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The Paralytic shellfish neurotoxins (PST) are of increasing interest for biomedical applications. The chemical synthesis is often complex and expensive that’s why the purification by membrane filtration of PST from Alexandrium minutum dinoflagellate was investigated. Disrupted micro-alga cells by ultrasonic treatment were diafiltred to let pass toxins through an ultrafiltration membrane. Then, the mean permeate was concentrated and diafiltrated by nanofiltration. Mean permeate fluxes equal to 187, 135 and 135 L.h–1.m-2 were obtained during the first diafiltration, the concentration step and the final diafiltration respectively. Up to 57 % (mol/mol) and 78 % (mol/mol) of organic matters and salts were removed respectively. Divalent ions were sparsely removed contrary to monovalent ones. C1 and C2 toxins were successfully purified since more than 75 % (mol/mol) were recovered. However, only 27 to 50 % (mol/mol) of GTX2, GTX3 and STX were recovered.

The dinoflagellate Alexandrium minutum is a toxic bloom-forming species distributed worldwide. The mechanisms driving and promoting the species blooms and their toxicity are studied and presented here. Most previously published work focuses on local and/or short-term scales. In this study, a broad temporal and spatial approach is addressed using time series covering several sites over several years and combining environmental variables and A. minutum abundances from the French English Channel − Atlantic coasts. Data were explored by means of phenology and threshold analysis. The A. minutum bloom characteristics are defined. Only one bloom per year is measured and it may reach more than a million of cells L−1. Bloom period extends from April to October and the bloom length ranges from two weeks to six months. In the ecosystems studied, water temperature and river flow, as regional and local factors respectively, are the main environmental drivers influencing the magnitude, growth rate and length of the blooms. Bloom toxicity is linked to the bloom maximum abundance and river flow. This work provides new knowledge for further managing tools for A. minutum blooms in the ecosystems studied.

Dinoflagellates of the genus Alexandrium are a major cause of harmful algal blooms (HABs) that have increasingly disrupted coastal ecosystems for the last several decades. Microalgae from the genus Alexandrium are known to produce paralytic shellfish toxins (PST) but also bioactive extracellular compounds (BEC) that can display cytotoxic, allelopathic, ichtyotoxic or haemolytic effects upon marine organisms. The objective of this experimental study was to assess the effects of PST and BEC produced by A. minutum upon juvenile great scallops Pecten maximus. Scallops were exposed for one week to two different strains of A. minutum, the first producing both PST and BEC and the second producing only BEC. Escape response to starfish, daily shell growth, histological effects, and accumulation of PST were recorded after one week of exposure, and after two subsequent weeks of recovery. Daily shell growth was delayed three days in scallops exposed to the BEC-producing A. minutum strain, probably during the three first days of exposure. An increase of reaction time to predators was observed in scallops exposed to the BEC condition, suggesting that BEC may have altered sensing processes. Scallops exposed to PST displayed a less-efficient escape response and muscular damage which could reflect the effects of paralytic toxins upon the nervous system of scallops. This study demonstrates contrasting effects of the distinct toxic compounds produced by A. minutum upon marine bivalves, thus highlighting the importance to better characterize these extracellular, bioactive compounds to better understand responses of other marine organisms.

The dinoflagellates of Alexandrium genus are known to be producers of paralytic shellfish toxins that regularly impact the shellfish aquaculture industry and fisheries. Accurate detection of Alexandrium including A. minutum is crucial for environmental monitoring and sanitary issues. In this study, we firstly developed a quantitative lateral flow immunoassay (LFIA) using super-paramagnetic nanobeads for A. minutum whole cells. This dipstick assay relies on two distinct monoclonal antibodies used in a sandwich format and directed against surface antigens of this organism. No sample preparation is required. Either frozen or live cells can be detected and quantified. The specificity and sensitivity are assessed by using phytoplankton culture and field samples spiked with a known amount of cultured A. minutum cells. This LFIA is shown to be highly specific for A. minutum and able to detect reproducibly 105 cells/L within 30 min. The test is applied to environmental samples already characterized by light microscopy counting. No significant difference is observed between the cell densities obtained by these two methods. This handy super-paramagnetic lateral flow immnunoassay biosensor can greatly assist water quality monitoring programs as well as ecological research.

Harmful algal blooms of Alexandrium spp. dinoflagellates regularly occur in French coastal waters contaminating shellfish. Studies have demonstrated that toxic Alexandrium spp. disrupt behavioural and physiological processes in marine filter-feeders, but molecular modifications triggered by phycotoxins are less well understood. This study analyzed the mRNA levels of 7 genes encoding antioxidant/detoxifying enzymes in gills of Pacific oysters (Crassostrea gigas) exposed to a cultured, toxic strain of A. minutum, a producer of paralytic shellfish toxins (PST) or fed Tisochrysis lutea (T. lutea, formerly Isochrysis sp., clone Tahitian (T. iso)), a non-toxic control diet, in four repeated experiments. Transcript levels of sigma-class glutathione S-transferase (GST), glutathione reductase (GR) and ferritin (Fer) were significantly higher in oysters exposed to A. minutum compared to oysters fed T. lutea. The detoxification pathway based upon glutathione (GSH)-conjugation of toxic compounds (phase II) is likely activated, and catalyzed by GST. This system appeared to be activated in gills probably for the detoxification of PST and/or extra-cellular compounds, produced by A. minutum. GST, GR and Fer can also contribute to antioxidant functions to prevent cellular damage from increased reactive oxygen species (ROS) originating either from A. minutum cells directly, from oyster hemocytes during immune response, or from other gill cells as by-products of detoxification.

In this paper, a simple detection of a toxic algae, Alexandrium minutum, was developed using highly sensitive quartz crystal microbalance. In terms of performance, compared with other conventional analytical tools, the main interest of our immunosensor is based on a fast and direct detection of these living cells. This system requires the use of one monoclonal antibody directed against the surface antigen of A. minutum. We demonstrate that the whole living and motile algae are caught and detected. The high specificity of the biosensor is also demonstrated by testing several other dinoflagellate species. The frequency shift is correlated to the A. minutum cells concentration. This simple system is potentially promising for environmental monitoring purposes.

This study assessed the effects of an artificial bloom of the toxin-producing dinoflagellate, Alexandrium minutum, upon nutrition related processes and reproductive output of the Pacific oyster, Crassostrea gigas. Oysters were exposed to A. minutum, Paralytic Shellfish Toxins (PST) producer and compared to a control batch of oysters fed Isochrysis galbana clone Tahitian (T.Iso). The experiment was performed in June 2008, when oysters were found ripe. Several physiological variables of oysters, such as PSTs accumulation, digestive gland and histological observations as well as spermatozoa quality, were measured at the end of the exposure. Results indicate that the digestive gland was greatly impacted upon A. minutum exposure. Monoacylglycerol and diacylglycerol contents as well as phospholipids (mainly phosphatidylcholine) drastically decreased in the digestive gland of oysters exposed to A. minutum compared to control oysters. At the same time, many oysters exposed to the harmful microalga presented a strong inflammatory response in different tissues of the digestive gland: in intestine as well as in digestive ducts and tubules. Spermatozoa in oysters exposed to A. minutum were morphologically and functionally modified compared to spermatozoa of control oysters. Indeed, spermatozoa were less motile and had lower ATP content in oysters exposed to A. minutum. Meanwhile, spermatozoa produced by control oysters showed higher percentage of mortality and relative DNA content than those produced by A. minutum exposed oysters. Finally, the characteristics of the mitochondria of spermatozoa also appeared to be modified upon A. minutum exposure. The results of this study suggests that an exposure of oysters to A. minutum, reducing energy status and motility of spermatozoa associated to morphological changes at the cellular and sub-cellular levels, can have consequences on spermatozoa fertility and reproduction success. (C) 2010 Elsevier B.V. All rights reserved.

The effects of an artificial bloom of the toxin-producing dinoflagellate, Alexandrium minutum, upon physiological parameters of the Pacific oyster, Crassostrea gigas, were assessed. Diploid and triploid oysters were exposed to cultured A. minutum and compared to control diploid and triploid oysters fed T. Isochrysis. Experiments were repeated twice, in April and mid-May 2007, to investigate effects of maturation stage on oyster responses to A. minutum exposure. Oyster maturation stage, Paralytic Shellfish Toxin (PST) accumulation, as well as several digestive gland and hematological variables, were assessed at the ends of the exposures. In both experiments, triploid oysters accumulated more PSTs (approximately twice) than diploid oysters. Significant differences, in terms of phenoloxidase activity (PO) and reactive oxygen species (ROS) production of hemocytes, were observed between A. minutum-exposed and non-exposed oysters. PO in hemocytes was lower in oysters exposed to A. minutum than in control oysters in an early maturation stage (diploids and triploids in April experiment and triploids in May experiment), but this contrast was reversed in ripe oysters (diploids in May experiment). In the April experiment, granulocytes of oysters exposed to A. minutum produced more ROS than those of control oysters; however, in the May experiment, ROS production of granulocytes was lower in A. minutum-exposed oysters. Moreover, significant decreases in free fatty acid, monoacylglycerol, and diacylglycerol contents in digestive glands of oysters exposed to A. minutum were observed. Concurrently, the ratio of reserve lipids (triacylglycerol, ether glycerides and sterol esters) to structural lipids (sterols) decreased upon A. minutum exposure in both experiments. Also, several physiological responses to A. minutum exposure appeared to be modulated by maturation stage as well as ploidy of the oysters. (C) 2009 Elsevier B.V. All rights reserved.