scholarly journals Characterization of the Domoic Acid Uptake Mechanism of the Mussel (Mytilus galloprovincialis) Digestive Gland

Toxins ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 458
Author(s):  
Juan Blanco ◽  
Carmen Mariño ◽  
Helena Martín ◽  
Gonzalo Álvarez ◽  
Araceli E. Rossignoli
Keyword(s):  
Digestive Gland ◽  
Domoic Acid ◽  
Mytilus Galloprovincialis ◽  
Metabolic Inhibitor ◽  
Acid Uptake ◽  
Free Medium ◽  
Shellfish Poisoning ◽  
Uptake Mechanism ◽  
Amnesic Shellfish Poisoning

Cultures of the mussel Mytilus galloprovincialis are frequently affected by accumulation of the amnesic shellfish poisoning toxin domoic acid (DA). This species is characterized by a fast uptake and release of the toxin. In this work, the main characteristics of the uptake mechanism have been studied by incubation of digestive gland thin slices in media with different composition and DA concentration. DA uptake seems to follow Michaelis–Menten kinetics, with a very high estimated KM (1722 µg DA mL−1) and a Vmax of 71.9 µg DA g−1 h−1, which is similar to those found for other amino acids in invertebrates. Replacement of NaCl from the incubation media by Cl-choline (Na+-free medium) did not significantly reduce the uptake, but replacement by sorbitol (Na+-free and Cl−-depleted medium) did. A new experiment replacing all chlorides with their equivalent gluconates (Na+- and Cl−-free medium) showed an important reduction in the uptake that should be attributed to the absence of chloride, pointing to a Na+-independent, Cl− (or anion-) dependent transporter. In media with Na+ and Cl−, neither decreasing the pH nor adding cyanide (a metabolic inhibitor) had significant effect on DA uptake, suggesting that the transport mechanism is not H+- or ATP-dependent. In a chloride depleted medium, lowering pH or adding CN increased the uptake, suggesting that other anions could, at least partially, substitute chloride.

scholarly journals Rapid Domoic Acid Depuration in the Scallop Argopecten purpuratus and Its Transfer from the Digestive Gland to Other Organs

Toxins ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 698
Author(s):  
Gonzalo Álvarez ◽  
José Rengel ◽  
Michael Araya ◽  
Francisco Álvarez ◽  
Roberto Pino ◽  
...  
Keyword(s):  
Digestive Gland ◽  
Domoic Acid ◽  
Marine Resources ◽  
Small Decrease ◽  
Shellfish Poisoning ◽  
Natural Conditions ◽  
Amnesic Shellfish Poisoning ◽  
Argopecten Purpuratus ◽  
The One ◽  
Best Fit

Domoic acid (DA), the main toxin responsible for Amnesic Shellfish Poisoning, frequently affects the marine resources of Chile and other countries across the South Pacific, thus becoming a risk for human health. One of the affected resources is the scallop Argopecten purpuratus. Even though this species has a high commercial importance in Northern Chile and Peru, the characteristics of its DA depuration are not known. In this work, the DA depuration was studied by means of two experiments: one in controlled (laboratory) and another in natural conditions. All organs of A. purpuratus depurated the toxin very quickly in both experiments. In some organs, an increase or a very small decrease of toxin was detected in the early depuration steps. Several models were used to describe this kinetics. The one that included toxin transfer between organs and independent depuration from each organ was the model that best fit the data. It seems, therefore, that the DA in this species is quickly transferred from the digestive gland to all other organs, which release it into the environment. Physiological differences in the two experiments have been shown to have some effect on the depuration from each organ but the actual reasons are still unknown.

scholarly journals Long Term Memory Outcome of Repetitive, Low-Level Dietary Exposure to Domoic Acid in Native Americans

2021 ◽  
Vol 18 (8) ◽  
pp. 3955
Author(s):  
Lynn M. Grattan ◽  
Laura Kaddis ◽  
J. Kate Tracy ◽  
John Glenn Morris
Keyword(s):  
Native Americans ◽  
Domoic Acid ◽  
Dietary Exposure ◽  
Long Term Memory ◽  
Shellfish Poisoning ◽  
Amnesic Shellfish Poisoning ◽  
The Pacific ◽  
Small Decline ◽  
Speed And Accuracy

Domoic acid (DA) is a marine-based neurotoxin that, if ingested via tainted shellfish, is associated with Amnesic Shellfish Poisoning (ASP). These acute effects of elevated DA exposure in humans have been well described. In contrast, the long-term impacts of lower level, repetitive, presumably safe doses of DA (less than 20 ppm) are minimally known. Since Native Americans (NA) residing in coastal communities of the Pacific NW United States are particularly vulnerable to DA exposure, this study focuses on the long-term, 8-year memory outcome associated with their repeated dietary consumption of the neurotoxin. Measures of razor clam consumption, memory, clerical speed and accuracy, and depression were administered over eight years to 500 randomly selected adult NA men and women ages 18–64. Data were analyzed using GEE analyses taking into consideration the year of study, demographic factors, and instrumentation in examining the association between dietary exposure and outcomes. Findings indicated a significant but small decline in total recall memory within the context of otherwise stable clerical speed and accuracy and depression scores. There is reason to believe that a continuum of memory difficulties may be associated with DA exposure, rather than a unitary ASP syndrome.

scholarly journals Monitoring the presence of domoic acid in the production areas of bivalve molluscs

2016 ◽  
Vol 5 (4) ◽  
Author(s):  
Rachele Rossi ◽  
Olga Arace ◽  
Maria Giovanna Buonomo ◽  
Daniela Capozzo ◽  
Vincenzo Castellano ◽  
...  
Keyword(s):  
Health Care ◽  
Experimental Method ◽  
Domoic Acid ◽  
Food Poisoning ◽  
Chemical Compounds ◽  
Official Method ◽  
Shellfish Poisoning ◽  
Bivalve Molluscs ◽  
Amnesic Shellfish Poisoning ◽  
Production Areas

Algal biotoxins, chemical compounds produced by some microscopic algae, constitute the <em>phytoplankton</em>. The mussels, feeding on phytoplankton, can accumulate these compounds to become themselves toxic. There have been several cases of food poisoning by consumption of contaminated shellfish. Such food poisoning have pushed our health care system to provide monitoring of shellfish in the framework of the monitoring plans carried out by AASSLL. In this paper we report the results obtained monitoring the presence of ASP (Amnesic Shellfish Poisoning) biotoxins, like domoic acid (DA) and its isomers, produced by <em>Pseudonitzschia</em> algae. The analysis were carried out by using both the HPLC-UV official method and an experimental method performed with a Time of Flight mass spectrometer (ESI-TOF). The 100% of samples analysed by the official method have always been below the limits of sensitivity (except one sample), the 65% of samples analysed by ESI-TOF, showed the presence of domoic acid.

scholarly journals Distribution of Domoic Acid in the Digestive Gland of the King Scallop Pecten maximus

Toxins ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 371
Author(s):  
Juan Blanco ◽  
Aida Mauríz ◽  
Gonzalo Álvarez
Keyword(s):  
Digestive Gland ◽  
Domoic Acid ◽  
Time Course ◽  
Cell Types ◽  
Individual Variability ◽  
Small Cells ◽  
Intestinal Loop ◽  
Pecten Maximus ◽  
Shellfish Poisoning ◽  
Long Time

The king scallop Pecten maximus retains the amnesic shellfish poisoning toxin, domoic acid (DA), for a long time. Most of the toxin is accumulated in the digestive gland, but this organ contains several cell types whose contribution to the accumulation of the toxin is unknown. Determining the time-course of the depuration by analyzing whole organs is difficult because the inter-individual variability is high. A sampling method, using biopsies of the digestive gland, has been developed. This method allows for repetitive sampling of the same scallop, but the representativeness of the samples obtained in this way needs to be validated. In this work, we found that the distribution of DA in the digestive gland of the scallops is mostly homogeneous. Only the area closest to the gonad, and especially its outer portion, had a lower concentration than the other ones, probably due to a transfer of the toxin to the intestinal loop. Samples obtained by biopsies can therefore be considered to be representative. Most of the toxin was accumulated in large cells (mostly digestive cells), which could be due to differences during the toxin absorption or to the preferential depuration of the toxin from the small cells (mostly secretory).

scholarly journals High-Throughput Analysis of Amnesic Shellfish Poisoning Toxins in Shellfish by Ultra-Performance Rapid Resolution LC-MS/MS

2011 ◽  
Vol 94 (2) ◽  
pp. 555-564 ◽  
Author(s):  
Pablo De La Iglesia ◽  
Esther Barber ◽  
Gemma Giménez ◽  
María Luisa Rodríguez-Velasco ◽  
Adriano Villar-González ◽  
...  
Keyword(s):  
Domoic Acid ◽  
Proficiency Test ◽  
Direct Analysis ◽  
Z Score ◽  
Shellfish Poisoning ◽  
High Throughput Analysis ◽  
Fast Analysis ◽  
Rapid Resolution ◽  
Crude Extracts ◽  
Amnesic Shellfish Poisoning

Abstract The application of ultra-performance rapid resolution LC on a 1.8 m particle-size column coupled with tandem MS (RRLC-MS/MS) is described for the analysis of amnesic shellfish poisoning (ASP) toxins in shellfish. Complete resolution among domoic acid (DA) and the isomers was achieved in less than 3 min. The method was intralaboratory validated for direct analysis of crude extracts without further cleanup. It showed LODs ranging from 0.05 to 0.09 mg/kg and a working range that complied with the current regulatory level for DA of 20 mg/kg, and with the level of 4.5 mg/kg recently proposed by the European Food Safety Authority. Confirmatory capabilities were demonstrated according to the Commission Decision 2002/657/EC criteria. The results obtained by RRLC-MS/MS agreed with those provided by the reference LC-UV method, both intralaboratory for the analysis of blind samples (R2 0.9751) and interlaboratory through participation in the proficiency test for ASP toxins during 2009 (z-score 0.962 and 0.177 for low- and high-contaminated samples, respectively). RRLC-MS/MS provided fast analysis and additional confirmatory capabilities for direct analysis of crude extracts while the performance and reliability of the results were maintained, even in very complex matrixes.

scholarly journals Determination of Domoic Acid in Japanese Mussels by Enzyme Immunoassay

2000 ◽  
Vol 83 (6) ◽  
pp. 1384-1386 ◽  
Author(s):  
Kentaro Kawatsu ◽  
Yonekazu Hamano ◽  
Tamao Noguchi
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Domoic Acid ◽  
Immunoaffinity Chromatography ◽  
Shellfish Poisoning ◽  
Mussel Tissue ◽  
Blue Mussels ◽  
Amnesic Shellfish Poisoning ◽  
Low Concentrations

Abstract Ten samples of commercial blue mussels (Mytilus edulis) from Japan were analyzed for domoic acid by an indirect competitive enzyme immunoassay (idc–EIA) based on an anti-domoic acid monoclonal antibody. Domoic acid was found in all samples at low concentrations (0.11–1.81 ng/g mussel tissue). The presence of domoic acid was confirmed by liquid chromatography coupled with immunoaffinity chromatography using an anti-domoic acid monoclonal antibody as ligand. To our knowledge, this is the first reported detection of domoic acid, a causative agent of amnesic shellfish poisoning, in Japanese mussels.

scholarly journals Unbiased high-throughput characterization of mussel transcriptomic responses to sublethal concentrations of the biotoxin okadaic acid

PeerJ ◽  
2015 ◽  
Vol 3 ◽  
pp. e1429 ◽  
Author(s):  
Victoria Suarez-Ulloa ◽  
Juan Fernandez-Tajes ◽  
Vanessa Aguiar-Pulido ◽  
M. Veronica Prego-Faraldo ◽  
Fernanda Florez-Barros ◽  
...  
Keyword(s):  
Digestive Gland ◽  
Okadaic Acid ◽  
Algal Blooms ◽  
Molecular Transport ◽  
Shellfish Poisoning ◽  
Transcriptomic Response ◽  
Custom Made ◽  
Transcriptomic Responses ◽  
Sublethal Concentrations

Background.Harmful Algal Blooms (HABs) responsible for Diarrhetic Shellfish Poisoning (DSP) represent a major threat for human consumers of shellfish. The biotoxin Okadaic Acid (OA), a well-known phosphatase inhibitor and tumor promoter, is the primary cause of acute DSP intoxications. Although several studies have described the molecular effects of high OA concentrations on sentinel organisms (e.g., bivalve molluscs), the effect of prolonged exposures to low (sublethal) OA concentrations is still unknown. In order to fill this gap, this work combines Next-Generation sequencing and custom-made microarray technologies to develop an unbiased characterization of the transcriptomic response of mussels during early stages of a DSP bloom.Methods.Mussel specimens were exposed to a HAB episode simulating an early stage DSP bloom (200 cells/L of the dinoflagellateProrocentrum limafor 24 h). The unbiased characterization of the transcriptomic responses triggered by OA was carried out using two complementary methods of cDNA library preparation: normalized and Suppression Subtractive Hybridization (SSH). Libraries were sequenced and read datasets were mapped to Gene Ontology and KEGG databases. A custom-made oligonucleotide microarray was developed based on these data, completing the expression analysis of digestive gland and gill tissues.Results.Our findings show that exposure to sublethal concentrations of OA is enough to induce gene expression modifications in the musselMytilus. Transcriptomic analyses revealed an increase in proteasomal activity, molecular transport, cell cycle regulation, energy production and immune activity in mussels. Oppositely, a number of transcripts hypothesized to be responsive to OA (notably the Serine/Threonine phosphatases PP1 and PP2A) failed to show substantial modifications. Both digestive gland and gill tissues responded similarly to OA, although expression modifications were more dramatic in the former, supporting the choice of this tissue for future biomonitoring studies.Discussion.Exposure to OA concentrations within legal limits for safe consumption of shellfish is enough to disrupt important cellular processes in mussels, eliciting sharp transcriptional changes as a result. By combining the study of cDNA libraries and a custom-made OA-specific microarray, our work provides a comprehensive characterization of the OA-specific transcriptome, improving the accuracy of the analysis of expresion profiles compared to single-replicated RNA-seq methods. The combination of our data with related studies helps understanding the molecular mechanisms underlying molecular responses to DSP episodes in marine organisms, providing useful information to develop a new generation of tools for the monitoring of OA pollution.

scholarly journals Aequipecten opercularis) Digestive Gland after Exposure to Domoic Acid-Producing Pseudo-nitzschia

Toxins ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 97 ◽  
Author(s):  
Pablo Ventoso ◽  
Antonio J. Pazos ◽  
M. Luz Pérez-Parallé ◽  
Juan Blanco ◽  
Juan C. Triviño ◽  
...  
Keyword(s):  
Digestive Gland ◽  
Ribosomal Proteins ◽  
Domoic Acid ◽  
De Novo ◽  
Transcriptional Response ◽  
Major Facilitator Superfamily ◽  
Functional Enrichment ◽  
Shellfish Poisoning ◽  
Aequipecten Opercularis ◽  
Glutathione S Transferases

Some species of the genus Pseudo-nitzschia produce the toxin domoic acid, which causes amnesic shellfish poisoning (ASP). Given that bivalve mollusks are filter feeders, they can accumulate these toxins in their tissues. To elucidate the transcriptional response of the queen scallop Aequipecten opercularis after exposure to domoic acid-producing Pseudo-nitzschia, the digestive gland transcriptome was de novo assembled using an Illumina HiSeq 2000 platform. Then, a differential gene expression analysis was performed. After the assembly, 142,137 unigenes were obtained, and a total of 10,144 genes were differentially expressed in the groups exposed to the toxin. Functional enrichment analysis found that 374 Pfam (protein families database) domains were significantly enriched. The C1q domain, the C-type lectin, the major facilitator superfamily, the immunoglobulin domain, and the cytochrome P450 were among the most enriched Pfam domains. Protein network analysis showed a small number of highly connected nodes involved in specific functions: proteasome components, mitochondrial ribosomal proteins, protein translocases of mitochondrial membranes, cytochromes P450, and glutathione S-transferases. The results suggest that exposure to domoic acid-producing organisms causes oxidative stress and mitochondrial dysfunction. The transcriptional response counteracts these effects with the up-regulation of genes coding for some mitochondrial proteins, proteasome components, and antioxidant enzymes (glutathione S-transferases, thioredoxins, glutaredoxins, and copper/zinc superoxide dismutases).

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