The association of bacterial C9-based TTX-like compounds with Prorocentrum minimum opens new uncertainties about shellfish seafood safety

Rodríguez, Inés; Alfonso, Amparo; Albrecht, Eva; Rubiolo, Juan A.; Roel, María; Vlamis, Aristidis; Katikou, Panagiota; Jackson, Stephen A.; Margassery, Lekha Menon; Dobson, Alan D. W.; Botana, Luís M.

doi:10.1038/srep40880

Cited by 48 publications

(52 citation statements)

References 41 publications

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“…Unlike many other marine toxins, which are of microalgal origin, TTX production is thought to be produced by bacteria of the genera Pseudomonas, Shewanella, Alteromonas or Vibrio [7], in symbiosis with certain animals [8]. Recently, the marine dinoflagellate Prorocentrum minimum has been described to produce TTXs in cultures, with possible implication of endosymbiotic bacteria [9].…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels

Reverté

Rambla-Alegre

Leonardo

et al. 2018

Talanta

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The recent detection of tetrodotoxins (TTXs) in puffer fish and shellfish in Europe highlights the necessity to monitor the levels of TTXs in seafood by rapid, specific, sensitive and reliable methods in order to protect human consumers. A previous immunoassay for TTX detection in puffer fish, based on the use of self-assembled monolayers (SAMs) for the immobilization of TTX on maleimide plates (mELISA), has been modified and adapted to the analysis of oyster and mussel samples. Changing dithiol for cysteamine-based SAMs enabled reductions in the assay time and cost, while maintaining the sensitivity of the assay. The mELISA showed high selectivity for TTX since the antibody did not cross-react with co-occurring paralytic shellfish poisoning (PSP) toxins and no interferences were observed from arginine (Arg). Moreover, TTX-coated maleimide plates stored for 3 months at -20°C and 4°C were stable, thus when pre-prepared, the time to perform the assay is reduced. When analyzing shellfish samples, matrix effects and toxin recovery values strongly depended on the shellfish type and the sample treatment. Blank oyster extracts could be directly analyzed without solid-phase extraction (SPE) clean-up, whereas blank mussel extracts showed strong matrix effects and SPE and subsequent solvent evaporation were required for removal. However, the SPE clean-up and evaporation resulted in toxin loss. Toxin recovery values were taken as correction factors (CFs) and were applied to the quantification of TTX contents in the analysis of naturally-contaminated shellfish samples by mELISA. The lowest effective limits of detection (eLODs) were about 20 and 50µg/kg for oyster extracts without and with SPE clean-up, respectively, and about 30µg/kg for mussel extracts with both protocols, all of them substantially below the eLOD attained in the previous mELISA for puffer fish (230µg/kg). Analysis of naturally-contaminated samples by mELISA and comparison with LC-MS/MS quantifications demonstrated the viability of the approach. This mELISA is a selective and sensitive tool for the rapid detection of TTX in oyster and mussel samples showing promise to be implemented in routine monitoring programs to protect human health.

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Section: Introductionmentioning

confidence: 99%

Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels

Reverté

Rambla-Alegre

Leonardo

et al. 2018

Talanta

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“…However, a similar trend between TTX in shellfish and P. minimum was not observed in samples from the UK [4]. More recently, TTX-like compounds were identified in cultures of P. minimum [11].…”

Section: Introductionmentioning

confidence: 75%

“…The presence of Vibrio parahaemolyticus, Vibrio alginolyticus and Pseudomonas spp. were confirmed in TTX contaminated mussels from the Greek and UK coasts [3,11]. In addition, the hypothesis of phytoplankton species, namely Prorocentrum minimum, as a source of TTX to bivalves, has been raised by [12] after correlating TTX in shellfish and algae occurrence in the Greek coast.…”

Section: Introductionmentioning

confidence: 90%

Evaluation of the Occurrence of Tetrodotoxin in Bivalve Mollusks from the Portuguese Coast

Rodrigues

Pinto

Oliveira

et al. 2019

JMSE

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Occurrence of tetrodotoxin (TTX), a potent natural neurotoxin, in marine organisms and seafood from the European Union has been of increasing concern due to its relatively recent detection in bivalve mollusks and gastropods. Following a request of the European Food Safety Authority (EFSA) to EU Member States to obtain more data on TTX occurrence, this study collected 117 samples of bivalve mollusks, including mussels (Mytilus galloprovincialis), oysters (Crassostrea gigas) and clams (Ruditapes philippinarum and Donax spp.), from the South and Southwest Portuguese coast between May and October 2018, for TTX determination and microbiological analysis. The analyses carried out by hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC-MS/MS) did not detect TTX in any sample, and microbiological analysis did not reveal high concentrations of Vibrio spp., which has been linked to the presence of TTX. Although preliminary, results from this study, the first investigating the presence of TTX in bivalve mollusks from the Portuguese coast, suggest that TTX may not represent a risk for human consumption of bivalve mollusks.

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“…Rodríguez et al [62] identified several Vibrio and Pseudomonas bacteria, which had previously been reported in TTX production, in contaminated shellfish samples collected in Greece. Most of the TTX-producing strains found in the puffer fish were isolated from the intestine and ovary (63%); others were isolated from the liver (13%), skin and slime on the skin surface (17%), and the total extract of the internal organs (7%).…”

Section: Ecology Of Marine Ttx-producing Bacteriamentioning

confidence: 99%

Tetrodotoxin-Producing Bacteria: Detection, Distribution and Migration of the Toxin in Aquatic Systems

Magarlamov

Melnikova

Chernyshev

2017

Toxins

129

105

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This review is devoted to the marine bacterial producers of tetrodotoxin (TTX), a potent non-protein neuroparalytic toxin. In addition to the issues of the ecology and distribution of TTX-producing bacteria, this review examines issues relating to toxin migration from bacteria to TTX-bearing animals. It is shown that the mechanism of TTX extraction from toxin-producing bacteria to the environment occur through cell death, passive/active toxin excretion, or spore germination of spore-forming bacteria. Data on TTX microdistribution in toxic organs of TTX-bearing animals indicate toxin migration from the digestive system to target organs through the transport system of the organism. The role of symbiotic microflora in animal toxicity is also discussed: despite low toxin production by bacterial strains in laboratory conditions, even minimal amounts of TTX produced by intestinal microflora of an animal can contribute to its toxicity. Special attention is paid to methods of TTX detection applicable to bacteria. Due to the complexity of toxin detection in TTX-producing bacteria, it is necessary to use several methods based on different methodological approaches. Issues crucial for further progress in detecting natural sources of TTX investigation are also considered.

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The association of bacterial C9-based TTX-like compounds with Prorocentrum minimum opens new uncertainties about shellfish seafood safety

Cited by 48 publications

References 41 publications

Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels

Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels

Evaluation of the Occurrence of Tetrodotoxin in Bivalve Mollusks from the Portuguese Coast

Tetrodotoxin-Producing Bacteria: Detection, Distribution and Migration of the Toxin in Aquatic Systems

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