Silva M et al. (2013), New invertebrate vectors for PST, spirolides an...

ECB-ART-50094

Mar Drugs 2013 Jun 05;116:1936-60. doi: 10.3390/md11061936.

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New invertebrate vectors for PST, spirolides and okadaic acid in the North Atlantic.

Silva M , Barreiro A , Rodriguez P , Otero P , Azevedo J , Alfonso A , Botana LM , Vasconcelos V .

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The prevalence of poisoning events due to harmful algal blooms (HABs) has declined during the last two decades through monitoring programs and legislation, implemented mainly for bivalves. However, new toxin vectors and emergent toxins pose a challenge to public health. Several locations on the Portuguese coast were surveyed between 2009 and 2010 for three distinct biotoxin groups [saxitoxin (PST), spirolide (SPX) and okadaic acid (OA)], in 14 benthic species of mollusks and echinoderms. Our main goals were to detect new vectors and unravel the seasonal and geographical patterns of these toxins. PSTs were analyzed by the Lawrence method, SPXs by LC-MS/MS, and OA by LC-MS/MS and UPLC-MS/MS. We report 16 new vectors for these toxins in the North Atlantic. There were differences in toxin contents among species, but no significant geographical or seasonal patterns were found. Our results suggest that legislation should be adjusted to extend the monitoring of marine toxins to a wider range of species besides edible bivalves.

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References [+] :

Aasen, Detection and identification of spirolides in norwegian shellfish and plankton. 2005, Pubmed

	Figure 1. Paralytic shellfish toxin (PST) content in saxitoxin (STX) 2 HCl equivalent ng/g fresh weight (fw) in all groups: (A) bivalves—Mytilus galloprovincialis; (B) echinoderms—Marthasterias glacialis and Paracentrotus lividus; (C) gastropods—Nucella lapillus; Charonia lampas; Monodonta turbinata; Monodonta lineata; Gibbula umbilicalis and Aplysia depilans. Limit value for PSTs is 800 ng SXT equivalents/g shellfish meat.
	Figure 2. Box and whisker plots for PST concentrations [SXT HCl 2 equivalents (ng/g fw)] found in each species (C. la.—C. lampas; G. um.—G. umbilicalis; M. ga.—M. galloprovincialis; M. gl.—M. glacialis; M. li.—M. lineata; M. tu.—M. turbinata; N. la.—N. lapillus; P. in.—P. intermedia; P. li.—P. lividus). Limit value for PSTs is 800 ng SXT equivalents/g shellfish meat.
	Figure 3. PST profiles in each group screened.
	Figure 4. Okadaic acid (OA) positive results (ng/g fw) for all sampled groups of organisms: (A) bivalves; (B) echinoderms; (C) gastropods. Y-axis in logarithmic scale. Limit value (LV) established in Europe is 160 ng OA equivalents (eq)/g shellfish meat (s.m.).
	Figure 5. Box and whisker plots of OA concentrations (ng/g fw) found in each species (C. la.—C. lampas; G. um.—G. umbilicalis; M. ga.—M. galloprovincialis; M. gl.—M. glacialis; M. li.—M. lineata; M. tu.—M. turbinata; N. la.—N. lapillus; P. in.—P. intermedia; P. li.—P. lividus). Limit value (LV) established in Europe is 160 ng OA equivalents (eq)/g shellfish meat (s.m.).
	Figure 6. Trophic relation between M. galloprovincialis, N. lapillus and M. glacialis.
	Figure 7. SPX positive results (ng/g fw) for all sampled groups of organisms.
	Figure 8. Box and whisker plots of SPX concentration (ng/g fw) found in each species (C. la.—C. lampas; G. um.—G. umbilicalis; M. ga.—M. galloprovincialis; M. gl.—M. glacialis; M. li.—M. lineata; M. tu.—M. turbinata; N. la.—N. lapillus; P. in.—P. intermedia; P. li.—P. lividus).
	Figure 9. Location of the sampling points on the Atlantic Portuguese coast: 1: Viana do Castelo; 2: Esposende; 3: Póvoa do Varzim; 4: Angeiras; 5: Memória; 6: Valadares; 7: Aguda; 8: São Martinho do Porto; 9: São Torpes; 10: Porto Côvo; 11: Monte Clérigos; 12: Vila Nova de Milfontes; 13: Almograve.
	Figure 10. Chromatograms of the PSTs standards and a positive sample for M. glacialis, hydroxylated toxins are oxidized with periodate and non-hydroxylated toxins with peroxide. (A) Standards injected with periodate oxidation, peak GTX1,4(1) and GTX1,4(3) are secondary oxidation products of GTX1,4. Peaks NEO(1) and NEO(3) are secondary oxidation products of NEO. (B) Standards injected with peroxide oxidation. (C) Chromatogram of a positive sample in M. glacialis injected with periodate oxidation. (D) Chromatogram of a positive sample in M. glacialis injected with peroxide oxidation.
	Figure 11. Mass chromatograms of the LC-ESI-CID-MS/MS obtained under multiple reaction monitoring (MRM): (A) extracted ion chromatogram (XIC) of OA standard (200 ng/mL), m/z 803.5 > 255.5/113.5; (B) XIC of 13-desmethyl SPX C standard (200 ng/mL), m/z 692.4 > 674.4/444.4; (C) XIC of a positive sample for OA in N. lapillus (m/z 803.5 > 255.5/113.5); (D) XIC of a positive sample for 13-desmethyl SPX C in N. lapillus (m/z 692.4 > 674.4/444.4).
	Figure 12. Mass chromatograms of the UPLC-MS/MS obtained under MRM operation: (A) total ion chromatogram (TIC) of OA standard 200 ng/mL, m/z 803.5 > 255.5/113.5; (B) TIC of a positive sample for OA in M. glacialis, m/z 803.5 > 255.5/113.5.