Catenatum Blooms Research Articles

Resting Cysts of the Toxic Dinoflagellate Gymnodinium catenatum (Dinophyceae) Ubiquitously Distribute along the Entire Coast of China, with Higher Abundance in Bloom-Prone Areas

Blooms of Gymnodinium catenatum have occurred occasionally in different areas of China and caused tremendous economic loss and even threatened human health. Not only is G. catenatum an important harmful-algal-bloom (HAB)-causing species, but also the only gymnodinioid dinoflagellate known to produce paralytic shellfish poisoning toxins (PSTs). Due to the germination of resting cysts, which often initiates blooms, the distribution and abundance of cysts in sediments and particularly the confirmation of cyst beds are important information for understanding and predicting dinoflagellate blooms. In this research, 199 sediment samples were collected from China’s coastal seas, ranging from the Beidaihe in the Bohai Sea (BS) to the southernmost sample from the Nansha Islands of the South China Sea (SCS). TaqMan quantitative PCR (qPCR) assays with species-specific primers and probes were developed to specifically detect the distribution and abundance of cysts in the 199 samples. The detection revealed that G. catenatum cysts were widely present in the sediments (126 of the 199 samples), with 93.55%, 74.65%, 42.37%, and 50% of the samples detected positively from the BS, YS, ECS and SCS, respectively, and covering the vast sea area from Nansha Islands to the Beidaihe area. The single-cyst morpho-molecular identification in the samples from Beidaihe confirmed the existence of G. catenatum cysts in the BS, and the positive detections of G. catenatum cysts using the qPCR methods. While G. catenatum cysts were widely distributed in all four seas of China, the average abundance was relatively low (1.0 cyst per gram of wet sediment). Three samples from the East China Sea (ECS), however, contained G. catenatum cysts at a relatively higher level (23 cysts g−1 wet sediment) than other sea areas, suggesting a pertinence of cyst abundance to the frequent occurrences of G. catenatum blooms in the area during recent years. Collectively, for G. catenatum being such an important toxic and HAB-causing species globally, the ubiquitous distribution of its cysts along the coastal waters of China and higher abundance in the bloom-prone areas warns us of a risk that cyst beds, although currently low in abundance, may seed HABs in any and many sea areas of China at any forthcoming year, and particularly those areas with records of frequent HABs outbreaks in the past.

Two decades of data on the neurotoxic Gymnodinium catenatum dynamics and paralytic shellfish toxins contamination of molluscs in the southwestern Mediterranean: What have we learned?

Harmful algal blooms have been documented in the Moroccan Western Mediterranean region since 1993, primarily associated with the presence of paralytic shellfish toxins (PSTs) produced by the dinoflagellate Gymnodinium catenatum. The proliferation of this neurotoxic species has led to recurring bans on the harvesting of molluscs, resulting in significant socio-economic repercussions and threats to human health. In the present study, we examine the dynamics of G. catenatum and mollusc PST contamination patterns over a 20-year period (2002−2021) in two distinct marine ecosystems: M'diq Bay and the Oued Laou Estuary. For the PST contamination, we considered two commercially important shellfish species: the smooth clam, Callista Chione, and the cockle, Acanthocardia tuberculata. The highest G. catenatum abundances were consistently observed from November to February in both sites. Our data revealed inter-annual variations in G. catenatum abundance, peaking at 91,840 cells.L−1 in November 2011. PST contamination levels in A. tuberculata were significantly higher than those observed in C. chione. Furthermore, we identified a significant correlation (Pearson, P-value <0.05) between PST contamination of smooth clams and the abundance of G. catenatum. The contamination of A. tuberculata by PSTs reached very high levels, with up to 13,500 μg STX di-HCl eq. kg−1 of shellfish meat, exceeding the established safety thresholds by 16-fold. Additionally, there has been an increase in the prevalence and incidence of PSTs over the years. Notably, we observed a substantial increase in G. catenatum blooms and PST events in the Western Mediterranean during the last decade (2010−2021). The examined data suggest that rainfall could play a pivotal role in G. catenatum bloom dynamics by enriching marine waters with nutrients. The statistical model selection approaches indicated that nutrient concentrations (i.e., nitrate and phosphorus) were the most significant parameters for G. catenatum blooms in the studied area.

Expansion risk of the toxic dinoflagellate Gymnodinium catenatum blooms in Chinese waters under climate change

The paralytic shellfish poison toxin (PST)-producing dinoflagellate, Gymnodinium catenatum, frequently blooms in China, posing a threat to food safety and human health. To understand the drivers of G. catenatum blooms and predict potential habitats for G. catenatum under climate change, samples from occurrence localities and environmental datasets from multiple agencies were aggregated and used to model the habitat suitability of G. catenatum in the China Sea using a maximum entropy model (Maxent). The accumulated variable contributions for the Maxent model were defined to measure the importance of key predictors in the model. The most important environmental variables were distance to the coastline, depth of seawater, and long-term average of the minimum annual temperature. This highlights the main reasons why G. catenatum blooms always occur in coastal waters. Occurrence probabilities higher than 0.66 were defined as habitats with high suitability for shellfish management and aquaculture. Projected habitats with high suitability in Haizhou Bay, coastal waters along the western Taiwan Strait, and Bohai Bay remained stable with increasing temperature by 2100, regardless of the IPCC Representative Concentration Pathways (RCPs). However, those in the China Sea would be reduced overall, leading to a northward movement of the center of integrated habitats. Habitats with a spatial area of >6000 km2 in the Bohai Sea, Yellow Sea, and South China Sea and >23,000 km2 in the East China Sea would be exposed to high risk under low greenhouse gas emission scenarios (RCP2.6).

Gymnodinium catenatum Paralytic Shellfish Toxin Production and Photobiological Responses under Marine Heat Waves

Marine heatwaves (MHWs) have doubled in frequency since the 1980s and are projected to be exacerbated during this century. MHWs have been shown to trigger harmful algal blooms (HABs), with severe consequences to marine life and human populations. Within this context, this study aims to understand, for the first time, how MHWs impact key biological and toxicological parameters of the paralytic shellfish toxin (PST) producer Gymnodinium catenatum, a dinoflagellate inhabiting temperate and tropical coastal waters. Two MHW were simulated-category I (i.e., peak: 19.9 °C) and category IV (i.e., peak: 24.1 °C)-relative to the estimated baseline in the western coast of Portugal (18.5 °C). No significant changes in abundance, size, and photosynthetic efficiency were observed among treatments. On the other hand, chain-formation was significantly reduced under category IV MHW, as was PSP toxicity and production of some PST compounds. Overall, this suggests that G. catenatum may have a high tolerance to MHWs. Nevertheless, some sublethal effects may have occurred since chain-formation was affected, suggesting that these growth conditions may be sub-optimal for this population. Our study suggests that the increase in frequency, intensity, and duration of MHWs may lead to reduced severity of G. catenatum blooms.

Twenty-Five Years of PSP Toxicity in Galician (NW Spain) Bivalves: Spatial, Temporal, and Interspecific Variations.

Twenty-five years of paralytic shellfish poisoning (PSP) toxicity in Galician bivalves have been studied. PSP was detected in 4785 out of 73,740 samples of the commercially important bivalve species analyzed from 1995 to 2020. Its general prevalence in the area was 6.5%. Only 1.6% of all samples tested were over the regulatory limit (incidence). The maximum level of PSP in the area, 40,800 µg STX 2HCl-eq kg-1, was recorded in raft mussels from Bueu (PON-II, Pontevedra) in December 2005. The highest maximum PSP values were found in mussels, which were mostly affected by Gymnodinium catenatum, but not those of prevalence and incidence which were recorded in clams, mostly affected by Alexandrium. Average levels in mussels were higher than in any other studied species. Spatially, in general, the prevalence, incidence, maximum, and average PSP toxicity during episodes tend to decrease from south to northeast, but some hot points with high levels can be identified. PCA analysis separates the southern rías, associated to G. catenatum blooms, from the middle and northern ones, associated to Alexandrium blooms. Along the year, two main peaks of the four variables are observed, the first one in late autumn-winter and the other in summer, the summer peak being much more important for the infaunal species than for raft mussels. In the seasonal pattern obtained by time series analysis of the average PSP toxicity, the autumn-winter peak was only maintained (and very reduced) in the southern rías, indicating that this peak is seasonally much less important than the summer peak. The observed seasonality is expected based on the timing of the blooms of the two PSP-producing phytoplankton groups present in the area. Over the 25 years of monitoring, large differences in PSP toxicity have been observed. Apart from some special years, an ascending trend in prevalence and incidence seems to be present from 2011 to 2020. No trend seems to exist during the same period for average or maximum toxicity.

Wind-driven development and transport of Gymnodinium catenatum blooms along the coast of Fujian, China

Gymnodinium catenatum is a cosmopolitan, bloom-forming dinoflagellate known to produce a suite of potent paralytic shellfish poisoning (PSP) toxins. Here, we revisit two major blooms of G. catenatum along the Fujianese Coast, China, in 2017 and 2018. The impact area of the 2017 bloom was larger than that of the 2018 event. Field sampling and remote satellite sensing revealed that alongshore transport driven by the southwest wind, as well as physical accumulation driven by the northeast wind, played important roles in the development and distribution of the two bloom events. The relationship between wind-induced hydrodynamic conditions and the unprecedented HAB events established in this study adds greatly to our understanding of algal bloom dynamics along the Fujianese coast. These results improve our ability to detect, track, and forecast G. catenatum blooms, thereby potentially minimizing the negative impacts of future HAB events.

Bacterioplankton assembly and interspecies interactions follow trajectories of Gymnodinium−diatom bloom

The underlying mechanisms of bacterioplankton community assembly and interspecies interactions during harmful algal blooms remain largely unclear. Using 16S rRNA gene amplicon sequencing, we analyzed the bacterioplankton communities over the continuous course of saxitoxin-producing Gymnodinium catenatum blooms and two diatom (i.e., Skeletonema costatum and Chaetoceros curvisetus) blooms in an anthropogenically controlled and eutrophic bay, East China Sea. The succession of bacterioplankton communities correlated with changes in the dynamics of algal species. Deterministic versus stochastic bacterioplankton community assemblage processes were quantified, demonstrating that stochastic processes increased when algal blooms happened. The occurrence of algal blooms caused weaker bacterioplankton interspecies interactions and higher degrees of cooperative activities, changed keystone taxa and diminished the stability of bacterial communities. These findings consequently have important implications for our understanding of bacterioplankton community ecology during algal blooms.

A Carbamoylase-Based Bioassay for the Detection of Paralytic Shellfish Poisoning Toxins.

Out of control proliferation of toxic phytoplankton, called harmful algal blooms (HABs), have a significant economic impact on bivalve aquaculture and harvesting in coastal waters. Some phytotoxins, such as paralytic shellfish toxins (PSTs), are of concern due to the life-threatening symptoms they can cause. Development of rapid and low-cost screening tools would be a welcome addition to the laboratory methodologies employed in routine monitoring programs. However, most of the assays and biosensors for the screening of PSTs, are restricted to a single target, saxitoxin (STX), which is the most potent PST. The present study aimed at developing an assay for the detection of N-sulfocarbamoyl PST—GTX5, which is one of the most abundant toxins in bivalves during G. catenatum blooms as found on the Portuguese coast. Enzymatic assay employing PSTs’ transforming enzyme—carbamoylase—was proposed. Carbamoylase was extracted and purified from the surf clam S. solida. Carbamoylase displayed similar specificity to both carbamate (STX) and N-sulfocarbamate toxins (GTX5 and C1+2) converting them into decarbamoyl saxitoxin (dcSTX) and decarbamoyl gonyautoxins 2+3 (dcGTX2+3), respectively. The enzymatic assay involved hydrolysis of GTX5 by carbamoylase and quantification of the product of enzymatic reaction, dcSTX, using a potentiometric chemical sensor. A potentiometric sensor with plasticized PVC membrane that displayed sensitivity to dcSTX and selectivity in the presence of GTX5 was employed. Enzymatic assay allowed determination of GTX5 in the concentration range from 0.43 to 3.30 µmolL−1, which encompasses levels of GTX5 in contaminated bivalve extracts with toxicities above PSTs regulatory limits. The feasibility of the carbamoylase-based potentiometric assay for detection of GTX5 was demonstrated.

Succession of bacterioplankton communities over complete Gymnodinium-diatom bloom cycles

Shifts in bacterioplankton communities during algal blooms have been widely investigated, but our understanding of their succession over the continuous course of paralytic shellfish poisoning producing Gymnodinium catenatum blooms and diatom (Skeletonema costatum and Chaetoceros curvisetus) blooms in natural bays is highly understudied. Here, we used high-throughput sequencing of bacterioplankton 16S rRNA genes to investigate the composition and successional patterns of bacterioplankton communities during Gymnodinium-diatom bloom cycles. Changes in community compositional patterns were then evaluated in context of environmental and phytoplankton community variation. Bacterioplankton α-diversity significantly decreased during the emergence of the algal blooms, with Flavobacteriaceae, Rhodobacteraceae, Cryomorphaceae, and Saprospiraceae as the dominant bacterial families in waters during the blooms. Bacterioplankton community compositions could be separated into three successive stages according to bloom dynamics, wherein the succession of bacterioplankton communities was correlated with changes in algal species. Environmental variables, and particularly pH, salinity, and nutrient concentrations (e.g., of nitrite, nitrate, and ammonium) were strongly associated with variation in bacterioplankton community structures. Variance partitioning analysis indicated that phytoplankton effects alone could explain more variance than only environmental effects. Moreover, LEfSe analysis was used to identify special bacterioplankton genera as “biomarkers” for bloom stages, such as Tepidisphaera and Pseudarcicella, whose abundances were significantly associated with different stages of the phytoplankton blooms. The phylotype “biomarkers” that were identified hold significant potential as indicators for phytoplankton bloom successional dynamics. Overall, these results may contribute to the understanding of the ecological processes shaping microbial communities during successive Gymnodinium-diatom blooms.

Accumulation and Elimination Dynamics of the Hydroxybenzoate Saxitoxin Analogues in Mussels Mytilus galloprovincialis Exposed to the Toxic Marine Dinoflagellate Gymnodinium catenatum.

Paralytic shellfish poisoning (PSP) is a severe food-borne illness, caused by the ingestion of seafood containing paralytic shellfish toxins (PST), which are naturally produced by marine dinoflagellates and accumulate in shellfish during algae blooms. Novel PST, designated as hydroxybenzoate analogues (also known as GC toxins), was relatively recently discovered in Gymnodinium catenatum strains worldwide. However, to date, there have been no studies examining their accumulation in shellfish. In this study, mussels (Mytilus galloprovincialis) were exposed to G. catenatum for five days and then exposed to a non-toxic diet for 24 h, to investigate the toxin’s accumulation/elimination dynamics. As determined by UHPLC-HILIC-MS/MS, the hydroxybenzoate analogues, GC1 to GC6, comprised 41% of the algae toxin profile and only 9% in mussels. Elimination of GC toxins after 24 h was not evident. This study highlights that a relevant fraction of PST in mussels are not routinely analysed in monitoring programs and that there is a need to better understand the toxicological potential of the hydroxybenzoate analogues, in order to properly address the risk of G. catenatum blooms.

Toxin profile of Gymnodinium catenatum (Dinophyceae) from the Portuguese coast, as determined by liquid chromatography tandem mass spectrometry.

The marine dinoflagellate Gymnodinium catenatum has been associated with paralytic shellfish poisoning (PSP) outbreaks in Portuguese waters for many years. PSP syndrome is caused by consumption of seafood contaminated with paralytic shellfish toxins (PSTs), a suite of potent neurotoxins. Gymnodinium catenatum was frequently reported along the Portuguese coast throughout the late 1980s and early 1990s, but was absent between 1995 and 2005. Since this time, G. catenatum blooms have been recurrent, causing contamination of fishery resources along the Atlantic coast of Portugal. The aim of this study was to evaluate the toxin profile of G. catenatum isolated from the Portuguese coast before and after the 10-year hiatus to determine changes and potential impacts for the region. Hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC-MS/MS) was utilized to determine the presence of any known and emerging PSTs in sample extracts. Several PST derivatives were identified, including the N-sulfocarbamoyl analogues (C1–4), gonyautoxin 5 (GTX5), gonyautoxin 6 (GTX6), and decarbamoyl derivatives, decarbamoyl saxitoxin (dcSTX), decarbamoyl neosaxitoxin (dcNeo) and decarbamoyl gonyautoxin 3 (dcGTX3). In addition, three known hydroxy benzoate derivatives, G. catenatum toxin 1 (GC1), GC2 and GC3, were confirmed in cultured and wild strains of G. catenatum. Moreover, two presumed N-hydroxylated analogues of GC2 and GC3, designated GC5 and GC6, are reported. This work contributes to our understanding of the toxigenicity of G. catenatum in the coastal waters of Portugal and provides valuable information on emerging PST classes that may be relevant for routine monitoring programs tasked with the prevention and control of marine toxins in fish and shellfish.

Profiles of paralytic shellfish toxins in bivalves of low and elevated toxicities following exposure to Gymnodinium catenatum blooms in Portuguese estuarine and coastal waters

Profiles of paralytic shellfish toxins (PSTs) were examined in 405 composite samples of Mytilus spp., Cerastoderma edule, Donax trunculus and Spisula solida collected between 2007 and 2012 from natural production areas in two estuaries (Aveiro and Mondego), two coastal lagoons (Óbidos and Formosa), and three open coastal areas (Aguda, Comporta and Culatra). Toxin concentrations were obtained from the biotoxin monitoring programme database. Episodes of PST toxicity in Portugal have been associated with Gymnodinium catenatum blooms. Toxin profiles for each species showed no trend over the surveyed years. In general, profiles differ only slightly among areas, except for Óbidos. However, toxin profiles in bivalves varied between low and elevated toxicities, corresponding to below and above the PST regulatory limit, respectively. The ratio R1=(C1+2):B1, which were the main toxins produced by G. catenatum cells, decreased considerably between elevated and low toxicity cockles, indicating the elimination of C1+2 or conversion of compounds into B1. R2=[(dcSTX)+(dcGTX2+3)]:[(C1+2)+(B1)], which represents the ratio of minor to major toxins in G. catenatum cells, increased substantially in wedge clams (D. trunculus) of low toxicity and less markedly in cockles (C. edule) and mussels (Mytilus spp.). These differences are interpreted as the predominance of a biotransformation phase after exposure to the algal bloom. The toxin profile of surf clams (S. solida) was dominated by decarbamoyl compounds, reflecting intense biotransformation during exposure to blooms. The higher ratio R2 in low toxicity samples suggests that elimination of the produced decarbamoyl toxins was slower than biotransformation.

Modeling the occurrence of shellfish poisoning outbreaks caused by Gymnodinium catenatum (Dinophyceae) through electromagnetic signal triggering

Accumulation of paralytic shellfish poisoning toxins (PSTs) in bivalves attributed to Gymnodinium catenatum blooms at the NW Portuguese coast was previously associated with periods of low solar activity (measured by the radio flux [R]), or low geomagnetic A(a) index. It was also observed that reduction of R preceded the occurrence of toxin accumulation, while A(a) index increase could be related to its absence during periods of low activity. For modeling toxin accumulation, the monthly decrease in R was studied along the decade 2003-2012. A match that helped explaining the highly toxic years of 2007 and 2008 was obtained by plotting the formula: ΔR = (R(n-1) - R(n))/(R(n) - 65)2, where 65 represented the lowest radio activity known to date. The complex denominator was required to take into account the sunspot cycle. A 1-2 month lag was observed between maximal relative decline and maximal PSTs accumulation. PSTs in bivalves from the Portuguese south coast were related with natural electromagnetic cycles for the first time, and were not statistically associated with low R. A statistically significant association with low A(a) index also was not achieved, due to the low number of occurrences, although the 25-75 percentile was restricted to low Aa indexes in a similar way to that found for the NW coast. PSTs accumulation outside solar minima could be triggered by a steep decline in the A(a) index (ΔA), but no lag was observed in this case. While ΔR amplitude helped explaining the highly toxic years of 2007 and 2008 at the NW coast, the amplitude of ΔA was not related to the severity of the accumulation. Other kind of local electromagnetic signaling was investigated resorting to the occurrence of seismologic phenomena, because these events can trigger electric activities. No statistical association was found between seism number or magnitude and PSTs at the south coast, located near the boundary between the African and Eurasian plates, and marked by moderate seismicity.

BMAA in shellfish from two Portuguese transitional water bodies suggests the marine dinoflagellate Gymnodinium catenatum as a potential BMAA source

The neurotoxin β-N-methylamino-l-alanine (BMAA) and its putative role in multiple neurodegenerative diseases have been intensely studied since 2005 when the toxin was discovered to be produced by worldwide-distributed cyanobacterial species inhabiting terrestrial, marine, brackish, and freshwater ecosystems. Recently, BMAA production was also associated with one eukaryotic group, namely, diatoms, raising questions about its production by other phytoplanktonic groups. To test for BMAA bioavailability in ecosystems where abundant phytoplanktonic blooms regularly occur, samples of filter-feeding shellfish were collected in two Portuguese transitional water bodies. BMAA content in cockles (Cerastoderma edule) collected weekly between September and November 2009 from Ria de Aveiro and at least once a month from May to November from Ria Formosa, fluctuated from 0.079±0.055 to 0.354±0.066μg/g DW and from below the limit of detection to 0.434±0.110μg/g DW, respectively. Simultaneously to BMAA occurrence in cockles, paralytic shellfish toxins were detected in shellfish as a result of Gymnodinium catenatum blooms indicating a possible link between this marine dinoflagellate and BMAA production. Moreover, considerable high BMAA levels, 0.457±0.186μg/g DW, were then determined in a laboratory grown culture of G. catenatum. This work reveals for the first time the presence of BMAA in shellfish from Atlantic transitional water bodies and consubstantiate evidences of G. catenatum as one of the main sources of BMAA in these ecosystems.

Can solar/geomagnetic activity restrict the occurrence of some shellfish poisoning outbreaks? The example of PSP caused by Gymnodinium catenatum at the Atlantic Portuguese coast

Cyclic outbreaks of accumulation of paralytic shellfish poisoning (PSP) toxins in mussels attributed to Gymnodinium catenatum blooms displayed several of the highest inter-annual maxima coincidental with minima of the 11-year solar sunspot number (SSN) cycle. The monthly distribution of PSP was associated with low levels of the solar radio flux, a more quantitative approach than SSN for fluctuations in solar activity. A comparison between monthly distribution of PSP and other common biotoxins (okadaic acid (OA), dinophysistoxin-2 (DTX2) and amnesic shellfish poisoning (ASP) toxins) demonstrated that only PSP was significantly associated with low levels of radio flux (p < 0.01). PSP occurrence suggests a prior decline in solar activity could be required to act as a trigger, in a similar manner to a photoperiodic signal. The seasonal frequency increased towards autumn during the study period, which might be related to the progressive atmospheric cut-off of deleterious radiation associated with the seasonal change in solar declination, and might play an additional role in seasonal signal-triggering. PSP distribution was also associated with low levels of the geomagnetic index Aa. A comparison between monthly distribution of PSP and other common biotoxins, also demonstrated that only PSP was significantly associated with low levels of the Aa index (p < 0.01). In some years of SSN minima no significant PSP-outbreaks in mussels were detected. This was attributed to a steady rise in geomagnetic activity that could disrupt the triggering signal. Global distribution patterns show that hotspots for G. catenatum blooms are regions with deficient crustal magnetic anomalies. In addition to the variable magnetic field mostly of solar origin, static fields related to magnetized rocks in the crust and upper mantle might play a role in restricting worldwide geographic distribution.

A bloom-forming freshwater diatom: &lt;i&gt;Achnanthidium catenatum&lt;/i&gt;

作为区域重要饮用水源的浙江绍兴汤浦水库于2010年5月期间暴发以硅藻和蓝藻为主的藻类水华.对库尾、库中和坝前3处浮游植物样品进行采集和分析,结果表明,汤浦水库浮游植物密度在1.13×108~1.56×108cells/L之间,水华优势种之一为我国大陆地区首次报道的羽纹纲硅藻——链状弯壳藻(Achnanthidium catenatum),另一种为丝状蓝藻——湖泊假鱼腥藻(Pseudanabaena limnetica),两者的平均相对丰度分别为65.6%和28.2%;链状弯壳藻是弯壳藻属中唯一适应浮游生长的种类,具有独特的带面观,能以壳面相连形成2~3个细胞的短链状群体;对国内外3次链状弯壳藻为优势种的水华案例进行分析,发现该藻为广温性种类,适应低光强和低磷浓度生长;水华生消机理则需进一步研究.;In May 2010, algal bloom dominated by diatom and blue-green algae broke out in Tangpu Reservoir,which is the drinking water resource of Shao Xing,Zhejiang Province.Phytoplankton samples collected from 3 sites(Kuwei,Kuzhong,and Baqian) along the flow direction were investigated.The phytoplankton density ranged between 1.13×108 and 1.56×108 cells/L.The dominant species Achnanthidium catenatum(Bílý et Marvan) is a pennatae diatom,which was first reported from mainland in China.Another dominant species Pseudanabaena limnetica belongs to filamentous cyanobacteria,and their average relative abundances were 65.6% and 28.2%,respectively.Achnanthidium catenatum is the only planktonic species of the genus Achnanthidium showing distinctive girdle view and colonies consisting of a maximum of 3 frustules.Case studied of three different Achnanthidium catenatum blooms indicated that this species was eurythermal and adapted to lower light intensity and phosphorus concentration.Further study should shed more lights on the formation and disappearance mechanisms of this algal bloom.

Hydroxybenzoate paralytic shellfish toxins induce transient GST activity depletion and chromosomal damage in white seabream (Diplodus sargus)

Fish are routinely exposed to harmful algal blooms that produce noxious compounds and impact the marine food web. This study investigates the role of phase I and II detoxification enzymes on metabolism of the novel paralytic shellfish toxins (PSTs), the hydroxybenzoate analogues recently discovered in Gymnodinium catenatum strains, in the liver of white seabream, assessing ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST) activities. Additionally, the genotoxic potential of hydroxybenzoate PSTs was examined through the erythrocytic nuclear abnormality (ENA) assay. Fish were injected with hydroxybenzoate PSTs into the coelomic cavity and sacrificed 2 and 6 days later for biochemical and cytogenetic analyses. While the activity of EROD was unresponsive to toxins, a significant GST activity decrease was observed at 2 days after injection indicating an impairment of this line of the detoxification system. The genotoxic potential of PSTs was demonstrated by the induction of clastogenic/aneugenic effects at 2 days, as measured by the ENA assay. Overall, this study contributes to better understand the impact of toxins produced by G. catenatum blooms in fish, revealing effects that, even transitory, point out a risk associated to hydroxybenzoate analogues.

Biotransformation modulation and genotoxicity in white seabream upon exposure to paralytic shellfish toxins produced by Gymnodinium catenatum

Fish are recurrently exposed to paralytic shellfish toxins (PSTs) produced by Gymnodinium catenatum. Nevertheless, the knowledge regarding metabolism of PSTs and their toxic effects in fish is scarce. Consequently, the current study aims to investigate the role of phase I and II detoxification enzymes on PST metabolism in the liver of white seabream ( Diplodus sargus), assessing ethoxyresorufin- O-deethylase (EROD) and glutathione S-transferase (GST) activities. Moreover, the genotoxic potential of PSTs was examined through the erythrocytic nuclear abnormality (ENA) assay. Fish were intracoelomically (IC) injected with a nominal dose (expressed as saxitoxin equivalents) of 1.60 μg STXeq kg −1 semipurified from a G. catenatum cell culture with previously determined toxin profile. Fish were sacrificed 2 and 6 days after IC injection. PST levels determined in fish liver were15.2 and 12.2 μg STXeq kg −1, respectively, at 2 and 6 days after the injection. Though several PSTs were administered, only dcSTX was detected in the liver after 2 and 6 days. This was regarded as an evidence that most of the N-sulfocarbamoyl and decarbamoyl toxins were rapidly biotransformed in D. sargus liver and/or eliminated. This was corroborated by a hepatic GST activity induction at 2 days after injection. Hepatic EROD activity was unresponsive to PSTs, suggesting that these toxins enter phase II of biotransformation directly. The genotoxic potential of PSTs was also demonstrated; these toxins were able to induce cytogenetic damage, such as chromosome (or chromatid) breaks or loss and segregational anomalies, measured by the ENA assay. Overall, this study pointed out the ecological risk associated with the contamination of fish with PSTs generated by G. catenatum blooms, providing the necessary first data for a proper interpretation of biomonitoring programs aiming to assess the impact of phytoplankton blooms in fish.

Characterization of paralytic shellfish toxins in seawater and sardines (Sardina pilchardus) during blooms of Gymnodinium catenatum

The re-emergence of Gymnodinum catenatum blooms after a 10 year hiatus of absence initiated the present investigation. This study aims to evaluate the exposure of small pelagic fishes to paralytic shellfish toxins (PST) during blooms of G. catenatum. Sardines (Sardina pilchardus) were selected as a representative fish species. In order to assess toxin availability to fish, both intracellular PSTs (toxin retained within the algal cells) and extracellular PSTs (toxin found in seawater outside algal cells) were quantified, as well as toxin levels within three fish tissue matrices (viscera, muscle and brain). During the study period, the highest cell densities of G. catenatum reached 2.5 × 104 cells l−1 and intracellular PST levels ranged from 3.4 to 398 ng STXeq l−1 as detected via an enzyme linked immunosorbent assay (ELISA). Measurable extracellular PSTs were also detected in seawater (0.2–1.1 μg STXeq l−1) for the first time in Atlantic waters. The PST profile in G. catenatum was determined via high performance liquid chromatography with fluorescence detection (HPLC-FLD) and consisted mostly of sulfocarbamoyl (C1+2, B1) and decarbamoyl (dcSTX, dcGTX2+3, dcNEO) toxins. The observed profile was similar to that reported previously in G. catenatum blooms in this region before the 10-year hiatus. Sardines, planktivorous fish that ingest a large number of phytoplankton cells, were found to contain PSTs in the viscera, reaching a maximum of 531 μg STXeq kg−1. PSTs were not detected in corresponding muscle or brain tissues. The PST profile characterized in sardine samples consisted of the same sulfocarbamoyl and decarbamoyl toxins found in the algal prey with minor differences in relative abundance of each toxin. Overall, the data suggest that significant biotransformation of PSTs does not occur in sardines. Therefore, planktivorous fish may be a good tracer for the occurrence of offshore G. catenatum blooms and the associated PSTs produced by these algae.

Bloom dynamics and life cycle strategies of two toxic dinoflagellates in a coastal upwelling system (NW Iberian Peninsula)

A study of Gymnodinium catenatum and Alexandrium minutum blooms on the Galician coast was conducted from 2005 to 2007 in order to increase knowledge of the mechanisms governing recurrent blooms of these species. Considerable differences in their bloom dynamics were observed. G. catenatum blooms occurred in autumn and winter, following the pattern previously reported in the literature: they began off-shore and were advected to the Galician rias when a relaxation of the coastal upwelling occurred. On the other hand, A. minutum blooms developed inside embayments in spring and summer during the upwelling season and were associated with water stability and stratification. Both the vegetative population and the cyst distribution of A. minutum were related to less saline water from freshwater river outputs, which support a saline-gradient relationship postulated herein for this species. Dinoflagellates may produce both long-term double-walled cysts (resting) and short-term pellicle cysts. Resting cyst deposition and distribution in sediments showed that seeding occurred during the blooms of both species. However, the relationship between the cyst distribution in the sediments in Baiona Bay and the intensity and occurrence of G. catenatum blooms, suggests that the latter are not directly related to resting cyst germination. Moreover, the results presented in the present study point to other difference between the two species, such as the detection of pellicle cysts only for A. minutum. Finally, we discuss how the life cycle strategies of these two species may help to explain the different mechanisms of bloom formation reported herein.