Outbreaks Of Paralytic Shellfish Poisoning Research Articles

Biogeochemical conditions controlling the intensity of paralytic shellfish poisoning (PSP) outbreak caused by Alexandrium blooms: Results from 6-year field observations in Jinhae-Masan Bay, Korea

Previous field observations from 2018 to 2019 revealed that paralytic shellfish poisoning (PSP) caused by the blooms of toxic dinoflagellate Alexandrium species occurred under low concentrations of dissolved inorganic nitrogen (DIN) and high concentrations of dissolved organic nitrogen (DON) and humic-like fluorescent dissolved organic matter (FDOMH) in Jinhae-Masan Bay, Korea. In this study, we obtained more data for DIN, DON, FDOMH, and Alexandrium cell density from 2020 to 2023 to further validate environmental conditions for the PSP outbreak. We also measured total hydrolyzed amino acids (THAA) to determine the bioavailability of DON fueling the PSP outbreak. Over the 6-year observations, there was a consistent pattern of low DIN concentrations and high DON and FDOMH concentrations during the PSP outbreak periods. The Alexandrium cell densities, together with the PSP toxin concentrations, increased rapidly under this environmental condition. The PSP outbreak occurs when a large amount of DIN originating from the stream waters near the upstream sites is transformed into DON by biological production before entering the PSP outbreak area. The produced DON is characterized by high bioavailability based on the various AA-derived indices (enantiomeric ratio, degradation index, non-protein AA mole%, and nitrogen-normalized AA yield). In addition, the intensities of PSP outbreaks are mainly dependent on the conversion stage of DIN to DON and enhanced FDOMH. We found that the strong PSP outbreak occurred consistently under a low level of DIN (<1.0 μM) and high levels of DON (>9.0 μM) and FDOMH (>1.5 R.U.). Thus, our results suggest that the monitoring data of environmental conditions can be used to predict the PSP outbreak in the coastal oceans.

Development of saxitoxin biosynthesis gene sxtB-targeted qPCR assay for the quantification of toxic dinoflagellates Alexandrium catenella (group I) and A. pacificum (group IV) occurring in the Korean coast

Toxic dinoflagellate Alexandrium can produce saxitoxins (STXs) and cause paralytic shellfish poisoning (PSP), and thus they are monitored for environmental safety management. Microscopic discrimination of dinoflagellates is difficult to distinguish between toxic and non-toxic species due to their similar morphology. Meanwhile, an alternative quantitative PCR (qPCR) assay is sensitive, rapid, and cost-effective for harmful species monitoring. Herein, we developed a novel qPCR assay to detect the STXs biosynthesis gene sxtB of Alexandrium catenella and A. pacificum, the leading cause of PSP outbreaks in Asian coasts and worldwide. The newly designed sxtB TaqMan probes target the species without any positive signal in other relative dinoflagellates. Deming regression analysis revealed that the sxtB copy number of A. catenella and A. pacificum was 3.6 and 4.1 copies per cell, respectively. During the blooming periods (April 13th–14th, 2020), only A. catenella cells were detected through the qPCR assay, ranging from 5.0 × 10 to 2.5 × 104 eq cells L−1. In addition, sxtB qPCR quantified more accurately compared to large subunit (LSU) rRNA targeting qPCR assay that overestimate cell density. Besides, the sensitivity of sxtB was higher compared to the microscope when the species were rarely present (5.0 × 102 cells L−1). These suggest that the sxtB qPCR assay can be applied to toxic Alexandrium monitoring in the Korean coast, even in the early stage of bloomings.

Spatiotemporal distribution of paralytic shellfish poisoning (PSP) toxins in Beagle Channel (South America) during 2005–2017

Toxigenic algal blooms have been reported since 1886 in the Beagle Channel, at the southern limit of South America, mainly associated with the occurrence of paralytic shellfish poisoning (PSP) toxins produced by dinoflagellates of the genus Alexandrium. Toxic outbreaks have serious impacts on marine wildlife and ecosystem services, particularly on fisheries and mussel aquaculture. Within the framework of the Shellfish Toxicity Monitoring Program carried out close to Almanza in Beagle Channel eastern coastal waters, here we analyzed the spatiotemporal patterns of PSP toxins in the Magellan mussel Aulacomya ater and the blue mussel Mytilus edulis, measured by the mouse bioassay for 12 years, between 2005 and 2017. PSP outbreaks occurred mostly during summer and their duration was significantly longer than for outbreaks detected during spring and winter. A marked interannual variability in their magnitude was also evident. The largest outbreaks were detected in summers 2010–2011, in extreme warm and low-wind conditions, reaching up to 5600 µg saxitoxin equivalent per 100 g of tissue, causing a closure of shellfish fisheries of up to 200 days. Small scale spatial variability in PSP was also found, the highest values being observed in the main mussel culture areas, and the lowest in the natural mussel beds. PSP toxicities for the Magellan mussel were significantly higher than for the blue mussel. Natural detoxification in mussels from Beagle Channel followed an exponential decay, with mean detoxification rates of 3.5% toxin day−1, lower than those from elsewhere in the world for similar PSP toxin concentrations, which means a longer time to attain concentration limits safe for human consumption. Understanding PSP dynamics is important to avoid large economic losses related to halting mussel production.

Latitudinal Variation in the Toxicity and Sexual Compatibility of Alexandrium catenella Strains from Southern Chile.

The bloom-forming toxic dinoflagellate Alexandrium catenella was first detected in southern Chile (39.5–55° S) 50 years ago and is responsible for most of the area’s cases of paralytic shellfish poisoning (PSP). Given the complex life history of A. catenella, which includes benthic sexual cysts, in this study, we examined the potential link between latitude, toxicity, and sexual compatibility. Nine clones isolated from Chilean Patagonia were used in self- and out-crosses in all possible combinations (n = 45). The effect of latitude on toxicity, reproductive success indexes, and cyst production was also determined. Using the toxin profiles for all strains, consisting of C1, C2, GTX4, GTX1, GTX3, and NeoSTX, a latitudinal gradient was determined for their proportions (%) and content per cell (pg cell−1), with the more toxic strains occurring in the north (−40.6° S). Reproductive success also showed a latitudinal tendency and was lower in the north. None of the self-crosses yielded resting cysts. Rather, the production of resting cysts was highest in pairings of clones separated by distances of 1000–1650 km. Our results contribute to a better understanding of PSP outbreaks in the region and demonstrate the importance of resting cysts in fueling new toxic events. They also provide additional evidence that the introduction of strains from neighboring regions is a cause for concern.

Drivers of dinoflagellate benthic cyst assemblages in the NW Patagonian Fjords System and its adjacent oceanic shelf, with a focus on harmful species

In recent decades, the alteration of coastal food webs (via aquaculture, fishing, and leisure activities), nutrient loading, and an expansion of monitoring programs have prompted an apparent worldwide rise in Harmful Algae Blooms (HABs). Over this time, a parallel increase in HABs has also been observed in the Chilean southern austral region (Patagonia fjords). HAB species like Alexandrium catenella—responsible for Paralytic Shellfish Poisoning (PSP)—are of great public concern due to their negative socioeconomic impacts and significant northward geographical range expansion. Many toxic dinoflagellate species (like A. catenella) produce benthic resting cysts, yet a holistic understanding of the physical-chemical and biological conditions influencing the distributions of cysts in this region is lacking. In this study, we measured a combination of hydrographic (temperature, salinity, and dissolved oxygen) and sediment physical-chemical properties (temperature, pH and redox potential), in addition to meiofaunal abundances –as sediment bioturbators and potential cyst predators– to determine the factors influencing dinoflagellate cyst distribution, with emphasis on A. catenella in and around a “hotspot” area of southern Chile. An analysis of similarities (ANOSIM) test revealed significant differences (p < 0.011) in cyst assemblages between the fjords and oceanic environments. Permutational Analysis of Variance (PERMANOVA) showed significant effects of sediment temperature and silt proportion in explaining differences in the cyst assemblages. A generalized linear model (GLM) indicated that sediment temperature, silt/sand, anoxic conditions, and low abundances of Harpacticoida —a meiofauna herbivore group and potential bioturbator— are associated with the higher resting cyst abundances of the harmful species A. catenella. The implications for A. catenella resting cysts dynamics are discussed, highlighting physical-chemical and biological interactions and their potential for PSP outbreak initiation.

Conditions of nutrients and dissolved organic matter for the outbreaks of Paralytic Shellfish Poisoning (PSP) in Jinhae Bay, Korea

We measured the concentrations of nutrients, fluorescent dissolved organic matter (FDOM), and photosynthetic pigments in seawater during the springs of 2018 and 2019 in Jinhae Bay, Korea. The samplings were carried out during the severe and weak outbreaks of paralytic shellfish poisoning (PSP) in April 2018 and March 2019, respectively. The additional sampling campaigns were carried out before and after the PSP outbreak for the comparison. During the severe PSP outbreak, lower salinities, higher organic and total nutrients, and higher humic-like FDOM were observed. Although the environmental condition of April 2018 is favorable for the growth of dinoflagellates, the lowest peridinin (dinoflagellate index) and highest fucoxanthin (diatom index) concentrations were observed amongst all sampling periods. Thus, our results suggest that PSP could be more effectively produced by dinoflagellates in the course of the ecological shift by interspecific competition under the environmental condition favorable for dinoflagellates.

Spatiotemporal distribution of paralytic shellfish poisoning (PSP) toxins in shellfish from Argentine Patagonian coast

Harmful algal blooms (HABs) have been recorded in the Chubut Province, Argentina, since 1980, mainly associated with the occurrence of paralytic shellfish poisoning (PSP) toxins produced by dinoflagellates of the genus Alexandrium. PSP events in this area impact on fisheries management and are also responsible for severe human intoxications by contaminated shellfish. Within the framework of a HAB monitoring program carried out at several coastal sites along the Chubut Province, we analyzed spatiotemporal patterns of PSP toxicity in shellfish during 2000–2011. The highest frequency of mouse bioassays exceeding the regulatory limit for human consumption was detected in spring and summer, with average values of up to ≈70% and 50%, respectively. By contrast, a lower percentage of positive bioassays (2–8%) or no toxicity at all was usually detected during autumn and winter. The most intense PSP events were usually observed between November and January, with values of up to 4,000 μg STX eq 100 g−1, and showed a marked interannual variability both in their magnitude and location. In addition, a severe PSP outbreak was recorded during autumn, 2009, at Camarones Bay, with toxicity values of up to 14,000 μg STX eq 100 g−1. The scallop Aequipecten tehuelchus showed significantly higher toxicity values compared to other shellfish species in SJG and SMG, suggesting a lower detoxification capacity. Our results contribute to the understanding of HABs dynamics on the Argentine Patagonian coast.

Pyrodinium bahamense One the Most Significant Harmful Dinoflagellate in Mexico

Pyrodinium bahamense produces saxitoxins and can cause paralytic shellfish poisoning (PSP). This species has caused more human illnesses and fatalities than any other toxic dinoflagellate in Mexico. The distribution of dinoflagellate cysts with their vegetative stage is wide, mainly along Mexican Pacific coasts, from the central Gulf of California to Chiapas; and, in the southern Gulf of Mexico, and the Mexican Caribbean Sea on the Atlantic coast. Cysts, from southern, Gulf of California exhibit thermophilic (20°C to 35°C) and euryhaline characteristics. Blooms occurred typically during summer rainy season, inside of restricted shallow lagoons surrounded by mangrove forests. The local strain toxicity has only been corroborated in one isolate from the southern Gulf of California, which exhibited a high saxitoxin concentration of 95 pg STX eq cell-1. After dinoflagellate blooms, the PSP outbreaks linked with P. bahamense in the Gulf of Tehuantepec, caused at least ~267 human intoxications, and ~21fatalities from 1989 to 2010. This mini-review ends with a viewpoint of management and research strategies to better understand the factors that play essential roles in the bloom dynamics and toxicity of this species.

Biogeography of resistance to paralytic shellfish toxins in softshell clam, Mya arenaria (L.), populations along the Atlantic coast of North America

Blooms of Alexandrium spp., the causative agent of paralytic shellfish poisoning (PSP), recur with varying frequency and intensity on the Northwest Atlantic coast of North America, from New York, USA, to northern Canadian waters. Along this latitudinal range blooms co-occur with abundant, intertidal populations of softshell clams, Mya arenaria. Prior work identified a naturally-occurring genetic mutation in Domain II α-subunit of the clams’ voltage-gated sodium channels (NaV), which significantly reduces the binding affinity of the paralytic shellfish toxin, saxitoxin (STX). This mutation provides clams with resistance to the deleterious effects of STX, allowing them to continue feeding during Alexandrium spp. blooms and attain very high tissue toxicities. This study used genetic sequencing of the NaV mutation locus in clams from four coastal regions of the Bay of Fundy-Gulf of Maine and the mid-Atlantic to determine the percentage of clams in each region that possess the resistant NaV mutation. The genotype composition was related to the occurrence and magnitude of PSP outbreaks based on shellfish toxicity, primarily that of mussels, Mytilus edulis, used as a proxy for the prevalence and severity of Alexandrium blooms in each region. As hypothesized, the proportion of clams bearing the resistant mutation generally matched up well with the historical incidence and intensity of Alexandrium spp. blooms. The highest percentage of homozygote resistant clams (RR = 70.0%), and the lowest percentage of sensitive clams (SS = 4.5%) were found in eastern Gulf of Maine populations. Exceptions at a few sites where anomalously high numbers of M. arenaria with the resistant mutation were found despite the absence of blooms, may be attributable to larval gene flow. There was no evidence that Alexandrium blooms occurring in Northport Harbor, Long Island, have resulted in a shift in genotypic composition of the local clam population, presumably due to their low cell toxicity. Seasonal mismatch of highly vulnerable M. arenaria postset with toxic blooms at this latitude may also partly explain this result. This study provides strong supporting evidence that Alexandrium blooms can select for resistance to PSP-toxins in M. arenaria populations and proposes a mechanism for the persistence of the sensitive allele throughout the region. Implications for clam aquaculture (seeding) efforts, as well as for shellfish toxicity monitoring are discussed.

Which species, Alexandrium catenella (Group I) or A. pacificum (Group IV), is really responsible for past paralytic shellfish poisoning outbreaks in Jinhae-Masan Bay, Korea?

Paralytic shellfish poisoning (PSP) caused the deaths of four people in coastal area of Korea, mainly Jinhae-Masan Bay and adjacent areas, in April 1986 and in 1996. The PSP outbreaks were caused by the consumption of mussels, Mytilus edulis. The organism that caused PSP was identified, from morphological data only, as Alexandrium tamarense which is recently renamed as A. catenella, however recent studies have shown that the morphological diagnostic characteristics used to identify Alexandrium species have uncertainties and molecular tools and other criteria should be considered as well. The organism that caused past PSP outbreaks and incidents in Korea therefore need to be carefully reconsidered. The aim of this study was to re-evaluate the species really responsible for past outbreaks of PSP in Jinhae-Masan Bay, Korea. The temporal production and fluxes of the resting cysts of Alexandrium species were investigated for one year (from March 2011 to February 2012) using a sediment trap, and the morphology and phylogeny of vegetative cells germinated from the resting cysts were analysed. The production of Alexandrium species peaked in August and November, when temporal discrepancies were found in the water temperature (22.4 and 22.7°C in August, 19.1 and 19.6°C in November) and salinity (29.5 and 26.1 psu in August, 30.5 and 31.8 psu in November). The morphological data revealed that Alexandrium species germinated from resting cysts collected in August have a ventral pore on the 1′ plate, whereas the 1′ plate in Alexandrium species germinated from resting cysts collected in November lacks a ventral pore. Molecular phylogenetic data for the vegetative cells from the germination experiments allowed the August and November peaks to be assigned to Alexandrium catenella (Group I) and A. pacificum (Group IV), respectively. This indicates that the production of resting cysts of A. catenella can be enhanced by relatively high water temperature. This result is not consistent with those of previous studies that A. catenella responsible for PSP outbreaks was found at relatively low water temperature. In addition, large subunit ribosomal sequences data revealed that A. pacificum isolates from Korea were closely related to those from Australia, Japan and New Zealand where the PSP toxicity of shellfish and blooms occurred in the 1990s, indicating that the introduction of toxic dinoflagellates were related to ballast water from bulk-cargo shipping. Based on these results, we concluded that past PSP outbreaks in Jinhae-Masan Bay of Korea could have been caused by A. pacificum rather than by A. catenella.

Morphological identification of Alexandrium species (Dinophyceae) from Jinhae-Masan Bay, Korea

Outbreaks of paralytic shellfish poisoning (PSP) and dense blooms caused by Alexandrium species in Jinhae-Masan Bay, Korea have been nearly annual events for many years. However, excluding some Alexandrium species responsible for PSP, there are no critical reports on the morphology of Alexandrium species in this bay. To identify the Alexandrium species based on detailed morphological features, vegetative cells collected water samples and established by the incubation of resting cysts isolated from sediment trap samples were analyzed. Four species of Alexandrium were identified: Alexandrium affine, A. fundyense, A. catenella, and A. insuetum. Morphological features of these species were basically consistent with those outlined in previous studies. However, the ventral pore and the connecting pore on the sulcal plate, which have been accepted as diagnostic characteristics for the identification of A. fundyense and A. catenella, need to be reevaluated, indicating that useful morphological features for identifying these two species should be recommended to avoid confusion in the classification of species in genus Alexandrium.

Lethal paralytic shellfish poisoning from consumption of green mussel broth, Western Samar, Philippines, August 2013.

In July 2013, the Philippines' Event-Based Surveillance & Response Unit received a paralytic shellfish poisoning (PSP) report from Tarangnan, Western Samar. A team from the Department of Health conducted an outbreak investigation to identify the implicated source and risk factors in coastal villages known for green mussel production and exportation. A case was defined as a previously well individual from Tarangan, Western Samar who developed gastrointestinal symptoms and any motor and/or sensory symptoms after consumption of shellfish from 29 June to 4 July 2013 in the absence of any known cause. The team reviewed medical records, conducted active case finding and a case-control study. Relatives of cases who died were interviewed. Sera and urine specimens, green mussel and seawater samples were tested for saxitoxin levels using high performance liquid chromatography. Thirty-one cases and two deaths were identified. Consumption of > 1 cup of green mussel broth was associated with being a case. Seawater sample was positive for Pyrodinium bahamense var. compressum and green mussel samples were positive for saxitoxin. Inspection revealed villagers practice open defecation and improper garbage disposal. This PSP outbreak was caused by the consumption of the green mussel broth contaminated by saxitoxin. As a result of this outbreak, dinoflagellate and saxitoxin surveillance was established, and since the outbreak, there have been no harmful algal blooms event or PSP case reported since. A "Save Cambatutay Bay" movement, focusing on proper waste disposal practice and clean-up drives has been mobilized.

Paralytic Shellfish Toxins in the Marine GastropodsZidona dufresneiandAdelomelon beckiifrom Argentina: Toxicity and Toxin Profiles

ABSTRACT The marine waters around Argentina are periodically affected by a range of phytoplankton species that are known producers of paralytic shellfish poisoning (PSP) toxins. The toxic dinoflagellates Alexandrium tamarense and Gymnodinium catenatum have been associated with PSP outbreaks in bivalve molluscs such as mussels. These bivalves are used as a food source by a number of species of carnivorous gastropods, including 2 species of marine snails, Zidona dufresnei and Adelomelon beckii. There is, consequently, a significant risk to consumers of the snails, which can accumulate PSP toxins after predation on the bivalves. Forty-one snail samples of carnivorous snails harvested between 1986 and 2012, representing 2 species (Z. dufresnei and A. beckii), were analyzed for PSP by the reference mouse bioassay (MBA) method and an alternative liquid chromatography with fluorescence detection (LC-FLD) method based on AOAC 2005.06. Sample toxicities determined by the 2 methods correlated well, with no statisti...

Spatial distribution and viability of Alexandrium tamarense resting cysts in surface sediments from the St. Lawrence Estuary, Eastern Canada

The dinoflagellate Alexandrium tamarense Group 1 (as defined by Lilly et al., 2007) is responsible for recurrent outbreaks of paralytic shellfish poisoning (PSP) in the St. Lawrence Estuary (SLE), Eastern Canada. In August 2008, a major bloom of A. tamarense developed in the SLE and caused major mortality of fish, seabirds and marine mammals notably in the vicinity of a marine park. Eleven months later, surface (0–5 cm) and deeper (5–10 cm) sediments were sampled to determine resting cysts concentrations, locate prospective cyst seedbeds and examine if these had changed following this major bloom. This information is thought to be important to understand inter-annual patterns in algal toxicity, cyst abundance being a good predictor of subsequent bloom magnitude in some regions. Surface cyst distribution was heterogeneous and it confirmed the location of the cyst seedbed previously reported on the north shore near the Manicouagan/aux-Outardes Rivers (>500 cysts cm−3). A zone of cyst accumulation was also observed on the south shore of the SLE (maximum of 1200 cysts cm−3), with higher concentrations relative to previous cyst mapping in the 1980s. A mismatch was observed between the zones with high surface cyst concentrations and those where the highest PSP toxins were detected (used as a proxy for vegetative cells in the water column). Cyst concentrations were negatively correlated with PSP levels from the same sites, suggesting that cysts were formed and deposited away from the major sites of toxicity. Deposition likely took place near the end of the bloom, once it had reached the eastern boundary of the SLE. PSP toxicity was worse near the peak of the bloom, which occurred westward of this region. This highlights the dynamic behaviour of local blooms, influenced by the estuarine and mesoscale circulation. Interestingly, the major bloom of August 2008 was not followed by particularly large cyst deposition or by any major bloom in 2009 in this region. Cyst viability was also examined, using Sytox Green, and found to be highest (nearly 100%) in the sites where cyst concentrations were maximum. Our results call for further investigation of the cyst mapping hypothesis in the St. Lawrence, where the local circulation seems to have an overriding influence on cyst deposition patterns.

A method for predicting the occurrence of paralytic shellfish poisoning along the coast of Hokkaido in the Okhotsk Sea in summer

To design a method for predicting outbreaks of paralytic shellfish poisoning (PSP) in scallop fishing grounds, the relationship between the distribution of the toxic dinoflagellate Alexandrium tamarense and the dynamics of the Soya Warm Current (SWC) was examined in the Okhotsk Sea off Hokkaido. Surveys were conducted from May to June to clarify the transportation mechanism of A. tamarense from the oceanic area to the coastal area. The sea-level difference (SLD) between Wakkanai and Abashiri was monitored as an index of the strength of the SWC southeastward flow in an alongshore belt to examine the possible occurrence of A. tamarense in the coastal area during temporal weakening of the SWC. A bottom-mounted acoustic Doppler current profiler (ADCP) was used for direct observations of the SWC. The results indicated that PSP occurred when low-salinity water contaminated with A. tamarense extended to the coast during temporal weakening of the SWC due to a decrease of the SLD. Our results strongly indicate that predictions can be realized by monitoring the decrease of SLD as an index of temporal weakening of the SWC after surveys of the distribution of A. tamarense in the oceanic area before the period of PSP occurrence.

Cluster analysis of toxins profile pattern as a tool for tracing shellfish contaminated with PSP-toxins

Paralytic shellfish poisoning (PSP) is one of the most lethal biotoxin-induced diseases worldwide, which may pose serious public health threat and potential devastating economic damage on fisheries industry in the affected region(s). To prevent the importation of PSP contaminated shellfish to a community, detailed documentation on the supply chain and routine surveillance systems are, in principle, crucial measures to protect people from this intoxication. However, difficulties have always been encountered on the traceability of the source/origin of contaminated shellfish.In the present study, we reported the potential application of PSP-toxins profiles with similarity analysis that can be used to identify epidemiological linkage between shellfish samples collected from markets and patients during a PSP outbreak. PSP-toxins were identified and quantified by ion-pair chromatographic separation followed by post-column oxidation to fluorescent imino purine derivatives. Samples from a PSP incident and other surveillance samples collected in our past 7-year record were also compared for their similarity in PSP-toxins profiles patterns. Molar distributions (nmol%) of 10 PSP-toxins were analyzed by Unweighted Pair Group Method with Arithmetric averages (UPGMA). Three prominent clusters emerged with similarity levels reaching over 80% for each, suggesting that each group of samples probably originated from a same source/batch. The PSP-toxins profiles and toxicities determined from surveillance samples could provide premonitory clues on the occurrences of PSP incident and outbreak with corresponding toxin profiles in the later time. Due to species-specific characteristics of PSP-toxins composition and profile in shellfish under varieties of environmental and physiological conditions, PSP-toxins profile can be a specific and useful biochemical indicator for tracing PSP contaminated shellfish provided that spatio-temporal occurrence patterns of toxins profiles are available in a databank for inter-laboratory comparison and standardized methodologies such as consentaneous toxins extraction and identification criteria are used for analysis and comparison.

Bacterial diversity of Gymnodinium catenatum and its relationship to dinoflagellate toxicity

Gymnodinium catenatum Graham (Dinophyceae) is one of several marine dinoflagel- lates responsible for outbreaks of paralytic shellfish poisoning (PSP), a problem that is considered to be increasing globally. Bacteria associated with these dinoflagellates have been implicated as poten- tially involved with the production of PSP toxins, and this study sought to identify whether there was a link between the toxicity of G. catenatum laboratory cultures and the diversity of the associated bacterial community. Bacterial 16S rRNA gene clone libraries were constructed and sequenced to identify the bacterial diversity of 7 G. catenatum cultures of 2 contrasting toxicity levels. Phylogenetic membership and community structure were examined, including the use of UniFrac, FST and LIB- COMPARE. No statistically significant differences that distinguished between toxic and low-toxicity G. catenatum cultures were identified in the bacterial community membership or structure. Further- more, no coherent phylogenetic group of bacteria was observed to co-associate with culture toxicity. However, observed variation in bacterial diversity and community structure was based on the geo- graphic origin of the G. catenatum cultures. Overall, while it was not possible to identify an apparent link between bacterial diversity and the toxicity of G. catenatum cultures, we suggest, on the balance of this study and others, that bacterial influence on PSP toxin production may be indirect and medi- ated by the effects of the bacterial community on algal physiology.

From homothally to heterothally: Mating preferences and genetic variation within clones of the dinoflagellate Gymnodinium catenatum

The chain-forming dinoflagellate Gymnodinium catenatum Graham is responsible for outbreaks of paralytic shellfish poisoning (PSP), a human health threat in coastal waters. Sexuality in this species is of great importance in its bloom dynamics, and has been shown to be very complex but lacks an explanation. For this reason, we tested if unreported homothallic behavior and rapid genetic changes may clarify the sexual system of this alga. To achieve this objective, 12 clonal strains collected from the Spanish coast were analyzed for the presence of sexual reproduction. Mating affinity results, self-compatibility studies, and genetic fingerprinting (amplified fragment length polymorphism, AFLP) analysis on clonal strains, showed three facts not previously described for this species: (i) That there is a continuous mating system within G. catenatum, with either self-compatible strains (homothallic), or strains that needed to be outcrossed (heterothallic), and with a range of differences in cyst production among the crosses. (ii) There was intraclonal genetic variation, i.e. genetic variation within an asexual lineage. Moreover, the variability among homothallic clones was smaller than among the heterothallic ones. (iii) Sibling strains (the two strains established by the germination of one cyst) increased their intra- and inter-sexual compatibility with time. To summarize, we have found that G. catenatum's sexual system is much more complex than previously described, including complex homothallic/heterothallic behaviors. Additionally, high rates of genetic variability may arise in clonal strains, although explanations for the mechanisms responsible are still lacking.

New life-cycle stages of Gymnodinium catenatum (Dinophyceae): laboratory and field observations

The chain-forming dinoflagellate Gymnodinium catenatum is responsible for outbreaks of paralytic shellfish poisoning (PSP); however, the relative importance of benthic-planktonic life- cycle transitions in the appearance of blooms of this species needs to be clarified. By coupling field and laboratory experiments, the present study is the first to analyze the dynamics of vegetative cells and sexual stages during a bloom of G. catenatum. In natural samples, the sexual stages of G. cate- natum were associated with several different cellular behaviors and morphologies. This confirmed laboratory evidence for the reversibility of the species' sexual processes and for the ability of zygotes to either bypass or shorten the route to resting-cyst formation. Moreover, chains of up to 4 viable cysts with differing morphologies occurred and these have never been reported previously for this species. At least two of the cysts had reticulated surfaces, a feature related to sexual reproduction in previous studies; this observation suggests the involvement of sexual processes in mechanisms that cannot be explained by any known life cycle route depicted for this species. Morphological variability and abundance of the sexual stages during the bloom indicated the complexity of the G. catenatum sex- ual cycle and the important role of sexual reproduction in the ecological succession of this species. However, the lack of a dormancy period in the sexual resting stage (evidenced by the large number of germinated cysts in sediments sampled 3 mo after the bloom) indicated that the advection of off- shore populations shoreward, rather than 'seed beds', is the main mechanism explaining G. catena- tum bloom formation in the Galician rias.

Discrimination of Alexandrium andersoni and A. minutum (Dinophyceae) using LSU rRNA-targeted oligonucleotide probes and fluorescent whole-cell hybridization

N. Touzet and R. Raine. 2007. Discrimination of Alexandrium andersoni and A. minutum (Dinophyceae) using LSU rRNA-targeted oligonucleotide probes and fluorescent whole-cell hybridization. Phycologia 46: 168–177. DOI: 10.2216/06-11.1Toxic marine dinoflagellates from the genus Alexandrium have been responsible for paralytic shellfish poisoning (PSP) throughout the world. Their monitoring relies on spatial and temporal sampling strategies and requires the reliable identification and enumeration of vegetative stages in order to enable the development of early warning policies. The accurate discrimination between Alexandrium species is labour intensive and requires taxonomic expertise as the genus contains morphologically similar toxic and non-toxic species. In Ireland, PSP outbreaks so far have been limited to Cork Harbour, a retentive inlet located on the south coast of the country, where the causative organism has been identified as A. minutum. Recently, the non-toxic and morphologically similar species A. andersoni has been detected on the south west coast of Ireland. In routine monitoring, Alexandrium spp. are identified on the basis of morphological features by conventional light microscopy, a method which does not allow their characterization at the species level. The development and application of large subunit (LSU) rRNA-targeted oligonucleotide probes for the detection and quantification of A. minutum (Global clade) and A. andersoni by whole-cell fluorescent in situ hybridization (FISH) is reported. The specificity and sensitivity of the two probe sets (MinA and AndA′ +C) were evaluated against Alexandrium species, including A. tamarense, A. tamutum and A. ostenfeldi, and a range of common dinoflagellates usually co-occurring with Alexandrium in Irish coastal waters. No cross-reactivity was observed with any of the strains tested or with phytoplankton species present in field samples rich in dinoflagellates. The format of the assay overcame possible matrix effects, such as probe adsorption, and allowed the reliable labelling of at least 1000 cells. Furthermore, the simultaneous use of calcofluor during the assays permitted the confirmation of the probe diagnostics by examining the general plate structure of the thecae of labelled cells.