Abstract

BackgroundMicrocystins are emerging marine biotoxins, produced by potentially toxic cyanobacteria. Their presence has been reported in aquatic animals in Greek freshwater, while data are few in marine environments. Since the climate change induces eutrophication and harmful algal blooms in coastal marine ecosystems affecting the public health, further research on microcystins’ presence in marine waters is required. The aim of this study was to examine the potential presence of microcystins in mussels Mytilus galloprovincialis in the largest farming areas in Thermaikos gulf, in Northern Greece, and to investigate their temporal and spatial distribution, adding to the knowledge of microcystins presence in Greek Mediterranean mussels.ResultsA 4-year microcystins’ assessment was conducted from 2013 to 2016, in farmed Mediterranean mussels M. galloprovincialis, in five sampling areas in Thermaikos gulf, in northern Greece, where the 90% of the Greek mussels’ farming activities is located. The isolation of potentially toxic cyanobacteria was confirmed by molecular methods. An initial screening was performed with a qualitative and quantitative direct monoclonal (DM) ELISA and results above 1 ng g−1 were confirmed for the occurrence of the most common microcystins-RR, -LR and -YR, by Ultra High Performance Liquid Chromatography (UHPLC) coupled with a high- resolution mass spectrometer (HRMS) (Orbitrap analyzer). Microcystin-RR and microcystin-LR were detected, while the intensity of microcystin-YR was below the method detection limit. Most samples that exhibited concentrations above 1 ng g-1 were detected during the warm seasons of the year and especially in spring. Results indicated an overestimation of the ELISA method, since concentrations ranged between 0.70 ± 0.15 ng g−1 and 53.90 ± 3.18 ng g−1, while the confirmation denoted that the levels of microcystins were 6 to 22 times lower.ConclusionsMicrocystin-RR and microcystin-LR were detected for the first time in mussel M. galloprovincialis, harvested from farms in Thermaikos gulf, in Central Macedonia, Greece. Their presence was linked to potentially toxic cyanobacteria. Bioaccumulation was observed in digestive gland, while the concentrations in muscles were found extremely low. Samples with levels above 1 ng g−1 were observed mostly during spring, confirming the seasonal distribution of microcystins. The comparison of the results by the ELISA and the LC-Orbitrap MS method indicated an overestimation of concentration by the ELISA method.

Highlights

  • Microcystins are emerging marine biotoxins, produced by potentially toxic cyanobacteria

  • The identification was based on the presence of at least one peak corresponding to the pseudomolecular ion of the microcystin, at a retention time (­tR) matching to that of the certified standards with a drift < 0.2 min, as well as the determination of the exact mass with a mass error

  • Our study indicates that microcystins can be bioaccumulated in mussels, a finding that is in accordance with the literature [42,43,44,45,46]

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Summary

Introduction

Microcystins are emerging marine biotoxins, produced by potentially toxic cyanobacteria Their presence has been reported in aquatic animals in Greek freshwater, while data are few in marine environments. Microcystins (MCs) are the most common group of cyanotoxins, with high toxicity [2] and they are considered as emerging toxins [3] They are produced by different genera of freshwater cyanobacteria, such as Microcystis, Anabaena (Dolichospermum), Nostoc, Planktothrix, Chroococcus [4], as well as by marine picoplanktonic species, such as Synechococcus and Synechocystis [5]. The detection of microcystins has been reported mainly in freshwater, as well as in brackish and marine environments [1] Their presence in food web and especially in seafood such as shellfish, fish and crustaceans is considered hazardous for the public health [1]. Regarding the presence of microcystins in marine aquatic organisms, there is only one report in Mediterranean mussels Mytilus galloprovincialis from

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