Abstract

Okadaic acid (OA) and dinophysistoxins (DTXs) are the main toxins responsible for diarrhetic shellfish poisoning (DSP) intoxications during harmful algal blooms (HABs). Although the genotoxic and cytotoxic responses to OA have been evaluated in vitro, the in vivo effects of these toxins have not yet been fully explored. The present work fills this gap by evaluating the in vivo effects of the exposure to the DSP-toxin-producing dinoflagellate Prorocentrum lima during the simulation of an early HAB episode in the mussel Mytilus galloprovincialis. The obtained results revealed that in vivo exposure to this toxic microalgae induced early genotoxicity in hemocytes, as a consequence of oxidative DNA damage. In addition, the DNA damage observed in gill cells seems to be mainly influenced by exposure time and P. lima concentration, similarly to the case of the oxidative damage found in hemocytes exposed in vitro to OA. In both cell types, the absence of DNA damage at low toxin concentrations is consistent with the notion suggesting that this level of toxicity does not disturb the antioxidant balance. Lastly, in vivo exposure to growing P. lima cell densities increased apoptosis but not necrosis, probably due to the presence of a high number of protein apoptosis inhibitors in molluscs. Overall, this work sheds light into the in vivo genotoxic and cytotoxic effects of P. lima. In doing so, it also demonstrates for the first time the potential of the modified (OGG1) comet assay for assessing oxidative DNA damage caused by marine toxins in marine invertebrates.

Highlights

  • Harmful Algal Blooms (HABs) constitute a major environmental threat for marine organisms and human consumers of shellfish

  • Since these levels are well below the limit allowed by the European Commission Regulation for harvesting and sale (160 μg of Okadaic acid (OA) equivalent/kg dry weight), mussel specimens were considered as being exposed to an early HAB stage and were used to evaluate the resulting genotoxic and cytotoxic effects

  • The obtained results revealed that: (1) low P. lima cell densities can be used to recreate early stages of HAB episodes in laboratory conditions; (2) the in vivo exposure to extremely low dinoflagellate concentrations (1,000 cells/L) did not produce significant genotoxic effects in hemocytes or gill cells; (3) the DNA damage observed in gill cells exposed in vivo to P. lima was predominantly influenced by exposure time and P. lima cell density; (4) the oxidative DNA damage of hemocytes exposed in vitro to OA was dependent on toxin concentration, while in the case of hemocytes exposed in vivo to P. lima damage was only observed after 24 h exposure to the highest dinoflagellate concentration studied

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Summary

Introduction

Harmful Algal Blooms (HABs) constitute a major environmental threat for marine organisms and human consumers of shellfish. Okadaic acid (OA) and its derivatives, the dinophysistoxins (DTXs), constitute the main lipophilic toxins produced during HABs in the Atlantic coast of Europe [3]. During these episodes, large amounts of these toxins are produced by dinoflagellates from the genera. Their subsequent accumulation by marine organisms is responsible for the Diarrhetic Shellfish Poisoning (DSP) syndrome [4], a disorder causing vomiting, diarrhea, and abdominal pains, among other symptoms, in human consumers of contaminated shellfish [5].

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