Skip to main content

Depuration of Tetrodotoxin and Changes in Bacterial Communities in Pleurobranchea maculata Adults and Egg Masses Maintained in Captivity

Journal of Chemical Ecology volume 38, pages 1342–1350 (2012)Cite this article

Abstract

Depuration of tetrodotoxin (TTX) was investigated in adult grey side-gilled sea slugs, Pleurobranchaea maculata, maintained in captivity on a TTX-free diet. Three adults were harvested every 21 days for 126 days, and TTX concentrations were measured in organs/tissues and egg masses. Automated rRNA intergenic spacer analysis (ARISA) was used to investigate bacterial community structure in selected samples. Linear modeling of adult data demonstrated a decline (P < 0.001) in average total TTX concentrations over time. Temporal data obtained from a wild population showed similar depuration rates, indicating that once adults reach a certain size, or sexual maturity, TTX is no longer produced or acquired substantially. Depuration rates differed among organs, with concentrations in the heart declining the fastest. The gonads had the slowest and least significant depuration rate indicating, at most, weak depuration of this tissue. There was a strong correlation (R 2 = 0.66) between TTX concentrations in the first-laid egg masses and total TTX in the corresponding adult. These data suggest that adult P. maculata transfer TTX to their offspring, and presumably that functions as a chemical defense. ARISA data showed a shift in bacterial community structure within 3 weeks of introduction to captivity. Based on the combined data, the exact origin of TTX in P. maculata is unclear, with evidence both in favor and against a dietary source, and endogenous or bacterial production.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  • Abdo, Z., Schüette, U., Bent, S., Williams, C., Forney, L., and Joyce, P. 2006. Statistical methods for characterizing diversity of microbial communities by analysis of terminal restriction fragment length polymorphisms of 16S rRNA genes. Environ. Microbiol. 8:929–938.

    PubMed  Article  Google Scholar 

  • Dhanasiri, A. K., Brunvold, L., Brinchmann, M. F., Korsnes, K., Bergh, Ø., and Kiron, V. 2011. Changes in the intestinal microbiota of wild atlantic cod Gadus morhua L. upon captive rearing. Microbial. Ecol. 61:20–30.

    Article  Google Scholar 

  • Cardalll, B. L., Brodie JR, E. D., Brodie III, E. D., and Hanifin, C. T. 2004. Secretion and regeneration of tetrodotoxin in the rough-skin newt (Taricha granulosa). Toxicon 44:933–938.

    Article  Google Scholar 

  • Cardinale, M., Brusetti, L., Quatrini, P., Borin, S., Puglia, A., Rizzi, A., Zanardini, E., Sorlini, C., Corselli, C., and Daffonchio, D. 2004. Comparison of different primer sets for use in automated ribosomal intergenic spacer analysis of complex bacterial communities. Appl. Environ. Microbiol. 70:6147–6156.

    PubMed  Article  CAS  Google Scholar 

  • Chau, R., Kalaitzis, J. A., and Neilan, B. A. 2011. On the origins and biosynthesis of tetrodotoxin. Aquat. Toxicol. 104:61–72.

    PubMed  Article  CAS  Google Scholar 

  • Dao, H. V., Yoshinobu, T., Sato, S., Fukuyo, Y., and Kodama, M. 2009. Frequent occurrence of the tetrodotoxin-bearing horseshoe crab Carcinoscorpius rotundicauda in Vietnam. Fish. Sci. 75:435–438.

    Article  CAS  Google Scholar 

  • Fisher, M. M. and Triplett, E. W. 1999. Automated approach for ribosomal intergenic spacer analysis of microbial diversity and its application to freshwater bacterial communities. Appl. Environ. Microbiol. 65:4630–4636.

    PubMed  CAS  Google Scholar 

  • Gibson, G. D. 2003. Larval development and metamorphosis in Pleurobranchaea maculata, with a review of development in the Notaspidea (Opisthobranchia). Biol. Bull. (Woods Hole) 205:121–132.

    Article  Google Scholar 

  • Hanifin, C. T. 2010. The chemical and evolutionary ecology of tetrodotoxin (TTX) toxicity in terrestrial vertebrates. Mar. Drugs 8:577–593.

    PubMed  Article  CAS  Google Scholar 

  • Hanifin, C. T., Brodie III, E. D., and Brodie Jr., E. D. 2002. Tetrodotoxin levels of the rough-skin newt, Taricha granulosa, increase in long-term captivity. Toxicon 40:1149–1153.

    PubMed  Article  CAS  Google Scholar 

  • Hanifin, C. T., Brodie III, E. D., and Brodie Jr., E. D. 2003. Tetrodotoxin levels in eggs of the rough-skin newt, Taricha granulosa, are correlated with female toxicity. J. Chem. Ecol. 25:1729–1739.

  • Hanifin, C. T., Yotu-Yamashita, M., Yasumoto, T., Brodie III, E. D., and Brodie JR, E. D. 1999. Toxicity of dangerous prey: Variation of tetrodotoxin levels within and among populations of the newt Taricha granulosa. J. Chem. Ecol. 25:2161–2175.

    Article  CAS  Google Scholar 

  • Hwang, D. E., Lu, S. C., and Jeng, S. S. 1991. Occurrence of tetrodotoxin in the gastropods Rapana rapiformis and R. venosa venosa. Mar. Biol. 111:65–69.

    Article  CAS  Google Scholar 

  • Isaacs, L. T., Kan, J., Nguyen, L., Videau, P., Anderson, M. A., Wright, T. L., and Hill, R. T. 2009. Comparison of the bacterial communities of wild and captive sponge Clathria prolifera from the Chesapeake Bay. Mar. Biotechnol. 6:758–70.

    Article  Google Scholar 

  • Lehman, E. M., Brodie, E. D. J. R., and Brodie III, E. D. 2004. No evidence for an endosymbiotic bacterial origin of tetrodotoxin in the newt Taricha granulosa. Toxicon 44:243–249.

    PubMed  Article  CAS  Google Scholar 

  • Matsui, T., Yamamori, K., Furukawa, K., and Kono, M. 2000. Purification and some properties of a tetrodotoxin binding protein from the blood plasma of kusafugu, Takifugu niphobles. Toxicon 38:463–468.

    PubMed  Article  CAS  Google Scholar 

  • Matsumura, K. 2001. No ability to produce tetrodotoxin in bacteria. Appl. Environ. Microbiol. 67:2393–2394.

    PubMed  Article  CAS  Google Scholar 

  • Matsumura, K. 1995. Re-examination of tetrodotoxin production by bacteria. Appl. Environ. Microbiol. 61:3468–3470.

    PubMed  CAS  Google Scholar 

  • McNabb, P., Selwood, A. I., Munday, R., Wood, S. A., Taylor, D. I., Mackenzie, L. A., van Ginkel, R., Rhodes, L. L., Cornelisen, C., Heasman, K., Holland, P. T., and King, C. 2010. Detection of tetrodotoxin from the grey side-gilled sea slug - Pleurobranchaea maculata, and associated dog neurotoxicosis on beaches adjacent to the Hauraki Gulf, Auckland, New Zealand. Toxicon 56:466–473.

    PubMed  Article  CAS  Google Scholar 

  • Nagashima, Y., Mataki, I., Toyoda, M., Nakajima, H., Tsumotoo, K., Shimakura, K., and Shioma, K. 2010. Change in tetrodotoxin content of the puffer fish Takifugu rubripes during seed production from fertilized eggs to juveniles. Food Hyg. Saf. Sci. 51:48–51.

    Article  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  • Noguchi, T., Arakawa, O., and Takatani, T. 2006. Toxicity of pufferfish Takifugu rubripes cultured in net cages at sea or aquaria on land. Comp. Biochem. Phys. D. 153–157.

  • Noguchi, T. and Ebesu, J. S. M. 2001. Puffer poisoning: epidemiology and treatment. J. Toxicol. Toxin Rev. 20:1–10.

    CAS  Google Scholar 

  • Pawlik, J. R., Kernan, M. R., Molinski, T. F., Harper, M. K., and Faulkner, D. J. 1988. Defensive chemicals of the Spanish dancer nudibranch Hexabranchus sanguineus and its egg ribbons: macrolides derived from a sponge diet. J. Exp. Mar. Biol. Ecol. 119:99–109.

    Article  CAS  Google Scholar 

  • Schroeder, F. C., Gonzalez, A., Eisner, T., and Meinwald, J. 1999. Miriamin, a defensive diterpene from the eggs of a land slug (Arion sp.). Proc. Nat. Acad. Sci. USA 96:13620–13625.

    PubMed  Article  CAS  Google Scholar 

  • Snedecor, G. W. and Cochran, W. O. 1980. pp. 503, Statistical methods. Iowa State University Press, Ames.

    Google Scholar 

  • Willan, R. C. 1984. A review of diets in the Notaspidea (Mollusca: Opisthobranchia). J. Mal. Soc. Aust. 6:125–142.

    Google Scholar 

  • Williams, B. L. and Caldwell, R. L. 2009. Intra‑organismal distribution of tetrodotoxin in two species of blue‑ringed octopuses (Hapalochlaena fasciata and H. lunulata). Toxicon 54:345–353.

    PubMed  Article  CAS  Google Scholar 

  • Williams, B. L., Hanifin, C. T., Brodie Jr., E. D., and Caldwell, R. L. 2011. Ontogenty of tetrodtoxin levels in blue-ringed octopuses: maternal investment and apparent independent production in offspring of Hapalochlaena lunulata. J. Chem. Ecol. 37:10–17.

    PubMed  Article  CAS  Google Scholar 

  • Wood, S. A., Taylor, D. I., McNabb, P., Walker, J., Adamson, J., and Cary, S. C. 2012. Tetrodotoxin concentrations in Pleurobranchaea maculata: temporal, spatial and individual variability from New Zealand populations. Mar. Drugs. 10:163–176.

    PubMed  Article  CAS  Google Scholar 

  • Yotsu-Yamashita, M., Sugimoto, A., Terakawa, T., Shoji, Y., Miyazawa, T., and Yasumoto, T. 2001. Purification, characterization, and cDNA cloning of a novel soluble saxitoxin and tetrodotoxin binding protein from plasma of the puffer fish, Fugu pardalis. Eur. J. Biochem. 268:5937–5946.

    PubMed  Article  CAS  Google Scholar 

  • Yotsu-Yamashita, M., Gilhen, J., Russell, R. W., Krysko, K. L., Melaun, C., Kurz, A., Kauferstein, S., Kordis, D., and Mebs, D. 2012. Variability of tetrodotoxin and of its analogues in the red-spotted newt, Notophthalmus viridescens (Amphibia: Urodela: Salamandridae). Toxicon 59:257–264.

    PubMed  Article  CAS  Google Scholar 

  • Yotsu-Yamashita, M., Mebs, D., and Yasumoto, T. 1992. Tetrodotoxin and its analogues in extracts from the toad Atelopus oxyrhynchus (family: bufonidae). Toxicon 30:1489–1492.

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was funded by the Marsden fund of the Royal Society of New Zealand (UOW1002) and Ngā Pae o Te Māramatanga (Project: 10RF18). We thank the Auckland Harbour Master and Jarrod Walker from the Auckland Council for assistance during P. maculata collections. P.M. is supported by a New Zealand Ministry for Science and Innovation Te Tipu Pūtaiao Ph.D. fellowship (CAWX0905). We are grateful to Janet Adamson (Cawthron) for technical assistance and Eric Bottos (Waikato University) and Eric Goodwin (Cawthron) for assistance with statistical analysis.

Author information

Authors and Affiliations

  1. Cawthron Institute, Nelson, New Zealand

    Susanna A. Wood, Margaux Casas, David I. Taylor, Paul McNabb & Shaun Ogilvie

  2. Department of Biological Sciences, University of Waikato, Hamilton, New Zealand

    Susanna A. Wood, Lauren Salvitti & S. Craig Cary

  3. Institut Polytechnique LaSalle, Beauvais, France

    Margaux Casas

  4. Department of Chemistry, Otago University, Dunedin, New Zealand

    Paul McNabb

  5. Eco Research Associates Ltd, Christchurch, New Zealand

    Shaun Ogilvie

Authors
  1. Susanna A. Wood
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Margaux Casas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. David I. Taylor
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paul McNabb
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lauren Salvitti
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shaun Ogilvie
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S. Craig Cary
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Susanna A. Wood.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wood, S.A., Casas, M., Taylor, D.I. et al. Depuration of Tetrodotoxin and Changes in Bacterial Communities in Pleurobranchea maculata Adults and Egg Masses Maintained in Captivity. J Chem Ecol 38, 1342–1350 (2012). https://doi.org/10.1007/s10886-012-0212-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10886-012-0212-9

Keywords

  • Chemical defense
  • Depuration
  • Egg mass toxicity
  • Pleurobranchaea maculata
  • Tetrodotoxin