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The bays of Tongoy and Guanaqueros are located in the Humboldt Current system, where Argopecten purpuratus has been the subject of intense aquaculture development. These bays lie in one of the most productive marine ecosystems on Earth and are dominated by permanent coastal upwelling at Lengua de Vaca Point and Choros Point, one of the three upwelling centers on the Chilean coast. Significantly, this productive system experiences a high recurrence of harmful algal bloom (HAB) events. This paper examines 9-year (2010-2018) samples of three toxic microalgal species collected in different monitoring programs and research projects. During this period, nine HAB events were detected in Guanaqueros Bay and 14 in Tongoy Bay. Among these, three HAB events were produced simultaneously in both bays by Pseudo-nitzschia australis, and two events produced simultaneously were detected in one bay by Alexandrium spp. and the other by Dinophysis acuminata. Before El Niño 2015-16, there were more HAB events of longer duration by the three species. Since El Niño, the number and duration of events were reduced and only produced by P. australis. HAB events were simulated with the FVCOM model and a virtual particle tracker model to evaluate the dynamics of bays and their relationship with HAB events. The results showed retention in bays during the relaxation conditions of upwelling and low connectivity between bays, which explains why almost no simultaneous events were recorded.

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To expand knowledge of Pseudo-nitzschia species in the Southeast Pacific, we isolated specimens from coastal waters of central Chile (36°S-30°S), the Gulf of Corcovado, and the oceanic Robinson Crusoe Island (700 km offshore) and grew them into monoclonal strains. A total of 123 Pseudo-nitzschia strains were identified to 11 species based on sequencing of the ITS region of the nuclear rDNA and on ultrastructural and morphometric analyses of the frustule in selected representatives of each clade: P. australis, P. bucculenta, P. cf. chiniana, P. cf. decipiens, P. fraudulenta, P. hasleana, P. multistriata, P. plurisecta, P. cf. sabit, the new species P. dampieri sp. nov., and one undescribed species. Partial 18S and 28S rDNA sequences, including the hypervariable V4 and D1-D3 regions used for barcoding, were gathered from representative strains of each species to facilitate future metabarcoding studies. Results showed different levels of genetic, and at times ultrastructural, diversity among the above-mentioned entities, suggesting morphological variants (P. bucculenta), rapidly radiating complexes with ill-defined species boundaries (P. cf. decipiens and P. cf. sabit), and the presence of new species (P. dampieri sp. nov., Pseudo-nitzschia sp. 1, and probably P. cf. chiniana). Domoic acid (DA) was detected in 18 out of 82 strains tested, including those of P. australis, P. plurisecta, and P. multistriata. Toxicity varied among species mostly corresponding to expectations from previous reports, with the prominent exception of P. fraudulenta; DA was not detected in any of its 10 strains tested. In conclusion, a high diversity of Pseudo-nitzschia exists in Chilean waters, particularly offshore.

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: Despite its small area, Araçá Bay (AB) holds cultural, historical, and economic value and displays great benthic biodiversity. Thus, it is crucial to monitor its environmental health, including knowing the main groups of phytoplankton and their temporal variability. The shallow waters of Araçá Bay are continuously modified by the complex hydrography of the adjacent São Sebastião channel (SSC), challenging standard experimental designs for phytoplankton collection. Here we report changes in phytoplankton composition at intervals of five to six weeks from September 2013 to August 2014 in both Araçá Bay and SSC. Samples were collected twice daily for three consecutive days to increase taxonomic resolution. Our goal was to provide an inventory of species occurrences to aid future public policies and environmental management of the area. Analyses revealed high species richness and 166 different phytoplankton taxa. Diatoms and dinoflagellates were always numerically dominant, but taxa occurrence changed markedly. Diatoms of the genera Pseudo-nitzschia were abundant during spring and summer concurrently to signatures of South Atlantic Central Water in the SSC, while Thalassiosira occurred when waters displayed relatively lower salinity. The inventory demonstrated several potentially harmful species of microalgae and cyanobacteria, strongly suggesting investments in monitoring programs in this area that currently experience an increase in population.

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The Beagle Channel is a remote subantarctic environment where mussel aquaculture initiatives have existed since the early 1990s. Here we analyze phytoplankton biomass and composition, and the occurrence of harmful microalgae species and their toxins at three sites during the period 2015-2016. The occurrence of potentially harmful algae was observed throughout the study period, including toxigenic dinoflagellates such as Alexandrium catenella (Group I of the A. tamarense complex), A. ostenfeldii, Dinophysis acuminata, Gonyaulax spinifera, Azadinium sp., and the diatoms Pseudo-nitzschia australis and P. fraudulenta. Toxic dinoflagellates were detected in low densities whereas a Pseudo-nitzschia bloom was observed in late February. Isolates of A. catenella and P. delicatissima sensu stricto were phylogenetically characterized. The toxin profile of A. catenella was dominated by GTX4, while P. delicatissima sensu stricto showed no production of the neurotoxin domoic acid in culture conditions. The results provide base-line information for the management of harmful algal blooms in this little explored subantarctic area.

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Domoic acid (DA) or Amnesic Shellfish Poisoning (ASP) produced by the genus Pseudo-nitzschia diatom was investigated in two seasonal periods in fishing areas of Katsuwonus pelamis in the South Atlantic Ocean. Higher DA concentrations were found in spring compared to winter. Pseudo-nitzschia spp. more quantified in winter than in spring, while P. pungens, a species among the most reported for an AD toxic potential, was only found in spring.

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A somewhat disparate, yet temporally cohesive, set of phytoplankton abundance, microphytobenthos, including the diatom Pseudo-nitzschia, benthic infauna, and sediment toxin data were used to develop a theory for the transfer of domoic acid (DA) from the toxic diatom to the benthos in the highly productive waters of the northern Gulf of Mexico near the Mississippi River plume. Archived samples and new data were used to test the theory that DA is likely to be incorporated into benthic consumers. High spring abundances of potentially toxic Pseudo-nitzschia diatoms were simultaneously present in the surface waters, bottom waters and on the seafloor. Examination of the gut contents of a typical deposit-feeding and suspension-feeding polychaete, Paraprionospio pinnata, during similar periods of high Pseudo-nitzschia abundance in surface water indicated consumption of the diatoms. Demersal fishes, particularly Atlantic croaker, are known to consume these polychaetes, with a potential for transfer of DA to even higher trophic levels. These findings warrant a theory to be tested with further studies about the trophic linkage of a phytoplankton toxin into the benthic food web.

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The gulfs that surround Península Valdés (PV), Golfo Nuevo and Golfo San José in Argentina, are important calving grounds for the southern right whale Eubalaena australis. However, high calf mortality events in recent years could be associated with phycotoxin exposure. The present study evaluated the transfer of domoic acid (DA) from Pseudo-nitzschia spp., potential producers of DA, to living and dead right whales via zooplanktonic vectors, while the whales are on their calving ground at PV. Phytoplankton and mesozooplankton (primary prey of the right whales at PV and potential grazers of Pseudo-nitzschia cells) were collected during the 2015 whale season and analyzed for species composition and abundance. DA was measured in plankton and fecal whale samples (collected during whale seasons 2013, 2014 and 2015) using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The genus Pseudo-nitzschia was present in both gulfs with abundances ranging from 4.4×102 and 4.56×105 cell l-1. Pseudo-nitzschia australis had the highest abundance with up to 4.56×105 cell l-1. DA in phytoplankton was generally low, with the exception of samples collected during a P. australis bloom. No clear correlation was found between DA in phytoplankton and mesozooplankton samples. The predominance of copepods in mesozooplankton samples indicates that they were the primary vector for the transfer of DA from Pseudo-nitzschia spp. to higher trophic levels. High levels of DA were detected in four whale fecal samples (ranging from 0.30 to 710µgg-1 dry weight of fecal sample or from 0.05 and 113.6µgg-1 wet weight assuming a mean water content of 84%). The maximum level of DA detected in fecal samples (710µg DA g-1 dry weight of fecal sample) is the highest reported in southern right whales to date. The current findings demonstrate for the first time that southern right whales, E. australis, are exposed to DA via copepods as vectors during their calving season in the gulfs of PV.

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The marine diatom genus Pseudo-nitzschia, the major known producer of the neurotoxin domoic acid (DA) responsible for the amnesic shellfish poisoning (ASP) syndrome in humans and marine mammals, is globally distributed. The genus presents high species richness in the Argentine Sea and DA has been frequently detected in the last few years in plankton and shellfish samples, but the species identity of the producers remains unclear. In the present work, the distribution and abundance of Pseudo-nitzschia species and DA were determined from samples collected on two oceanographic cruises carried out through the Argentine Sea (∼39-47°S) during summer and spring 2013. Phytoplankton composition was analysed by light and electron microscopy while DA was determined by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The genus Pseudo-nitzschia was recorded in 71 and 86% of samples collected in summer and spring, respectively, whereas DA was detected in only 42 and 21% of samples, respectively. Microscopic analyses revealed at least five potentially toxic species (P. australis, P. brasiliana, P. fraudulenta, P. pungens, P. turgidula), plus putatively non-toxigenic P. dolorosa, P. lineola, P. turgiduloides and unidentified specimens of the P. pseudodelicatissima complex. The species P. australis showed the highest correlation with DA occurrence (r=0.55; p<0.05), suggesting its importance as a major DA producer in the Argentine Sea. In the northern area and during summer, DA was associated with the presence of P. brasiliana, a species recorded for the first time in the Argentine Sea. By contrast, high concentrations of P. fraudulenta, P. pungens and P. turgidula did not correspond with DA occurrence. This study represents the first successful attempt to link toxigenicity with Pseudo-nitzschia diversity and cell abundance in field plankton populations in the south-western Atlantic.

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AbstractSince 1983, San Pedro Bay in the Philippines had been reported to be the site of episodic Pyrodinium bahamense var. compressum blooms that caused paralytic shellfish poisoning in its nearby coastal communities. This bay is also subjected to numerous storms; the strongest was super typhoon Haiyan in November 8, 2013. For the first time, the seasonal dynamics of potentially toxic and harmful phytoplankton in this bay is elucidated. This is also the first record of a bloom of the cyanobacteria, Trichodesmium erythraeum that reached 70 000 colonies/L in April 2013 in this area. There were other 19 potentially toxic and harmful phytoplankton encountered during the sampling period. These consisted of a haptophyte, Phaeocystis globosa, the diatom Pseudonitzschia and 17 dinoflagellates. Seven of these harmful algae had densities high enough to be traced through time. Normally, diatoms abound during the dry season. But Pseudo-nitzschia increased in abundance during the wet season of 2012 and 2013. The dinoflagellates and Phaeocystis globosa behaved as expected and exhibited a relative increase in cell density during the rainy season of both years too. High nutrient availability during this season must have influenced the behavior of the phytoplankton despite differences in temperature and light intensity among seasons. Other notable but rare harmful species found only in plankton net tows during the study were Pyrodinium bahamense var. compressum, Alexandrium tamiyavanichii, Cochlodiniumpolykrikoides, and Noctiluca scintillans. Rev. Biol. Trop. 64 (2): 897-911. Epub 2016 June 01.

ResumenDesde 1983, la Bahía de San Pedro en Filipinas ha sido reportada como un sitio de proliferación de Pyrodinium bahamense var. compressum que causó intoxicación paralítica en sus comunidades costeras cercanas. Esta bahía también está sometida a numerosas tormentas; entre las más fuertes se presentó un súper tifón en Haiyan, el 8 de noviembre 2013. Por primera vez, se explica la dinámica estacional del fitoplancton potencialmente tóxico y dañino en esta bahía. Este es también el primer registro en esta área de una proliferación de cianobacterias (Trichodesmium erythraeum) que alcanzó 70 000 colonias/L en abril 2013. Durante el periodo de muestreo se presentaron otras 19 proliferaciones de fitoplancton potencialmente tóxicas y dañinas. Estos consistían en una haptófita, Phaeocystis globosa, la diatomea Pseudo-nitzschia y 17 dinoflagelados. Siete de estas algas nocivas tenían densidades suficientemente altas como para ser rastreadas a través del tiempo. Normalmente, las diatomeas abundan durante la estación seca. Pero Pseudo-nitzschia aumentó en abundancia durante la temporada de lluvias de 2012 y 2013. Los dinoflagelados y Phaeocystis globosa se comportaron como se esperaba y exhibieron un aumento relativo de la densidad celular durante la temporada de lluvias en los dos años. La alta disponibilidad de nutrientes durante esta temporada debe haber influido en el comportamiento del fitoplancton a pesar de las diferencias en la temperatura y la intensidad de la luz entre estaciones. Otras especies nocivas notables, pero raras que se encontraron sólo en las redes de arrastre de plancton durante el estudio fueron: Pyrodinium bahamense var. compressum, Alexandriumtamiyavanichii, Cochlodinium polykrikoides y Noctiluca scintillans.

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Multiple species in the toxic marine diatom genus Pseudo-nitzschia have been identified in the Northwestern Atlantic region encompassing the Gulf of Maine (GOM), including the Bay of Fundy (BOF). To gain further knowledge of the taxonomic composition and toxicity of species in this region, Pseudo-nitzschia isolates (n=146) were isolated from samples collected during research cruises that provided broad spatial coverage across the GOM and the southern New England shelf, herein referred to as the GOM region, during 2007-2008. Isolates, and cells in field material collected at 38 stations, were identified using electron microscopy (EM). Eight species (P. americana, P. fraudulenta, P. subpacifica, P. heimii, P. pungens, P. seriata, P. delicatissima and P. turgidula), and a novel form, Pseudo-nitzschia sp. GOM, were identified. Species identity was confirmed by sequencing the large subunit of the ribosomal rDNA (28S) and the internal transcribed spacer 2 (ITS2) for six species (36 isolates). Phylogenetic analyses (including neighbor joining, maximum parsimony, and maximum likelihood estimates and ITS2 secondary structure analysis) and morphometric data supported the placement of P. sp. GOM in a novel clade that includes morphologically and genetically similar isolates from Australia and Spain and is genetically most similar to P. pseudodelicatissima and P. cuspidata. Seven species (46 isolates) were grown in nutrient-replete batch culture and aliquots consisting of cells and growth medium were screened by Biosense ASP ELISA to measure total domoic acid (DA) produced (intracellular + extracellular); P. americana and P. heimii were excluded from all toxin analyses as they did not persist in culture long enough for testing. All 46 isolates screened produced DA in culture and total DA varied among species (e.g., 0.04 to 320 ng ml-1 for P. pungens and P. sp. GOM isolates, respectively) and among isolates of the same species (e.g., 0.24 - 320 ng ml-1 for P. sp. GOM). The 15 most toxic isolates corresponded to P. seriata, P. sp. GOM and P. pungens, and fg DA cell-1 was determined for whole cultures (cells and medium) using ELISA and liquid chromatography (LC) with fluorescence detection (FLD); for seven isolates, toxin levels were also estimated using LC - with mass spectrometry and ultraviolet absorbance detection. Pseudo-nitzschia seriata was the most toxic species (up to 3,500 fg cell-1) and was observed in the GOM region during all cruises (i.e., during the months of April, May, June and October). Pseudo-nitzschia sp. GOM, observed only during September and October 2007, was less toxic (19 - 380 fg cell-1) than P. seriata but more toxic than P. pungens var. pungens (0. 4 fg cell-1). Quantitation of DA indicated that concentrations measured by LC and ELISA were positively and significantly correlated; the lower detection limit of the ELISA permitted quantification of toxicity in isolates that were found to be nontoxic with LC methods. The confirmation of at least seven toxic species and the broad spatial and temporal distribution of toxic Pseudo-nitzschia spp. have significant implications for the regional management of nearshore and offshore shellfisheries resources.

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Most pennate diatoms are allogamous, and various types of mating systems have been described. In Pseudo-nitzschia, reproductive stages have been identified in some species, and it is generally accepted that the genus is mainly heterothallic. Here we report homothallic auxosporulation of Pseudo-nitzschia brasiliana Lundholm, Hasle et G. A. Fryxell. To our knowledge, this is the first verified description of homothallic sexual reproduction in the genus. Auxospore formation was observed in all 16 subclones derived from three initial clonal cultures of P. brasiliana. Pairing was followed by production of two gametes per gametangium, which fused to give two zygotes. Each zygote (early auxospore) was initially spherical and adhered to one girdle band of the parental frustule. The two auxospores tended to expand parallel to each other and perpendicular to the parental frustule. Elongation was synchronous, slightly asynchronous, or totally asynchronous. The entire process of sexual reproduction, from gamete formation to the appearance of the initial vegetative cells, took 2-4 d. The occurrence of sex in a homothallic species seems an advantageous life strategy for this species in that any encounter between cells of the right size class is potentially sexual.