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First Report of Ciguatoxins in Two Starfish Species: Ophidiaster ophidianus and Marthasterias glacialis.

Silva M, Rodriguez I, Barreiro A, Kaufmann M, Isabel Neto A, Hassouani M, Sabour B, Alfonso A, Botana LM, Vasconcelos V - Toxins (Basel) (2015)

Bottom Line: We detected differences regarding uptake values by organisms and geographical location.Toxin amounts were significant, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, in order to better access human health risk.In addition, we suggest monitoring of these toxins should be extended to other vectors, starfish being a good alternative for protecting and accessing human health risk.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Faculty of Sciences, University of Porto, Rua do Campo Alegre, Porto 4619-007, Portugal. marisasilva17@gmail.com.

ABSTRACT
Ciguatera fish poisoning (CFP) is a syndrome caused by the ingestion of fish contaminated with Ciguatoxins (CTXs). These phycotoxins are produced mainly by dinoflagellates that belong to the genus Gambierdiscus that are transformed in more toxic forms in predatory fish guts, and are more present in the Indo-Pacific and Caribbean areas. It is estimated that CFP causes per year more than 10,000 intoxications worldwide. With the rise of water temperature and anthropogenic intervention, it is important to study the prevalence of CFP in more temperate waters. Through inter- and subtidal sampling, 22 species of organisms were collected, in Madeira and Azores archipelagos and in the northwestern Moroccan coast, during September of 2012 and June and July of 2013. A total of 94 samples of 22 different species of bivalves, gastropods, echinoderms and crustaceans where analyzed by Ultra Performance Liquid Chromatography-Mass Spectometry-Ion Trap-Time of Flight (UPLC-MS-IT-TOF) and Ultra Performance Chromatography- Mass Spectrometry (UPLC-MS). Our main aim was to detect new vectors and ascertain if there were some geographical differences. We detected for the first time putative CTXs in echinoderms, in two starfish species-M. glacialis and O. ophidianus. We detected differences regarding uptake values by organisms and geographical location. Toxin amounts were significant, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, in order to better access human health risk. In addition, we suggest monitoring of these toxins should be extended to other vectors, starfish being a good alternative for protecting and accessing human health risk.

No MeSH data available.


Related in: MedlinePlus

CTX purification scheme.
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toxins-07-03740-f005: CTX purification scheme.

Mentions: The Otero et al. (2010) extraction protocol was followed [45]. The efficiency of the method was studied by analyzing the extracts discarded in each stage of the protocol. Data showed no loss of toxin in each step. The results agree with the efficiency achieved in the method previously described (>95% for P-CTX-1B) [46]. Animals were dissected and homogenized with a blender (A320R1, 700 W, Moulinex, Lisbon, Portugal) in pooled groups in order to obtain 2 g of tissue, with the exception of Aplysia depilans, Charonia lampas, Diadema africanum, Holothuria (Platyperona) sanctori, Marthasterias glacialis, Ophidiaster ophidianus, Paracentrotus lividus, Sphaerechinus granularis, and Umbraculum umbraculum. In these cases, each animal was treated separately since they had enough extractable biomass. The homogenized tissue was cooked for 20 min at 70 °C, then homogenized with 8 mL of Methanol/Hexane (3:1), sonicated (1 min, 70 Hz, Vibra Cell, Sonic & Materials, Newtown, CT, USA), and subsequently centrifuged at 4000 rpm for 20 min. The upper hexane layer was discarded, and the lower methanol phase was filtered through a 0.45-µm filter (Millipore Ultrafree-MC centrifugal filter units, Bedford, MA, USA). The resulting filtered was diluted into methanol water (50:50). Thereafter Solid Phase Extraction (SPE) was performed using C18 SPE cartridges (500 mg/3 mL volume from Supelco, Bellefonte, PA, USA). Cartridges were previously conditioned with 4 mL of milliQ water, then samples were loaded and washed with 65% Methanol, and finally samples were eluted in 80% Methanol. Thereafter, samples were mixed with 4.2 mL of 1 M NaCl and 6.7 mL of Chloroform and centrifuged for 4 min at 2000 rpm (Centrifugal-Legend RT, Sorvall, Waltham, MA, USA). The upper methanolic layer was discarded and the lower organic layer was evaporated to dryness in a rotary evaporator (Büchi, Flawil, Switzerland) and dissolved in 4 mL chloroform. For reducing matrix interference, another cleanup procedure was done with Silica Sep-Pak cartridges (Waters, Milford, CT, USA). After loading the sample cartridges were conditioned with chloroform, samples were washed with chloroform and eluted 90% of chloroform. Extract was concentrated to dryness and then re-suspended in methanol. In Figure 5 is displayed the totality of the purification procedure. Before UPLC-MS analysis, positive samples were confirmed and the exact mass was obtained by UPLC-MS-IT-TOF.


First Report of Ciguatoxins in Two Starfish Species: Ophidiaster ophidianus and Marthasterias glacialis.

Silva M, Rodriguez I, Barreiro A, Kaufmann M, Isabel Neto A, Hassouani M, Sabour B, Alfonso A, Botana LM, Vasconcelos V - Toxins (Basel) (2015)

CTX purification scheme.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591666&req=5

toxins-07-03740-f005: CTX purification scheme.
Mentions: The Otero et al. (2010) extraction protocol was followed [45]. The efficiency of the method was studied by analyzing the extracts discarded in each stage of the protocol. Data showed no loss of toxin in each step. The results agree with the efficiency achieved in the method previously described (>95% for P-CTX-1B) [46]. Animals were dissected and homogenized with a blender (A320R1, 700 W, Moulinex, Lisbon, Portugal) in pooled groups in order to obtain 2 g of tissue, with the exception of Aplysia depilans, Charonia lampas, Diadema africanum, Holothuria (Platyperona) sanctori, Marthasterias glacialis, Ophidiaster ophidianus, Paracentrotus lividus, Sphaerechinus granularis, and Umbraculum umbraculum. In these cases, each animal was treated separately since they had enough extractable biomass. The homogenized tissue was cooked for 20 min at 70 °C, then homogenized with 8 mL of Methanol/Hexane (3:1), sonicated (1 min, 70 Hz, Vibra Cell, Sonic & Materials, Newtown, CT, USA), and subsequently centrifuged at 4000 rpm for 20 min. The upper hexane layer was discarded, and the lower methanol phase was filtered through a 0.45-µm filter (Millipore Ultrafree-MC centrifugal filter units, Bedford, MA, USA). The resulting filtered was diluted into methanol water (50:50). Thereafter Solid Phase Extraction (SPE) was performed using C18 SPE cartridges (500 mg/3 mL volume from Supelco, Bellefonte, PA, USA). Cartridges were previously conditioned with 4 mL of milliQ water, then samples were loaded and washed with 65% Methanol, and finally samples were eluted in 80% Methanol. Thereafter, samples were mixed with 4.2 mL of 1 M NaCl and 6.7 mL of Chloroform and centrifuged for 4 min at 2000 rpm (Centrifugal-Legend RT, Sorvall, Waltham, MA, USA). The upper methanolic layer was discarded and the lower organic layer was evaporated to dryness in a rotary evaporator (Büchi, Flawil, Switzerland) and dissolved in 4 mL chloroform. For reducing matrix interference, another cleanup procedure was done with Silica Sep-Pak cartridges (Waters, Milford, CT, USA). After loading the sample cartridges were conditioned with chloroform, samples were washed with chloroform and eluted 90% of chloroform. Extract was concentrated to dryness and then re-suspended in methanol. In Figure 5 is displayed the totality of the purification procedure. Before UPLC-MS analysis, positive samples were confirmed and the exact mass was obtained by UPLC-MS-IT-TOF.

Bottom Line: We detected differences regarding uptake values by organisms and geographical location.Toxin amounts were significant, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, in order to better access human health risk.In addition, we suggest monitoring of these toxins should be extended to other vectors, starfish being a good alternative for protecting and accessing human health risk.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Faculty of Sciences, University of Porto, Rua do Campo Alegre, Porto 4619-007, Portugal. marisasilva17@gmail.com.

ABSTRACT
Ciguatera fish poisoning (CFP) is a syndrome caused by the ingestion of fish contaminated with Ciguatoxins (CTXs). These phycotoxins are produced mainly by dinoflagellates that belong to the genus Gambierdiscus that are transformed in more toxic forms in predatory fish guts, and are more present in the Indo-Pacific and Caribbean areas. It is estimated that CFP causes per year more than 10,000 intoxications worldwide. With the rise of water temperature and anthropogenic intervention, it is important to study the prevalence of CFP in more temperate waters. Through inter- and subtidal sampling, 22 species of organisms were collected, in Madeira and Azores archipelagos and in the northwestern Moroccan coast, during September of 2012 and June and July of 2013. A total of 94 samples of 22 different species of bivalves, gastropods, echinoderms and crustaceans where analyzed by Ultra Performance Liquid Chromatography-Mass Spectometry-Ion Trap-Time of Flight (UPLC-MS-IT-TOF) and Ultra Performance Chromatography- Mass Spectrometry (UPLC-MS). Our main aim was to detect new vectors and ascertain if there were some geographical differences. We detected for the first time putative CTXs in echinoderms, in two starfish species-M. glacialis and O. ophidianus. We detected differences regarding uptake values by organisms and geographical location. Toxin amounts were significant, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, in order to better access human health risk. In addition, we suggest monitoring of these toxins should be extended to other vectors, starfish being a good alternative for protecting and accessing human health risk.

No MeSH data available.


Related in: MedlinePlus