawa K, Noguchi T (2001) Distribution and origin of tetrodotoxin. J Toxicol-Toxin Rev 20:11–33

    Article  CAS  Google Scholar 

  • Narahashi T (2001) Pharmacology of tetrodotoxin. J Toxicol-Toxin Rev 20:67–84

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Noguchi T, Arakawa O, Takatani T (2006) TTX accumulation in pufferfish. Comp Biochem Physiol D 1:145–152

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Numano S, Kudo Y, Cho Y, Konoki K, Yotsu-Yamashita M (2019) Temporal variation of the profile and concentrations of paralytic shellfish toxins and tetrodotoxin in the scallop, Patinopecten yessoensis, cultured in a bay of east Japan. Mar Drugs 17:653

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ogilvie S, Taylor D, McNabb P, Hamon D, Nathan P, Anderson A (2012) Tetrodotoxin in Kaimoana: Science and Mātauranga mitigating health risks from a lethal neurotoxin. Cawthron Institute, Report No. 2219:1–46

  • Okabe T, Oyama H, Kashitani M, Ishimaru Y, Suo R, Sugita H, Itoi S (2019) Toxic flatworm egg plates serve as a possible source of tetrodotoxin for pufferfish. Toxins 11:402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Okabe T, Saito R, Yamamoto K, Watanabe R, Kaneko Y, Yanaoka M, Furukoshi S, Yasukawa S, Ito M, Oyama H, Suo R, Suzuki M, Takatani T, Arakawa O, Sugita H, Itoi S (2021) The role of toxic planocerid flatworm larvae on tetrodotoxin accumulation in marine bivalves. Aquat Toxicol 237:105908

  • Oshima Y, Yasumoto T, Kodama M, Ogata T, Fukuyo Y, Matsuura F (1982) Features of paralytic shellfish poison occurring in Tohoku district. Bull Jpn Soc Sci Fish 48:525–530

    Article  Google Scholar 

  • Suo R, Tanaka M, Oyama H, Kojima Y, Yui K, Sakakibara R, Nakahigashi R, Adachi M, Nishikawa T, Sugita H, Itoi S (2022) Tetrodotoxins in the flatworm Planocera multitentaculata. Toxicon 216:168–173

    Article  Google Scholar 

  • Teshirogi W, Ishida S, Jatani K (1981) On the early development of some Japanese polyclads. Rep Fukaura Marine Biol Lab Hirosaki Univ 9:2–31

    Google Scholar 

  • Tsuda K, Ikuma S, Kawamura M, Tachikawa R, Sakai K (1964) Tetrodotoxin. VII. On the structure of tetrodotoxin and its derivatives. Chem Pharm Bull 12:1357–1374

    Article  CAS  Google Scholar 

  • Turner AD, Dhanji-Rapkova M, Coates L, Bickerstaff L, Milligan S, O’Neill A, Faulkner D, McEneny H, Baker-Austin C, Lees DN (2017) Detection of tetrodotoxin shellfish poisoning (TSP) toxins and causative factors in bivalve molluscs from the UK. Mar Drugs 15:277

    Article  PubMed  PubMed Central  Google Scholar 

  • Turner AD, Powell A, Schofield A, Lees DN, Baker-Austin C (2015) Detection of the pufferfish toxin tetrodotoxin in European bivalves, England, 2013 to 2014. Euro Surveill 20:21009

    Article  PubMed  Google Scholar 

  • Vlamis A, Katikou P, Rodriguez I, Rey V, Alfonso A, Papazachariou A, Zacharaki T, Botana AM, Botana LM (2015) First detection of tetrodotoxin in Greek shellfish by UPLC-MS/MS potentially linked to the presence of the dinoflagellate Prorocentrum minimum. Toxins 7:1779–1807

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Woodward RB (1964) The structure of tetrodotoxin. Pure Appl Chem 86:49–74

    Article  Google Scholar 

  • Xiao J, Ford SE, Yang H, Zhang G, Zhang F, Guo X (2005) Studies on mass summer mortality of cultured zhikong scallops (Chlamys farreri Jones et Preston) in China. Aquaculture 250:602–615

    Article  Google Scholar 

  • Yamada R, Tsunashima T, Takei M, Sato T, Wajima Y, Kawase M, Oshikiri S, Kajitani Y, Kosoba K, Ueda H, Abe K, Itoi S, Sugita H (2017) Seasonal changes in the tetrodotoxin content of the flatworm Planocera multitentaculata. Mar Drugs 15:56

    Article  PubMed  PubMed Central  Google Scholar 

  • Yasumoto T, Oshima Y, Yamaguchi M (1978) Occurrence of a new type of shellfish poisoning in the Tohoku district. Bull Jpn Soc Sci Fish 44:1249–1255

    Article  Google Scholar 

  • Yuan Y, Kanno M, Kijima A (2012) Genetic diversity of wild populations of Chlamys farreri in Japan and their genetic relationship with cultured stocks in China. Aquaculture 370–371:109–122

    Article  Google Scholar 

  • Zhu M-Y, Zou Y-L, Wu R-J, Hall S (2003) Accumulation and depuration of paralytic shellfish poisons (PSP) in Chinese scallop Chlamys farreri. Acta Oceanol Sin 25:75–83 (in Chinese with English abstract)

    CAS  Google Scholar 

Download references

Acknowledgements

We express sincere thanks to Mr. Shunsuke Noguchi, Kyoto Prefecture Sea-Farming Association, for collecting samples. This study was partly performed under the cooperative research program of Institute of Nature and Environmental Technology, Kanazawa University (Accept No. 22037).

Funding

This study was supported in part by the Grant-in-Aid for Challenging Research (Exploratory) from Japanese Society for the Promotion of Science (JSPS) (18 K19238, S.I.) and Grant-in-Aid for Scientific Research (A) from JSPS (19H00954, S.I.).

Author information

Authors and Affiliations

  1. Department of Marine Science and Resources, Nihon University, Fujisawa, Kanagawa, 252-0880, Japan

    Shino Yasukawa, Kyoko Shirai, Kaho Namigata, Masaaki Ito, Mei Tsubaki, Hikaru Oyama, Yukino Fujita, Taiki Okabe, Rei Suo, Haruo Sugita & Shiro Itoi

  2. Noto Marine Laboratory, Institute of Nature and Environmental Technology, Division of Marine Environmental Studies, Kanazawa University, Ogi, Noto-Cho, Ishikawa , 927-0553, Japan

    Shouzo Ogiso, Yukina Watabe & Nobuo Suzuki

  3. Noto Center for Fisheries Science and Technology, Kanazawa University, Ossaka, Noto-Cho, Ishikawa , 927-0552, Japan

    Hajime Matsubara

  4. Graduate School of Integrated Sciences for Life, Hiroshima University, 1-4-4 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8528, Japan

    Makoto Hirayama

Authors
  1. Shino Yasukawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kyoko Shirai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kaho Namigata
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Masaaki Ito
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mei Tsubaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hikaru Oyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yukino Fujita
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Taiki Okabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Rei Suo
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Shouzo Ogiso
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yukina Watabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Hajime Matsubara
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Nobuo Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Makoto Hirayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Haruo Sugita
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Shiro Itoi
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Study conception and design: ShinoYasukawa, Rei Suo, Haruo Sugita, and Shiro Itoi. Sample collection: Kyoko Shirai, Masaaki Ito, Hikaru Oyama, Shouzo Ogiso, Yukina Watabe, Hajime Matsubara, Nobuo Suzuki, Makoto Hirayama, and Shiro Itoi. TTX extraction and LC–MS/MS analysis: Shino Yasukawa, Kyoko Shirai, Kaho Namigata, Masaaki Ito, Mei Tsubaki, Hikaru Oyama, Yukino Fujita, Taiki Okabe, and Rei Suo. Statistical analysis: Shino Yasukawa and Shiro Itoi. Manuscript drafting: Shino Yasukawa. Critical revision: Shiro Itoi. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Shiro Itoi.

Ethics declarations

Conflict of Interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Special Issue - JSMB Marine Biotechnology 2022 Conference

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 153 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yasukawa, S., Shirai, K., Namigata, K. et al. Tetrodotoxin Detection in Japanese Bivalves: Toxification Status of Scallops. Mar Biotechnol 25, 666–676 (2023). https://doi.org/10.1007/s10126-023-10199-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10126-023-10199-3

Keywords

Search

Navigation