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Sample records for alexandrium catenella whedon

  1. Exposure to the Paralytic Shellfish Toxin Producer Alexandrium catenella Increases the Susceptibility of the Oyster Crassostrea gigas to Pathogenic Vibrios

    PubMed Central

    Abi-Khalil, Celina; Lopez-Joven, Carmen; Abadie, Eric; Savar, Veronique; Amzil, Zouher; Laabir, Mohamed; Rolland, Jean-Luc

    2016-01-01

    The multifactorial etiology of massive Crassostrea gigas summer mortalities results from complex interactions between oysters, opportunistic pathogens and environmental factors. In a field survey conducted in 2014 in the Mediterranean Thau Lagoon (France), we evidenced that the development of the toxic dinoflagellate Alexandrium catenella, which produces paralytic shellfish toxins (PSTs), was concomitant with the accumulation of PSTs in oyster flesh and the occurrence of C. gigas mortalities. In order to investigate the possible role of toxic algae in this complex disease, we experimentally infected C. gigas oyster juveniles with Vibrio tasmaniensis strain LGP32, a strain associated with oyster summer mortalities, after oysters were exposed to Alexandrium catenella. Exposure of oysters to A. catenella significantly increased the susceptibility of oysters to V. tasmaniensis LGP32. On the contrary, exposure to the non-toxic dinoflagellate Alexandrium tamarense or to the haptophyte Tisochrysis lutea used as a foraging alga did not increase susceptibility to V. tasmaniensis LGP32. This study shows for the first time that A. catenella increases the susceptibility of Crassostrea gigas to pathogenic vibrios. Therefore, in addition to complex environmental factors explaining the mass mortalities of bivalve mollusks, feeding on neurotoxic dinoflagellates should now be considered as an environmental factor that potentially increases the severity of oyster mortality events. PMID:26784228

  2. Role of resting cysts in Chilean Alexandrium catenella dinoflagellate blooms revisited.

    PubMed

    Mardones, Jorge I; Bolch, Chris; Guzmán, Leonardo; Paredes, Javier; Varela, Daniel; Hallegraeff, Gustaaf M

    2016-05-01

    The detection of sparse Alexandrium catenella-resting cysts in sediments of southern Chilean fjords has cast doubts on their importance in the recurrence of massive toxic dinoflagellate blooms in the region. The role of resting cysts and the existence of different regional Chilean populations was studied by culturing and genetic approaches to define: (1) cyst production; (2) dormancy period; (3) excystment success; (4) offspring viability and (5) strain mating compatibility. This study newly revealed a short cyst dormancy (minimum 69 days), the role of key abiotic factors (in decreasing order salinity, irradiance, temperature and nutrients) controlling cyst germination (max. 60%) and germling growth rates (up to 0.36-0.52div.day(-1)). Amplified fragment length polymorphism (AFLP) characterization showed significant differences in genetic distances (GD) among A. catenella populations that were primarily determined by the geographical origin of isolates and most likely driven by oceanographic dispersal barriers. A complex heterothallic mating system pointed to variable reproductive compatibility (RCs) among Chilean strains that was high among northern (Los Lagos/North Aysén) and southern populations (Magallanes), but limited among the genetically differentiated central (South Aysén) populations. Field cyst surveys after a massive 2009 bloom event revealed the existence of exceptional high cyst densities in particular areas of the fjords (max. 14.627cystscm(-3)), which contrast with low cyst concentrations (<221.3cystscm(-3)) detected by previous oceanographic campaigns. In conclusion, the present study suggests that A. catenella resting cysts play a more important role in the success of this species in Chilean fjords than previously thought. Results from in vitro experiments suggest that pelagic-benthic processes can maintain year-round low vegetative cell concentrations in the water column, but also can explain the detection of high cysts aggregations after the

  3. Genome size of Alexandrium catenella and Gracilariopsis lemaneiformis estimated by flow cytometry

    NASA Astrophysics Data System (ADS)

    Du, Qingwei; Sui, Zhenghong; Chang, Lianpeng; Wei, Huihui; Liu, Yuan; Mi, Ping; Shang, Erlei; Zeeshan, Niaz; Que, Zhou

    2016-08-01

    Flow cytometry (FCM) technique has been widely applied to estimating the genome size of various higher plants. However, there is few report about its application in algae. In this study, an optimized procedure of FCM was exploited to estimate the genome size of two eukaryotic algae. For analyzing Alexandrium catenella, an important red tide species, the whole cell instead of isolated nucleus was studied, and chicken erythrocytes were used as an internal reference. The genome size of A. catenella was estimated to be 56.48 ± 4.14 Gb (1C), approximately nineteen times larger than that of human genome. For analyzing Gracilariopsis lemaneiformis, an important economical red alga, the purified nucleus was employed, and Arabidopsis thaliana and Chondrus crispus were used as internal references, respectively. The genome size of Gp. lemaneiformis was 97.35 ± 2.58 Mb (1C) and 112.73 ± 14.00 Mb (1C), respectively, depending on the different internal references. The results of this research will promote the related studies on the genomics and evolution of these two species.

  4. Exposure to the toxic dinoflagellate Alexandrium catenella modulates juvenile oyster Crassostrea gigas hemocyte variables subjected to different biotic conditions.

    PubMed

    Lassudrie, Malwenn; Soudant, Philippe; Nicolas, Jean-Louis; Miner, Philippe; Le Grand, Jacqueline; Lambert, Christophe; Le Goïc, Nelly; Hégaret, Hélène; Fabioux, Caroline

    2016-04-01

    The Pacific oyster Crassostrea gigas is an important commercial species cultured throughout the world. Oyster production practices often include transfers of animals into new environments that can be stressful, especially at young ages. This study was undertaken to determine if a toxic Alexandrium bloom, occurring repeatedly in French oyster beds, could modulate juvenile oyster cellular immune responses (i.e. hemocyte variables). We simulated planting on commercial beds by conducting a cohabitation exposure of juvenile, "specific pathogen-free" (SPF) oysters (naïve from the environment) with previously field-exposed oysters to induce interactions with new microorganisms. Indeed, toxic Alexandrium spp. exposures have been reported to modulate bivalve interaction with specific pathogens, as well as physiological and immunological variables in bivalves. In summary, SPF oysters were subjected to an artificial bloom of Alexandrium catenella, simultaneously with a cohabitation challenge. Exposure to A. catenella, and thus to the paralytic shellfish toxins (PSTs) and extracellular bioactive compounds produced by this alga, induced higher concentration, size, complexity and reactive oxygen species (ROS) production of circulating hemocytes. Challenge by cohabitation with field-exposed oysters also activated these hemocyte responses, suggesting a defense response to new microorganism exposure. These hemocyte responses to cohabitation challenge, however, were partially inhibited by A. catenella exposure, which enhanced hemocyte mortality, suggesting either detrimental effects of the interaction of both stressors on immune capacity, or the implementation of an alternative immune strategy through apoptosis. Indeed, no infection with specific pathogens (herpesvirus OsHV-1 or Vibrio aesturianus) was detected. Additionally, lower PST accumulation in challenged oysters suggests a physiological impairment through alteration of feeding-related processes. Overall, results of this

  5. Examination of the Seasonal Dynamics of the Toxic Dinoflagellate Alexandrium catenella at Redondo Beach, California, by Quantitative PCR▿

    PubMed Central

    Garneau, Marie-Ève; Schnetzer, Astrid; Countway, Peter D.; Jones, Adriane C.; Seubert, Erica L.; Caron, David A.

    2011-01-01

    The presence of neurotoxic species within the genus Alexandrium along the U.S. coastline has raised concern of potential poisoning through the consumption of contaminated seafood. Paralytic shellfish toxins (PSTs) detected in shellfish provide evidence that these harmful events have increased in frequency and severity along the California coast during the past 25 years, but the timing and location of these occurrences have been highly variable. We conducted a 4-year survey in King Harbor, CA, to investigate the seasonal dynamics of Alexandrium catenella and the presence of a particulate saxitoxin (STX), the parent compound of the PSTs. A quantitative PCR (qPCR) assay was developed for quantifying A. catenella in environmental microbial assemblages. This approach allowed for the detection of abundances as low as 12 cells liter−1, 2 orders of magnitude below threshold abundances that can impact food webs. A. catenella was found repeatedly during the study, particularly in spring, when cells were detected in 38% of the samples (27 to 5,680 cells liter−1). This peak in cell abundances was observed in 2006 and corresponded to a particulate STX concentration of 12 ng liter−1, whereas the maximum STX concentration of 26 ng liter−1 occurred in April 2008. Total cell abundances and toxin levels varied strongly throughout each year, but A. catenella was less abundant during summer, fall, and winter, when only 2 to 11% of the samples yielded positive qPCR results. The qPCR method developed here provides a useful tool for investigating the ecology of A. catenella at subbloom and bloom abundances. PMID:21926210

  6. Three-dimensional (3-D) fluorescence spectroscopy analysis of the fluorescent dissolved organic matter released by the marine toxic dinoflagellate Alexandrium catenella exposed to metal stress by zinc or lead.

    PubMed

    Herzi, Faouzi; Jean, Natacha; Sakka Hlaili, Asma; Mounier, Stéphane

    2014-08-01

    We investigated the effects of zinc or lead on growth and on exudation of fluorescent dissolved organic matter (FDOM) by the marine toxic dinoflagellate Alexandrium catenella (Whedon & Kofoid) Balech. The species was exposed to increasing free zinc (1.34 × 10(-7) M-3.98 × 10(-6) M) or lead (5.13 × 10(-9) M-1.82 × 10(-7) M) concentra-tions. Low metal levels ([Zn(2+) ] = 1.34 × 10(-7) M; [Pb(2+) ] = 5.13 × 10(-9) M) had no effect on cell growth. Toxic effects were observed from higher metal contamination ([Zn(2+) ] = 3.98 × 10(-6) M; [Pb(2+) ] = 6.54 × 10(-8) M), as a conversion of vegetative cells into cysts. Analysis of the released FDOM by three-dimensional (3-D) fluorescence spectroscopy was achieved, using the parallel factor analysis (PARAFAC). The PARAFAC modeling revealed four components associated with two contributions: one related to the biological activity; the other linked to the organic matter decomposition in the culture medium. The C1 component combined a tryptophan peak and characteristics of humic substances, whereas the C2 component was considered as a tryptophan protein fluorophore. The two others C3 and C4 components were associated with marine organic matter production. Relea-sed fluorescent substances were induced by low ([Zn(2+) ]= 1.34 × 10(-7) M; [Pb(2+) ] = 5.13 × 10(-9) M) and moderate ([Zn(2+) ] = 6.21 × 10(-7) M; [Pb(2+) ] = 2.64× 10(-9) M) metal concentrations, suggesting the activation of cellular mechanisms in response to metal stress, to exudate FDOM that could complex metal cations and reduce their toxicity toward A. catenella cells.

  7. Comparative Transcriptome Analysis of a Toxin-Producing Dinoflagellate Alexandrium catenella and Its Non-Toxic Mutant

    PubMed Central

    Zhang, Yong; Zhang, Shu-Fei; Lin, Lin; Wang, Da-Zhi

    2014-01-01

    The dinoflagellates and cyanobacteria are two major kingdoms of life producing paralytic shellfish toxins (PSTs), a large group of neurotoxic alkaloids causing paralytic shellfish poisonings around the world. In contrast to the well elucidated PST biosynthetic genes in cyanobacteria, little is known about the dinoflagellates. This study compared transcriptome profiles of a toxin-producing dinoflagellate, Alexandrium catenella (ACHK-T), and its non-toxic mutant form (ACHK-NT) using RNA-seq. All clean reads were assembled de novo into a total of 113,674 unigenes, and 66,812 unigenes were annotated in the known databases. Out of them, 35 genes were found to express differentially between the two strains. The up-regulated genes in ACHK-NT were involved in photosynthesis, carbon fixation and amino acid metabolism processes, indicating that more carbon and energy were utilized for cell growth. Among the down-regulated genes, expression of a unigene assigned to the long isoform of sxtA, the initiator of toxin biosynthesis in cyanobacteria, was significantly depressed, suggesting that this long transcript of sxtA might be directly involved in toxin biosynthesis and its depression resulted in the loss of the ability to synthesize PSTs in ACHK-NT. In addition, 101 putative homologs of 12 cyanobacterial sxt genes were identified, and the sxtO and sxtZ genes were identified in dinoflagellates for the first time. The findings of this study should shed light on the biosynthesis of PSTs in the dinoflagellates. PMID:25421324

  8. Exposure to the Neurotoxic Dinoflagellate, Alexandrium catenella, Induces Apoptosis of the Hemocytes of the Oyster, Crassostrea gigas

    PubMed Central

    Medhioub, Walid; Ramondenc, Simon; Vanhove, Audrey Sophie; Vergnes, Agnes; Masseret, Estelle; Savar, Veronique; Amzil, Zouher; Laabir, Mohamed; Rolland, Jean Luc

    2013-01-01

    This study assessed the apoptotic process occurring in the hemocytes of the Pacific oyster, Crassostrea gigas, exposed to Alexandrium catenella, a paralytic shellfish toxins (PSTs) producer. Oysters were experimentally exposed during 48 h to the toxic algae. PSTs accumulation, the expression of 12 key apoptotic-related genes, as well as the variation of the number of hemocytes in apoptosis was measured at time intervals during the experiment. Results show a significant increase of the number of hemocytes in apoptosis after 29 h of exposure. Two pro-apoptotic genes (Bax and Bax-like) implicated in the mitochondrial pathway were significantly upregulated at 21 h followed by the overexpression of two caspase executor genes (caspase-3 and caspase-7) at 29 h, suggesting that the intrinsic pathway was activated. No modulation of the expression of genes implicated in the cell signaling Fas-Associated protein with Death Domain (FADD) and initiation-phase (caspase-2) was observed, suggesting that only the extrinsic pathway was not activated. Moreover, the clear time-dependent upregulation of five (Bcl2, BI-1, IAP1, IAP7B and Hsp70) inhibitors of apoptosis-related genes associated with the return to the initial number of hemocytes in apoptosis at 48 h of exposure suggests the involvement of strong regulatory mechanisms of apoptosis occurring in the hemocytes of the Pacific oyster. PMID:24317471

  9. A feedback mechanism to control apoptosis occurs in the digestive gland of the oyster crassostrea gigas exposed to the paralytic shellfish toxins producer Alexandrium catenella.

    PubMed

    Rolland, Jean-Luc; Medhioub, Walid; Vergnes, Agnes; Abi-Khalil, Celina; Savar, Véronique; Abadie, Eric; Masseret, Estelle; Amzil, Zouher; Laabir, Mohamed

    2014-09-25

    To better understand the effect of Paralytic Shellfish Toxins (PSTs) accumulation in the digestive gland of the Pacific oyster, Crassostrea gigas, we experimentally exposed individual oysters for 48 h to a PSTs producer, the dinoflagellate Alexandrium catenella. In comparison to the effect of the non-toxic Alexandrium tamarense, on the eight apoptotic related genes tested, Bax and BI.1 were significantly upregulated in oysters exposed 48 h to A. catenella. Among the five detoxification related genes tested, the expression of cytochrome P450 (CYP1A) was shown to be correlated with toxin concentration in the digestive gland of oysters exposed to the toxic dinoflagellate. Beside this, we observed a significant increase in ROS production, a decrease in caspase-3/7 activity and normal percentage of apoptotic cells in this tissue. Taken together, these results suggest a feedback mechanism, which may occur in the digestive gland where BI.1 could play a key role in preventing the induction of apoptosis by PSTs. Moreover, the expression of CYP1A, Bax and BI.1 were found to be significantly correlated to the occurrence of natural toxic events, suggesting that the expression of these genes together could be used as biomarker to assess the biological responses of oysters to stress caused by PSTs.

  10. A Feedback Mechanism to Control Apoptosis Occurs in the Digestive Gland of the Oyster Crassostrea gigas Exposed to the Paralytic Shellfish Toxins Producer Alexandrium catenella

    PubMed Central

    Rolland, Jean-Luc; Medhioub, Walid; Vergnes, Agnes; Abi-khalil, Celina; Savar, Véronique; Abadie, Eric; Masseret, Estelle; Amzil, Zouher; Laabir, Mohamed

    2014-01-01

    To better understand the effect of Paralytic Shellfish Toxins (PSTs) accumulation in the digestive gland of the Pacific oyster, Crassostrea gigas, we experimentally exposed individual oysters for 48 h to a PSTs producer, the dinoflagellate Alexandrium catenella. In comparison to the effect of the non-toxic Alexandrium tamarense, on the eight apoptotic related genes tested, Bax and BI.1 were significantly upregulated in oysters exposed 48 h to A. catenella. Among the five detoxification related genes tested, the expression of cytochrome P450 (CYP1A) was shown to be correlated with toxin concentration in the digestive gland of oysters exposed to the toxic dinoflagellate. Beside this, we observed a significant increase in ROS production, a decrease in caspase-3/7 activity and normal percentage of apoptotic cells in this tissue. Taken together, these results suggest a feedback mechanism, which may occur in the digestive gland where BI.1 could play a key role in preventing the induction of apoptosis by PSTs. Moreover, the expression of CYP1A, Bax and BI.1 were found to be significantly correlated to the occurrence of natural toxic events, suggesting that the expression of these genes together could be used as biomarker to assess the biological responses of oysters to stress caused by PSTs. PMID:25257788

  11. Copper and cadmium effects on growth and extracellular exudation of the marine toxic dinoflagellate Alexandrium catenella: 3D-fluorescence spectroscopy approach.

    PubMed

    Herzi, Faouzi; Jean, Natacha; Zhao, Huiyu; Mounier, Stéphane; Mabrouk, Hassine Hadj; Hlaili, Asma Sakka

    2013-10-01

    In this study, metal contamination experiments were conducted to investigate the effects of copper and cadmium on the growth of the marine toxic dinoflagellate Alexandrium catenella and on the production of dissolved organic matter (Dissolved Organic Carbon: DOC; Fluorescent Dissolved Organic Matter: FDOM). This species was exposed to increasing concentrations of Cu(2+) (9.93 × 10(-10)-1.00 × 10(-7)M) or Cd(2+) (1.30 × 10(-8)-4.38 × 10(-7)M), to simulate polluted environments. The drastic effects were observed at pCu(2+)=7.96 (Cu(2+): 1.08 × 10(-8)M) and pCd(2+)=7.28 (Cd(2+): 5.19 × 10(-8)M), where cyst formation occurred. Lower levels of Cu(2+) (pCu(2+)>9.00) and Cd(2+) (pCd(2+)>7.28) had no effect on growth. However, when levels of Cu(2+) and Cd(2+) were beyond 10(-7)M, the growth was totally inhibited. The DOC released per cell (DOC/Cell) was different depending on the exposure time and the metal contamination, with higher DOC/Cell values in response to Cu(2+) and Cd(2+), comparatively to the control. Samples were also analyzed by 3D-fluorescence spectroscopy, using the Parallel Factor Analysis (PARAFAC) algorithm to characterize the FDOM. The PARAFAC analytical treatment revealed four components (C1, C2, C3 and C4) that could be associated with two contributions: one, related to the biological activity; the other, linked to the decomposition of organic matter. The C1 component combined a tryptophan peak and a characteristic humic substances response, and the C2 component was considered as a tryptophan protein fluorophore. The C3 and C4 components were associated to marine organic matter production.

  12. Preliminary identification of three new isolates in genus Alexandrium (Dinophyceae) from China sea area

    NASA Astrophysics Data System (ADS)

    Fan, Lijing; Sui, Zhenghong; Mao, Yunxiang; Guo, Hao

    2007-01-01

    The 5.8S ribosomal DNA sequences (5.8S rDNA) and their flanking regions, internal transcribed spacer 1 and spacer 2 (ITS1 and ITS2) of three new isolates in genus Alexandrium ( Alexandrium sp. qd1, Alexandrium sp. qd2, Alexandrium sp. gz) from China were amplified, sequenced, and subjected to phylogenetic analysis. Alexandrium sp. gz and Alexandrium sp. qd1 were grouped with high bootstrap values with four strains/species, i.e., A. catenella South Korea strain, A. catenella Japan strain, and two from China, Alexandrium sp. AC03 and Alexandrium sp. AN01 being proposed to be A. catenalla in a previous study. Then Alexandrium sp. gz and Alexandrium sp. qd1 were identified as Alexandrium catenella. As A. catenella was isolated from Qingdao and Guangzhou sea areas, it supposedly distributed at least in these two areas and was genetically different. Alexandrium sp. qd2 differed greatly from species in Alexandrium. It clustered with Symbiodinium californium, Symbiodinium sp. G15 and Gymnodinium sp. Zhao 01 with 100% bootstrap value; so Alexandrium sp. qd2 affiliates to genus Symbiodinium, and is probably a free-living Symbiodinium species.

  13. Detection and quantification of cultured marine Alexandrium species by real-time PCR.

    PubMed

    Zhang, Fengli; Li, Zhiyong

    2012-12-01

    The occurrence of harmful algal blooms (HABs) throughout the world has increased and poses a large threat to human health, fishery resources and tourism industries. The genus Alexandrium includes a number of toxic species associated with HABs. Therefore, it is very important to rapidly detect and monitor the harmful algae, such as Alexandrium genus. In this study, a standard curve of plasmid containing 18S rDNA-28S rDNA region from Alexandrium catenella was constructed and 5.8S rDNA sequence served as the primer of the real-time PCR. Cultured A. catenella, Alexandrium affine, Alexandrium lusitanicum and Alexandrium minutum samples were analyzed by real-time PCR using the same set of primers simultaneously. Using microscopy cells counts, 5.8S rDNA copies per cell and total DNA per cell were estimated. This assay method is promising for rapid detection of large number of Alexandrium samples.

  14. Distribution of the genus Alexandrium (Halim) and paralytic shellfish toxins along the coastline of New South Wales, Australia.

    PubMed

    Farrell, Hazel; Brett, Steve; Ajani, Penelope; Murray, Shauna

    2013-07-15

    Blooms of Alexandrium species, in particular the species Alexandrium catenella, accounted for more than 50% of algal related, shellfish aquaculture harvest zone closures in New South Wales (NSW) Australia since 2005. While there are indications that species of Alexandrium are more abundant than they were formerly, there is little data available on the spatial and temporal distribution and abundance of the genus in NSW. A six and a half year dataset comprising a total of 8649 fortnightly samples from 31 estuaries spread over 2000 km of NSW coastline was analysed. The greatest abundances of Alexandrium spp. were observed during the austral Spring and Summer, in estuaries in the mid and southern latitudes of the state. In identifying these high risk zones, we propose variables such as season, temperature, rainfall and estuarine flushing to be targeted in intensive site specific studies, to support the development of predictive tools for resource managers.

  15. Molecular detection and species identification of Alexandrium (Dinophyceae) causing harmful algal blooms along the Chilean coastline

    PubMed Central

    Jedlicki, Ana; Fernández, Gonzalo; Astorga, Marcela; Oyarzún, Pablo; Toro, Jorge E.; Navarro, Jorge M.; Martínez, Víctor

    2012-01-01

    Background and aims On the basis of morphological evidence, the species involved in South American Pacific coast harmful algal blooms (HABs) has been traditionally recognized as Alexandrium catenella (Dinophyceae). However, these observations have not been confirmed using evidence based on genomic sequence variability. Our principal objective was to accurately determine the species of Alexandrium involved in local HABs in order to implement a real-time polymerase chain reaction (PCR) assay for its rapid and easy detection on filter-feeding shellfish, such as mussels. Methodology For species-specific determination, the intergenic spacer 1 (ITS1), 5.8S subunit, ITS2 and the hypervariable genomic regions D1–D5 of the large ribosomal subunit of local strains were sequenced and compared with two data sets of other Alexandrium sequences. Species-specific primers were used to amplify signature sequences within the genomic DNA of the studied species by conventional and real-time PCR. Principal results Phylogenetic analysis determined that the Chilean strain falls into Group I of the tamarensis complex. Our results support the allocation of the Chilean Alexandrium species as a toxic Alexandrium tamarense rather than A. catenella, as currently defined. Once local species were determined to belong to Group I of the tamarensis complex, a highly sensitive and accurate real-time PCR procedure was developed to detect dinoflagellate presence in Mytilus spp. (Bivalvia) samples after being fed (challenged) in vitro with the Chilean Alexandrium strain. The results show that real-time PCR is useful to detect Alexandrium intake in filter-feeding molluscs. Conclusions It has been shown that the classification of local Alexandrium using morphological evidence is not very accurate. Molecular methods enabled the HAB dinoflagellate species of the Chilean coast to be assigned as A. tamarense rather than A. catenella. Real-time PCR analysis based on A. tamarense primers allowed the

  16. Modulation of ecdysal cyst and toxin dynamics of two Alexandrium (Dinophyceae) species under small-scale turbulence

    NASA Astrophysics Data System (ADS)

    Bolli, L.; Llaveria, G.; Garcés, E.; Guadayol, Ó.; van Lenning, K.; Peters, F.; Berdalet, E.

    2007-03-01

    In some dinoflagellate species, physiological processes appear to be altered by exposure to certain turbulent conditions. Here we investigated how two levels of turbulent kinetic energy dissipation rates (ɛ = 0.4 and 27 cm2 s-3) affected the toxin and ecdysal cyst dynamics of two bloom forming species, Alexandrium minutum and A. catenella. The most striking responses were observed at the high ɛ generated by an orbital shaker. In A. catenella, lower cellular toxin content was measured in cultures shaken for more than 4 days. The same trend was observed in A. minutum, although variability masked statistical significance. For the two species, inhibition of ecdysal cyst production occurred immediately and during the period of exposure of the cultures to stirring (4 or more days) at any time during their growth curve. Recovery of cyst abundances was always observed when turbulence stopped. When turbulence persisted for more than 4 days the net growth rate significantly decreased and the final biomass yield was lower than in the unshaken cultures. This study suggests that high levels of small-scale turbulence would contribute to the modulation of the harmful bloom dynamics through the interaction at the level of toxin and encystment processes.

  17. Occurrence of PSP-producing dinoflagellate Alexandrium tamiyavanichii in Bingo-Nada, the central coastal water of the Seto Inland Sea, Hiroshima Prefecture, Japan.

    PubMed

    Beppu, Rieko; Nojima, Kanako; Tsuruda, Shintaro; Gomez-Delan, Gloria; Barte-Quilantang, Mercy; Taniyama, Shigeto; Sagara, Takefumi; Nishio, Sachio; Takayama, Haruyoshi; Miyazawa, Keisuke; Asakawa, Manabu

    2008-04-01

    During surveillance of the distribution of the paralytic shellfish poison (PSP)-producing dinoflagellate in 2003, 2004 and 2005 along the coastlines of the Seto Inland Sea, Hiroshima Prefecture, Japan, some species of toxic phytoplankton were isolated from the eastern coasts, Bingo-Nada, the central regions of the Seto Inland Sea. It was rather unexpectedly revealed from the basis of the morphological characteristics that they were unambiguously identified as Alexandrium tamiyavanichii and Alexandrium catenella. Two strains (ATY041106, ATY051018) of A. tamiyavanichii showed a specific toxicity of 38.7 x 10(-6) and 111.5 x 10(-6)MU/cell, respectively. These values seemed to be several times or much higher than that of A. catenella (AC030816, AC040614), having a specific toxicity of 4.5 x 10(-6) and 4.1 x 10(-6)MU/cell, respectively, isolated in the same area. From the results of HPLC-furuorometric analysis, it revealed that the toxins in ATY041106 exist almost exclusively as beta-epimers (C2, GTX3, GTX4), which accounted for 72.7 mol%. The toxin profiles of this strain are featured by the presence of a large amount of GTX3 (59.1 mol%) and a small amount (20.6%) of C1 and 2 in comparison with the PSP compositions of A. tamarense, which is isolated as the main responsible species in Hiroshima Bay, a western part of coastal sea in Hiroshima Prefecture. On the other hand, it revealed that the toxin profiles of two strains (AC030816, AC040614) of A. catenella exist almost exclusively as beta-epimers (C2, GTX3, GTX4), which accounted for 81.8 and 56.5 mol%, as the same manner. The toxin profiles of these two strains are featured by the presence of a large amount of C2 (80.5 and 46.3 mol%) in comparison with the PSP compositions of A. tamiyavanichii. To our knowledge, this is the first record to show the distribution and harmful influence of A. tamiyavanichii and A. catenella in Bingo-Nada in Hiroshima Prefecture. Though contamination of bivalves with these PSP

  18. [The red tide caused by the dinoflagellate Alexandrium tamarense in the Colombian Pacific coast (2001)].

    PubMed

    García-Hansen, Ingrid; Cortés-Altamirano, Roberto; Sierra-Beltrán, Arturo P

    2004-09-01

    From April 26th to May 15th 2001, a large algae bloom was observed off Tumaco Bay on the Pacific coast of Colombia. This was the first harmful algae bloom (HAB) reported in the region, and reached Gorgona Island, about 120 km north. A year later, starting March 2002, an offshore HAB developed from Cabo Corrientes North to Solano Bay. The typical abundance during the blooms reached 7.5 x 10(6) cells l(-1) for the 2001 event and 1.6 x 10(6) cells l(-1) for the 2002 event. During both events, low temperature and high salinity were recorded. Typical measurements in the area are 27-27.5 degrees C and 30-31.5 psu. Values observed during the two events were 24-24.6 degrees C and 33-34 psu; 3 degrees C below normal and more than 2.5 psu above average values. These conditions are indicative of local upwelling processes at the time of the events. On both occasions, cells corresponding to the Alexandrium catenella/fundeyense/tamarense complex represented 99-100% of the biomass. It was difficult to differentiate the cells from A. catenella, but the presence of short chains of only 4 cells (single cells represented most of the biomass) was suggestive of A. tamarense. Shape, dimensions, and detailed structure of the apical pore complex, first apical plate, posterior sulcal plate, and position of the ventral pore on plate 1' of cells were consistent with the description of A. tamarense, which has not been reported in the tropical East Pacific. The Control Center of Pacific Contamination of the Maritime General Direction of the Colombian Navy has been monitoring the area since 1994 without finding this species or HABs. This leads us to consider the two events as caused by recently introduced species, where local upwelling processes favor permanent and cyclic HABs. However, during these two events, there were no reports of effects on marine biota or of human poisoning, probably because the blooms occurred some distance offshore and far from exploited shellfish beds.

  19. Modulation of ecdysal cyst and toxin dynamics of two Alexandrium (Dinophyceae) species under small-scale turbulence

    NASA Astrophysics Data System (ADS)

    Bolli, L.; Llaveria, G.; Garcés, E.; Guadayol, Ò.; van Lenning, K.; Peters, F.; Berdalet, E.

    2007-08-01

    Some dinoflagellate species have shown different physiological responses to certain turbulent conditions. Here we investigate how two levels of turbulent kinetic energy dissipation rates (ɛ = 0.4 and 27 cm² s-3) affect the PSP toxins and ecdysal cyst dynamics of two bloom forming species, Alexandrium minutum and A. catenella. The most striking responses were observed at the high ɛ generated by an orbital shaker. In the cultures of the two species shaken for more than 4 days, the cellular GTX(1+4) toxin contents were significantly lower than in the still control cultures. In A. minutum this trend was also observed in the C(1+2) toxin content. For the two species, inhibition of ecdysal cyst production occurred during the period of exposure of the cultures to stirring (4 or more days) at any time during their growth curve. Recovery of cyst abundances was always observed when turbulence stopped. When shaking persisted for more than 4 days, the net growth rate significantly decreased in A. minutum (from 0.25±0.01 day-1 to 0.19±0.02 day-1) and the final cell numbers were lower (ca. 55.4%) than in the still control cultures. In A. catenella, the net growth rate was not markedly modified by turbulence although under long exposure to shaking, the cultures entered earlier in the stationary phase and the final cell numbers were significantly lower (ca. 23%) than in the control flasks. The described responses were not observed in the experiments performed at the low turbulence intensities with an orbital grid system, where the population development was favoured. In those conditions, cells appeared to escape from the zone of the influence of the grids and concentrated in calmer thin layers either at the top or at the bottom of the containers. This ecophysiological study provides new evidences about the sensitivity to high levels of small-scale turbulence by two life cycle related processes, toxin production and encystment, in dinoflagellates. This can contribute to the

  20. Formal Revision of the Alexandrium tamarense Species Complex (Dinophyceae) Taxonomy: The Introduction of Five Species with Emphasis on Molecular-based (rDNA) Classification

    PubMed Central

    John, Uwe; Litaker, R. Wayne; Montresor, Marina; Murray, Shauna; Brosnahan, Michael L.; Anderson, Donald M.

    2015-01-01

    The Alexandrium tamarense species complex is one of the most studied marine dinoflagellate groups due to its ecological, toxicological and economic importance. Several members of this complex produce saxitoxin and its congeners – potent neurotoxins that cause paralytic shellfish poisoning. Isolates from this complex are assigned to A. tamarense, A. fundyense, or A. catenella based on two main morphological characters: the ability to form chains and the presence/absence of a ventral pore between Plates 1′ and 4′. However, studies have shown that these characters are not consistent and/or distinctive. Further, phylogenies based on multiple regions in the rDNA operon indicate that the sequences from morphologically indistinguishable isolates partition into five clades. These clades were initially named based on their presumed geographic distribution, but recently were renamed as Groups I–V following the discovery of sympatry among some groups. In this study we present data on morphology, ITS/5.8S genetic distances, ITS2 compensatory base changes, mating incompatibilities, toxicity, the sxtA toxin synthesis gene, and rDNA phylogenies. All results were consistent with each group representing a distinct cryptic species. Accordingly, the groups were assigned species names as follows: Group I, A. fundyense; Group II, A. mediterraneum; Group III, A. tamarense; Group IV, A. pacificum; Group V, A. australiense. PMID:25460230

  1. Alexandrium minutum growth controlled by phosphorus . An applied model

    NASA Astrophysics Data System (ADS)

    Chapelle, A.; Labry, C.; Sourisseau, M.; Lebreton, C.; Youenou, A.; Crassous, M. P.

    2010-11-01

    Toxic algae are a worldwide problem threatening aquaculture, public health and tourism. Alexandrium, a toxic dinoflagellate proliferates in Northwest France estuaries (i.e. the Penzé estuary) causing Paralytic Shellfish Poisoning events. Vegetative growth, and in particular the role of nutrient uptake and growth rate, are crucial parameters to understand toxic blooms. With the goal of modelling in situ Alexandrium blooms related to environmental parameters, we first try to calibrate a zero-dimensional box model of Alexandrium growth. This work focuses on phosphorus nutrition. Our objective is to calibrate Alexandrium minutum as well as Heterocapsa triquetra (a non-toxic dinoflagellate) growth under different rates of phosphorus supply, other factors being optimal and constant. Laboratory experiments are used to calibrate two growth models and three uptake models for each species. Models are then used to simulate monospecific batch and semi-continuous experiments as well as competition between the two algae (mixed cultures). Results show that the Droop growth model together with linear uptake versus quota can represent most of our observations, although a power law uptake function can more accurately simulate our phosphorus uptake data. We note that such models have limitations in non steady-state situations and cell quotas can depend on a variety of factors, so care must be taken in extrapolating these results beyond the specific conditions studied.

  2. Distribution and toxicity of Alexandrium ostenfeldii (Dinophyceae) in the Gulf of Maine, USA

    NASA Astrophysics Data System (ADS)

    Gribble, Kristin E.; Keafer, Bruce A.; Quilliam, Michael A.; Cembella, Allan D.; Kulis, David M.; Manahan, Abigail; Anderson, Donald M.

    2005-09-01

    Alexandrium ostenfeldii is a thecate, mixotrophic dinoflagellate recently linked to a novel suite of toxins called spirolides. This study provides the first description of the regional distribution of A. ostenfeldii in the Gulf of Maine (GOM), and the first report and analysis of spirolide toxicity in A. ostenfeldii in waters south of Nova Scotia. Morphological examination of cells in field samples and of clonal cultures isolated from several stations in the GOM confirmed the presence of A. ostenfeldii. A genus-specific antibody probe, and an A. ostenfeldii species-specific oligonucleotide probe labeled these cells; a probe specific for the North American A. fundyense/tamarense/catenella species complex did not label A. ostenfeldii cells. Cell size ranged from 20 to nearly 90 μm, and most cells contained food vacuoles, with a total vacuole size from 1 to 48 μm. The hydrographic forcings controlling the distribution of A. ostenfeldii in the GOM are quite similar to those acting on the A. fundyense population at the same time of the year. The highest concentrations of A. ostenfeldii were observed nearshore, to the east of Penobscot Bay, at times with an offshore-turning branch of high cell concentration to the south of Penobscot Bay. Casco Bay appears to be an area of accumulation for A. ostenfeldii cells advected toward shore from the core of the population to the northeast. Concentrations of A. ostenfeldii were generally higher at the surface than deeper, except at locations where the pooling of lower-salinity water at the surface may have led to the subduction of the population flowing in from the east. PSP toxins were detected in field populations containing A. ostenfeldii and A. fundyense, but not in A. ostenfeldii cultures isolated from the GOM. Spirolide toxins were found in 36 of 60 field samples. More than 83% of samples containing A. ostenfeldii cells had one or more of spirolide congeners A, B, C2 and D2. The total concentration of spirolides per cell at

  3. SxtA gene sequence analysis of dinoflagellate Alexandrium minutum

    NASA Astrophysics Data System (ADS)

    Norshaha, Safida Anira; Latib, Norhidayu Abdul; Usup, Gires; Yusof, Nurul Yuziana Mohd

    2015-09-01

    The dinoflagellate Alexandrium minutum is typically known for the production of potent neurotoxins such as saxitoxin, affecting the health of human seafood consumers via paralytic shellfish poisoning (PSP). These phenomena is related to the harmful algal blooms (HABs) that is believed to be influenced by environmental and nutritional factors. Previous study has revealed that SxtA gene is a starting gene that involved in the saxitoxin production pathway. The aim of this study was to analyse the sequence of the sxtA gene in A. minutum. The dinoflagellates culture was cultured at temperature 26°C with 16:8-hour light:dark photocycle. After the samples were harvested, RNA was extracted, complementary DNA (cDNA) was synthesised and amplified by polymerase chain reaction (PCR). The PCR products were then purified and cloned before sequenced. The SxtA sequence obtained was then analyzed in order to identify the presence of SxtA gene in Alexandrium minutum.

  4. Seasonal variation in the biochemical composition of red seaweed ( Catenella repens) from Gangetic delta, northeast coast of India

    NASA Astrophysics Data System (ADS)

    Banerjee, Kakoli; Ghosh, Rajrupa; Homechaudhuri, Sumit; Mitra, Abhijit

    2009-10-01

    The biochemical composition of red seaweeds, Catenella repens was investigated in this present study along with subsequent analysis of relevant physico-chemical variables. In this study, the relationship between the nutritive components of this species and the ambient environmental parameters was established. Protein content varied from 2.78 ± 0.30% of dry weight (stn.3) to 16.03 ± 0.96% of dry weight (stn.1) with highest values during monsoon. The protein levels were positively correlated with dissolved nitrate content and negatively correlated with water temperature (except stn.3) and salinity. Carbohydrate content of this species varied significantly ( p < 0.05) during pre-monsoon between stations and the values showed positive relationship with salinity and surface water temperature. In contrast to carbohydrate, lipid concentration was lowest in values and varied very slightly between seasons and stations. Astaxanthin content of the seaweed species was greater in pre-monsoon than monsoon and post-monsoon in all the selected stations. Compared with the three seasons, samples of red seaweed collected in pre-monsoon has high carbohydrate-astaxanthin in contrast to protein-lipid which showed high values during monsoon. Statistical analysis computed among the environmental and biochemical parameters suggests the potential role played by the abiotic parameters on biosynthetic pathways of seaweed. This paper also highlights the influence of the nutritional quality of water that can be used for mass cultivation of Catenella repens.

  5. Suspended Alexandrium spp. hypnozygote cysts in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Kirn, Sarah L.; Townsend, David W.; Pettigrew, Neal R.

    2005-09-01

    The life cycle of dinoflagellates of the genus Alexandrium includes sexual reproduction followed by the formation of a dormant hypnozygote cyst, which serves as a resting stage. Negatively buoyant cysts purportedly fall to the benthos where they undergo a mandatory period of quiescence. Previous reports of cysts in the surficial sediments of the Gulf of Maine, where Alexandrium blooms are well documented, show a broad distribution of cysts, with highest concentrations generally in sediments below 100 m depth. We report here an exploration of cysts suspended in the water column, where they would be better positioned to inoculate springtime Alexandrium populations. During cruises in February, April, and June of 2000, water samples were collected at depths just off the bottom (within 5 m), at the top of the bottom nepheloid layer, and near the surface (1 m) and examined for cyst concentrations. Suspended cysts were found throughout the Gulf of Maine and westernmost Bay of Fundy. Planktonic cyst densities were generally greater in near-bottom and top of the bottom nepheloid layer samples than in near-surface water samples; densities were of the order of 10 2 cysts m -3 in surface waters, and 10 2-10 3 cysts m -3 at near-bottom depths. Temporally, they were most abundant in February and least abundant in April. Reports by earlier workers of cysts in the underlying sediments were on the order of 10 3 cysts cm -3. We present calculations that demonstrate the likelihood of cyst resuspension from bottom sediments forced by swell and tidal currents, and propose that such resuspended cysts are important in inoculating the seasonal bloom. We estimate that suspended cysts may contribute significantly to the annual vegetative cell population in the Gulf of Maine.

  6. Termination of a toxic Alexandrium bloom with hydrogen peroxide.

    PubMed

    Burson, Amanda; Matthijs, Hans C P; de Bruijne, Wilco; Talens, Renee; Hoogenboom, Ron; Gerssen, Arjen; Visser, Petra M; Stomp, Maayke; Steur, Kees; van Scheppingen, Yvonne; Huisman, Jef

    2014-01-01

    The dinoflagellate Alexandrium ostenfeldii is a well-known harmful algal species that can potentially cause paralytic shellfish poisoning (PSP). Usually A. ostenfeldii occurs in low background concentrations only, but in August of 2012 an exceptionally dense bloom of more than 1millioncellsL(-1) occurred in the brackish Ouwerkerkse Kreek in The Netherlands. The A. ostenfeldii bloom produced both saxitoxins and spirolides, and is held responsible for the death of a dog with a high saxitoxin stomach content. The Ouwerkerkse Kreek routinely discharges its water into the adjacent Oosterschelde estuary, and an immediate reduction of the bloom was required to avoid contamination of extensive shellfish grounds. Previously, treatment of infected waters with hydrogen peroxide (H2O2) successfully suppressed cyanobacterial blooms in lakes. Therefore, we adapted this treatment to eradicate the Alexandrium bloom using a three-step approach. First, we investigated the required H2O2 dosage in laboratory experiments with A. ostenfeldii. Second, we tested the method in a small, isolated canal adjacent to the Ouwerkerkse Kreek. Finally, we brought 50mgL(-1) of H2O2 into the entire creek system with a special device, called a water harrow, for optimal dispersal of the added H2O2. Concentrations of both vegetative cells and pellicle cysts declined by 99.8% within 48h, and PSP toxin concentrations in the water were reduced below local regulatory levels of 15μgL(-1). Zooplankton were strongly affected by the H2O2 treatment, but impacts on macroinvertebrates and fish were minimal. A key advantage of this method is that the added H2O2 decays to water and oxygen within a few days, which enables rapid recovery of the system after the treatment. This is the first successful field application of H2O2 to suppress a marine harmful algal bloom, although Alexandrium spp. reoccurred at lower concentrations in the following year. The results show that H2O2 treatment provides an effective emergency

  7. Factors regulating excystment of Alexandrium in Puget Sound, WA, USA

    PubMed Central

    Moore, Stephanie K.; Bill, Brian D.; Hay, Levi R.; Emenegger, Jennifer; Eldred, Kiara C.; Greengrove, Cheryl L.; Masura, Julie E.; Anderson, Donald M.

    2015-01-01

    Factors regulating excystment of a toxic dinoflagellate in the genus Alexandrium were investigated in cysts from Puget Sound, Washington State, USA. Experiments were carried out in the laboratory using cysts collected from benthic seedbeds to determine if excystment is controlled by internal or environmental factors. The results suggest that the timing of germination is not tightly controlled by an endogenous clock, though there is a suggestion of a cyclical pattern. This was explored using cysts that had been stored under cold (4 °C), anoxic conditions in the dark and then incubated for 6 weeks at constant favorable environmental conditions. Excystment occurred during all months of the year, with variable excystment success ranging from 31–90%. When cysts were isolated directly from freshly collected sediments every month and incubated at the in situ bottom water temperature, a seasonal pattern in excystment was observed that was independent of temperature. This pattern may be consistent with secondary dormancy, an externally modulated pattern that prevents excystment during periods that are not favorable for sustained vegetative growth. However, observation over more annual cycles is required and the duration of the mandatory dormancy period of these cysts must be determined before the seasonality of germination can be fully characterized in Alexandrium from Puget Sound. Both temperature and light were found to be important environmental factors regulating excystment, with the highest rates of excystment observed for the warmest temperature treatment (20 °C) and in the light. PMID:26109923

  8. Impact of zooplankton grazing on Alexandrium blooms in the offshore Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Turner, Jefferson T.; Borkman, David G.

    2005-09-01

    Zooplankton grazing was investigated by shipboard experiments during natural blooms of Alexandrium spp. in the offshore Gulf of Maine in spring and/or summer of 1998, 2000, and 2001. Grazing studies were done in conjunction with studies of accumulation of Alexandrium toxins in the zooplankton, as part of the ECOHAB-Gulf of Maine regional program. Several species of copepods, marine cladocerans, and appendicularians were allowed to graze upon natural phytoplankton assemblages, at ambient temperatures (14-17 °C). Grazing was measured by quantitative microscopic analyses of disappearance of phytoplankton cells in initial, control, and experimental food suspensions. Thus, we were able to examine grazing upon Alexandrium in comparison to grazing on other co-occurring phytoplankton taxa. Even during Alexandrium "blooms," this dinoflagellate was a minor component of the overall phytoplankton assemblage. It was present at stations where grazing experiments were conducted at levels of 0.12-7.57×10 3 cells l -1, or 0.03-3.93% of total phytoplankton cells. Maximum ingestion of Alexandrium accounted for only up to 3.2% of total cells ingested. Phytoplankton assemblages were dominated by athecate microflagellates, and to a lesser extent by diatoms and non-toxic dinoflagellates. Microflagellates were present at abundances of 159.62-793.93 cells ml -1, or 60.6-95.56% of total cells. Grazing on microflagellates accounted for 35.59-98.21% of total grazing. Grazing on Alexandrium spp. and microflagellates was generally non-selective, with these taxa being ingested in similar proportions to their availability in food assemblages. Grazing on diatoms was selective, with diatoms being disproportionately ingested, compared to their proportions in food assemblages. There were no apparent adverse effects of Alexandrium on grazers during incubations of 18-24 h, and grazer survival was 100%. Estimated daily zooplankton grazing impact on Alexandrium spp. field populations by field

  9. The globally distributed genus Alexandrium: multifaceted roles in marine ecosystems and impacts on human health

    PubMed Central

    Alpermann, Tilman J.; Cembella, Allan D.; Collos, Yves; Masseret, Estelle; Montresor, Marina

    2011-01-01

    The dinoflagellate genus Alexandrium is one of the major harmful algal bloom (HAB) genera with respect to the diversity, magnitude and consequences of blooms. The ability of Alexandrium to colonize multiple habitats and to persist over large regions through time is testimony to the adaptability and resilience of this group of species. Three different families of toxins, as well as an as yet incompletely characterized suite of allelochemicals are produced among Alexandrium species. Nutritional strategies are equally diverse, including the ability to utilize a range of inorganic and organic nutrient sources, and feeding by ingestion of other organisms. Many Alexandrium species have complex life histories that include sexuality and often, but not always, cyst formation, which is characteristic of a meroplanktonic life strategy and offers considerable ecological advantages. Due to the public health and ecosystem impacts of Alexandrium blooms, the genus has been extensively studied, and there exists a broad knowledge base that ranges from taxonomy and phylogeny through genomics and toxin biosynthesis to bloom dynamics and modeling. Here we present a review of the genus Alexandrium, focusing on the major toxic and otherwise harmful species. PMID:22308102

  10. Effect of Alexandrium tamarense on three bloom-forming algae

    NASA Astrophysics Data System (ADS)

    Yin, Juan; Xie, Jin; Yang, Weidong; Li, Hongye; Liu, Jiesheng

    2010-07-01

    We investigated the allelopathic properties of Alexandrium tamarense (Laboar) Balech on the growth of Prorocentrum donghaiense Lu, Chattonella marina (Subrahmanyan) Hara et Chihara and Heterosigma akashiwo (Hada) Hada in a laboratory experiment. We examined the growth of A. tamarense, C. marina, P. donghaiense and H. Akashiwo in co-cultures and the effect of filtrates from A. tamarense cultures in various growth phases, on the three harmful algal bloom (HAB)-forming algae. In co-cultures with A. tamarense, both C. marina and H. akashiwo were dramatically suppressed at high cell densities; in contrast, the growth of P. donghaiense varied in different inoculative ratios of A. tamarense and P. donghaiense. When the ratio was 1:1 ( P. donghaiense: A. tamarense), growth of P. donghaiense was inhibited considerably, while the growth of P. donghaiense was almost the same as that of the control when the ratio was 9:1. The growth difference of P. donghaiense, C. marina and H. akashiwo when co-cultured with A. tamarense indicated that the allelopathic effect may be one of the important factors in algal competition and phytoplankton succession involving A. tamarense. In addition, the filtrate from A. tamarense culture had negative impacts on these three HAB algae, and such inhibition varied with different growth phases of A. tamarense in parallel with reported values of PSP toxin content in Alexandrium cells. This implied that PSP toxin was possibly involved in allelopathy of A. tamarense. However, the rapid decomposition and inactivation of PSP toxin above pH 7 weakened this possibility. Further studies on the allelochemicals responsible for the allelopathy of A. tamarense need to be carried out in future.

  11. A Molecular and Co-Evolutionary Context for Grazer Induced Toxin Production in Alexandrium tamarense

    PubMed Central

    Wohlrab, Sylke; Iversen, Morten H.; John, Uwe

    2010-01-01

    Marine dinoflagellates of the genus Alexandrium are the proximal source of neurotoxins associated with Paralytic Shellfish Poisoning. The production of these toxins, the toxin biosynthesis and, thus, the cellular toxicity can be influenced by abiotic and biotic factors. There is, however, a lack of substantial evidence concerning the toxins' ecological function such as grazing defense. Waterborne cues from copepods have been previously found to induce a species-specific increase in toxin content in Alexandrium minutum. However, it remains speculative in which context these species-specific responses evolved and if it occurs in other Alexandrium species as well. In this study we exposed Alexandrium tamarense to three copepod species (Calanus helgolandicus, Acartia clausii, and Oithona similis) and their corresponding cues. We show that the species-specific response towards copepod-cues is not restricted to one Alexandrium species and that co-evolutionary processes might be involved in these responses, thus giving additional evidence for the defensive role of phycotoxins. Through a functional genomic approach we gained insights into the underlying molecular processes which could trigger the different outcomes of these species-specific responses and consequently lead to increased toxin content in Alexandrium tamarense. We propose that the regulation of serine/threonine kinase signaling pathways has a major influence in directing the external stimuli i.e. copepod-cues, into different intracellular cascades and networks in A. tamarense. Our results show that A. tamarense can sense potential predating copepods and respond to the received information by increasing its toxin production. Furthermore, we demonstrate how a functional genomic approach can be used to investigate species interactions within the plankton community. PMID:21124775

  12. A molecular and co-evolutionary context for grazer induced toxin production in Alexandrium tamarense.

    PubMed

    Wohlrab, Sylke; Iversen, Morten H; John, Uwe

    2010-11-29

    Marine dinoflagellates of the genus Alexandrium are the proximal source of neurotoxins associated with Paralytic Shellfish Poisoning. The production of these toxins, the toxin biosynthesis and, thus, the cellular toxicity can be influenced by abiotic and biotic factors. There is, however, a lack of substantial evidence concerning the toxins' ecological function such as grazing defense. Waterborne cues from copepods have been previously found to induce a species-specific increase in toxin content in Alexandrium minutum. However, it remains speculative in which context these species-specific responses evolved and if it occurs in other Alexandrium species as well. In this study we exposed Alexandrium tamarense to three copepod species (Calanus helgolandicus, Acartia clausii, and Oithona similis) and their corresponding cues. We show that the species-specific response towards copepod-cues is not restricted to one Alexandrium species and that co-evolutionary processes might be involved in these responses, thus giving additional evidence for the defensive role of phycotoxins. Through a functional genomic approach we gained insights into the underlying molecular processes which could trigger the different outcomes of these species-specific responses and consequently lead to increased toxin content in Alexandrium tamarense. We propose that the regulation of serine/threonine kinase signaling pathways has a major influence in directing the external stimuli i.e. copepod-cues, into different intracellular cascades and networks in A. tamarense. Our results show that A. tamarense can sense potential predating copepods and respond to the received information by increasing its toxin production. Furthermore, we demonstrate how a functional genomic approach can be used to investigate species interactions within the plankton community.

  13. Georges Bank: A leaky incubator of Alexandrium fundyense blooms

    NASA Astrophysics Data System (ADS)

    McGillicuddy, D. J.; Townsend, D. W.; Keafer, B. A.; Thomas, M. A.; Anderson, D. M.

    2014-05-01

    A series of oceanographic surveys on Georges Bank document variability of populations of the toxic dinoflagellate Alexandrium fundyense on time scales ranging from synoptic to seasonal to interannual. Blooms of A. fundyense on Georges Bank can reach concentrations on the order of 104 cells l-1, and are generally bank-wide in extent. Georges Bank populations of A. fundyense appear to be quasi-independent of those in the adjacent coastal Gulf of Maine, insofar as they occupy a hydrographic niche that is colder and saltier than their coastal counterparts. In contrast to coastal populations that rely on abundant resting cysts for bloom initiation, very few cysts are present in the sediments on Georges Bank. Bloom dynamics must therefore be largely controlled by the balance between growth and mortality processes, which are at present largely unknown for this population. Based on correlations between cell abundance and nutrient distributions, ammonium appears to be an important source of nitrogen for A. fundyense blooms on Georges Bank.

  14. The role of resting cysts in Alexandrium minutum population dynamics

    NASA Astrophysics Data System (ADS)

    Estrada, Marta; Solé, Jordi; Anglès, Sílvia; Garcés, Esther

    2010-02-01

    The role of resting cysts on the development of Alexandrium minutum blooms in a typical Mediterranean semi-enclosed water body (Arenys de Mar Harbor, NW Mediterranean) was studied by means of matrix and dynamic population models. We used a series of scenarios, constrained when possible by experimentally measured parameters to test whether excystment and encystment fluxes and changes in the dormancy period had a major effect on bloom intensity and duration. The results of the simulations highlighted the importance of knowing not only the magnitude and variability of growth and life-cycle transition rates, but also those of loss rates (both in the water column and in the sediment) due to physical or biological factors. Given the maximum encystment rates determined for A. minutum in the study area (0.01 d -1), this process contributed to reduce the peak concentrations of vegetative cells but did not have a dominant effect on bloom termination. Excystment fluxes could contribute to enhance population densities of vegetative cells during times or low or negative net growth rate and during the initial phases of a bloom, but once exponential growth had started, additional excystment had negligible effect on bloom magnitude. However, even if cysts did not contribute to larger blooms, they could represent a safety mechanism for reintroduction of the species when the vegetative cell population went extinct due to unfavorable environmental conditions. Increasing the dormancy time exposed newly formed cysts to a longer period of losses in the sediment that reduced the concentration of excystment-ready sediment cysts and decreased excystment fluxes. More complex models will be needed to explore the implications of different life-cycle strategies in a wider natural ecological context.

  15. Alexandrium minutum resting cyst distribution dynamics in a confined site

    NASA Astrophysics Data System (ADS)

    Anglès, Sílvia; Jordi, Antoni; Garcés, Esther; Basterretxea, Gotzon; Palanques, Albert

    2010-02-01

    The life cycle of the toxic dinoflagellate Alexandrium minutum consists of an asexual stage, characterized by motile vegetative cells, and a sexual stage, a resting cyst that once formed remains dormant in the sediment. Insight into the factors that determine the distribution and abundance of resting cysts is essential to understanding the dynamics of the vegetative phase. In investigations carried out between January 2005 and January 2008 in Arenys de Mar harbor (northwestern Mediterranean Sea), the spatial and temporal distribution patterns of A. minutum resting cysts and of the sediments were studied during different bloom stages of the vegetative population. Maximum cyst abundance was recorded mainly in the innermost part of the harbor while the lowest abundance always occurred near the harbor entrance, consistent with the distribution of silt-clay sediment fractions. The tendency of cysts in sediments to increase after bloom periods was clearly associated with new cyst formation, while cyst abundance decreased during non-bloom periods. Exceptions to this trend were observed in stations dominated by the deposition of coarse sediments. High correlation between the presence of cysts and clays during non-bloom periods indicates that cysts behave as passive sediment particles and are influenced by the same hydrodynamic processes as clays. In Arenys de Mar, the main physical forcing affecting sediment resuspension is the seiche, which was studied using in situ measurements and numerical models to interpret the observed distribution patterns. During non-bloom periods, cyst losses were smaller when the seiche was more active and at the station where the seiche-induced current was larger. Thus, seiche-forced resuspension appears to reduce cyst losses by reallocating cysts back to the sediment surface such that their burial in the sediment is avoided. The observed vertical profiles of the cysts were consistent with this process.

  16. THE RED-TIDE DINOFLAGELLATE, ALEXANDRIUM MONILATUM, SUPPRESSES GROWTH OF MIXED NATURAL PHYTOPLANKTON

    EPA Science Inventory

    Alexandrium monilatum is a large, chain-forming, autotrophic dinoflagellate associated with red-tides and fish kills along the US Gulf of Mexico coast. When cultured inocula of A. monilatum were added to nutrient-amended seawater samples, growth rates and biomass yields of the na...

  17. GROWTH RATES, PHYSIOLOGICAL INDICATORS AND ELEMENTAL COMPOSITION OF THE RED-TIDE DINOFLAGELLATE, ALEXANDRIUM MONILATUM

    EPA Science Inventory

    Alexandrium monilatum is a thecate, autotrophic, bioluminescent and chain-forming dinoflagellate. Although it has been known to be associated with red tides and fish kills along the US Gulf of Mexico coast for almost 50 years, little basic physiological information is available f...

  18. GROWTH RATES AND ELEMENTAL COMPOSITION OF ALEXANDRIUM MONILATUM, A REDTIDE DINOFLAGELLATE

    EPA Science Inventory

    The combined effects of temperature and salinity on growth of Alexandrium monilatum were studied in laboratory cultures. This toxic, red-tide dinoflagellate grew faster with higher temperatures, up to a maximum of approximately 1 division d-1 at 31 C. Salinities above 15 psu had ...

  19. IDENTIFICATION AND TOXICITY OF ALEXANDRIUM TAMARENSE (DINOPHYCEAE) IN SCOTTISH WATERS(1).

    PubMed

    Collins, C; Graham, J; Brown, L; Bresnan, E; Lacaze, J-P; Turrell, E A

    2009-06-01

    Contamination of shellfish with paralytic shellfish poisoning (PSP) toxins produced by Alexandrium species poses a potential threat to the sustainability of the Scottish aquaculture industry. Routine LM analysis of water samples from around the Scottish coast has previously identified Alexandrium (Dinophyceae) as a regular part of the spring and summer phytoplankton communities in Scottish coastal waters. In this study, Alexandrium tamarense (M. Lebour) Balech isolated from sediment and water samples was established in laboratory culture. Species identification of these isolates was confirmed using thecal plate dissections and by molecular characterization based on their LSU and, in some cases, ITS rDNA sequence. Molecular characterization and phylogenetic analysis showed the presence of two ribotypes of A. tamarense: Group I (North American ribotype) and Group III (Western European ribotype). Assessment of PSP toxin production using hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) showed that A. tamarense Group I produced a complex array of toxins (∼2,000 fg STX equivalents · cell(-1) ) with the major toxins being C2, neosaxitoxin (NEO), saxitoxin (STX), gonyautoxin-4 (GTX-4), and GTX-3, while A. tamarense Group III did not produce toxins. Historically, it was considered that all Alexandrium species occurring in Scottish waters produce potent PSP toxins. This study has highlighted the presence of both PSP toxin-producing and benign species of A. tamarense and questions the ecological significance of this finding.

  20. Alexandrium fundyense cyst dynamics in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Anderson, Donald M.; Stock, Charles A.; Keafer, Bruce A.; Bronzino Nelson, Amy; Thompson, Brian; McGillicuddy, Dennis J.; Keller, Maureen; Matrai, Patricia A.; Martin, Jennifer

    2005-09-01

    The flux of cells from germinated cysts is critical in the population dynamics of many dinoflagellates. Here, data from a large-scale cyst survey are combined with surveys in other years to yield an Alexandrium fundyense cyst distribution map for the Gulf of Maine that is massive in geographic extent and cyst abundance. The benthic cyst population extends nearly 500 km alongshore. Embedded within it are several distinct accumulation zones or "seedbeds," each 3000-5000 km 2 in area. Maximal cyst abundances range from 2-20×10 6 cysts m -2. Cysts are equally or more abundant in deeper sediment layers; nearshore, cysts are fewer by a factor of 10 or more. This cyst distribution reflects sedimentary dynamics and the location of blooms in overlying surface waters. The flux of germinated cells from sediments was estimated using a combination of laboratory measurements of cyst germination and autofluorescence and observations of cyst autofluorescence in the field. These measurements constrained a germination function that, when applied to the cyst distribution map, provided an estimate of the germination inoculum for a physical/biological numerical model. In the laboratory studies, virtually all cysts incubated at different temperatures and light regimes became autofluorescent, but the rate of development was slower at lower temperatures, with no difference between light and dark incubations. Germination rates were highest at elevated temperatures, and were 2-fold greater in the light than in the dark. Laboratory and field fluorescence measurements suggest that>70% of the cysts in the top cm of sediment would germinate over a 60-90 day period in offshore waters. The combination of laboratory germination experiments and numerical modeling predicts nearly 100% germination of cysts in the top cm of sediment and resulting early season cell concentrations that are comparable in magnitude to observed cell distributions. It cannot account for late-season peaks in cell abundance

  1. Nutrient conditions during Alexandrium fundyense blooms in the western Gulf of Maine, USA

    NASA Astrophysics Data System (ADS)

    Love, Rebecca C.; Loder, Theodore C.; Keafer, Bruce A.

    2005-09-01

    Inorganic nutrients and organic nitrogen were measured in April-June of 1998 and 2000 near Casco Bay, Maine and the adjacent coastal waters as part of the Ecology and Oceanography of Harmful Algal Blooms—Gulf of Maine (ECOHAB-GOM) program. The samples were collected during development of toxic Alexandrium fundyense blooms [ Keafer, B.A., Churchill, J.H., Anderson, D.M., 2005. Blooms of the toxic dinoflagellate, Alexandrium fundyense in the Casco Bay region of the western Gulf of Maine: advection from offshore source populations and interactions with the Kennebec River plume. Deep Sea Research II, this issue [ doi:10.1016/j.dsr2.2005.06.017

  2. Warm temperature acclimation impacts metabolism of paralytic shellfish toxins from Alexandrium minutum in commercial oysters.

    PubMed

    Farrell, Hazel; Seebacher, Frank; O'Connor, Wayne; Zammit, Anthony; Harwood, D Tim; Murray, Shauna

    2015-09-01

    Species of Alexandrium produce potent neurotoxins termed paralytic shellfish toxins and are expanding their ranges worldwide, concurrent with increases in sea surface temperature. The metabolism of molluscs is temperature dependent, and increases in ocean temperature may influence both the abundance and distribution of Alexandrium and the dynamics of toxin uptake and depuration in shellfish. Here, we conducted a large-scale study of the effect of temperature on the uptake and depuration of paralytic shellfish toxins in three commercial oysters (Saccostrea glomerata and diploid and triploid Crassostrea gigas, n = 252 per species/ploidy level). Oysters were acclimated to two constant temperatures, reflecting current and predicted climate scenarios (22 and 27 °C), and fed a diet including the paralytic shellfish toxin-producing species Alexandrium minutum. While the oysters fed on A. minutum in similar quantities, concentrations of the toxin analogue GTX1,4 were significantly lower in warm-acclimated S. glomerata and diploid C. gigas after 12 days. Following exposure to A. minutum, toxicity of triploid C. gigas was not affected by temperature. Generally, detoxification rates were reduced in warm-acclimated oysters. The routine metabolism of the oysters was not affected by the toxins, but a significant effect was found at a cellular level in diploid C. gigas. The increasing incidences of Alexandrium blooms worldwide are a challenge for shellfish food safety regulation. Our findings indicate that rising ocean temperatures may reduce paralytic shellfish toxin accumulation in two of the three oyster types; however, they may persist for longer periods in oyster tissue.

  3. Circannual excystment of resting cysts of Alexandrium spp. from eastern Gulf of Maine populations

    NASA Astrophysics Data System (ADS)

    Matrai, P.; Thompson, B.; Keller, M.

    2005-09-01

    Species of the marine dinoflagellate Alexandrium, present in most of the Gulf of Maine (GOM), Bay of Fundy and Gulf of St. Lawrence as well as in many other areas of the world, are known to cause toxicity to marine organisms and humans alike. Excystment of Alexandrium fundyense from the eastern region of the GOM (Penobscot Bay to Bay of Fundy) was followed through four germination cycles (4 years). An annual, free-running oscillation in germination was observed under constant environmental conditions, indicating control by an endogenous clock for these eastern cysts, as shown earlier for cysts from the western region of the GOM. This circannual endogenous clock had an average period of 11 months. The phase of germination remained constant for cysts from all three stations sampled. Cysts did not germinate, despite favorable growth conditions, in summer-to-fall and this timing was consistent among cysts from all stations. The timing of cyst germination is highly relevant to modeling of Alexandrium sp. bloom initiation and depletion, as there are cyst "seed beds" near shore and offshore in the eastern and western regions of the GOM.

  4. Mechanisms of inorganic carbon acquisition in two estuarine Rhodophyceans: Bostrychia scorpioides (Hudson) ex Kützing Montagne and Catenella caespitosa (Withering) L. M. Irvine.

    PubMed

    Ruiz-Nieto, Miriam; Fernández, José A; Niell, F Xavier; Carmona, Raquel

    2014-09-01

    Marine macroalgae possess a range of mechanisms to increase the availability of CO2 for fixation by ribulose-1,5-bisphosphate carboxylase/oxygenase. Of these, possession of a periplasmic or external carbonic anhydrase and the ability to use bicarbonate ions is widely distributed. The mechanisms of carbon acquisition were studied in two estuarine red macroalgae Bostrychia scorpioides and Catenella caespitosa using a range of techniques. pH-drift and CO2-depletion experiments at constant pH suggested that CO2 is the main source of inorganic carbon in both species. Inhibitors indicated that internal and external carbonic anhydrase were present in both species. Inhibitors also suggested that uptake of bicarbonate is unlikely to be present (P < 0.05).

  5. Diel vertical distributions of the red tide dinoflagellate Alexandrium fundyense in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Townsend, David W.; Bennett, Stephanie L.; Thomas, Maura A.

    2005-09-01

    Two 24-h experiments, designed to test whether the toxic dinoflagellate Alexandrium fundyense exhibited significant changes in vertical distribution, were performed in offshore waters of the Gulf of Maine in June 2000. Standard hydrocasts with a CTD/carousel water sampler package were made hourly on-station while following a drogue set at 25 m depth. Continuous measurements of T, S, and chlorophyll fluorescence were made on each down cast, and discrete water samples were collected at 5-m intervals from 55 m depth to the surface on the up cast, for analyses of extracted phytoplankton chlorophyll, inorganic nutrients and cell densities of Alexandrium In the first experiment we observed a bimodal vertical distribution of cells, with relatively high cell densities near the surface (<15 m depth) and a second peak of relatively high cell densities at depths between 25 and 40 m, coincident with the depth of the pycnocline and nitricline. Internal waves of 10-15 m amplitude appeared to exert control over the depth distribution of the deep population. Approximately 12 h into the first experiment, a relatively warm surface water mass with low Alexandrium cell densities intruded over the drogue station, leaving only the deep population. In the second experiment overall cell densities of Alexandrium were much lower, but again we observed initially a bimodal depth distribution of cells. As in the first experiment, the surface population effectively disappeared after a few hours, leaving only the deep population; in this case, however, there was some evidence of an initial downward movement of the surface population prior to its complete disappearance. Evidence for intrusion of a surface-water layer was not as clear in the second experiment as in the first. In addition to higher-frequency internal waves, as was observed in the first experiment, we also observed a low-frequency internal tidal wave of greater than 20 m amplitude that controlled the vertical distribution of the

  6. Interannual variability of Alexandrium fundyense abundance and shellfish toxicity in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    McGillicuddy, D. J.; Anderson, D. M.; Solow, A. R.; Townsend, D. W.

    2005-09-01

    Six years of oceanographic surveys of Alexandrium fundyense concentrations in the Gulf of Maine are combined with shellfish toxicity records from coastal monitoring stations to assess covariations of these quantities on seasonal to interannual time scales. Annual mean gulf-wide cell abundance varies by less than one order of magnitude during the time interval examined (1993-2002). Fluctuations in gulf-wide annual mean cell abundance and shellfish toxicity are not related in a consistent manner. This suggests that interannual variations in toxicity may be regulated by transport and delivery of offshore cell populations, rather than the absolute abundance of the source populations themselves.

  7. Genome Sequence of Bacillus sp. Strain UMTAT18 Isolated from the Dinoflagellate Alexandrium tamiyavanichii Found in the Straits of Malacca

    PubMed Central

    Ming, Gan Han; Mohd Noor, Mohd Ezhar; Sung, Yeong Yik; Usup, Gires

    2016-01-01

    Bacillus sp. strain UMTAT18 was isolated from the harmful dinoflagellate Alexandrium tamiyavanichii. Its genome consists of 5,479,367 bp with 5,546 open reading frames, 102 tRNAs, and 29 rRNAs. Gene clusters for biosynthesis of nonribosomal peptides, bacteriocin, and lantipeptide were identified. It also contains siderophore and genes related to stress tolerance. PMID:27795265

  8. Transcriptomic profiling of Alexandrium fundyense during physical interaction with or exposure to chemical signals from the parasite Amoebophrya.

    PubMed

    Lu, Yameng; Wohlrab, Sylke; Groth, Marco; Glöckner, Gernot; Guillou, Laure; John, Uwe

    2016-03-01

    Toxic microalgae have their own pathogens, and understanding the way in which these microalgae respond to antagonistic attacks may provide information about their capacity to persist during harmful algal bloom events. Here, we compared the effects of the physical presence of the parasite Amoebophrya sp. and exposure to waterborne cues from cultures infected with this parasite, on gene expression by the toxic dinoflagellates, Alexandrium fundyense. Compared with control samples, a total of 14,882 Alexandrium genes were differentially expressed over the whole-parasite infection cycle at three different time points (0, 6 and 96 h). RNA sequencing analyses indicated that exposure to the parasite and parasitic waterborne cues produced significant changes in the expression levels of Alexandrium genes associated with specific metabolic pathways. The observed upregulation of genes associated with glycolysis, the tricarboxylic acid cycle, fatty acid β-oxidation, oxidative phosphorylation and photosynthesis suggests that parasite infection increases the energy demand of the host. The observed upregulation of genes correlated with signal transduction indicates that Alexandrium could be sensitized by parasite attacks. This response might prime the defence of the host, as indicated by the increased expression of several genes associated with defence and stress. Our findings provide a molecular overview of the response of a dinoflagellate to parasite infection.

  9. Comparative gene expression in toxic versus non-toxic strains of the marine dinoflagellate Alexandrium minutum

    PubMed Central

    2010-01-01

    Background The dinoflagellate Alexandrium minutum typically produces paralytic shellfish poisoning (PSP) toxins, which are known only from cyanobacteria and dinoflagellates. While a PSP toxin gene cluster has recently been characterized in cyanobacteria, the genetic background of PSP toxin production in dinoflagellates remains elusive. Results We constructed and analysed an expressed sequence tag (EST) library of A. minutum, which contained 15,703 read sequences yielding a total of 4,320 unique expressed clusters. Of these clusters, 72% combined the forward-and reverse reads of at least one bacterial clone. This sequence resource was then used to construct an oligonucleotide microarray. We analysed the expression of all clusters in three different strains. While the cyanobacterial PSP toxin genes were not found among the A. minutum sequences, 192 genes were differentially expressed between toxic and non-toxic strains. Conclusions Based on this study and on the lack of identified PSP synthesis genes in the two existent Alexandrium tamarense EST libraries, we propose that the PSP toxin genes in dinoflagellates might be more different from their cyanobacterial counterparts than would be expected in the case of a recent gene transfer. As a starting point to identify possible PSP toxin-associated genes in dinoflagellates without relying on a priori sequence information, the sequences only present in mRNA pools of the toxic strain can be seen as putative candidates involved in toxin synthesis and regulation, or acclimation to intracellular PSP toxins. PMID:20403159

  10. Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens

    PubMed Central

    Hartmann, Anja; Becker, Kathrin; Karsten, Ulf; Remias, Daniel; Ganzera, Markus

    2015-01-01

    Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R2 > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline. PMID:26473886

  11. Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens.

    PubMed

    Hartmann, Anja; Becker, Kathrin; Karsten, Ulf; Remias, Daniel; Ganzera, Markus

    2015-10-09

    Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R² > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline.

  12. Spatial and temporal variability of Alexandrium cyst fluxes in the Gulf of Maine: Relationship to seasonal particle export and resuspension

    NASA Astrophysics Data System (ADS)

    Pilskaln, C. H.; Anderson, D. M.; McGillicuddy, D. J.; Keafer, B. A.; Hayashi, K.; Norton, K.

    2014-05-01

    Quantification of Alexandrium cyst fluxes through the Gulf of Maine water column is central to understanding the linkage between the source and fate of annual Alexandrium blooms in the offshore waters. These blooms often lead to paralytic shellfish poisoning (PSP) and extensive closures of shellfish beds. We report here on time-series sediment trap deployments completed at four offshore locations in the gulf between 2005 and 2010 as components of two ECOHAB-GOM field programs. Data presented documents the substantial spatial and temporal fluctuations in Alexandrium fundyense cyst fluxes in the gulf. Cyst delivery out of the euphotic zone peaked primarily between July and August following annual spring-summer Alexandrium blooms and was greatest in the western gulf. At all sites, cyst flux maxima to the subsurface waters were rarely coincident with seasonal peaks in the total mass export of particulate material indicating that cyst delivery was primarily via individually sinking cysts. Where persistent benthic nepheloid layers (BNLs) exist, significant sediment resuspension input of cysts to the near-bottom water column was evidenced by deep cyst fluxes that were up to several orders of magnitude greater than that measured above the BNL. The largest cyst fluxes in the BNL were observed in the eastern gulf, suggesting greater resuspension energy and BNL cyst inventories in this region. Temporal similarities between peak cyst export out of the upper ocean and peak cyst fluxes in the BNL were observed and document the contribution of seasonal, newly formed cysts to the BNL. The data however also suggest that many Alexandrium cells comprising the massive, short-lived blooms do not transition into cysts. Time-series flow measurements and a simple 1D model demonstrate that the BNL cyst fluxes reflect the combined effects of tidal energy-maintained resuspension, deposition, and input of cysts from the overlying water column.

  13. Spatial and temporal variability of Alexandrium cyst fluxes in the Gulf of Maine: Relationship to seasonal particle export and resuspension

    PubMed Central

    Pilskaln, C.H.; Anderson, D.M.; McGillicuddy, D.J.; Keafer, B.A.; Hayashi, K.; Norton, K.

    2014-01-01

    Quantification of Alexandrium cyst fluxes through the Gulf of Maine water column is central to understanding the linkage between the source and fate of annual Alexandrium blooms in the offshore waters. These blooms often lead to paralytic shellfish poisoning (PSP) and extensive closures of shellfish beds. We report here on time-series sediment trap deployments completed at four offshore locations in the gulf between 2005 and 2010 as components of two ECOHAB–GOM field programs. Data presented documents the substantial spatial and temporal fluctuations in Alexandrium fundyense cyst fluxes in the gulf. Cyst delivery out of the euphotic zone peaked primarily between July and August following annual spring–summer Alexandrium blooms and was greatest in the western gulf. At all sites, cyst flux maxima to the subsurface waters were rarely coincident with seasonal peaks in the total mass export of particulate material indicating that cyst delivery was primarily via individually sinking cysts. Where persistent benthic nepheloid layers (BNLs) exist, significant sediment resuspension input of cysts to the near-bottom water column was evidenced by deep cyst fluxes that were up to several orders of magnitude greater than that measured above the BNL. The largest cyst fluxes in the BNL were observed in the eastern gulf, suggesting greater resuspension energy and BNL cyst inventories in this region. Temporal similarities between peak cyst export out of the upper ocean and peak cyst fluxes in the BNL were observed and document the contribution of seasonal, newly formed cysts to the BNL. The data however also suggest that many Alexandrium cells comprising the massive, short-lived blooms do not transition into cysts. Time-series flow measurements and a simple 1D model demonstrate that the BNL cyst fluxes reflect the combined effects of tidal energy-maintained resuspension, deposition, and input of cysts from the overlying water column. PMID:25431527

  14. Spatial and temporal variability of Alexandrium cyst fluxes in the Gulf of Maine: Relationship to seasonal particle export and resuspension.

    PubMed

    Pilskaln, C H; Anderson, D M; McGillicuddy, D J; Keafer, B A; Hayashi, K; Norton, K

    2014-05-01

    Quantification of Alexandrium cyst fluxes through the Gulf of Maine water column is central to understanding the linkage between the source and fate of annual Alexandrium blooms in the offshore waters. These blooms often lead to paralytic shellfish poisoning (PSP) and extensive closures of shellfish beds. We report here on time-series sediment trap deployments completed at four offshore locations in the gulf between 2005 and 2010 as components of two ECOHAB-GOM field programs. Data presented documents the substantial spatial and temporal fluctuations in Alexandrium fundyense cyst fluxes in the gulf. Cyst delivery out of the euphotic zone peaked primarily between July and August following annual spring-summer Alexandrium blooms and was greatest in the western gulf. At all sites, cyst flux maxima to the subsurface waters were rarely coincident with seasonal peaks in the total mass export of particulate material indicating that cyst delivery was primarily via individually sinking cysts. Where persistent benthic nepheloid layers (BNLs) exist, significant sediment resuspension input of cysts to the near-bottom water column was evidenced by deep cyst fluxes that were up to several orders of magnitude greater than that measured above the BNL. The largest cyst fluxes in the BNL were observed in the eastern gulf, suggesting greater resuspension energy and BNL cyst inventories in this region. Temporal similarities between peak cyst export out of the upper ocean and peak cyst fluxes in the BNL were observed and document the contribution of seasonal, newly formed cysts to the BNL. The data however also suggest that many Alexandrium cells comprising the massive, short-lived blooms do not transition into cysts. Time-series flow measurements and a simple 1D model demonstrate that the BNL cyst fluxes reflect the combined effects of tidal energy-maintained resuspension, deposition, and input of cysts from the overlying water column.

  15. The effects of elevated CO2 on the growth and toxicity of field populations and cultures of the saxitoxin-producing dinoflagellate, Alexandrium fundyense

    PubMed Central

    Hattenrath-Lehmann, Theresa K.; Smith, Juliette L.; Wallace, Ryan B.; Merlo, Lucas; Koch, Florian; Mittelsdorf, Heidi; Goleski, Jennifer A.; Anderson, Donald M.; Gobler, Christopher J.

    2016-01-01

    The effects of coastal acidification on the growth and toxicity of the saxitoxin-producing dinoflagellate Alexandrium fundyense were examined in culture and ecosystem studies. In culture experiments, Alexandrium strains isolated from Northport Bay NY, USA, and the Bay of Fundy, Canada, grew significantly faster (16 -190%; p<0.05) when exposed to elevated levels of pCO2 (~ 800- 1900μatm) compared to lower levels (~390μatm). Exposure to higher levels of pCO2 also resulted in significant increases (71 – 81%) in total cellular toxicity (fg STX eq. cell−1) in the Northport Bay strain, while no changes in toxicity were detected in the Bay of Fundy strain. The positive relationship between pCO2 enhancement and elevated growth was reproducible using natural populations from Northport; Alexandrium densities were significantly and consistently enhanced when natural populations were incubated at 1500 μatm pCO2, a value at the upper range of those recorded in Northport Bay, 390 – 1500 µatm. During natural Alexandrium blooms in Northport Bay, pCO2 concentrations increased over the course of a bloom to more than 1700μatm and were highest in regions with the greatest Alexandrium abundances, suggesting Alexandrium may be further exacerbating acidification or be especially adapted to these extreme, acidified conditions. The co-occurrence of Alexandrium blooms and elevated pCO2 represents a previously unrecognized, compounding environmental threat to coastal ecosystems. The ability of elevated pCO2 to enhance the growth and toxicity of Alexandrium indicates that acidification promoted by eutrophication or climate change can intensify these, and perhaps other, harmful algal blooms. PMID:27721521

  16. Transcriptomic responses of the calanoid copepod Calanus finmarchicus to the saxitoxin producing dinoflagellate Alexandrium fundyense

    NASA Astrophysics Data System (ADS)

    Roncalli, Vittoria; Cieslak, Matthew C.; Lenz, Petra H.

    2016-05-01

    In the Gulf of Maine, the copepod Calanus finmarchicus co-occurs with the neurotoxin-producing dinoflagellate, Alexandrium fundyense. The copepod is resistant to this toxic alga, but little is known about other effects. Gene expression profiles were used to investigate the physiological response of females feeding for two and five days on a control diet or a diet containing either a low or a high dose of A. fundyense. The physiological responses to the two experimental diets were similar, but changed between the time points. At 5-days the response was characterized by down-regulated genes involved in energy metabolism. Detoxification was not a major component of the response. Instead, genes involved in digestion were consistently regulated, suggesting that food assimilation may have been affected. Thus, predicted increases in the frequency of blooms of A. fundyense could affect C. finmarchicus populations by changing the individuals’ energy budget and reducing their ability to build lipid reserves.

  17. Direct and fast detection of Alexandrium minutum algae by using high frequency microbalance.

    PubMed

    Sousa, Célia; Compère, Chantal; Dreanno, Catherine; Crassous, Marie-Pierre; Gas, Fabienne; Baus, Beatrice; Perrot, Hubert

    2014-09-01

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

  18. Thiol- and biotin-labeled probes for oligonucleotide quartz crystal microbalance biosensors of microalga alexandrium minutum.

    PubMed

    Lazerges, Mathieu; Perrot, Hubert; Rabehagasoa, Niriniony; Compère, Chantal

    2012-07-04

    Two quartz crystal microbalance oligonucleotide biosensors of a toxic microalga gene sequence (Alexandrium Minutum) have been designed. Grafting on a gold surface of 20-base thiol- or biotin-labeled probe, and selective hybridization with the complementary 20-base target, have been monitored in situ with a 27 MHz quartz crystal microbalance under controlled hydrodynamic conditions. The frequency of the set up is stable to within a few hertz, corresponding to the nanogram scale, for three hour experiments. DNA recognition by the two biosensors is efficient and selective. Hybridization kinetic curves indicate that the biosensor designed with the thiol-labeled probe is more sensitive, and that the biosensor designed with the biotin-labeled probe has a shorter time response and a higher hybridization efficiency.

  19. Thiol- and Biotin-Labeled Probes for Oligonucleotide Quartz Crystal Microbalance Biosensors of Microalga Alexandrium Minutum

    PubMed Central

    Lazerges, Mathieu; Perrot, Hubert; Rabehagasoa, Niriniony; Compère, Chantal

    2012-01-01

    Two quartz crystal microbalance oligonucleotide biosensors of a toxic microalga gene sequence (Alexandrium Minutum) have been designed. Grafting on a gold surface of 20-base thiol- or biotin-labeled probe, and selective hybridization with the complementary 20-base target, have been monitored in situ with a 27 MHz quartz crystal microbalance under controlled hydrodynamic conditions. The frequency of the set up is stable to within a few hertz, corresponding to the nanogram scale, for three hour experiments. DNA recognition by the two biosensors is efficient and selective. Hybridization kinetic curves indicate that the biosensor designed with the thiol-labeled probe is more sensitive, and that the biosensor designed with the biotin-labeled probe has a shorter time response and a higher hybridization efficiency. PMID:25585927

  20. Rapid detection and quantification of the marine toxic algae, Alexandrium minutum, using a super-paramagnetic immunochromatographic strip test.

    PubMed

    Gas, Fabienne; Baus, Béatrice; Queré, Julien; Chapelle, Annie; Dreanno, Catherine

    2016-01-15

    The dinoflagellates of Alexandrium genus are known to be producers of paralytic shellfish toxins that regularly impact the shellfish aquaculture industry and fisheries. Accurate detection of Alexandrium including Alexandrium minutum is crucial for environmental monitoring and sanitary issues. In this study, we firstly developed a quantitative lateral flow immunoassay (LFIA) using super-paramagnetic nanobeads for A. minutum whole cells. This dipstick assay relies on two distinct monoclonal antibodies used in a sandwich format and directed against surface antigens of this organism. No sample preparation is required. Either frozen or live cells can be detected and quantified. The specificity and sensitivity are assessed by using phytoplankton culture and field samples spiked with a known amount of cultured A. minutum cells. This LFIA is shown to be highly specific for A. minutum and able to detect reproducibly 10(5)cells/L within 30min. The test is applied to environmental samples already characterized by light microscopy counting. No significant difference is observed between the cell densities obtained by these two methods. This handy super-paramagnetic lateral flow immnunoassay biosensor can greatly assist water quality monitoring programs as well as ecological research.

  1. Molecular analysis of complete ssu to lsu rdna sequence in the harmful dinoflagellate alexandrium tamarense (korean isolate, HY970328M)

    NASA Astrophysics Data System (ADS)

    Ki, Jang-Seu; Han, Myung-Soo

    2005-09-01

    New PCR primers (N=18) were designed for the isolation of complete SSU to LSU rDNA sequences from the dinoflagellate Alexandrium tamarense. Standard PCR, employing each primer set selected for amplifications of less than 1.5 kb, successfully amplified the expected rDNA regions of A. tamarense (Korean isolate, HY970328M). Complete SSU, LSU rDNAs and ITS sequences, including 5.8S rDNA, were recorded at 1,800 bp, 520 bp and 3,393 bp, respectively. The LSU rDNA sequence was the first report in Alexandrium genus. No intron was found in the LSU rRNA coding region. Twelve D-domains within the LSU rDNA were put together into 1,879 bp (44.4% G+C), and cores into 1514 bp (42.8% G+C). The core sequence was significantly different (0.0867 of genetic distance, 91% sequence similarity) in comparison with Prorocentrum micans (GenBank access. no. X16108). The D2 region was the longest in length (300 bp) and highly variable among the 12 D-domains. In a phylogenetic analysis using complete LSU rDNA sequences of a variety of phytoplankton, A tamarense was clearly separated with high resolution against other species. The result suggests that the sequence may resolve the taxonomic ambiguities of Alexandrium genus, particularly of the tamarensis complex.

  2. A red tide of Alexandrium fundyense in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    McGillicuddy, D. J.; Brosnahan, M. L.; Couture, D. A.; He, R.; Keafer, B. A.; Manning, J. P.; Martin, J. L.; Pilskaln, C. H.; Townsend, D. W.; Anderson, D. M.

    2014-05-01

    In early July 2009, an unusually high concentration of the toxic dinoflagellate Alexandrium fundyense occurred in the western Gulf of Maine, causing surface waters to appear reddish brown to the human eye. The discolored water appeared to be the southern terminus of a large-scale event that caused shellfish toxicity along the entire coast of Maine to the Canadian border. Rapid-response shipboard sampling efforts together with satellite data suggest the water discoloration in the western Gulf of Maine was a highly ephemeral feature of less than two weeks in duration. Flow cytometric analysis of surface samples from the red water indicated the population was undergoing sexual reproduction. Cyst fluxes downstream of the discolored water were the highest ever measured in the Gulf of Maine, and a large deposit of new cysts was observed that fall. Although the mechanisms causing this event remain unknown, its timing coincided with an anomalous period of downwelling-favorable winds that could have played a role in aggregating upward-swimming cells. Regardless of the underlying causes, this event highlights the importance of short-term episodic phenomena on regional population dynamics of A. fundyense.

  3. Toxic dinoflagellate Alexandrium tamarense induces oxidative stress and apoptosis in hepatopancreas of shrimp ( Fenneropenaeus chinensis)

    NASA Astrophysics Data System (ADS)

    Liang, Zhongxiu; Li, Jian; Li, Jitao; Tan, Zhijun; Ren, Hai; Zhao, Fazhen

    2014-12-01

    This study investigated the inductive effect of Alexandrium tamarense, a toxic dinoflagellate producing paralytic shellfish poison, on oxidative stress and apoptosis in hepatopancreas of Chinese shrimp, Fenneropenaeus chinensis. The individuals of F. chinensis were exposed to 200 and 1000 cells mL-1 of A. tamarense with their superoxide dismutase (SOD), glutathione S-transferase (GST) activities, malonyldialdehyde (MDA) concentration, and caspase gene ( FcCasp) expression in hepatopancreas determined at 12, 24, 48, 72 and 96 h. In addition, apoptosis in hepatopancreas of F. chinensis at 96 h after exposure was determined through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. The hepatopancreatic SOD and GST activities of F. chinensis exposed to 1000 cells mL-1 of A. tamarense showed a bell-shaped response to exposure time. The hepatopancreatic MDA concentration of F. chinensis exposed to 1000 cells mL-1 of A. tamarense increased gradually from 48 to 96 h, and such a trend corresponded to the decrease of GST activity. The hepatopancreatic FcCasp transcript abundance of F. chinensis exposed to 1000 cells mL-1 of A. tamarense was positively and linearly correlated to MDA concentration. Results of TUNEL assay showed that exposure to 1000 cells mL-1 of A. tamarense induced apoptosis in the hepatopancreas of F. chinensis. Our study revealed that A. tamarense exposure influenced the antioxidative status of F. chinensis and caused lipid peroxidation and apoptosis in the hepatopancreas of shrimp.

  4. A red tide of Alexandrium fundyense in the Gulf of Maine

    PubMed Central

    McGillicuddy, D.J.; Brosnahan, M.L.; Couture, D.A.; He, R.; Keafer, B.A.; Manning, J.P.; Martin, J.L.; Pilskaln, C.H.; Townsend, D.W.; Anderson, D.M.

    2013-01-01

    In early July 2009, an unusually high concentration of the toxic dinoflagellate Alexandrium fundyense occurred in the western Gulf of Maine, causing surface waters to appear reddish brown to the human eye. The discolored water appeared to be the southern terminus of a large-scale event that caused shellfish toxicity along the entire coast of Maine to the Canadian border. Rapid-response shipboard sampling efforts together with satellite data suggest the water discoloration in the western Gulf of Maine was a highly ephemeral feature of less than two weeks in duration. Flow cytometric analysis of surface samples from the red water indicated the population was undergoing sexual reproduction. Cyst fluxes downstream of the discolored water were the highest ever measured in the Gulf of Maine, and a large deposit of new cysts was observed that fall. Although the mechanisms causing this event remain unknown, its timing coincided with an anomalous period of downwelling-favorable winds that could have played a role in aggregating upward-swimming cells. Regardless of the underlying causes, this event highlights the importance of short-term episodic phenomena on regional population dynamics of A. fundyense. PMID:25170191

  5. Rapid growth and concerted sexual transitions by a bloom of the harmful dinoflagellate Alexandrium fundyense (Dinophyceae)

    PubMed Central

    Velo‐Suárez, Lourdes; Ralston, David K.; Fox, Sophia E.; Sehein, Taylor R.; Shalapyonok, Alexi; Sosik, Heidi M.; Olson, Robert J.; Anderson, Donald M.

    2015-01-01

    Abstract Transitions between life cycle stages by the harmful dinoflagellate Alexandrium fundyense are critical for the initiation and termination of its blooms. To quantify these transitions in a single population, an Imaging FlowCytobot (IFCB), was deployed in Salt Pond (Eastham, Massachusetts), a small, tidally flushed kettle pond that hosts near annual, localized A. fundyense blooms. Machine‐based image classifiers differentiating A. fundyense life cycle stages were developed and results were compared to manually corrected IFCB samples, manual microscopy‐based estimates of A. fundyense abundance, previously published data describing prevalence of the parasite Amoebophrya, and a continuous culture of A. fundyense infected with Amoebophrya. In Salt Pond, a development phase of sustained vegetative division lasted approximately 3 weeks and was followed by a rapid and near complete conversion to small, gamete cells. The gametic period (∼3 d) coincided with a spike in the frequency of fusing gametes (up to 5% of A. fundyense images) and was followed by a zygotic phase (∼4 d) during which cell sizes returned to their normal range but cell division and diel vertical migration ceased. Cell division during bloom development was strongly phased, enabling estimation of daily rates of division, which were more than twice those predicted from batch cultures grown at similar temperatures in replete medium. Data from the Salt Pond deployment provide the first continuous record of an A. fundyense population through its complete bloom cycle and demonstrate growth and sexual induction rates much higher than are typically observed in culture. PMID:27667858

  6. Genomic Insights into Processes Driving the Infection of Alexandrium tamarense by the Parasitoid Amoebophrya sp.

    PubMed Central

    Wohlrab, Sylke; Glöckner, Gernot; Guillou, Laure; John, Uwe

    2014-01-01

    The regulatory circuits during infection of dinoflagellates by their parasites are largely unknown on the molecular level. Here we provide molecular insights into these infection dynamics. Alexandrium tamarense is one of the most prominent harmful algal bloom dinoflagellates. Its pathogen, the dinoflagellate parasitoid Amoebophrya sp., has been observed to infect and control the blooms of this species. We generated a data set of transcripts from three time points (0, 6, and 96 h) during the infection of this parasite-host system. Assembly of all transcript data from the parasitoid (>900,000 reads/313 Mbp with 454/Roche next-generation sequencing [NGS]) yielded 14,455 contigs, to which we mapped the raw transcript reads of each time point of the infection cycle. We show that particular surface lectins are expressed at the beginning of the infection cycle which likely mediate the attachment to the host cell. In a later phase, signal transduction-related genes together with transmembrane transport and cytoskeleton proteins point to a high integration of processes involved in host recognition, adhesion, and invasion. At the final maturation stage, cell division- and proliferation-related genes were highly expressed, reflecting the fast cell growth and nuclear division of the parasitoid. Our molecular insights into dinoflagellate parasitoid interactions point to general mechanisms also known from other eukaryotic parasites, especially from the Alveolata. These similarities indicate the presence of fundamental processes of parasitoid infection that have remained stable throughout evolution within different phyla. PMID:25239978

  7. ANALYSIS OF ALEXANDRIUM TAMARENSE (DINOPHYCEAE) GENES REVEALS THE COMPLEX EVOLUTIONARY HISTORY OF A MICROBIAL EUKARYOTE1

    PubMed Central

    Chan, Cheong Xin; Soares, Marcelo B.; Bonaldo, Maria F.; Wisecaver, Jennifer H.; Hackett, Jeremiah D.; Anderson, Donald M.; Erdner, Deana L.; Bhattacharya, Debashish

    2012-01-01

    Microbial eukaryotes may extinguish much of their nuclear phylogenetic history due to endosymbiotic/horizontal gene transfer (E/HGT). We studied E/HGT in 32,110 contigs of expressed sequence tags (ESTs) from the dinoflagellate Alexandrium tamarense (Dinophyceae) using a conservative phylogenomic approach. The vast majority of predicted proteins (86.4%) in this alga are novel or dinoflagellate-specific. We searched for putative homologs of these predicted proteins against a taxonomically broadly sampled protein database that includes all currently available data from algae and protists and reconstructed a phylogeny from each of the putative homologous protein sets. Of the 2,523 resulting phylogenies, 14-17% are potentially impacted by E/HGT involving both prokaryote and eukaryote lineages, with 2-4% showing clear evidence of reticulate evolution. The complex evolutionary histories of the remaining proteins, many of which may also have been affected by E/HGT, cannot be interpreted using our approach with currently available gene data. We present empirical evidence of reticulate genome evolution that combined with inadequate or highly complex phylogenetic signal in many proteins may impede genome-wide approaches to infer the tree of microbial eukaryotes. PMID:23066170

  8. A red tide of Alexandrium fundyense in the Gulf of Maine.

    PubMed

    McGillicuddy, D J; Brosnahan, M L; Couture, D A; He, R; Keafer, B A; Manning, J P; Martin, J L; Pilskaln, C H; Townsend, D W; Anderson, D M

    2014-05-01

    In early July 2009, an unusually high concentration of the toxic dinoflagellate Alexandrium fundyense occurred in the western Gulf of Maine, causing surface waters to appear reddish brown to the human eye. The discolored water appeared to be the southern terminus of a large-scale event that caused shellfish toxicity along the entire coast of Maine to the Canadian border. Rapid-response shipboard sampling efforts together with satellite data suggest the water discoloration in the western Gulf of Maine was a highly ephemeral feature of less than two weeks in duration. Flow cytometric analysis of surface samples from the red water indicated the population was undergoing sexual reproduction. Cyst fluxes downstream of the discolored water were the highest ever measured in the Gulf of Maine, and a large deposit of new cysts was observed that fall. Although the mechanisms causing this event remain unknown, its timing coincided with an anomalous period of downwelling-favorable winds that could have played a role in aggregating upward-swimming cells. Regardless of the underlying causes, this event highlights the importance of short-term episodic phenomena on regional population dynamics of A. fundyense.

  9. 2D-PAGE protein analysis of dinoflagellate Alexandrium minutum based on three different temperatures

    NASA Astrophysics Data System (ADS)

    Latib, Norhidayu Abdul; Norshaha, Safida Anira; Usup, Gires; Yusof, Nurul Yuziana Mohd

    2015-09-01

    Harmful algae bloom or red tide seems to be considered as threat to ecosystem, especially to human consumption because of the production of neurotoxin by dinoflagellates species such as Alexandrium minutum which can lead to paralytic shellfish poisoning. The aim of this study is to determine the most suitable method for protein extraction of A. minutum followed by determination of differential protein expression of A. minutum on three different temperatures (15°C, 26°C and 31.5°C). After the optimization, the protein extract was subjected to two-dimensional polyacrylamide gel electrophoresis (2-DE) to compare the intensity and distribution of the protein spots. Based on quantitative and qualitative protein assessment, use of Trizol reagent is the most suitable method to extract protein from A. minutum. 2-DE analysis of the samples results in different distribution and intensity of the protein spots were compared between 15°C, 26°C and 31.5°C.

  10. Distribution of Alexandrium fundyense (Dinophyceae) cysts in Greenland and Iceland, with an emphasis on viability and growth in the Arctic

    PubMed Central

    Richlen, Mindy L.; Zielinski, Oliver; Holinde, Lars; Tillmann, Urban; Cembella, Allan; Lyu, Yihua; Anderson, Donald M.

    2016-01-01

    The bloom-forming dinoflagellate Alexandrium fundyense has been extensively studied due its toxin-producing capabilities and consequent impacts to human health and economies. This study investigated the prevalence of resting cysts of A. fundyense in western Greenland and Iceland to assess the historical presence and magnitude of bloom populations in the region, and to characterize environmental conditions during summer, when bloom development may occur. Analysis of sediments collected from these locations showed that Alexandrium cysts were present at low to moderate densities in most areas surveyed, with highest densities observed in western Iceland. Additionally, laboratory experiments were conducted on clonal cultures established from isolated cysts or vegetative cells from Greenland, Iceland, and the Chukchi Sea (near Alaska) to examine the effects of photoperiod interval and irradiance levels on growth. Growth rates in response to the experimental treatments varied among isolates, but were generally highest under conditions that included both the shortest photoperiod interval (16h:8h light:dark) and higher irradiance levels (~146–366 μmol photons m−2 s−1), followed by growth under an extended photoperiod interval and low irradiance level (~37 μmol photons m−2 s−1). Based on field and laboratory data, we hypothesize that blooms in Greenland are primarily derived from advected Alexandrium populations, as low bottom temperatures and limited light availability would likely preclude in situ bloom development. In contrast, the bays and fjords in Iceland may provide more favorable habitat for germling cell survival and growth, and therefore may support indigenous, self-seeding blooms. PMID:27721528

  11. Toxin variability in natural populations of Alexandrium fundyense in Casco Bay, Maine—evidence of nitrogen limitation

    NASA Astrophysics Data System (ADS)

    Poulton, N. J.; Keafer, B. A.; Anderson, D. M.

    2005-09-01

    The dinoflagellate Alexandrium fundyense is a common, recurring harmful algal bloom (HAB) species in the Gulf of Maine. To date, most physiological measurements of phytoplankton in the field provide data on the entire community, yet efforts to obtain species-specific data are particularly important for understanding the ecological and physiological dynamics of HAB species, such as, Alexandrium. Alexandrium spp., do not usually dominate the planktonic community in the Gulf of Maine, but are of great interest due to the potent toxins produced. In order to determine the nutritional status of Alexandrium spp. in natural populations, indicators of nutrient deprivation need to be identified that are specific to that one species. To date, the saxitoxin content of A. fundyense is known to vary under different environmental conditions such as nitrogen and phosphorous limitation. However, in batch culture the composition of the toxin (the relative amounts of each saxitoxin derivative per cell) appears to be a stable quantity and thus is sometimes viewed as a biochemical marker of individual strains. In more recent studies, toxin composition has been shown to vary during progressive N- and P- limitation, once the cells are given time to achieve steady state in semi-continuous, nutrient-limited cultures. Using both the absolute toxin concentrations and relative proportion (mole % total toxin) of each toxin derivative, N- and P-limitation can be distinguished based on the observed trends in the different saxitoxin derivatives. In this study, we examine the toxin content and composition in natural A. fundyense populations during a spring bloom in Casco Bay, ME from April-June of 1998. This allows us to examine whether A. fundyense populations in the western Gulf of Maine are sufficiently homogenous to permit the detection of toxin composition and toxin content differences through time and space, and if so, to determine whether those changes are indicative of a particular

  12. Benthic nepheloid layers in the Gulf of Maine and Alexandrium cyst inventories

    PubMed Central

    Pilskaln, C.H.; Hayashi, K.; Keafer, B.A.; Anderson, D.M.; McGillicuddy, D.J.

    2014-01-01

    Cysts residing in benthic nepheloid layers (BNLs) documented in the Gulf of Maine have been proposed as a possible source of inoculum for annual blooms of a toxic dinoflagellate in the region. Herein we present a spatially extensive data set of the distribution and thickness of benthic nepheloid layers in the Gulf of Maine and the abundance and inventories of suspended Alexandrium fundyense cysts within these near-bottom layers. BNLs are pervasive throughout the gulf and adjacent Bay of Fundy with maximum layer thicknesses of 50–60 m observed. Mean BNL thickness is 30 m in the eastern gulf and Bay of Fundy, and 20 m in the western gulf. Cyst densities in the near-bottom particle resuspension layers varied by three orders of magnitude across the gulf with maxima of 105 cysts m−3. An important interconnection of elevated BNL cyst densities is observed between the Bay of Fundy, the Maine Coastal Current and the south-central region of the gulf. BNL cyst inventories estimated for the eastern and western gulf are each on the order of 1015 cysts, whereas the BNL inventory in the Bay of Fundy is on the order of 1016 . Although BNL cyst inventories in the eastern and western gulf are 1–2 orders of magnitude smaller than the abundance of cysts in the upper 1 cm of sediment in those regions, BNL and sediment-bound cyst inventories are comparable in the Bay of Fundy. The existence of widespread BNLs containing substantial cyst inventories indicates that these near-bottom layers represent an important source of germinating A. fundyense cysts in the region. PMID:25419055

  13. Comprehensive insights into the response of Alexandrium tamarense to algicidal component secreted by a marine bacterium

    PubMed Central

    Lei, Xueqian; Li, Dong; Li, Yi; Chen, Zhangran; Chen, Yao; Cai, Guanjing; Yang, Xujun; Zheng, Wei; Zheng, Tianling

    2015-01-01

    Harmful algal blooms occur throughout the world, threatening human health, and destroying marine ecosystems. Alexandrium tamarense is a globally distributed and notoriously toxic dinoflagellate that is responsible for most paralytic shellfish poisoning incidents. The culture supernatant of the marine algicidal bacterium BS02 showed potent algicidal effects on A. tamarense ATGD98-006. In this study, we investigated the effects of this supernatant on A. tamarense at physiological and biochemical levels to elucidate the mechanism involved in the inhibition of algal growth by the supernatant of the strain BS02. Reactive oxygen species (ROS) levels increased following exposure to the BS02 supernatant, indicating that the algal cells had suffered from oxidative damage. The levels of cellular pigments, including chlorophyll a and carotenoids, were significantly decreased, which indicated that the accumulation of ROS destroyed pigment synthesis. The decline of the maximum photochemical quantum yield (Fv/Fm) and relative electron transport rate (rETR) suggested that the photosynthesis systems of algal cells were attacked by the BS02 supernatant. To eliminate the ROS, the activities of antioxidant enzymes, including superoxide dismutase (SOD) and catalase (CAT), increased significantly within a short period of time. Real-time PCR revealed changes in the transcript abundances of two target photosynthesis-related genes (psbA and psbD) and two target respiration-related genes (cob and cox). The transcription of the respiration-related genes was significantly inhibited by the treatments, which indicated that the respiratory system was disturbed. Our results demonstrate that the BS02 supernatant can affect the photosynthesis process and might block the PS II electron transport chain, leading to the production of excessive ROS. The increased ROS can further destroy membrane integrity and pigments, ultimately inducing algal cell death. PMID:25667582

  14. Bacterial diversity in toxic Alexandrium tamarense blooms off the Orkney Isles and the Firth of Forth

    NASA Astrophysics Data System (ADS)

    Wichels, Antje; Hummert, Christian; Elbrächter, Malte; Luckas, Bernd; Schütt, Christian; Gerdts, Gunnar

    2004-04-01

    The genetic diversity of the bacterial community associated with Alexandrium tamarense blooms was studied in blooms of the toxic dinoflagellates in the waters around the Orkney Isles and the Firth of Forth (Scotland). For toxin and molecular analysis of the bacterial communities associated with the toxic bloom, water samples were taken in 1998 and 1999 from A. tamarense blooms. The bacterial community structure, as determined by DGGE (denaturing gradient gel electrophoresis) showed clear differences between all three investigated size fractions (dinoflagellate-associated bacteria, attached bacteria and free-living bacteria), with high diversity within each sample. DNA sequence analysis of the dominant and most frequent DGGE bands revealed the dominance of α Proteobacteria, mainly of the Roseobacter clade, with similarities of 91-99%. Moreover, DGGE bands occurring at the same position in the gel throughout in most samples corroborate the presence of several specific α Proteobacteria of the Roseobacter clade. Overall, 500 bacteria were isolated from the bloom and partly phylogenetically analysed. They were members of two prokaryotic phyla, the Proteobacteria and the Bacteroidetes, related to Proteobacteria of the α and γ subdivisions (Alteromonas, Pseudoalteromonas and Colwellia). All bacteria were tested for the production of sodium channel blocking (SCB) toxins using mouse neuroblastoma assay. No production of SCB toxins was found and high performance liquid chromatography (HPLC) analysis confirmed these results. The content of total paralytic shellfish poisoning (PSP) toxin in the water samples, as measured within the toxic dinoflagellate blooms using HPLC, ranged from 53 to 2191 ng PSP l-1 in 1998 and from 0 to 478 ng PSP l-1 in 1999. Changes in PSP toxin content were not accompanied by changes of DGGE band patterns. We therefore presume that the bacterial groups identified in this study were not exclusively associated with toxic A. tamarense, but were

  15. The Hidden Sexuality of Alexandrium Minutum: An Example of Overlooked Sex in Dinoflagellates

    PubMed Central

    Figueroa, Rosa I.; Dapena, Carlos; Bravo, Isabel; Cuadrado, Angeles

    2015-01-01

    Dinoflagellates are haploid eukaryotic microalgae in which rapid proliferation causes dense blooms, with harmful health and economic effects to humans. The proliferation mode is mainly asexual, as the sexual cycle is believed to be rare and restricted to stressful environmental conditions. However, sexuality is key to explaining the recurrence of many dinoflagellate blooms because in many species the fate of the planktonic zygotes (planozygotes) is the formation of resistant cysts in the seabed (encystment). Nevertheless, recent research has shown that individually isolated planozygotes in the lab can enter other routes besides encystment, a behavior of which the relevance has not been explored at the population level. In this study, using imaging flow cytometry, cell sorting, and Fluorescence In Situ Hybridization (FISH), we followed DNA content and nuclear changes in a population of the toxic dinoflagellate Alexandrium minutum that was induced to encystment. Our results first show that planozygotes behave like a population with an “encystment-independent” division cycle, which is light-controlled and follows the same Light:Dark (L:D) pattern as the cycle governing the haploid mitosis. Resting cyst formation was the fate of just a small fraction of the planozygotes formed and was restricted to a period of strongly limited nutrient conditions. The diploid-haploid turnover between L:D cycles was consistent with two-step meiosis. However, the diel and morphological division pattern of the planozygote division also suggests mitosis, which would imply that this species is not haplontic, as previously considered, but biphasic, because individuals could undergo mitotic divisions in both the sexual (diploid) and the asexual (haploid) phases. We also report incomplete genome duplication processes. Our work calls for a reconsideration of the dogma of rare sex in dinoflagellates. PMID:26599692

  16. Population genetic structure and connectivity of the harmful dinoflagellate Alexandrium minutum in the Mediterranean Sea.

    PubMed

    Casabianca, Silvia; Penna, Antonella; Pecchioli, Elena; Jordi, Antoni; Basterretxea, Gotzon; Vernesi, Cristiano

    2012-01-07

    The toxin-producing microbial species Alexandrium minutum has a wide distribution in the Mediterranean Sea and causes high biomass blooms with consequences on the environment, human health and coastal-related economic activities. Comprehension of algal genetic differences and associated connectivity is fundamental to understand the geographical scale of adaptation and dispersal pathways of harmful microalgal species. In the present study, we combine A. minutum population genetic analyses based on microsatellites with indirect connectivity (C(i)) estimations derived from a general circulation model of the Mediterranean sea. Our results show that four major clusters of genetically homogeneous groups can be identified, loosely corresponding to four regional seas: Adriatic, Ionian, Tyrrhenian and Catalan. Each of the four clusters included a small fraction of mixed and allochthonous genotypes from other Mediterranean areas, but the assignment to one of the four clusters was sufficiently robust as proved by the high ancestry coefficient values displayed by most of the individuals (>84%). The population structure of A. minutum on this scale can be explained by microalgal dispersion following the main regional circulation patterns over successive generations. We hypothesize that limited connectivity among the A. minutum populations results in low gene flow but not in the erosion of variability within the population, as indicated by the high gene diversity values. This study represents a first and new integrated approach, combining both genetic and numerical methods, to characterize and interpret the population structure of a toxic microalgal species. This approach of characterizing genetic population structure and connectivity at a regional scale holds promise for the control and management of the harmful algal bloom events in the Mediterranean Sea.

  17. Toxic Alexandrium blooms in the western Gulf of Maine: The plume advection hypothesis revisited

    USGS Publications Warehouse

    Anderson, D.M.; Keafer, B.A.; Geyer, W.R.; Signell, R.P.; Loder, T.C.

    2005-01-01

    The plume advection hypothesis links blooms of the toxic dinoflagellate Alexandrium fundyense in the western Gulf of Maine (GOM) to a buoyant plume derived from river outflows. This hypothesis was examined with cruise and moored-instrument observations in 1993 when levels of paralytic shellfish poisoning (PSP) toxins were high, and in 1994 when toxicity was low. A coupled physical-biological model simulated hydrography and A. fundyense distributions. Initial A. fundyense populations were restricted to low-salinity nearshore waters near Casco Bay, but also occurred in higher salinity waters along the plume boundary. This suggests two sources of cells - those from shallow-water cyst populations and those transported to shore from offshore blooms in the eastern segment of the Maine coastal current (EMCC). Observations confirm the role of the plume in A. fundyense transport and growth. Downwelling-favorable winds in 1993 transported the plume and its cells rapidly alongshore, enhancing toxicity and propagating PSP to the south. In 1994, sustained upwelling moved the plume offshore, resulting in low toxicity in intertidal shellfish. A. fundyense blooms were likely nutrient limited, leading to low growth rates and moderate cell abundances. These observations and mechanisms were reproduced by coupled physical-biological model simulations. The plume advection hypothesis provides a viable explanation for outbreaks of PSP in the western GOM, but should be refined to include two sources for cells that populate the plume and two major pathways for transport: one within the low-salinity plume and another where A. fundyense cells originating in the EMCC are transported along the outer boundary of the plume front with the western segment of the Maine coastal current.

  18. Benthic nepheloid layers in the Gulf of Maine and Alexandrium cyst inventories.

    PubMed

    Pilskaln, C H; Hayashi, K; Keafer, B A; Anderson, D M; McGillicuddy, D J

    2014-05-01

    Cysts residing in benthic nepheloid layers (BNLs) documented in the Gulf of Maine have been proposed as a possible source of inoculum for annual blooms of a toxic dinoflagellate in the region. Herein we present a spatially extensive data set of the distribution and thickness of benthic nepheloid layers in the Gulf of Maine and the abundance and inventories of suspended Alexandrium fundyense cysts within these near-bottom layers. BNLs are pervasive throughout the gulf and adjacent Bay of Fundy with maximum layer thicknesses of 50-60 m observed. Mean BNL thickness is 30 m in the eastern gulf and Bay of Fundy, and 20 m in the western gulf. Cyst densities in the near-bottom particle resuspension layers varied by three orders of magnitude across the gulf with maxima of 10(5) cysts m(-3). An important interconnection of elevated BNL cyst densities is observed between the Bay of Fundy, the Maine Coastal Current and the south-central region of the gulf. BNL cyst inventories estimated for the eastern and western gulf are each on the order of 10(15) cysts, whereas the BNL inventory in the Bay of Fundy is on the order of 10(16) . Although BNL cyst inventories in the eastern and western gulf are 1-2 orders of magnitude smaller than the abundance of cysts in the upper 1 cm of sediment in those regions, BNL and sediment-bound cyst inventories are comparable in the Bay of Fundy. The existence of widespread BNLs containing substantial cyst inventories indicates that these near-bottom layers represent an important source of germinating A. fundyense cysts in the region.

  19. Paralytic shellfish toxin content is related to genomic sxtA4 copy number in Alexandrium minutum strains

    PubMed Central

    Stüken, Anke; Riobó, Pilar; Franco, José; Jakobsen, Kjetill S.; Guillou, Laure; Figueroa, Rosa I.

    2015-01-01

    Dinoflagellates are microscopic aquatic eukaryotes with huge genomes and an unusual cell regulation. For example, most genes are present in numerous copies and all copies seem to be obligatorily transcribed. The consequence of the gene copy number (CPN) for final protein synthesis is, however, not clear. One such gene is sxtA, the starting gene of paralytic shellfish toxin (PST) synthesis. PSTs are small neurotoxic compounds that can accumulate in the food chain and cause serious poisoning incidences when ingested. They are produced by dinoflagellates of the genera Alexandrium, Gymnodium, and Pyrodinium. Here we investigated if the genomic CPN of sxtA4 is related to PST content in Alexandrium minutum cells. SxtA4 is the 4th domain of the sxtA gene and its presence is essential for PST synthesis in dinoflagellates. We used PST and genome size measurements as well as quantitative PCR to analyze sxtA4 CPN and toxin content in 15 A. minutum strains. Our results show a strong positive correlation between the sxtA4 CPN and the total amount of PST produced in actively growing A. minutum cells. This correlation was independent of the toxin profile produced, as long as the strain contained the genomic domains sxtA1 and sxtA4. PMID:25983733

  20. Immunological and physiological responses of the periwinkle Littorina littorea during and after exposure to the toxic dinoflagellate Alexandrium minutum.

    PubMed

    Neves, Raquel A F; Figueiredo, Gisela M; Valentin, Jean Louis; da Silva Scardua, Patricia Mirella; Hégaret, Hélène

    2015-03-01

    Species of the dinoflagellate genus Alexandrium produce phycotoxins responsible for paralytic shellfish poisoning. Blooms of Alexandrium minutum reach very high concentrations of vegetative cells in the water column; and when these blooms occur, large numbers of toxic cysts can be produced and deposited on sediments becoming available to benthic species. The present study investigated the potential effect of exposure to toxic cysts of A. minutum on the periwinkle Littorinalittorea. Snails were exposed for nine days to pellicle cysts of toxic and non-toxic dinoflagellates, A. minutum and Heterocapsa triquetra, respectively, followed by six days of depuration while they were fed only H. triquetra. Toxin accumulation, condition index, immune and histopathological responses were analyzed. Histological alterations were also monitored in snails exposed to a harmful A. minutum bloom, which naturally occurred in the Bay of Brest. Snails exposed to toxic cysts showed abnormal behavior that seems to be toxin-induced and possibly related to muscle paralysis. Periwinkles accumulated toxins by preying on toxic cysts and accumulation appeared dependent on the time of exposure, increasing during intoxication period but tending to stabilize during depuration period. Toxic exposure also seemed to negatively affect hemocyte viability and functions, as ROS production and phagocytosis. Histological analyses revealed that toxic exposure induced damages on digestive organs of snails, both in laboratory and natural systems. This study demonstrates that an exposure to the toxic dinoflagellate A. minutum leads to sublethal effects on L. littorea, which may alter individual fitness and increase the susceptibility of snails to pathogens and diseases.

  1. Dynamics of co-occurring Alexandrium minutum (Global Clade) and A. tamarense (West European) (Dinophyceae) during a summer bloom in Cork Harbour, Ireland (2006)

    NASA Astrophysics Data System (ADS)

    Touzet, N.; Farrell, H.; Ní Rathaille, A.; Rodriguez, P.; Alfonso, A.; Botana, L. M.; Raine, R.

    2010-02-01

    The dinoflagellate genus Alexandrium contains neurotoxin-producing species, which have adversely affected the aquaculture industry and fisheries worldwide. Seasonal toxic blooms of Alexandrium spp. occur on an annual basis in the North Channel area of Cork Harbour, Ireland, where resident populations of non-toxic A. tamarense (West European ribotype) and PSP toxin-producing A. minutum (Global Clade) co-occur. Field surveys were carried out throughout a bloom of Alexandrium spp. in the summer of 2006. Taxa-specific fluorescently labelled probes were used in a dual whole-cell fluorescent in situ hybridization (WC-FISH) assay for the simultaneous discrimination and quantification of A. minutum and A. tamarense in the water column. The bloom occurred following a weak spring tide in early June and Alexandrium cell concentrations exceeded 3×10 4 cells L -1. A. minutum dominated numerically over A. tamarense throughout the sampling period (74% on average). The maximum cell concentration was ˜3.3×10 5 cells L -1 at the peak of the bloom and was localized at the eastern end of the North Channel. The bloom collapse coincided with increasing tidal flushing and significantly changing meteorological conditions (wind speed increase, lesser irradiance), which led to a water temperature drop of ˜3 °C within a period of 7 days. GTX3 was the dominant PSP toxin variant and C-toxins were at times observed in samples. Assuming that A. minutum was the only microorganism synthesising PSP toxins, the internal toxin quota was on average 13.4 fmol cell -1, a value similar to that observed in laboratory experiments. Monitoring of toxic Alexandrium species in Ireland will require the use of molecular methods for reliable discrimination and quantification.

  2. Alexandrium fundyense cyst viability and germling survival in light vs. dark at a constant low temperature.

    PubMed

    Vahtera, Emil; Crespo, Bibiana G; McGillicuddy, Dennis J; Olli, Kalle; Anderson, Donald M

    2014-05-01

    Both observations and models suggest that large-scale coastal blooms of Alexandrium fundyense in the Gulf of Maine are seeded by deep-bottom cyst accumulation zones ("seed beds") where cysts germinate from the sediment surface or the overlying near-bottom nepheloid layers at water depths exceeding 100 m. The germling cells and their vegetative progeny are assumed to be subject to mortality while in complete darkness, as they swim to illuminated surface waters. To test the validity of this assumption we conducted laboratory investigations of cyst viability and the survival of the germling cells and their vegetative progeny during prolonged exposure to darkness at a temperature of 6 °C, simulating the conditions in deep Gulf of Maine waters. We isolated cysts from bottom sediments collected in the Gulf of Maine under low red light and incubated them in 96-well tissue culture-plates in culture medium under a 10:14 h light:dark cycle and under complete darkness. Cyst viability was high, with excystment frequency reaching 90% in the illuminated treatment after 30 days and in the dark treatment after 50 days. Average germination rates were 0.062 and 0.038 d(-1) for light and dark treatments, respectively. The dark treatment showed an approximately 2-week time lag in maximum germination rates compared to the light treatment. Survival of germlings was considerably lower in the dark treatment. In the light treatments, 47% of germinated cysts produced germlings that were able to survive for 7 days and produce vegetative progeny, i.e., there were live cells in the well along with an empty cyst at least once during the experiment. In the dark treatments 12% of the cysts produced germlings that were able to survive for the same length of time. When dark treatments are scaled to take into account non-darkness related mortality, approximately 28% of the cysts produced germlings that were able to survive for at least 7 days. Even though cysts are able to germinate in darkness

  3. Experimental and computational studies on molecularly imprinted solid-phase extraction for gonyautoxins 2,3 from dinoflagellate Alexandrium minutum.

    PubMed

    Lian, Ziru; Li, Hai-Bei; Wang, Jiangtao

    2016-08-01

    An innovative and effective extraction procedure based on molecularly imprinted solid-phase extraction (MISPE) was developed for the isolation of gonyautoxins 2,3 (GTX2,3) from Alexandrium minutum sample. Molecularly imprinted polymer microspheres were prepared by suspension polymerization and and were employed as sorbents for the solid-phase extraction of GTX2,3. An off-line MISPE protocol was optimized. Subsequently, the extract samples from A. minutum were analyzed. The results showed that the interference matrices in the extract were obviously cleaned up by MISPE procedures. This outcome enabled the direct extraction of GTX2,3 in A. minutum samples with extraction efficiency as high as 83 %, rather significantly, without any need for a cleanup step prior to the extraction. Furthermore, computational approach also provided direct evidences of the high selective isolation of GTX2,3 from the microalgal extracts.

  4. Molecular phylogeny and PSP toxin profile of the Alexandrium tamarense species complex along the coast of China.

    PubMed

    Zou, Cheng; Ye, Rui-Min; Zheng, Jian-Wei; Luo, Zhao-He; Gu, Hai-Feng; Yang, Wei-Dong; Li, Hong-Ye; Liu, Jie-Sheng

    2014-12-15

    To explore the genetic diversity and paralytic shellfish poisoning (PSP) toxin profile of the Alexandrium tamarense species complex along the coast of China, 67 strains of A. tamarense from the China Sea were collected and genetic diversity were analyzed based on the rDNA sequences. In addition, PSP toxin compositions and contents were detected by HPLC. According to the 5.8S rDNA and ITS, and LSU rDNA D1-D2 sequence, A. tamarense in the China Sea comprises at least Group IV and Group I ribotypes. In these Chinese strains, the toxins with the highest concentration in the profile were C1/2, gonyautoxins 1/4 (GTX1/4) and neosaxitoxin (NEO). However, the toxin profiles were atypical and C1/2 toxins were not detected in some strains. No strict correlation was observed between the PSP toxins profile and the geographical distribution.

  5. Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum

    NASA Astrophysics Data System (ADS)

    Tang, Xianghai; Yu, Rencheng; Zhou, Mingjiang; Yu, Zhigang

    2012-03-01

    The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology, making identification difficult. In this study, species-specific rRNA probes were designed for whole-cell fluorescence in situ hybridization (FISH) to distinguish A. minutum from two phylogenetic clades. We acquired the complete SSU to LSU rDNA sequences (GenBank accession numbers JF906989-JF906999) of 11 Alexandrium strains and used these to design rRNA targeted oligonucleotide probes. Three ribotype-specific probes, M-GC-1, M-PC-2, and M-PC-3, were designed. The former is specific for the GC clade ("Global clade") of A. minutum, the majority of which have been found non-toxic, and the latter two are specific for the PSP (paralytic shellfish poisoning)-producing PC clade ("Pacific clade"). The specificity of these three probes was confirmed by FISH. All cells in observed fields of view were fluorescently labeled when probes and target species were incubated under optimized FISH conditions. However, the accessibility of rRNA molecules in ribosomes varied among the probe binding positions. Thus, there was variation in the distribution of positive signals in labeled cells within nucleolus and cytosol (M-GC-1, M-PC-3), or just nucleolus (M-PC-2). Our results provide a methodological basis for studying the biogeography and population dynamics of A. minutum, and providing an early warning of toxic HABs.

  6. Bioturbation, germination and deposition of Alexandrium fundyense cysts in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Shull, David H.; Kremp, Anke; Mayer, Lawrence M.

    2014-05-01

    Like many other dinoflagellate species, Alexandrium fundyense possesses a benthic resting cyst which enables long-term persistence and annual blooms of this species in the Gulf of Maine. The size and extent of these harmful algal blooms are associated with high cyst concentrations in the top 1 cm of sediment. Despite the importance of this resting stage in the life history of A. fundyense, little work has been done on bioturbation of cysts in the deep-water cyst beds of the western Gulf of Maine. Our work intensively examined one site within a major regional “seedbed” from February 2003 until August 2005, a time span that included an extraordinarily large bloom of A. fundyense in 2005. Over the course of 2 years we collected samples for benthic infauna and cyst profiles down to a depth of 30 cm. We also measured sediment porosity, organic carbon, 210Pb, and porewater dissolved oxygen. On several dates we measured depth profiles of cyst autofluorescence. Profiles of cysts revealed large subsurface maxima peaking between 10 and 15 cm depth with cyst concentrations declining strongly toward the sediment surface. On one sampling date (August 2004) we observed a cyst concentration peak at the sediment surface. Using these data we constructed a mechanistic model of cyst bioturbation, mortality, germination, and deposition. Modeled bioturbation was calibrated using 210Pb and modeled cyst profiles were compared to measured profiles. Model runs with constant and interannually-varying rates of cyst deposition produced similar time-averaged cyst profiles. Results indicate that the deeper portions of cyst profiles are determined primarily by bioturbation, germination and cyst mortality and less so by interannual variation in cyst depositional history. This is due to the relatively low sedimentation rate at the study site compared to the rate of bioturbation, and the fact that the number of cysts deposited each year tends to be a small fraction of the total inventory

  7. The death mechanism of the harmful algal bloom species Alexandrium tamarense induced by algicidal bacterium Deinococcus sp. Y35

    PubMed Central

    Li, Yi; Zhu, Hong; Lei, Xueqian; Zhang, Huajun; Cai, Guanjing; Chen, Zhangran; Fu, Lijun; Xu, Hong; Zheng, Tianling

    2015-01-01

    Harmful algal blooms (HABs) cause a variety of deleterious effects on aquatic ecosystems, especially the toxic dinoflagellate Alexandrium tamarense, which poses a serious threat to marine economic and human health based on releasing paralytic shellfish poison into the environment. The algicidal bacterium Deinococcus sp. Y35 which can induce growth inhibition on A. tamarense was used to investigate the functional mechanism. The growth status, reactive oxygen species (ROS) content, photosynthetic system and the nuclear system of algal cells were determined under algicidal activity. A culture of strain Y35 not only induced overproduction of ROS in algal cells within only 0.5 h of treatment, also decrease the total protein content as well as the response of the antioxidant enzyme. Meanwhile, lipid peroxidation was induced and cell membrane integrity was lost. Photosynthetic pigments including chlorophyll a and carotenoid decreased along with the photosynthetic efficiency being significantly inhibited. At the same time, photosynthesis-related gene expression showed down-regulation. More than, the destruction of cell nuclear structure and inhibition of proliferating cell nuclear antigen (PCNA) related gene expression were confirmed. The potential functional mechanism of the algicidal bacterium on A. tamarense was investigated and provided a novel viewpoint which could be used in HABs control. PMID:26441921

  8. The first evidence of deinoxanthin from Deinococcus sp. Y35 with strong algicidal effect on the toxic dinoflagellate Alexandrium tamarense.

    PubMed

    Li, Yi; Zhu, Hong; Lei, Xueqian; Zhang, Huajun; Guan, Chengwei; Chen, Zhangran; Zheng, Wei; Xu, Hong; Tian, Yun; Yu, Zhiming; Zheng, Tianling

    2015-06-15

    Harmful algal blooms (HABs) could be deemed hazardous materials in aquatic environment. Alexandrium tamarense is a toxic HAB causing alga, which causes serious economic losses and health problems. In this study, the bacterium Deinococcus xianganensis Y35 produced a new algicide, showing a high algicidal effect on A. tamarense. The algicidal compound was identified as deinoxanthin, a red pigment, based on high resolution mass spectrometry and NMR after the active compound was isolated and purified. Deinoxanthin exhibited an obvious inhibitory effect on algal growth, and showed algicidal activity against A. tamarense with an EC50 of 5.636 μg/mL with 12h treatment time. Based on the unique structure and characteristics of deinoxanthin, the content of reactive oxygen species (ROS) increased after 0.5h exposure, the structure of organelles including chloroplasts and mitochondria were seriously damaged. All these results firstly confirmed that deinoxanthin as the efficient and eco-environmental algicidal compound has potential to be used for controlling harmful algal blooms through overproduction of ROS.

  9. SxtA and sxtG Gene Expression and Toxin Production in the Mediterranean Alexandrium minutum (Dinophyceae)

    PubMed Central

    Perini, Federico; Galluzzi, Luca; Dell’Aversano, Carmela; Dello Iacovo, Emma; Tartaglione, Luciana; Ricci, Fabio; Forino, Martino; Ciminiello, Patrizia; Penna, Antonella

    2014-01-01

    The dinoflagellate Alexandrium minutum is known for the production of potent neurotoxins affecting the health of human seafood consumers via paralytic shellfish poisoning (PSP). The aim of this study was to investigate the relationship between the toxin content and the expression level of the genes involved in paralytic shellfish toxin (PST) production. The algal cultures were grown both in standard f/2 medium and in phosphorus/nitrogen limitation. In our study, LC-HRMS analyses of PST profile and content in different Mediterranean A. minutum strains confirmed that this species was able to synthesize mainly the saxitoxin analogues Gonyautoxin-1 (GTX1) and Gonyautoxin-4 (GTX4). The average cellular toxin content varied among different strains, and between growth phases, highlighting a decreasing trend from exponential to stationary phase in all culture conditions tested. The absolute quantities of intracellular sxtA1 and sxtG mRNA were not correlated with the amount of intracellular toxins in the analysed A. minutum suggesting that the production of toxins may be regulated by post-transcriptional mechanisms and/or by the concerted actions of alternative genes belonging to the PST biosynthesis gene cluster. Therefore, it is likely that the sxtA1 and sxtG gene expression could not reflect the PST accumulation in the Mediterranean A. minutum populations under the examined standard and nutrient limiting conditions. PMID:25341029

  10. The death mechanism of the harmful algal bloom species Alexandrium tamarense induced by algicidal bacterium Deinococcus sp. Y35.

    PubMed

    Li, Yi; Zhu, Hong; Lei, Xueqian; Zhang, Huajun; Cai, Guanjing; Chen, Zhangran; Fu, Lijun; Xu, Hong; Zheng, Tianling

    2015-01-01

    Harmful algal blooms (HABs) cause a variety of deleterious effects on aquatic ecosystems, especially the toxic dinoflagellate Alexandrium tamarense, which poses a serious threat to marine economic and human health based on releasing paralytic shellfish poison into the environment. The algicidal bacterium Deinococcus sp. Y35 which can induce growth inhibition on A. tamarense was used to investigate the functional mechanism. The growth status, reactive oxygen species (ROS) content, photosynthetic system and the nuclear system of algal cells were determined under algicidal activity. A culture of strain Y35 not only induced overproduction of ROS in algal cells within only 0.5 h of treatment, also decrease the total protein content as well as the response of the antioxidant enzyme. Meanwhile, lipid peroxidation was induced and cell membrane integrity was lost. Photosynthetic pigments including chlorophyll a and carotenoid decreased along with the photosynthetic efficiency being significantly inhibited. At the same time, photosynthesis-related gene expression showed down-regulation. More than, the destruction of cell nuclear structure and inhibition of proliferating cell nuclear antigen (PCNA) related gene expression were confirmed. The potential functional mechanism of the algicidal bacterium on A. tamarense was investigated and provided a novel viewpoint which could be used in HABs control.

  11. CHARACTERIZATION OF 17 NEW MICROSATELLITE MARKERS FOR THE DINOFLAGELLATE ALEXANDRIUM FUNDYENSE (DINOPHYCEAE), A HARMFUL ALGAL BLOOM SPECIES

    PubMed Central

    Sehein, Taylor; Richlen, Mindy L.; Nagai, Satoshi; Yasuike, Motoshige; Nakamura, Yoji; Anderson, Donald M.

    2016-01-01

    Alexandrium fundyense is the toxic marine dinoflagellate responsible for “red tide” events in temperate and sub-arctic waters worldwide. In the Gulf of Maine (GOM) and Bay of Fundy in the Northwest Atlantic, blooms of A. fundyense recur annually, and are associated with major health and ecosystem impacts. In this region, microsatellite markers have been used to investigate genetic structure and gene flow; however, the loci currently available for this species were isolated from populations from Japan and the North Sea, and only a subset are suitable for the analysis of A. fundyense populations in the Northwest Atlantic. To facilitate future studies of A. fundyense blooms, both in this region and globally, we isolated and characterized 17 polymorphic microsatellite loci from 31 isolates collected from the GOM and from the Nauset Marsh System, an estuary on Cape Cod, MA, USA. These loci yielded between two and 15 alleles per locus, with an average of 7.1. Gene diversities ranged from 0.297 to 0.952. We then analyzed these same 31 isolates using previously published markers for comparison. We determined the new markers are sufficiently variable and better suited for the investigation of genetic structure, bloom dynamics, and diversity in the Northwest Atlantic. PMID:27274617

  12. The effect of the toxic dinoflagellate Alexandrium fundyense on the fitness of the calanoid copepod Calanus finmarchicus

    PubMed Central

    Roncalli, Vittoria; Turner, Jefferson T.; Kulis, David; Anderson, Donald M.; Lenz, Petra H.

    2016-01-01

    Inshore and offshore waters of the Gulf of Maine (USA) have spring/summer harmful algal blooms (HABs) of the toxic dinoflagellate Alexandrium fundyense, which is responsible for paralytic shellfish poisoning (PSP) in humans. The calanoid copepod Calanus finmarchicus co-occurs with A. fundyense during the seasonal blooms. At that time, C. finmarchicus population abundances are high, dominated by immature copepods preparing for diapause, and by actively-reproducing adults. High survival has been reported for copepods exposed to toxic A. fundyense, but little is known about possible sublethal effects. In this study, C. finmarchicus adult females were fed either a control diet of non-toxic Rhodomonas spp. or one of two diets containing either low dose (LD) or high dose (HD) levels (50 and 200 cells mL−1, respectively) of toxic A. fundyense for a total of 7 days in two independent experiments. As expected, ingestion of the dinoflagellate had no effect on copepod survival and grazing activity. However, significant reductions of egg production and egg viability were observed in C. finmarchicus females fed on either experimental diet. After the 7-day experiment, total nauplius production by females on the LD and HD diets was reduced by 35% to 75% compared to the control females. These results suggest that blooms of A. fundyense in the Gulf of Maine may be an environmental challenge for C. finmarchicus populations, with a potential negative effect on copepod recruitment. PMID:27721677

  13. No evidence for induction or selection of mutant sodium channel expression in the copepod Acartia husdsonica challenged with the toxic dinoflagellate Alexandrium fundyense

    PubMed Central

    Finiguerra, Michael; Avery, David E; Dam, Hans G

    2014-01-01

    Some species in the dinoflagellate genus Alexandrium spp. produce a suite of neurotoxins that block sodium channels, known as paralytic shellfish toxins (PST), which have deleterious effects on grazers. Populations of the ubiquitous copepod grazer Acartia hudsonica that have co-occurred with toxic Alexandrium spp. are better adapted than naïve populations. The mechanism of adaptation is currently unknown. We hypothesized that a mutation in the sodium channel could account for the grazer adaptation. We tested two hypotheses: (1) Expression of the mutant sodium channel could be induced by exposure to toxic Alexandrium fundyense; (2) in the absence of induction, selection exerted by toxic A. fundyense would favor copepods that predominantly express the mutant isoform. In the copepod A. hudsonica, both isoforms are expressed in all individuals in varying proportions. Thus, in addition to comparing expression ratios of wild-type to mutant isoforms for individual copepods, we also partitioned copepods into three groups: those that predominantly express the mutant (PMI) isoform, the wild-type (PWI) isoform, or both isoforms approximately equally (EI). There were no differences in isoform expression between individuals that were fed toxic and nontoxic food after three and 6 days; induction of mutant isoform expression did not occur. Furthermore, the hypothesis that mutant isoform expression responds to toxic food was also rejected. That is, no consistent evidence showed that the wild-type to mutant isoform ratios decreased, or that the relative proportion of PMI individuals increased, due to the consumption of toxic food over four generations. However, in the selected line that was continuously exposed to toxic food sources, egg production rate increased, which suggested that adaptation occurred but was unrelated to sodium channel isoform expression. PMID:25535562

  14. A quantitative real-time PCR assay for the identification and enumeration of Alexandrium cysts in marine sediments

    NASA Astrophysics Data System (ADS)

    Erdner, D. L.; Percy, L.; Keafer, B.; Lewis, J.; Anderson, D. M.

    2010-02-01

    Harmful algal blooms (HABs) are a global problem that affects both human and ecosystem health. One of the most serious and widespread HAB poisoning syndromes is paralytic shellfish poisoning, commonly caused by Alexandrium spp. dinoflagellates. Like many toxic dinoflagellates, Alexandrium produces resistant resting cysts as part of its life cycle. These cysts play a key role in bloom initiation and decline, as well as dispersal and colonization of new areas. Information on cyst numbers and identity is essential for understanding and predicting blooms, yet comprehensive cyst surveys are extremely time- and labor-intensive. Here we describe the development and validation of a quantitative real-time PCR (qPCR) technique for the enumeration of cysts of A. tamarense of the toxic North American/Group I ribotype. The method uses a cloned fragment of the large subunit ribosomal RNA gene as a standard for cyst quantification, with an experimentally determined conversion factor of 28,402±6152 LSU ribosomal gene copies per cyst. Tests of DNA extraction and PCR efficiency show that mechanical breakage is required for adequate cyst lysis, and that it was necessary to dilute our DNA extracts 50-fold in order to abolish PCR inhibition from compounds co-extracted from the sediment. The resulting assay shows a linear response over 6 orders of magnitude and can reliably quantify ≥10 cysts/cm 3 sediment. For method validation, 129 natural sediment samples were split and analyzed in parallel, using both the qPCR and primulin-staining techniques. Overall, there is a significant correlation ( p<0.001) between the cyst abundances determined by the two methods, although the qPCR counts tend to be lower than the primulin values. This underestimation is less pronounced in those samples collected from the top 1 cm of sediment, and more pronounced in those derived from the next 1-3 cm of the core. These differences may be due to the condition of the cysts in the different layers, as the

  15. Relationships between satellite-measured thermal features and Alexandrium-imposed toxicity in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Luerssen, Remy M.; Thomas, Andrew C.; Hurst, John

    2005-09-01

    Relationships between satellite-derived sea-surface temperature (SST) patterns and the occurrence of paralytic shellfish poisoning (PSP) toxicity events caused by Alexandrium fundyense in the western Gulf of Maine are examined. Comparisons between surface A. fundyense cell distribution patterns and SST images indicate that highest cell concentrations are associated with colder waters of the eastern segment of the Gulf of Maine coastal current (EMCC) and that frontal zones at the edges of the EMCC often act as boundaries to surface distributions. Surface thermal patterns coincident with a May 2000 PSP toxic event and shellfish harvesting closure on the western Maine coast show enhanced connectivity between the EMCC and the western Gulf of Maine, suggesting transport linking A. fundyense cells in the EMCC to inshore areas of the western Gulf of Maine. Surface drifter data support such transport. Thirteen years (1990-2002) of toxicity data from eight monitoring sites along the coast of Maine and concurrent SST data show that in years of either large or very reduced toxicity, a consistent relationship exists between the timing and strength of fronts, taken as an indicator of alongshore connectivity, and the occurrence and strength of toxic events. Years with weak fronts and/or fronts that become established relatively late in the summer growing season are years of the strongest toxicity events in western Gulf of Maine. Years of early and strong fronts are years with few and/or weak toxicity events. Our results suggest that advective connections exist between cells present in the EMCC and toxicity along the western Gulf of Maine coast and that large-scale hydrographic processes, characterized here as surface thermal patterns, influence A. fundyense populations in the western Gulf of Maine, either through delivery of actual cells or advection of advantageous conditions into the region. These data point to the utility of satellite and other coastal observing system data

  16. Historic 2005 toxic bloom of Alexandrium fundyense in the western Gulf of Maine: 2. Coupled biophysical numerical modeling

    NASA Astrophysics Data System (ADS)

    He, Ruoying; McGillicuddy, Dennis J.; Keafer, Bruce A.; Anderson, Donald M.

    2008-07-01

    A coupled physical/biological modeling system was used to hindcast a massive Alexandrium fundyense bloom that occurred in the western Gulf of Maine in 2005 and to investigate the relative importance of factors governing the bloom's initiation and development. The coupled system consists of a state-of-the-art, free-surface primitive equation Regional Ocean Modeling System (ROMS) tailored for the Gulf of Maine (GOM) using a multinested configuration, and a population dynamics model for A. fundyense. The system was forced by realistic momentum and buoyancy fluxes, tides, river runoff, observed A. fundyense benthic cyst abundance, and climatological nutrient fields. Extensive comparisons were made between simulated (both physical and biological) fields and in situ observations, revealing that the hindcast model is capable of reproducing the temporal evolution and spatial distribution of the 2005 bloom. Sensitivity experiments were then performed to distinguish the roles of three major factors hypothesized to contribute to the bloom: (1) the high abundance of cysts in western GOM sediments; (2) strong `northeaster' storms with prevailing downwelling-favorable winds; and (3) a large amount of fresh water input due to abundant rainfall and heavy snowmelt. Model results suggest the following. (1) The high abundance of cysts in western GOM was the primary factor of the 2005 bloom. (2) Wind-forcing was an important regulator, as episodic bursts of northeast winds caused onshore advection of offshore populations. These downwelling favorable winds accelerated the alongshore flow, resulting in transport of high cell concentrations into Massachusetts Bay. A large regional bloom would still have happened, however, even with normal or typical winds for that period. (3) Anomalously high river runoff in 2005 resulted in stronger buoyant plumes/currents, which facilitated the transport of cell population to the western GOM. While affecting nearshore cell abundance in Massachusetts

  17. Physiological responses of Manila clams Venerupis (=Ruditapes) philippinarum with varying parasite Perkinsus olseni burden to toxic algal Alexandrium ostenfeldii exposure.

    PubMed

    Lassudrie, Malwenn; Soudant, Philippe; Richard, Gaëlle; Henry, Nicolas; Medhioub, Walid; da Silva, Patricia Mirella; Donval, Anne; Bunel, Mélanie; Le Goïc, Nelly; Lambert, Christophe; de Montaudouin, Xavier; Fabioux, Caroline; Hégaret, Hélène

    2014-09-01

    Manila clam stock from Arcachon Bay, France, is declining, as is commercial harvest. To understand the role of environmental biotic interactions in this decrease, effects of a toxic dinoflagellate, Alexandrium ostenfeldii, which blooms regularly in Arcachon bay, and the interaction with perkinsosis on clam physiology were investigated. Manila clams from Arcachon Bay, with variable natural levels of perkinsosis, were exposed for seven days to a mix of the nutritious microalga T-Iso and the toxic dinoflagellate A. ostenfeldii, a producer of spirolides, followed by seven days of depuration fed only T-Iso. Following sacrifice and quantification of protozoan parasite Perkinsus olseni burden, clams were divided into two groups according to intensity of the infection ("Light-Moderate" and "Moderate-Heavy"). Hemocyte and plasma responses, digestive enzyme activities, antioxidant enzyme activities in gills, and histopathological responses were analyzed. Reactive oxygen species (ROS) production in hemocytes and catalase (CAT) activity in gills increased with P. olseni intensity of infection in control clams fed T-Iso, but did not vary among A. ostenfeldii-exposed clams. Exposure to A. ostenfeldii caused tissue alterations associated with an inflammatory response and modifications in hemocyte morphology. In the gills, superoxide dismutase (SOD) activity decreased, and an increase in brown cell occurrence was seen, suggesting oxidative stress. Observations of hemocytes and brown cells in tissues during exposure and depuration suggest involvement of both cell types in detoxication processes. Results suggest that exposure to A. ostenfeldii disrupted the pro-/anti-oxidant response of clams to heavy P. olseni intensity. In addition, depressed mitochondrial membrane potential (MMP) in hemocytes of clams exposed to A. ostenfeldii suggests that mitochondrial functions are regulated to maintain homeostasis of digestive enzyme activity and condition index.

  18. Distinctly different behavioral responses of a copepod, Temora longicornis, to different strains of toxic dinoflagellates, Alexandrium spp.

    PubMed

    Xu, Jiayi; Hansen, Per Juel; Nielsen, Lasse Tor; Krock, Bernd; Tillmann, Urban; Kiørboe, Thomas

    2017-02-01

    Zooplankton responses to toxic algae are highly variable, even towards taxonomically closely related species or different strains of the same species. Here, the individual level feeding behavior of a copepod, Temora longicornis, was examined which offered 4 similarly sized strains of toxic dinoflagellate Alexandrium spp. and a non-toxic control strain of the dinoflagellate Protoceratium reticulatum. The strains varied in their cellular toxin concentration and composition and in lytic activity. High-speed video observations revealed four distinctly different strain-specific feeding responses of the copepod during 4h incubations: (i) the 'normal' feeding behavior, in which the feeding appendages were beating almost constantly to produce a feeding current and most (90%) of the captured algae were ingested; (ii) the beating activity of the feeding appendages was reduced by ca. 80% during the initial 60min of exposure, after which very few algae were captured and ingested; (iii) capture and ingestion rates remained high, but ingested cells were regurgitated; and (iv) the copepod continued beating its appendages and captured cells at a high rate, but after 60min, most captured cells were rejected. The various prey aversion responses observed may have very different implications to the prey and their ability to form blooms: consumed but regurgitated cells are dead, captured but rejected cells survive and may give the prey a competitive advantage, while reduced feeding activity of the grazer may be equally beneficial to the prey and its competitors. These behaviors were not related to lytic activity or overall paralytic shellfish toxins (PSTs) content and composition and suggest that other cues are responsible for the responses.

  19. Thirty years - Alexandrium fundyense cyst, bloom dynamics and shellfish toxicity in the Bay of Fundy, eastern Canada

    NASA Astrophysics Data System (ADS)

    Martin, Jennifer L.; LeGresley, Murielle M.; Hanke, Alex R.

    2014-05-01

    Sediment and water samples were collected for Alexandrium fundyense spatial and temporal distribution and abundance at more than 120 locations throughout the Bay of Fundy during the summers and winters of 1980-1984. These broad surveys have been repeated at various times through the past 30 years, with more regular sampling since 2004. In addition, A. fundyense abundance has been monitored at several locations within the Bay of Fundy at weekly intervals from April to November and monthly during the remaining months since 1988. Paralytic Shellfish Poisoning (PSP) toxins in shellfish (notably Mya arenaria) have also been monitored at multiple locations in the Bay of Fundy since 1943. The datasets were examined to determine relationships and roles between overwintering resting cysts, bloom initiation, bloom decline, motile cell dispersal and A. fundyense motile populations and resulting shellfish toxicity since 1980. Cysts are widely dispersed throughout the Bay of Fundy in the offshore, inshore and intertidal zones with the largest deposits located in the offshore in silt/clay sediments to the east and north of Grand Manan Island at depths of 60-180 m. Results show that there is a constant stable source of cysts in the Bay of Fundy with highest concentrations of cysts (9780 cysts cm-3) observed in 2010 and highest concentrations of A. fundyense motile cells (18×106 cells L-1) observed in 1980. Interannual changes in abundance in A. fundyense populations, resting cysts and the temporal trends in M. arenaria toxicity are discussed. Results show that there was no relationship between the abundance of overwintering cysts and the magnitude of A. fundyense blooms. The offshore seed beds appear to be relatively constant in cyst density among most years and serve as an important source for the motile cells that lead to initiation of major blooms and resulting shellfish toxicity throughout the Bay of Fundy.

  20. Temperature and residence time controls on an estuarine harmful algal bloom: Modeling hydrodynamics and Alexandrium fundyense in Nauset estuary.

    PubMed

    Ralston, David K; Brosnahan, Michael L; Fox, Sophia E; Lee, Krista; Anderson, Donald M

    2015-11-01

    A highly resolved, 3-d model of hydrodynamics and Alexandrium fundyense in an estuarine embayment has been developed to investigate the physical and biological controls on a recurrent harmful algal bloom. Nauset estuary on Cape Cod (MA, USA) consists of three salt ponds connected to the ocean through a shallow marsh and network of tidal channels. The model is evaluated using quantitative skill metrics against observations of physical and biological conditions during three spring blooms. The A. fundyense model is based on prior model applications for the nearby Gulf of Maine, but notable modifications were made to be consistent with the Nauset observations. The dominant factors controlling the A. fundyense bloom in Nauset were the water temperature, which regulates organism growth rates, and the efficient retention of cells due to bathymetric constraints, stratification, and cell behavior (diel vertical migration). Spring-neap variability in exchange altered residence times, but for cell retention to be substantially longer than the cell doubling time required both active vertical migration and stratification that inhibits mixing of cells into the surface layer by wind and tidal currents. Unlike in the Gulf of Maine, the model results were relatively insensitive to cyst distributions or germination rates. Instead, in Nauset, high apparent rates of vegetative cell division by retained populations dictated bloom development. Cyst germination occurred earlier in the year than in the Gulf of Maine, suggesting that Nauset cysts have different controls on germination timing. The model results were relatively insensitive to nutrient concentrations, due to eutrophic conditions in the highly impacted estuary or due to limitations in the spatial and temporal resolution of nutrient sampling. Cell loss rates were inferred to be extremely low during the growth phase of the bloom, but increased rapidly during the final phase due to processes that remain uncertain. The validated

  1. Temperature and residence time controls on an estuarine harmful algal bloom: Modeling hydrodynamics and Alexandrium fundyense in Nauset estuary

    PubMed Central

    Ralston, David K.; Brosnahan, Michael L.; Fox, Sophia E.; Lee, Krista; Anderson, Donald M.

    2015-01-01

    A highly resolved, 3-d model of hydrodynamics and Alexandrium fundyense in an estuarine embayment has been developed to investigate the physical and biological controls on a recurrent harmful algal bloom. Nauset estuary on Cape Cod (MA, USA) consists of three salt ponds connected to the ocean through a shallow marsh and network of tidal channels. The model is evaluated using quantitative skill metrics against observations of physical and biological conditions during three spring blooms. The A. fundyense model is based on prior model applications for the nearby Gulf of Maine, but notable modifications were made to be consistent with the Nauset observations. The dominant factors controlling the A. fundyense bloom in Nauset were the water temperature, which regulates organism growth rates, and the efficient retention of cells due to bathymetric constraints, stratification, and cell behavior (diel vertical migration). Spring-neap variability in exchange altered residence times, but for cell retention to be substantially longer than the cell doubling time required both active vertical migration and stratification that inhibits mixing of cells into the surface layer by wind and tidal currents. Unlike in the Gulf of Maine, the model results were relatively insensitive to cyst distributions or germination rates. Instead, in Nauset, high apparent rates of vegetative cell division by retained populations dictated bloom development. Cyst germination occurred earlier in the year than in the Gulf of Maine, suggesting that Nauset cysts have different controls on germination timing. The model results were relatively insensitive to nutrient concentrations, due to eutrophic conditions in the highly impacted estuary or due to limitations in the spatial and temporal resolution of nutrient sampling. Cell loss rates were inferred to be extremely low during the growth phase of the bloom, but increased rapidly during the final phase due to processes that remain uncertain. The validated

  2. Study of molecularly imprinted solid-phase extraction of gonyautoxins 2,3 in the cultured dinoflagellate Alexandrium tamarense by high-performance liquid chromatography with fluorescence detection.

    PubMed

    Lian, Zi-Ru; Wang, Jiang-Tao

    2013-11-01

    A highly selective sample cleanup procedure combined with molecularly imprinted solid-phase extraction (MISPE) was developed for the isolation of gonyautoxins 2,3 (GTX2,3) from Alexandrium tamarense sample. The molecularly imprinted polymer microspheres (MIPMs) were prepared by suspension polymerization using caffeine as the dummy template molecule, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker and polyvinyl alcohol as the dispersive reagent. The polymer microspheres were used as a selective sorbent for the solid-phase extraction of gonyautoxins 2,3. An off-line MISPE method followed by high-performance liquid chromatography (HPLC) with fluorescence detection for the analysis of gonyautoxins 2,3 was established. Finally, the extract samples from Alexandrium tamarense were analyzed. The results showed the imprinted polymer microspheres exhibited high affinity and selectivity for gonyautoxins 2,3. The interference matrix in the extract were obviously cleaned by MISPE and the extraction efficiency of gonyautoxins 2,3 in the sample ranged from 81.74% to 85.86%.

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

    NASA Astrophysics Data System (ADS)

    Gracia, Stéphanie; Roy, Suzanne; Starr, Michel

    2013-04-01

    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

  4. Germination fluctuation of toxic Alexandrium fundyense and A. pacificum cysts and the relationship with bloom occurrences in Kesennuma Bay, Japan.

    PubMed

    Natsuike, Masafumi; Yokoyama, Katsuhide; Nishitani, Goh; Yamada, Yuichiro; Yoshinaga, Ikuo; Ishikawa, Akira

    2017-02-01

    While cyst germination may be an important factor for the initiation of harmful/toxic blooms, assessments of the fluctuation in phytoplankton cyst germination, from bottom sediments to water columns, are rare in situ due to lack of technology that can detect germinated cells in natural bottom sediments. This study introduces a simple mesocosm method, modeled after previous in situ methods, to measure the germination of plankton resting stage cells. Using this method, seasonal changes in germination fluxes of toxic dinoflagellates resting cysts, specifically Alexandrium fundyense (A. tamarense species complex Group I) and A. pacificum (A. tamarense species complex Group IV), were investigated at a fixed station in Kesennuma Bay, northeast Japan, from April 2014 to April 2015. This investigation was conducted in addition to the typical samplings of seawater and bottom sediments to detect the dinoflagellates vegetative cells and resting cysts. Bloom occurrences of A. fundyense were observed June 2014 and February 2015 with maximum cell densities reaching 3.6×10(6) cells m(-2) and 1.4×10(7) cells m(-2), respectively. The maximum germination fluxes of A. fundyense cysts occurred in April 2014 and December 2014 and were 9.3×10(3) cells m(-2)day(-1) and 1.4×10(4) cells m(-2)day(-1), respectively. For A. pacificum, the highest cell density was 7.3×10(7) cells m(-2) during the month of August, and the maximum germination fluxes occurred in July and August, reaching 5.8×10(2) cells m(-2)day(-1). Thus, this study revealed the seasonal dynamics of A. fundyense and A. pacificum cyst germination and their bloom occurrences in the water column. Blooms occurred one to two months after peak germination, which strongly suggests that both the formation of the initial population by cyst germination and its continuous growth in the water column most likely contributed to toxic bloom occurrences of A. fundyense and A. pacificum in the bay.

  5. Zooplankton Community Grazing Impact on a Toxic Bloom of Alexandrium fundyense in the Nauset Marsh System, Cape Cod, Massachusetts, USA.

    PubMed

    Petitpas, Christian M; Turner, Jefferson T; Keafer, Bruce A; McGillicuddy, Dennis J; Anderson, Donald M

    2015-07-01

    Embayments and salt ponds along the coast of Massachusetts can host localized blooms of the toxic dinoflagellate Alexandrium fundyense. One such system, exhibiting a long history of toxicity and annual closures of shellfish beds, is the Nauset Marsh System (NMS) on Cape Cod. In order measure net growth rates of natural A. fundyense populations in the NMS during spring 2012, incubation experiments were conducted on seawater samples from two salt ponds within the NMS (Salt Pond and Mill Pond). Seawater samples containing natural populations of grazers and A. fundyense were incubated at ambient temperatures. Concentrations of A. fundyense after incubations were compared to initial abundances to determine net increases from population growth, or decreases presumed to be primarily due to grazing losses. Abundances of both microzooplankton (ciliates, rotifers, copepod nauplii and heterotrophic dinoflagellates) and mesozooplankton (copepodites and adult copepods, marine cladocerans, and meroplankton) grazers were also determined. This study documented net growth rates that were highly variable throughout the bloom, calculated from weekly bloom cell counts from the start of sampling to bloom peak in both ponds (Mill Pond range = 0.12 - 0.46 d(-1); Salt Pond range = -0.02 - 0.44 d(-1)). Microzooplankton grazers that were observed with ingested A. fundyense cells included polychaete larvae, rotifers, tintinnids, and heterotrophic dinoflagellates of the genera Polykrikos and Gymnodinium. Significant A. fundyense net growth was observed in two incubation experiments, and only a single experiment exhibited significant population losses. For the majority of experiments, due to high variability in data, net changes in A. fundyense abundance were not significant after the 24-hr incubations. However, experimental net growth rates through bloom peak were not statistically distinguishable from estimated long-term average net growth rates of natural populations in each pond (Mill Pond

  6. Zooplankton Community Grazing Impact on a Toxic Bloom of Alexandrium fundyense in the Nauset Marsh System, Cape Cod, Massachusetts, USA

    PubMed Central

    Petitpas, Christian M.; Turner, Jefferson T.; Keafer, Bruce A.; McGillicuddy, Dennis J.; Anderson, Donald M.

    2016-01-01

    Embayments and salt ponds along the coast of Massachusetts can host localized blooms of the toxic dinoflagellate Alexandrium fundyense. One such system, exhibiting a long history of toxicity and annual closures of shellfish beds, is the Nauset Marsh System (NMS) on Cape Cod. In order measure net growth rates of natural A. fundyense populations in the NMS during spring 2012, incubation experiments were conducted on seawater samples from two salt ponds within the NMS (Salt Pond and Mill Pond). Seawater samples containing natural populations of grazers and A. fundyense were incubated at ambient temperatures. Concentrations of A. fundyense after incubations were compared to initial abundances to determine net increases from population growth, or decreases presumed to be primarily due to grazing losses. Abundances of both microzooplankton (ciliates, rotifers, copepod nauplii and heterotrophic dinoflagellates) and mesozooplankton (copepodites and adult copepods, marine cladocerans, and meroplankton) grazers were also determined. This study documented net growth rates that were highly variable throughout the bloom, calculated from weekly bloom cell counts from the start of sampling to bloom peak in both ponds (Mill Pond range = 0.12 – 0.46 d−1; Salt Pond range = −0.02 – 0.44 d−1). Microzooplankton grazers that were observed with ingested A. fundyense cells included polychaete larvae, rotifers, tintinnids, and heterotrophic dinoflagellates of the genera Polykrikos and Gymnodinium. Significant A. fundyense net growth was observed in two incubation experiments, and only a single experiment exhibited significant population losses. For the majority of experiments, due to high variability in data, net changes in A. fundyense abundance were not significant after the 24-hr incubations. However, experimental net growth rates through bloom peak were not statistically distinguishable from estimated long-term average net growth rates of natural populations in each pond

  7. Identification and Characterization of Three Differentially Expressed Genes, Encoding S-Adenosylhomocysteine Hydrolase, Methionine Aminopeptidase, and a Histone-Like Protein, in the Toxic Dinoflagellate Alexandrium fundyense†

    PubMed Central

    Taroncher-Oldenburg, Gaspar; Anderson, Donald M.

    2000-01-01

    Genes showing differential expression related to the early G1 phase of the cell cycle during synchronized circadian growth of the toxic dinoflagellate Alexandrium fundyense were identified and characterized by differential display (DD). The determination in our previous work that toxin production in Alexandrium is relegated to a narrow time frame in early G1 led to the hypothesis that transcriptionally up- or downregulated genes during this subphase of the cell cycle might be related to toxin biosynthesis. Three genes, encoding S-adenosylhomocysteine hydrolase (Sahh), methionine aminopeptidase (Map), and a histone-like protein (HAf), were isolated. Sahh was downregulated, while Map and HAf were upregulated, during the early G1 phase of the cell cycle. Sahh and Map encoded amino acid sequences with about 90 and 70% similarity to those encoded by several eukaryotic and prokaryotic Sahh and Map genes, respectively. The partial Map sequence also contained three cobalt binding motifs characteristic of all Map genes. HAf encoded an amino acid sequence with 60% similarity to those of two histone-like proteins from the dinoflagellate Crypthecodinium cohnii Biecheler. This study documents the potential of applying DD to the identification of genes that are related to physiological processes or cell cycle events in phytoplankton under conditions where small sample volumes represent an experimental constraint. The identification of an additional 21 genes with various cell cycle-related DD patterns also provides evidence for the importance of pretranslational or transcriptional regulation in dinoflagellates, contrary to previous reports suggesting the possibility that translational mechanisms are the primary means of circadian regulation in this group of organisms. PMID:10788388

  8. Bloom development and transport of toxic Alexandrium fundyense populations within a coastal plume in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Keafer, Bruce A.; Churchill, James H.; McGillicuddy, Dennis J.; Anderson, Donald M.

    2005-09-01

    Toxic Alexandrium fundyense blooms in the western Gulf of Maine (GOM) are a common occurrence, causing paralytic shellfish poisoning (PSP) each spring. In contrast, high A. fundyense abundances and PSP toxicity commonly occur later in the summer in the eastern GOM and the Bay of Fundy. The objective of this study was to determine if the bloom dynamics of the two areas are linked early in the bloom season when initial outbreaks of toxicity are reported. A. fundyense cell abundance and hydrographic data were acquired during three cruises in May and June, 2001 spanning areas of the western and eastern GOM. Surface drifters also were released into the nearshore coastal flow of the eastern GOM. These data provide a coherent view of the springtime evolution of toxic A. fundyense blooms in the GOM and the influence of both small- and large-scale circulation. Early in the bloom season (May), the bulk of the A. fundyense population was consistently observed in an alongshore band of cells in the eastern GOM associated with relatively low-salinity water (<32) that likely originated from inputs further upstream in the coastal flow, predominantly the St. John River in the Bay of Fundy. At that time, the western GOM was virtually devoid of cells. In June, the population was bifurcated, with one branch extending alongshore into the western GOM and the other into the offshore waters of the interior GOM. This pattern was consistent with circulation models of the coastal GOM that unambiguously revealed a bifurcated flow with the branch nearest the coast directed alongshore to the western GOM. The most significant finding of this study is that A. fundyense populations along the eastern Maine coast were delivered along an "inside track" relative to the core of the eastern segment of the Maine Coastal Current. The transport pathway carried cells across the mouth of Penobscot Bay and into the western GOM coincident with outbreaks of nearshore PSP toxicity. The transport is influenced by

  9. Investigating the importance of sediment resuspension in Alexandrium fundyense cyst population dynamics in the Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Butman, Bradford; Aretxabaleta, Alfredo L.; Dickhudt, Patrick J.; Dalyander, P. Soupy; Sherwood, Christopher R.; Anderson, Donald M.; Keafer, Bruce A.; Signell, Richard P.

    2014-05-01

    Cysts of Alexandrium fundyense, a dinoflagellate that causes toxic algal blooms in the Gulf of Maine, spend the winter as dormant cells in the upper layer of bottom sediment or the bottom nepheloid layer and germinate in spring to initiate new blooms. Erosion measurements were made on sediment cores collected at seven stations in the Gulf of Maine in the autumn of 2011 to explore if resuspension (by waves and currents) could change the distribution of over-wintering cysts from patterns observed in the previous autumn; or if resuspension could contribute cysts to the water column during spring when cysts are viable. The mass of sediment eroded from the core surface at 0.4 Pa ranged from 0.05 kg m-2 near Grand Manan Island, to 0.35 kg m-2 in northern Wilkinson Basin. The depth of sediment eroded ranged from about 0.05 mm at a station with sandy sediment at 70 m water depth on the western Maine shelf, to about 1.2 mm in clayey-silt sediment at 250 m water depth in northern Wilkinson Basin. The sediment erodibility measurements were used in a sediment-transport model forced with modeled waves and currents for the period October 1, 2010 to May 31, 2011 to predict resuspension and bed erosion. The simulated spatial distribution and variation of bottom shear stress was controlled by the strength of the semi-diurnal tidal currents, which decrease from east to west along the Maine coast, and oscillatory wave-induced currents, which are strongest in shallow water. Simulations showed occasional sediment resuspension along the central and western Maine coast associated with storms, steady resuspension on the eastern Maine shelf and in the Bay of Fundy associated with tidal currents, no resuspension in northern Wilkinson Basin, and very small resuspension in western Jordan Basin. The sediment response in the model depended primarily on the profile of sediment erodibility, strength and time history of bottom stress, consolidation time scale, and the current in the water column

  10. Investigating the importance of sediment resuspension in Alexandrium fundyense cyst population dynamics in the Gulf of Maine

    USGS Publications Warehouse

    Butman, Bradford; Aretxabaleta, Alfredo L.; Dickhudt, Patrick J.; Dalyander, P. Soupy; Sherwood, Christopher R.; Anderson, Donald M.; Keafer, Bruce A.; Signell, Richard P.

    2014-01-01

    Cysts of Alexandrium fundyense, a dinoflagellate that causes toxic algal blooms in the Gulf of Maine, spend the winter as dormant cells in the upper layer of bottom sediment or the bottom nepheloid layer and germinate in spring to initiate new blooms. Erosion measurements were made on sediment cores collected at seven stations in the Gulf of Maine in the autumn of 2011 to explore if resuspension (by waves and currents) could change the distribution of over-wintering cysts from patterns observed in the previous autumn; or if resuspension could contribute cysts to the water column during spring when cysts are viable. The mass of sediment eroded from the core surface at 0.4 Pa ranged from 0.05 kg m−2 near Grand Manan Island, to 0.35 kg m−2 in northern Wilkinson Basin. The depth of sediment eroded ranged from about 0.05 mm at a station with sandy sediment at 70 m water depth on the western Maine shelf, to about 1.2 mm in clayey–silt sediment at 250 m water depth in northern Wilkinson Basin. The sediment erodibility measurements were used in a sediment-transport model forced with modeled waves and currents for the period October 1, 2010 to May 31, 2011 to predict resuspension and bed erosion. The simulated spatial distribution and variation of bottom shear stress was controlled by the strength of the semi-diurnal tidal currents, which decrease from east to west along the Maine coast, and oscillatory wave-induced currents, which are strongest in shallow water. Simulations showed occasional sediment resuspension along the central and western Maine coast associated with storms, steady resuspension on the eastern Maine shelf and in the Bay of Fundy associated with tidal currents, no resuspension in northern Wilkinson Basin, and very small resuspension in western Jordan Basin. The sediment response in the model depended primarily on the profile of sediment erodibility, strength and time history of bottom stress, consolidation time scale, and the current in the water

  11. Investigating the importance of sediment resuspension in Alexandrium fundyense cyst population dynamics in the Gulf of Maine

    PubMed Central

    Butman, Bradford; Aretxabaleta, Alfredo L.; Dickhudt, Patrick J.; Dalyander, P. Soupy; Sherwood, Christopher R.; Anderson, Donald M.; Keafer, Bruce A.; Signell, Richard P.

    2014-01-01

    Cysts of Alexandrium fundyense, a dinoflagellate that causes toxic algal blooms in the Gulf of Maine, spend the winter as dormant cells in the upper layer of bottom sediment or the bottom nepheloid layer and germinate in spring to initiate new blooms. Erosion measurements were made on sediment cores collected at seven stations in the Gulf of Maine in the autumn of 2011 to explore if resuspension (by waves and currents) could change the distribution of over-wintering cysts from patterns observed in the previous autumn; or if resuspension could contribute cysts to the water column during spring when cysts are viable. The mass of sediment eroded from the core surface at 0.4 Pa ranged from 0.05 kg m−2 near Grand Manan Island, to 0.35 kg m−2 in northern Wilkinson Basin. The depth of sediment eroded ranged from about 0.05 mm at a station with sandy sediment at 70 m water depth on the western Maine shelf, to about 1.2 mm in clayey–silt sediment at 250 m water depth in northern Wilkinson Basin. The sediment erodibility measurements were used in a sediment-transport model forced with modeled waves and currents for the period October 1, 2010 to May 31, 2011 to predict resuspension and bed erosion. The simulated spatial distribution and variation of bottom shear stress was controlled by the strength of the semi-diurnal tidal currents, which decrease from east to west along the Maine coast, and oscillatory wave-induced currents, which are strongest in shallow water. Simulations showed occasional sediment resuspension along the central and western Maine coast associated with storms, steady resuspension on the eastern Maine shelf and in the Bay of Fundy associated with tidal currents, no resuspension in northern Wilkinson Basin, and very small resuspension in western Jordan Basin. The sediment response in the model depended primarily on the profile of sediment erodibility, strength and time history of bottom stress, consolidation time scale, and the current in the water

  12. Investigating the importance of sediment resuspension in Alexandrium fundyense cyst population dynamics in the Gulf of Maine.

    PubMed

    Butman, Bradford; Aretxabaleta, Alfredo L; Dickhudt, Patrick J; Dalyander, P Soupy; Sherwood, Christopher R; Anderson, Donald M; Keafer, Bruce A; Signell, Richard P

    2014-05-01

    Cysts of Alexandrium fundyense, a dinoflagellate that causes toxic algal blooms in the Gulf of Maine, spend the winter as dormant cells in the upper layer of bottom sediment or the bottom nepheloid layer and germinate in spring to initiate new blooms. Erosion measurements were made on sediment cores collected at seven stations in the Gulf of Maine in the autumn of 2011 to explore if resuspension (by waves and currents) could change the distribution of over-wintering cysts from patterns observed in the previous autumn; or if resuspension could contribute cysts to the water column during spring when cysts are viable. The mass of sediment eroded from the core surface at 0.4 Pa ranged from 0.05 kg m(-2) near Grand Manan Island, to 0.35 kg m(-2) in northern Wilkinson Basin. The depth of sediment eroded ranged from about 0.05 mm at a station with sandy sediment at 70 m water depth on the western Maine shelf, to about 1.2 mm in clayey-silt sediment at 250 m water depth in northern Wilkinson Basin. The sediment erodibility measurements were used in a sediment-transport model forced with modeled waves and currents for the period October 1, 2010 to May 31, 2011 to predict resuspension and bed erosion. The simulated spatial distribution and variation of bottom shear stress was controlled by the strength of the semi-diurnal tidal currents, which decrease from east to west along the Maine coast, and oscillatory wave-induced currents, which are strongest in shallow water. Simulations showed occasional sediment resuspension along the central and western Maine coast associated with storms, steady resuspension on the eastern Maine shelf and in the Bay of Fundy associated with tidal currents, no resuspension in northern Wilkinson Basin, and very small resuspension in western Jordan Basin. The sediment response in the model depended primarily on the profile of sediment erodibility, strength and time history of bottom stress, consolidation time scale, and the current in the water

  13. Outbreeding lethality between toxic Group I and nontoxic Group III Alexandrium tamarense spp. isolates: Predominance of heterotypic encystment and implications for mating interactions and biogeography

    NASA Astrophysics Data System (ADS)

    Brosnahan, Michael L.; Kulis, David M.; Solow, Andrew R.; Erdner, Deana L.; Percy, Linda; Lewis, Jane; Anderson, Donald M.

    2010-02-01

    We report the zygotic encystment of geographically dispersed isolates in the dinoflagellate species complex Alexandrium tamarense, in particular, successful mating of toxic Group I and nontoxic Group III isolates. However, hypnozygotes produced in Group I/III co-cultures complete no more than three divisions after germinating. Previous reports have suggested a mate recognition mechanism whereby hypnozygotes produced in co-cultures could arise from either homotypic (inbred) or heterotypic (outbred) gamete pairs. To determine the extent to which each occurs, a nested PCR assay was developed to determine parentage of individual hypnozygotes. The vast majority of hypnozygotes from pairwise Group I/III co-cultures were outbred, so that inviability was a result of hybridization, not inbreeding. These findings support the assertion that complete speciation underlies the phylogenetic structure of the Alexandrium tamarense species complex. Additionally, the ribosomal DNA (rDNA) copy numbers of both hybrid and single ribotype hypnozygotes were reduced substantially from those of haploid motile cells. The destruction of rDNA loci may be crucial for the successful mating of genetically distant conjugants and appears integral to the process of encystment. The inviability of Group I/III hybrids is important for public health because the presence of hybrid cysts may indicate ongoing displacement of a nontoxic population by a toxic one (or vice versa). Hybrid inviability also suggests a bloom control strategy whereby persistent, toxic Group I blooms could be mitigated by introduction of nontoxic Group III cells. The potential for hybridization in nature was investigated by applying the nested PCR assay to hypnozygotes from Belfast Lough, Northern Ireland, a region where Group I and III populations co-occur. Two hybrid cysts were identified in 14 successful assays, demonstrating that Group I and III populations do interbreed in that region. However, an analysis of mating data

  14. Relationship between valve activity, microalgae concentration in the water and toxin accumulation in the digestive gland of the Pacific oyster Crassostrea gigas exposed to Alexandrium minutum.

    PubMed

    Haberkorn, Hansy; Tran, Damien; Massabuau, Jean-Charles; Ciret, Pierre; Savar, Véronique; Soudant, Philippe

    2011-06-01

    The complexity of the relationships between Alexandrium minutum (A.m.) concentration in the water ([A.m.](w)), Paralytic Shellfish Poisoning contamination in the digestive gland ([PSP](dg)) and valve behavior was explored in oysters Crassostrea gigas. Two experiments were conducted, during which oysters' valve behaviour were analyzed. Oysters, first acclimated for 10-days with the non harmful microalgae Heterocapsa triquetra (H.t.), were exposed to four microalgae mixtures at constant total concentrations of 10×10(3)cells ml(-1) (experiment-1) and 5×10(3)cells ml(-1) (experiment-2): 100% A.m.; 50% A.m.-50% H.t.; 25% A.m.-75% H.t.; 100% H.t. At the end of experiment-2, [PSP](dg) were measured. At 10×10(3)cells ml(-1), the microalgal ingestion decreased (p<0.05) with increasing [A.m.](w) but not at 5×10(3)cells ml(-1) (p>0.05). The frequency of microclosures specifically increased with [A.m.](w) (p<0.05) and the opening duration with [PSP](dg) (p<0.0001). Oysters exhibiting the maximum increase in opening duration also exhibited the highest [PSP](dg). The results are discussed in terms of oyster physiology and origin of the behavioral response.

  15. Discovery of an algicidal compound from Brevibacterium sp. BS01 and its effect on a harmful algal bloom-causing species, Alexandrium tamarense

    PubMed Central

    An, Xinli; Zhang, Bangzhou; Zhang, Huajun; Li, Yi; Zheng, Wei; Yu, Zhiming; Fu, Lijun; Zheng, Tianling

    2015-01-01

    Blooms of the dinoflagellate Alexandrium tamarense have become worldwide phenomena and have detrimental impacts on aquatic ecosystems and human health. In this study, a culture supernatant of the marine actinomycete BS01 exerted a strong algicidal effect on A. tamarense (ATGD98-006). The target algicide from BS01 was separated by adsorption chromatography and identified by MALDI-TOF-MS and NMR analysis. The results suggested that the purified algicidal component corresponded to a hydrophobic compound (2-isobutoxyphenyl)amine (C10H15NO) with a molecular weight of 165 Da, which exhibited a significant algicidal effect (64.5%) on A. tamarense. After incubation in 5 μg/mL of (2-isobutoxyphenyl)amine for 24 h, the algae lost mobility and sank to the bottom of the flasks, and 56.5% of the algae cells lost vitality at a concentration of 20 μg/mL (p < 0.01) despite having intact cell profiles. Morphological analysis revealed that the cell structure of A. tamarense was altered by (2-isobutoxyphenyl)amine resulting in cytoplasm degradation and the loss of organelle integrity. The images following propidium iodide staining suggested that the algal nucleus was also severely damaged and eventually degraded due to exposure to the algicidal compound. All of the results indicate that (2-isobutoxyphenyl)amine from the actinomycete might be a candidate for the control of bloom-forming A. tamarense. PMID:26594205

  16. Genome sequence of the Roseovarius mucosus type strain (DSM 17069T), a bacteriochlorophyll a-containing representative of the marine Roseobacter group isolated from the dinoflagellate Alexandrium ostenfeldii

    PubMed Central

    2015-01-01

    Roseovarius mucosus Biebl et al. 2005 is a bacteriochlorophyll a-producing representative of the marine Roseobacter group within the alphaproteobacterial family Rhodobacteraceae, which was isolated from the dinoflagellate Alexandrium ostenfeldii. The marine Roseobacter group was found to be abundant in the ocean and plays an important role for global and biogeochemical processes. Here we describe the features of the R. mucosus strain DFL-24T together with its genome sequence and annotation generated from a culture of DSM 17069T. The 4,247,724 bp containing genome sequence encodes 4,194 protein-coding genes and 57 RNA genes. In addition to the presence of four plasmids, genome analysis revealed the presence of genes associated with host colonization, DMSP utilization, cytotoxins, and quorum sensing that could play a role in the interrelationship of R. mucosus with the dinoflagellate A. ostenfeldii and other marine organisms. Furthermore, the genome encodes genes associated with mixotrophic growth, where both reduced inorganic compounds for lithotrophic growth and a photoheterotrophic lifestyle using light as additional energy source could be used. PMID:26203330

  17. Effects of adaptation, chance, and history on the evolution of the toxic dinoflagellate Alexandrium minutum under selection of increased temperature and acidification

    PubMed Central

    Flores-Moya, Antonio; Rouco, Mónica; García-Sánchez, María Jesús; García-Balboa, Camino; González, Raquel; Costas, Eduardo; López-Rodas, Victoria

    2012-01-01

    The roles of adaptation, chance, and history on evolution of the toxic dinoflagellate Alexandrium minutum Halim, under selective conditions simulating global change, have been addressed. Two toxic strains (AL1V and AL2V), previously acclimated for two years at pH 8.0 and 20°C, were transferred to selective conditions: pH 7.5 to simulate acidification and 25°C. Cultures under selective conditions were propagated until growth rate and toxin cell quota achieved an invariant mean value at 720 days (ca. 250 and ca. 180 generations for strains AL1V and AL2V, respectively). Historical contingencies strongly constrained the evolution of growth rate and toxin cell quota, but the forces involved in the evolution were not the same for both traits. Growth rate was 1.5–1.6 times higher than the one measured in ancestral conditions. Genetic adaptation explained two-thirds of total adaptation while one-third was a consequence of physiological adaptation. On the other hand, the evolution of toxin cell quota showed a pattern attributable to neutral mutations because the final variances were significantly higher than those measured at the start of the experiment. It has been hypothesized that harmful algal blooms will increase under the future scenario of global change. Although this study might be considered an oversimplification of the reality, it can be hypothesized that toxic blooms will increase but no predictions can be advanced about toxicity. PMID:22833798

  18. Near-bottom circulation and dispersion of sediment containing Alexandrium fundyense cysts in the Gulf of Maine during 2010–2011

    PubMed Central

    Aretxabaleta, Alfredo L.; Butman, Bradford; Signell, Richard P.; Dalyander, P. Soupy; Sherwood, Christopher R.; Sheremet, Vitalii A.; McGillicuddy, Dennis J.

    2015-01-01

    The life cycle of Alexandrium fundyense in the Gulf of Maine includes a dormant cyst stage that spends the winter predominantly in the bottom sediment. Wave-current bottom stress caused by storms and tides induces resuspension of cyst-containing sediment during winter and spring. Resuspended sediment could be transported by water flow to different locations in the Gulf and the redistribution of sediment containing A. fundyense cysts could alter the spatial and temporal manifestation of its spring bloom. The present study evaluates model near-bottom flow during storms, when sediment resuspension and redistribution are most likely to occur, between October and May when A. fundyense cells are predominantly in cyst form. Simulated water column sediment (mud) concentrations from representative locations of the Gulf are used to initialize particle tracking simulations for the period October 2010–May 2011. Particles are tracked in full three-dimensional model solutions including a sinking velocity characteristic of cyst and aggregated mud settling (0.1 mm s−1). Although most of the material was redeposited near the source areas, small percentages of total resuspended sediment from some locations in the western (~4%) and eastern (2%) Maine shelf and the Bay of Fundy (1%) traveled distances longer than 100 km before resettling. The redistribution changed seasonally and was sensitive to the prescribed sinking rate. Estimates of the amount of cysts redistributed with the sediment are small compared to the inventory of cysts in the upper few centimeters of sediment. PMID:26045635

  19. Near-bottom circulation and dispersion of sediment containing Alexandrium fundyense cysts in the Gulf of Maine during 2010-2011.

    PubMed

    Aretxabaleta, Alfredo L; Butman, Bradford; Signell, Richard P; Dalyander, P Soupy; Sherwood, Christopher R; Sheremet, Vitalii A; McGillicuddy, Dennis J

    2014-05-01

    The life cycle of Alexandrium fundyense in the Gulf of Maine includes a dormant cyst stage that spends the winter predominantly in the bottom sediment. Wave-current bottom stress caused by storms and tides induces resuspension of cyst-containing sediment during winter and spring. Resuspended sediment could be transported by water flow to different locations in the Gulf and the redistribution of sediment containing A. fundyense cysts could alter the spatial and temporal manifestation of its spring bloom. The present study evaluates model near-bottom flow during storms, when sediment resuspension and redistribution are most likely to occur, between October and May when A. fundyense cells are predominantly in cyst form. Simulated water column sediment (mud) concentrations from representative locations of the Gulf are used to initialize particle tracking simulations for the period October 2010-May 2011. Particles are tracked in full three-dimensional model solutions including a sinking velocity characteristic of cyst and aggregated mud settling (0.1 mm s(-1)). Although most of the material was redeposited near the source areas, small percentages of total resuspended sediment from some locations in the western (~4%) and eastern (2%) Maine shelf and the Bay of Fundy (1%) traveled distances longer than 100 km before resettling. The redistribution changed seasonally and was sensitive to the prescribed sinking rate. Estimates of the amount of cysts redistributed with the sediment are small compared to the inventory of cysts in the upper few centimeters of sediment.

  20. Blooms of the toxic dinoflagellate Alexandrium fundyense in the western Gulf of Maine in 1993 and 1994: A comparative modeling study

    NASA Astrophysics Data System (ADS)

    Stock, Charles A.; McGillicuddy, Dennis J.; Anderson, Donald M.; Solow, Andrew R.; Signell, Richard P.

    2007-11-01

    Blooms of the toxic dinoflagellate Alexandrium fundyense commonly occur in the western Gulf of Maine but the amount of toxin observed in coastal shellfish is highly variable. In this study, a coupled physical-biological model is used to investigate the dynamics underlying the observed A. fundyense abundance and shellfish toxicity in 1993 (a high toxicity year) and 1994 (low toxicity year). The physical model simulates the spring circulation, while the biological model estimates the germination and population dynamics of A. fundyense based on laboratory and field data. The model captures the large-scale aspects of the initiation and development of A. fundyense blooms during both years, but small-scale patchiness and the dynamics of bloom termination remain problematic. In both cases, the germination of resting cysts accounts for the magnitude of A. fundyense populations early in the spring. Simulations with low net A. fundyense growth rates capture the mean observed concentration during the bloom peak, which is of similar magnitude during both years. There is little evidence that large-scale changes in biological dynamics between 1993 and 1994 were a primary driver of the differences in shellfish toxicity. Results instead suggest that the persistent southwesterly flow of the western Maine Coastal Current led to A. fundyense populations of similar alongshore extent by late May of both years. This period coincides with peak cell abundance in the region. Variations in wind forcing (downwelling favorable in 1993, upwelling favorable in 1994) and subsequent cell transport (inshore in 1993, offshore in 1994) in early June then provides a plausible explanation for the dramatic mid-June differences in shellfish toxicity throughout the western Gulf of Maine.

  1. A Kinetic and Factorial Approach to Study the Effects of Temperature and Salinity on Growth and Toxin Production by the Dinoflagellate Alexandrium ostenfeldii from the Baltic Sea

    PubMed Central

    Salgado, Pablo; Vázquez, José A.; Riobó, Pilar; Franco, José M.; Figueroa, Rosa I.; Kremp, Anke; Bravo, Isabel

    2015-01-01

    Alexandrium ostenfeldii is present in a wide variety of environments in coastal areas worldwide and is the only dinoflagellate known species that produces paralytic shellfish poisoning (PSP) toxins and two types of cyclic imines, spirolides (SPXs) and gymnodimines (GYMs). The increasing frequency of A. ostenfeldii blooms in the Baltic Sea has been attributed to the warming water in this region. To learn more about the optimal environmental conditions favoring the proliferation of A. ostenfeldii and its complex toxicity, the effects of temperature and salinity on the kinetics of both the growth and the net toxin production of this species were examined using a factorial design and a response-surface analysis (RSA). The results showed that the growth of Baltic A. ostenfeldii occurs over a wide range of temperatures and salinities (12.5–25.5°C and 5–21, respectively), with optimal growth conditions achieved at a temperature of 25.5°C and a salinity of 11.2. Together with the finding that a salinity > 21 was the only growth-limiting factor detected for this strain, this study provides important insights into the autecology and population distribution of this species in the Baltic Sea. The presence of PSP toxins, including gonyautoxin (GTX)-3, GTX-2, and saxitoxin (STX), and GYMs (GYM-A and GYM-B/-C analogues) was detected under all temperature and salinity conditions tested and in the majority of the cases was concomitant with both the exponential growth and stationary phases of the dinoflagellate’s growth cycle. Toxin concentrations were maximal at temperatures and salinities of 20.9°C and 17 for the GYM-A analogue and > 19°C and 15 for PSP toxins, respectively. The ecological implications of the optimal conditions for growth and toxin production of A. ostenfeldii in the Baltic Sea are discussed. PMID:26636674

  2. The ability of the branchiopod, Artemia salina, to graze upon harmful algal blooms caused by Alexandrium fundyense, Aureococcus anophagefferens, and Cochlodinium polykrikoides

    NASA Astrophysics Data System (ADS)

    Marcoval, M. Alejandra; Pan, Jerónimo; Tang, Yingzhong; Gobler, Christopher J.

    2013-10-01

    We present experiments that examined the grazing and survivorship of zooplankton native (Acartia tonsa) and non-native (Artemia salina) to NY (USA) estuaries when exposed to blooms and cultures of the three harmful algae native to NY, Alexandrium fundyense, Aureococcus anophagefferens (strains CCMP 1850 and CCMP 1984) and Cochlodinium polykrikoides. During experiments with cultures of A. anophagefferens, clearance rates (CR) of A. salina were significantly greater than those of A. tonsa for both algal strains examined. A. salina fed on cultures of C. polykrikoides at higher rates than all phytoplankton species examined, including the control diet (Rhodomonas salina), and faster than rates of A. tonsa fed C. polykrikoides. During experiments with A. fundyense, A. salina actively grazed all cell concentrations (250-1500 cells ml-1) while A. tonsa did not feed at any concentration. Percent mortality of A. salina and A. tonsa fed A. fundyense for 48 h were 43 ± 7.7% and 72 ± 7.8%, respectively, percentages significantly higher than those of individuals fed all other algal diets. During 25 field experiments using natural blooms of the three HAB species performed across six NY estuaries, A. salina significantly (p < 0.05) reduced cell densities of A. anophagefferens, C. polykrikoides, and A. fundyense relative to the control treatments in all but one experiment. The sum of these findings demonstrates that a failure to graze these HABs by the indigenous copepod, A. tonsa, may permit blooms to occur. In addition, the ability of A. salina to graze these HABs at densities that were inhibitory to A. tonsa suggests that A. salina could, in some circumstances, be considered as a part of mitigation strategy for these events.

  3. A Kinetic and Factorial Approach to Study the Effects of Temperature and Salinity on Growth and Toxin Production by the Dinoflagellate Alexandrium ostenfeldii from the Baltic Sea.

    PubMed

    Salgado, Pablo; Vázquez, José A; Riobó, Pilar; Franco, José M; Figueroa, Rosa I; Kremp, Anke; Bravo, Isabel

    2015-01-01

    Alexandrium ostenfeldii is present in a wide variety of environments in coastal areas worldwide and is the only dinoflagellate known species that produces paralytic shellfish poisoning (PSP) toxins and two types of cyclic imines, spirolides (SPXs) and gymnodimines (GYMs). The increasing frequency of A. ostenfeldii blooms in the Baltic Sea has been attributed to the warming water in this region. To learn more about the optimal environmental conditions favoring the proliferation of A. ostenfeldii and its complex toxicity, the effects of temperature and salinity on the kinetics of both the growth and the net toxin production of this species were examined using a factorial design and a response-surface analysis (RSA). The results showed that the growth of Baltic A. ostenfeldii occurs over a wide range of temperatures and salinities (12.5-25.5°C and 5-21, respectively), with optimal growth conditions achieved at a temperature of 25.5°C and a salinity of 11.2. Together with the finding that a salinity > 21 was the only growth-limiting factor detected for this strain, this study provides important insights into the autecology and population distribution of this species in the Baltic Sea. The presence of PSP toxins, including gonyautoxin (GTX)-3, GTX-2, and saxitoxin (STX), and GYMs (GYM-A and GYM-B/-C analogues) was detected under all temperature and salinity conditions tested and in the majority of the cases was concomitant with both the exponential growth and stationary phases of the dinoflagellate's growth cycle. Toxin concentrations were maximal at temperatures and salinities of 20.9°C and 17 for the GYM-A analogue and > 19°C and 15 for PSP toxins, respectively. The ecological implications of the optimal conditions for growth and toxin production of A. ostenfeldii in the Baltic Sea are discussed.

  4. The role of a PSP-producing Alexandrium bloom in an unprecedented diamondback terrapin (Malaclemys terrapin) mortality event in Flanders Bay, New York, USA.

    PubMed

    Hattenrath-Lehmann, Theresa K; Ossiboff, Robert J; Burnell, Craig A; Rauschenberg, Carlton D; Hynes, Kevin; Burke, Russell L; Bunting, Elizabeth M; Durham, Kim; Gobler, Christopher J

    2017-04-01

    Diamondback terrapins (Malaclemys terrapin) are a threatened or endangered species in much of their range along the U.S. Atlantic and Gulf coasts. Over an approximately three-week period from late April to mid-May 2015, hundreds of adult diamondback terrapins were found dead on the shores of Flanders Bay, Long Island, New York, USA. Concurrent with the mortality event, elevated densities of the paralytic shellfish toxin (PST)-producing dinoflagellate, Alexandrium fundyense (>10(4) cells L(-1)) and high levels of PST in bivalves (maximal levels = 540 μg STX eq. 100 g(-1) shellfish tissue) were observed in the Flanders Bay region, resulting in shellfish bed closures in regional tributaries. Gross and histologic postmortem examinations of terrapins revealed no physical trauma to individuals or a common, underlying disease process to explain the deaths. PST compounds (0.2-12.5 μg STX eq. 100 g(-1)) were present in various M. terrapin tissues collected over the duration of the mortality event. High-throughput sequencing revealed that the ribbed mussel (Geukensia demissa, a PST vector) was present in the gastrointestinal tracks of all terrapin samples tested. While the potential of PST to cause mortality in chelonians has not been well-characterized, in the absence of other significant findings from necropsies and pathological analyses, we provide evidence that PST in shellfish was likely high enough to cause or contribute to the mortality in these small (<2.0 kg) animals.

  5. Photosynthetic Carbon Isotope Fractionation of the Marine Dinoflagellate Alexandrium tamarense: A Chemostat Investigation of Taxonomic and Physiological Controls on the Stable Carbon Isotope Record

    NASA Astrophysics Data System (ADS)

    Wilkes, E.; Carter, S. J.; Pearson, A.

    2015-12-01

    Interpretations of stable carbon isotope excursions in the sedimentary record are strengthened by laboratory culture studies investigating the photosynthetic carbon isotope fractionation (ɛp) of marine phytoplankton taxa with long geological records. These studies are essential for understanding organic matter δ13C signals in terms of environmental changes (e.g., atmospheric pCO2 and nutrient availability) or taxonomic changes (e.g., algal species succession and community composition). Dinoflagellates are among the most widespread and ecologically dominant primary producers in modern oceans and throughout the Mesozoic and Cenozoic. Compared to more recently evolved phytoplankton taxa, however, dinoflagellate carbon isotope fractionation has received relatively little mechanistic study. Several dilute batch culture experiments with dinoflagellates have investigated ɛp as a function of CO2 availability, but the influences of changing growth rates, nutrient limitation, pH, and irradiance require further systematic exploration. We investigated stable carbon isotope fractionation in the marine dinoflagellate Alexandrium tamarense under nitrate-limited conditions in a chemostat culture system in which full DIC system parameters, including the concentration and δ13C value of CO2, were determined. Growth rates were varied between experiments, and cells were grown under continuous light. Previously reported ɛp values for seven dinoflagellate species including A. tamarense ranged from approximately -1 to 14‰ in nutrient-replete batch culture studies ([CO2] = 0-50 µmol kg-1). In contrast, in chemostat conditions we measured ɛp values on the order of 20‰ ([CO2] = 20-30 µmol kg-1). These experiments provide an initial step toward understanding the physiological controls on ɛp in dinoflagellates and illuminating the role of algal taxonomy in shaping the Phanerozoic stable carbon isotope record.

  6. Extensive genetic diversity and rapid population differentiation during blooms of Alexandrium fundyense (Dinophyceae) in an isolated salt pond on Cape Cod, MA, USA

    PubMed Central

    Richlen, Mindy L; Erdner, Deana L; McCauley, Linda A R; Libera, Katie; Anderson, Donald M

    2012-01-01

    In Massachusetts, paralytic shellfish poisoning (PSP) is annually recurrent along the coastline, including within several small embayments on Cape Cod. One such system, the Nauset Marsh System (NMS), supports extensive marshes and a thriving shellfishing industry. Over the last decade, PSP in the NMS has grown significantly worse; however, the origins and dynamics of the toxic Alexandrium fundyense (Balech) populations that bloom within the NMS are not well known. This study examined a collection of 412 strains isolated from the NMS and the Gulf of Maine (GOM) in 2006–2007 to investigate the genetic characteristics of localized blooms and assess connectivity with coastal populations. Comparisons of genetic differentiation showed that A. fundyense blooms in the NMS exhibited extensive clonal diversity and were genetically distinct from populations in the GOM. In both project years, genetic differentiation was observed among temporal samples collected from the NMS, sometimes occurring on the order of approximately 7 days. The underlying reasons for temporal differentiation are unknown, but may be due, in part, to life-cycle characteristics unique to the populations in shallow embayments, or possibly driven by selection from parasitism and zooplankton grazing; these results highlight the need to investigate the role of selective forces in the genetic dynamics of bloom populations. The small geographic scale and limited connectivity of NMS salt ponds provide a novel system for investigating regulators of blooms, as well as the influence of selective forces on population structure, all of which are otherwise difficult or impossible to study in the adjacent open-coastal waters or within larger estuaries. PMID:23145343

  7. Near-bottom circulation and dispersion of sediment containing Alexandrium fundyense cysts in the Gulf of Maine during 2010-2011

    USGS Publications Warehouse

    Aretxabaleta, Alfredo L.; Butman, Bradford; Signell, Richard P.; Dalyander, P. Soupy; Sherwood, Christopher R.; Sheremet, Vitalii A.; McGillicuddy, Dennis J.

    2014-01-01

    The life cycle of Alexandrium fundyense in the Gulf of Maine includes a dormant cyst stage that spends the winter predominantly in the bottom sediment. Wave-current bottom stress caused by storms and tides induces resuspension of cyst-containing sediment during winter and spring. Resuspended sediment could be transported by water flow to different locations in the Gulf and the redistribution of sediment containing A. fundyense cysts could alter the spatial and temporal manifestation of its spring bloom. The present study evaluates model near-bottom flow during storms, when sediment resuspension and redistribution are most likely to occur, between October and May when A. fundyense cells are predominantly in cyst form. Simulated water column sediment (mud) concentrations from representative locations of the Gulf are used to initialize particle tracking simulations for the period October 2010–May 2011. Particles are tracked in full three-dimensional model solutions including a sinking velocity characteristic of cyst and aggregated mud settling (0.1 mm s−1). Although most of the material was redeposited near the source areas, small percentages of total resuspended sediment from some locations in the western (~4%) and eastern (2%) Maine shelf and the Bay of Fundy (1%) traveled distances longer than 100 km before resettling. The redistribution changed seasonally and was sensitive to the prescribed sinking rate. Estimates of the amount of cysts redistributed with the sediment were small compared to the inventory of cysts in the upper few centimeters of sediment but could potentially have more relevance immediately after deposition.

  8. Short-Term Behavioural Responses of the Great Scallop Pecten maximus Exposed to the Toxic Alga Alexandrium minutum Measured by Accelerometry and Passive Acoustics

    PubMed Central

    Coquereau, Laura; Jolivet, Aurélie; Hégaret, Hélène; Chauvaud, Laurent

    2016-01-01

    Harmful algal blooms produced by toxic dinoflagellates have increased worldwide, impacting human health, the environment, and fisheries. Due to their potential sensitivity (e.g., environmental changes), bivalves through their valve movements can be monitored to detect harmful algal blooms. Methods that measure valve activity require bivalve-attached sensors and usually connected cables to data transfers, leading to stress animals and limit the use to sessile species. As a non-intrusive and continuously deployable tool, passive acoustics could be an effective approach to detecting harmful algal blooms in real time based on animal sound production. This study aimed to detect reaction changes in the valve movements of adult Pecten maximus exposed to the toxic dinoflagellate Alexandrium minutum using both accelerometry and passive acoustic methods. Scallops were experimentally exposed to three ecologically relevant concentrations of A. minutum for 2 hours. The number of each type of valve movement and their sound intensity, opening duration, and valve-opening amplitude were measured. Four behaviours were identified: closures, expulsion, displacement, and swimming. The response of P. maximus to A. minutum occurred rapidly at a high concentration. The valve activity of P. maximus was different when exposed to high concentrations (500 000 cells L-1) of A. minutum compared to the non-toxic dinoflagellate Heterocapsa triquetra; the number of valve movements increased, especially closure and expulsion, which were detected acoustically. Thus, this study demonstrates the potential for acoustics and sound production changes in the detection of harmful algal blooms. However, field trials and longer duration experiments are required to provide further evidence for the use of acoustics as a monitoring tool in the natural environment where several factors may interfere with valve behaviours. PMID:27508498

  9. The intricacies of dinoflagellate pellicle cysts: The example of Alexandrium minutum cysts from a bloom-recurrent area (Bay of Baiona, NW Spain)

    NASA Astrophysics Data System (ADS)

    Bravo, Isabel; Isabel Figueroa, Rosa; Garcés, Esther; Fraga, Santiago; Massanet, Ana

    2010-02-01

    The terms "temporary", "pellicle", and "ecdysal" cyst have been employed arbitrarily in the literature of the dinoflagellate life cycle to describe a non-motile and single-layered-wall stage with no mandatory dormancy period, of asexual or sexual origin. These three terms have been used more or less synonymously, but more specific definitions, taking into account morphological and physiological aspects and their roles in dinoflagellate population dynamics, are still needed. To clarify the current terminology, we examine and discuss the usages and foundations of those terms. The background for this discussion is provided by a comparison of the morphology and germination times of three different types of Alexandrium minutum cysts collected during a seasonal bloom in the Bay of Baiona (NW Spain). The double-walled cysts were similar to the resting cysts reported for this species, but other, thin-walled and thecate cysts were also observed. These latter cyst types needed between 1 and 17 days to germinate and were therefore considered as short-term cysts, in contrast to the 1.5-month dormancy period of resting (hypnozygotic) cysts. Our results showed that the temporal distribution of these short-term cysts during the bloom period followed a pattern very similar to that of vegetative cells. However, resting cysts were only detected at the end of the bloom. In the context of our present knowledge regarding the dormancy and quiescence of dinoflagellate cysts, "temporary" is a very misleading and uncertain term and must be rejected. The term "ecdysal" has been used in reference to thin-walled cysts when ecdysis has been proven; however, ecdysis is not unique to this type of cysts as thick-walled zygotic cysts can be formed thorough ecdysis of a thecate planozygote. In conclusion, based on our current understanding of cysts, the term "pellicle" more appropriately describes single-layered-wall stages.

  10. Blooms of the toxic dinoflagellate Alexandrium fundyense in the western Gulf of Maine in 1993 and 1994: A comparative modeling study

    USGS Publications Warehouse

    Stock, C.A.; McGillicuddy, D.J.; Anderson, D.M.; Solow, A.R.; Signell, R.P.

    2007-01-01

    Blooms of the toxic dinoflagellate Alexandrium fundyense commonly occur in the western Gulf of Maine but the amount of toxin observed in coastal shellfish is highly variable. In this study, a coupled physical-biological model is used to investigate the dynamics underlying the observed A. fundyense abundance and shellfish toxicity in 1993 (a high toxicity year) and 1994 (low toxicity year). The physical model simulates the spring circulation, while the biological model estimates the germination and population dynamics of A. fundyense based on laboratory and field data. The model captures the large-scale aspects of the initiation and development of A. fundyense blooms during both years, but small-scale patchiness and the dynamics of bloom termination remain problematic. In both cases, the germination of resting cysts accounts for the magnitude of A. fundyense populations early in the spring. Simulations with low net A. fundyense growth rates capture the mean observed concentration during the bloom peak, which is of similar magnitude during both years. There is little evidence that large-scale changes in biological dynamics between 1993 and 1994 were a primary driver of the differences in shellfish toxicity. Results instead suggest that the persistent southwesterly flow of the western Maine Coastal Current led to A. fundyense populations of similar alongshore extent by late May of both years. This period coincides with peak cell abundance in the region. Variations in wind forcing (downwelling favorable in 1993, upwelling favorable in 1994) and subsequent cell transport (inshore in 1993, offshore in 1994) in early June then provides a plausible explanation for the dramatic mid-June differences in shellfish toxicity throughout the western Gulf of Maine. ?? 2007 Elsevier Ltd. All rights reserved.

  11. Mechanisms regulating large-scale seasonal fluctuations in Alexandrium fundyense populations in the Gulf of Maine: Results from a physical biological model

    NASA Astrophysics Data System (ADS)

    McGillicuddy, D. J.; Anderson, D. M.; Lynch, D. R.; Townsend, D. W.

    2005-09-01

    Observations of Alexandrium fundyense in the Gulf of Maine indicate several salient characteristics of the vegetative cell distributions: patterns of abundance are gulf-wide in geographic scope; their main features occur in association with the Maine Coastal Current; and the center of mass of the distribution shifts upstream from west to east during the growing season from April to August. The mechanisms underlying these aspects are investigated using coupled physical-biological simulations that represent the population dynamics of A. fundyense within the seasonal mean flow. A model that includes germination, growth, mortality, and nutrient limitation is qualitatively consistent with the observations. Germination from resting cysts appears to be a key aspect of the population dynamics that confines the cell distribution near the coastal margin, as simulations based on a uniform initial inoculum of vegetative cells across the Gulf of Maine produces blooms that are broader in geographic extent than is observed. In general, cells germinated from the major cyst beds (in the Bay of Fundy and near Penobscot and Casco Bays) are advected in the alongshore direction from east to west in the coastal current. Growth of the vegetative cells is limited primarily by temperature from April through June throughout the gulf, whereas nutrient limitation occurs in July and August in the western gulf. Thus the seasonal shift in the center of mass of cells from west to east can be explained by changing growth conditions: growth is more rapid in the western gulf early in the season due to warmer temperatures, whereas growth is more rapid in the eastern gulf later in the season due to severe nutrient limitation in the western gulf during that time period. A simple model of encystment based on nutrient limitation predicts deposition of new cysts in the vicinity of the observed cyst bed offshore of Casco and Penobscot Bays, suggesting a pathway of re-seeding the bed from cells advected

  12. Alexandrium fundyense cysts in the Gulf of Maine: long-term time series of abundance and distribution, and linkages to past and future blooms.

    PubMed

    Anderson, Donald M; Keafer, Bruce A; Kleindinst, Judith L; McGillicuddy, Dennis J; Martin, Jennifer L; Norton, Kerry; Pilskaln, Cynthia H; Smith, Juliette L; Sherwood, Christopher R; Butman, Bradford

    2014-05-01

    Here we document Alexandrium fundyense cyst abundance and distribution patterns over nine years (1997 and 2004-2011) in the coastal waters of the Gulf of Maine (GOM) and identify linkages between those patterns and several metrics of the severity or magnitude of blooms occurring before and after each autumn cyst survey. We also explore the relative utility of two measures of cyst abundance and demonstrate that GOM cyst counts can be normalized to sediment volume, revealing meaningful patterns equivalent to those determined with dry weight normalization. Cyst concentrations were highly variable spatially. Two distinct seedbeds (defined here as accumulation zones with > 300 cysts cm(-3)) are evident, one in the Bay of Fundy (BOF) and one in mid-coast Maine. Overall, seedbed locations remained relatively constant through time, but their area varied 3-4 fold, and total cyst abundance more than 10 fold among years. A major expansion of the mid-coast Maine seedbed occurred in 2009 following an unusually intense A. fundyense bloom with visible red-water conditions, but that feature disappeared by late 2010. The regional system thus has only two seedbeds with the bathymetry, sediment characteristics, currents, biology, and environmental conditions necessary to persist for decades or longer. Strong positive correlations were confirmed between the abundance of cysts in both the 0-1 and the 0-3 cm layers of sediments in autumn and geographic measures of the extent of the bloom that occurred the next year (i.e., cysts → blooms), such as the length of coastline closed due to shellfish toxicity or the southernmost latitude of shellfish closures. In general, these metrics of bloom geographic extent did not correlate with the number of cysts in sediments following the blooms (blooms → cysts). There are, however, significant positive correlations between 0-3 cm cyst abundances and metrics of the preceding bloom that are indicative of bloom intensity or vegetative cell abundance

  13. Alexandrium fundyense cysts in the Gulf of Maine: Long-term time series of abundance and distribution, and linkages to past and future blooms

    NASA Astrophysics Data System (ADS)

    Anderson, Donald M.; Keafer, Bruce A.; Kleindinst, Judith L.; McGillicuddy, Dennis J.; Martin, Jennifer L.; Norton, Kerry; Pilskaln, Cynthia H.; Smith, Juliette L.; Sherwood, Christopher R.; Butman, Bradford

    2014-05-01

    Here we document Alexandrium fundyense cyst abundance and distribution patterns over nine years (1997 and 2004-2011) in the coastal waters of the Gulf of Maine (GOM) and identify linkages between those patterns and several metrics of the severity or magnitude of blooms occurring before and after each autumn cyst survey. We also explore the relative utility of two measures of cyst abundance and demonstrate that GOM cyst counts can be normalized to sediment volume, revealing meaningful patterns equivalent to those determined with dry weight normalization. Cyst concentrations were highly variable spatially. Two distinct seedbeds (defined here as accumulation zones with>300 cysts cm-3) are evident, one in the Bay of Fundy (BOF) and one in mid-coast Maine. Overall, seedbed locations remained relatively constant through time, but their area varied 3-4 fold, and total cyst abundance more than 10 fold among years. A major expansion of the mid-coast Maine seedbed occurred in 2009 following an unusually intense A. fundyense bloom with visible red-water conditions, but that feature disappeared by late 2010. The regional system thus has only two seedbeds with the bathymetry, sediment characteristics, currents, biology, and environmental conditions necessary to persist for decades or longer. Strong positive correlations were confirmed between the abundance of cysts in both the 0-1 and the 0-3 cm layers of sediments in autumn and geographic measures of the extent of the bloom that occurred the next year (i.e., cysts→blooms), such as the length of coastline closed due to shellfish toxicity or the southernmost latitude of shellfish closures. In general, these metrics of bloom geographic extent did not correlate with the number of cysts in sediments following the blooms (blooms→cysts). There are, however, significant positive correlations between 0-3 cm cyst abundances and metrics of the preceding bloom that are indicative of bloom intensity or vegetative cell abundance (e

  14. Nutrients and water masses in the Gulf of Maine-Georges Bank region: Variability and importance to blooms of the toxic dinoflagellate Alexandrium fundyense

    NASA Astrophysics Data System (ADS)

    Townsend, D. W.; McGillicuddy, D. J.; Thomas, M. A.; Rebuck, N. D.

    2014-05-01

    We report here the results of ten oceanographic survey cruises carried out in the Gulf of Maine-Georges Bank region of the Northwest Atlantic during the late spring to summer period in 2007, 2008 and 2010, for which we examine and characterize relationships among dissolved inorganic nutrient fields, water mass dynamics and cell densities of the toxic dinoflagellate Alexandrium fundyense. Nutrients are supplied to continental shelf waters of the Gulf of Maine-Georges Bank region by inflows of deep offshore water masses; once in the Gulf they are transported with the residual circulation and mix with surface waters, both in the Gulf and on the Bank. Those fluxes of offshore water masses and their nutrient loads are the major source of nutrients for phytoplankton production in the region, including annual blooms of A. fundyense in the Gulf and on Georges Bank. This much is already known. We suggest here that the locations and magnitude of A. fundyense blooms are controlled in part by variable nutrient fluxes to the interior Gulf of Maine from offshore, and, those interior Gulf of Maine waters are, in turn, the main nutrient source to Georges Bank, which are brought onto the Bank by tidal pumping on the Northern Flank. We present evidence that nitrate is the initial form of nitrogenous nutrient for A. fundyense blooms, but it is quickly depleted to limiting concentrations of less than 0.5 μM, at which time continued growth and maintenance of the population is likely fueled by recycled ammonium. We also show that phosphate may be the limiting nutrient over much of Georges Bank in summer, allowing recycled ammonium concentrations to increase. Our temperature-salinity analyses reveal spatial and temporal (seasonal and interannual) variability in the relative proportions of two deep source waters that enter the Gulf of Maine at depth through the Northeast Channel: Warm Slope Water (WSW) and Labrador Slope Water (LSW). Those two source waters are known to vary in their

  15. Alexandrium fundyense cysts in the Gulf of Maine: long-term time series of abundance and distribution, and linkages to past and future blooms

    USGS Publications Warehouse

    Anderson, Donald M.; Keafer, Bruce A.; Kleindinst, Judith L.; McGillicuddy, Dennis J.; Martin, Jennifer L.; Norton, Kerry; Pilskaln, Cynthia H.; Smith, Juliette L.; Sherwood, Christopher R.; Butman, Bradford

    2014-01-01

    Here we document Alexandrium fundyense cyst abundance and distribution patterns over nine years (1997 and 2004–2011) in the coastal waters of the Gulf of Maine (GOM) and identify linkages between those patterns and several metrics of the severity or magnitude of blooms occurring before and after each autumn cyst survey. We also explore the relative utility of two measures of cyst abundance and demonstrate that GOM cyst counts can be normalized to sediment volume, revealing meaningful patterns equivalent to those determined with dry weight normalization. Cyst concentrations were highly variable spatially. Two distinct seedbeds (defined here as accumulation zones with>300 cysts cm−3) are evident, one in the Bay of Fundy (BOF) and one in mid-coast Maine. Overall, seedbed locations remained relatively constant through time, but their area varied 3–4 fold, and total cyst abundance more than 10 fold among years. A major expansion of the mid-coast Maine seedbed occurred in 2009 following an unusually intense A. fundyense bloom with visible red-water conditions, but that feature disappeared by late 2010. The regional system thus has only two seedbeds with the bathymetry, sediment characteristics, currents, biology, and environmental conditions necessary to persist for decades or longer. Strong positive correlations were confirmed between the abundance of cysts in both the 0–1 and the 0–3 cm layers of sediments in autumn and geographic measures of the extent of the bloom that occurred the next year (i.e., cysts→blooms), such as the length of coastline closed due to shellfish toxicity or the southernmost latitude of shellfish closures. In general, these metrics of bloom geographic extent did not correlate with the number of cysts in sediments following the blooms (blooms→cysts). There are, however, significant positive correlations between 0–3 cm cyst abundances and metrics of the preceding bloom that are indicative of bloom intensity or vegetative cell

  16. Complexities of bloom dynamics in the toxic dinoflagellate Alexandrium fundyense revealed through DNA measurements by imaging flow cytometry coupled with species-specific rRNA probes

    NASA Astrophysics Data System (ADS)

    Brosnahan, Michael L.; Farzan, Shahla; Keafer, Bruce A.; Sosik, Heidi M.; Olson, Robert J.; Anderson, Donald M.

    2014-05-01

    Measurements of the DNA content of different protist populations can shed light on a variety of processes, including cell division, sex, prey ingestion, and parasite invasion. Here, we modified an Imaging FlowCytobot (IFCB), a custom-built flow cytometer that records images of microplankton, to measure the DNA content of large dinoflagellates and other high-DNA content species. The IFCB was also configured to measure fluorescence from Cy3-labeled rRNA probes, aiding the identification of Alexandrium fundyense (syn. A. tamarense Group I), a photosynthetic dinoflagellate that causes paralytic shellfish poisoning (PSP). The modified IFCB was used to analyze samples from the development, peak and termination phases of an inshore A. fundyense bloom (Salt Pond, Eastham, MA, USA), and from a rare A. fundyense ‘red tide’ that occurred in the western Gulf of Maine, offshore of Portsmouth, NH (USA). Diploid or G2 phase (‘2C’) A. fundyense cells were frequently enriched at the near-surface, suggesting an important role for aggregation at the air-sea interface during sexual events. Also, our analysis showed that large proportions of A. fundyense cells in both the Salt Pond and red tide blooms were planozygotes during bloom decline, highlighting the importance of sexual fusion to bloom termination. At Salt Pond, bloom decline also coincided with a dramatic rise in infections by the parasite genus Amoebophrya. The samples that were most heavily infected contained many large cells with higher DNA-associated fluorescence than 2C vegetative cells, but these cells' nuclei were also frequently consumed by Amoebophrya trophonts. Neither large cell size nor increased DNA-associated fluorescence could be replicated by infecting an A. fundyense culture of vegetative cells. Therefore, we attribute these characteristics of the large Salt Pond cells to planozygote maturation rather than Amoebophrya infection, though an interaction between infection and planozygote maturation may

  17. Nutrients and water masses in the Gulf of Maine - Georges Bank region: Variability and importance to blooms of the toxic dinoflagellate Alexandrium fundyense.

    PubMed

    Townsend, D W; McGillicuddy, D J; Thomas, M A; Rebuck, N R

    2014-05-01

    We report here the results of ten oceanographic survey cruises carried out in the Gulf of Maine - Georges Bank region of the Northwest Atlantic during the late spring to summer period in 2007, 2008 and 2010, for which we examine and characterize relationships among dissolved inorganic nutrient fields, water mass dynamics and cell densities of the toxic dinoflagellate Alexandrium fundyense. Nutrients are supplied to continental shelf waters of the Gulf of Maine - Georges Bank region by inflows of deep offshore water masses; once in the Gulf they are transported with the residual circulation and mix with surface waters, both in the Gulf and on the Bank. Those fluxes of offshore water masses and their nutrient loads are the major source of nutrients for phytoplankton production in the region, including annual blooms of A. fundyense in the Gulf and on Georges Bank. This much is already known. We suggest here that the locations and magnitude of A. fundyense blooms are controlled in part by variable nutrient fluxes to the interior Gulf of Maine from offshore, and, those interior Gulf of Maine waters are, in turn, the main nutrient source to Georges Bank, which are brought onto the Bank by tidal pumping on the Northern Flank. We present evidence that nitrate is the initial form of nitrogenous nutrient for A. fundyense blooms, but it is quickly depleted to limiting concentrations of less than 0.5 μM, at which time continued growth and maintenance of the population is likely fueled by recycled ammonium. We also show that phosphate may be the limiting nutrient over much of Georges Bank in summer, allowing recycled ammonium concentrations to increase. Our temperature-salinity analyses reveal spatial and temporal (seasonal and interannual) variability in the relative proportions of two deep source waters that enter the Gulf of Maine at depth through the Northeast Channel: Warm Slope Water (WSW) and Labrador Slope Water (LSW). Those two source waters are known to vary in their

  18. Driving factors behind the distribution of dinocyst composition and abundance in surface sediments in a western Mediterranean coastal lagoon: report from a high resolution mapping study.

    PubMed

    Fertouna-Bellakhal, Mouna; Dhib, Amel; Béjaoui, Béchir; Turki, Souad; Aleya, Lotfi

    2014-07-15

    Species composition and abundance of dinocysts in relation to environmental factors were studied at 123 stations of surface sediment in Bizerte Lagoon. Forty-eight dinocyst types were identified, mainly dominated by Brigantidinium simplex, Votadinum spinosum, Alexandrium pseudogonyaulax, Alexandrium catenella, and Lingulodinum machaerophorum along with many round brown cysts and spiny round brown cysts. Cysts ranged from 1276 to 20126 cysts g(-1)dry weight sediment. Significant differences in cyst distribution pattern were recorded among the zones, with a higher cyst abundance occurring in the lagoon's inner areas. Redundancy analyses showed two distinct associations of dinocysts according to location and environmental variables. Ballast water discharges are potential introducers of non-indigenous species, especially harmful ones such as A. catenella and Polysphaeridium zoharyi, with currents playing a pivotal role in cyst distribution. Findings concerning harmful cyst species indicate potential seedbeds for initiation of future blooms and outbreaks of potentially toxic species in the lagoon.

  19. Evaluating hypotheses for the initiation and development of Alexandrium fundyense blooms in the western Gulf of Maine using a coupled physical biological model

    NASA Astrophysics Data System (ADS)

    Stock, Charles A.; McGillicuddy, Dennis J.; Solow, Andrew R.; Anderson, Donald M.

    2005-09-01

    A coupled physical/biological model and observations are used to investigate the factors governing the initiation and development of an Alexandrium fundyense bloom in the western Gulf of Maine (WGOM) during the spring of 1993 (March 19-June 6). The physical circulation is simulated using a 3D primitive equation model forced by climatological sea-surface elevation and observed winds, irradiance, and river outflow. This is coupled with a biological model constructed from laboratory and field data that estimates the germination and growth rates of A. fundyense as a function of environmental conditions. Four biological model structures of increasing complexity are considered, with each structure representing a hypothesis for factors controlling bloom initiation and development. The model/data fit is optimized over the uncertainty in the parameters to which the model is most sensitive. The significance of changes in the model/data fit between model structures is quantified using a maximum likelihood ratio test. The baseline biological model, which parameterizes growth as only a function of temperature, salinity, and light, severely over-estimates observed A. fundyense abundance in the late spring. It is thus rejected with greater than 99% confidence in favor of biological models that include a mortality term or a dependence of growth on dissolved inorganic nitrogen (DIN). The overall best-fit simulation uses both nitrogen dependence and mortality. However, simulations using one or the other of these factors could not be differentiated from the best-fit case with greater than 90% confidence. The best-fit model captures the general timing and magnitude of the observed bloom and some of its secondary features. However, considerable misfits may exist in the point-to-point comparison, and some regional misfits remain. Diagnosis of the cell budget suggests that germination from a large cyst bed offshore of Casco Bay provides the majority of cells comprising spring A

  20. Blooms of the toxic dinoflagellate, Alexandrium fundyense in the Casco Bay region of the western Gulf of Maine: Advection from offshore source populations and interactions with the Kennebec River plume

    NASA Astrophysics Data System (ADS)

    Keafer, Bruce A.; Churchill, James H.; Anderson, Donald M.

    2005-09-01

    The Casco Bay region, an embayment adjacent to the Kennebec River, has been suggested as a source region for Alexandrium fundyense bloom development in the western Gulf of Maine (GOM). In this study, shipboard observations were acquired within Casco Bay and the nearby coastal waters during the spring of 1998 and 2000. In the early bloom season, low A. fundyense abundances (<100 cells l -1) were observed within the bay, sometimes isolated from A. fundyense populations observed in adjacent coastal waters. When high abundances of A. fundyense (>500 cells l -1) were observed within Casco Bay, they were contiguous with coastal populations observed within the Kennebec/Penobscot river plume and within offshore waters of the western segment of the Maine Coastal Current (WMCC). This general distributional pattern occurred during both study years. Wind directly affected the pathway of the incoming coastal populations. Downwelling-favorable winds generally facilitated bloom formation (and outbreaks of shellfish toxicity) within Casco Bay by enhancing the connection with offshore populations via alongshore and onshore transport of cells from the upstream coastal waters. In contrast, persistent upwelling-favorable winds were associated with low A. fundyense cell abundances (and shellfish toxicity) in Casco Bay by slowing the advance of the coastal population and shifting it offshore with the Kennebec plume front. The striking difference between late season (June) population abundances of the two study years can be explained by a combination of the wind pre-history and interannual differences in large-scale (Gulf-wide) circulation patterns, as evidenced by higher salinities in the coastal waters in 2000 vs. 1998. Advection of A. fundyense cells into Casco Bay and retention, not local growth within the Bay, are likely the dominant processes that typically result in the accumulation of high populations and shellfish toxicity in the Bay. A variety of mechanisms (e.g., circulation

  1. Detection of harmful algal bloom causing microalgae using covalently immobilised capture oligonucleotide probes on glass and poly(dimethylsiloxane) surfaces

    NASA Astrophysics Data System (ADS)

    Bruce, Karen L.; Ellis, Amanda V.; Leterme, Sophie C.; Khodakov, Dmitriy A.; Lenehan, Claire E.

    2013-12-01

    Harmful algal bloom (HAB) events have been on the rise in the last few decades with some of the causative microalgae exhibiting toxic properties. Therefore, detection is essential in order to prevent mortality of aquatic life and poisoning events from consumption of these biotoxins. Here, oligonucleotide modified glass and poly(dimethylsiloxane) (PDMS) surfaces have been developed for the detection of the HAB causing microalgae, Alexandrium catenella, in a model system. Our preliminary studies show that the glass surface offers superior stability and analytical response when compared to those prepared from PDMS.

  2. Contrasting physiological responses of two populations of the razor clam Tagelus dombeii with different histories of exposure to paralytic shellfish poisoning (PSP).

    PubMed

    Navarro, Jorge M; González, Katerina; Cisternas, Barbara; López, Jorge A; Chaparro, Oscar R; Segura, Cristian J; Córdova, Marco; Suárez-Isla, Benjamín; Fernandez-Reiriz, María J; Labarta, Uxio

    2014-01-01

    This study describes the physiological performance of two populations of the razor clam Tagelus dombeii from two geographic areas with different histories of exposure to paralytic shellfish poisoning (PSP) linked to the toxic dinoflagellate Alexandrium catenella. Clams from Melinka-Aysén, which are frequently exposed to PSP, were not affected by the presence of toxins in the diet. However, clams from Corral-Valdivia, which have never been exposed to PSP, exhibited significantly reduced filtration activity and absorption, affecting the energy allocated to scope for growth (SFG). Ammonia excretion and oxygen uptake were not affected significantly by the presence of A. catenella in the diet. Measurements of energy acquisition and expenditure were performed during a 12-day intoxication period. According to three-way repeated measure ANOVAs, the origin of the clams had a highly significant effect on all physiological variables, and the interaction between diet and origin was significant for the clearance and absorption rates and for the scope for growth. The scope for growth index showed similar positive values for both the toxic and non-toxic individuals from the Melinka-Aysén population. However, it was significantly reduced in individuals from Corral-Valdivia when exposed to the diet containing A. catenella. The absence of differences between the physiological response of the toxic and non-toxic clams from Melinka-Aysén may be related to the frequent presence of A. catenella in the environment, indicating that this bivalve does not suffer negative consequences from PSP. By contrast, A. catenella has a negative effect on the physiological performance, primarily on the energy gained from the environment, on T. dombeii from Corral-Valdivia. This study supports the hypothesis that the history of PSP exposure plays an important role in the physiological performance and fitness of filter feeding bivalves.

  3. Contrasting Physiological Responses of Two Populations of the Razor Clam Tagelus dombeii with Different Histories of Exposure to Paralytic Shellfish Poisoning (PSP)

    PubMed Central

    Navarro, Jorge M.; González, Katerina; Cisternas, Barbara; López, Jorge A.; Chaparro, Oscar R.; Segura, Cristian J.; Córdova, Marco; Suárez-Isla, Benjamín; Fernandez-Reiriz, María J.; Labarta, Uxio

    2014-01-01

    This study describes the physiological performance of two populations of the razor clam Tagelus dombeii from two geographic areas with different histories of exposure to paralytic shellfish poisoning (PSP) linked to the toxic dinoflagellate Alexandrium catenella. Clams from Melinka-Aysén, which are frequently exposed to PSP, were not affected by the presence of toxins in the diet. However, clams from Corral-Valdivia, which have never been exposed to PSP, exhibited significantly reduced filtration activity and absorption, affecting the energy allocated to scope for growth (SFG). Ammonia excretion and oxygen uptake were not affected significantly by the presence of A. catenella in the diet. Measurements of energy acquisition and expenditure were performed during a 12-day intoxication period. According to three-way repeated measure ANOVAs, the origin of the clams had a highly significant effect on all physiological variables, and the interaction between diet and origin was significant for the clearance and absorption rates and for the scope for growth. The scope for growth index showed similar positive values for both the toxic and non-toxic individuals from the Melinka-Aysén population. However, it was significantly reduced in individuals from Corral-Valdivia when exposed to the diet containing A. catenella. The absence of differences between the physiological response of the toxic and non-toxic clams from Melinka-Aysén may be related to the frequent presence of A. catenella in the environment, indicating that this bivalve does not suffer negative consequences from PSP. By contrast, A. catenella has a negative effect on the physiological performance, primarily on the energy gained from the environment, on T. dombeii from Corral-Valdivia. This study supports the hypothesis that the history of PSP exposure plays an important role in the physiological performance and fitness of filter feeding bivalves. PMID:25153329

  4. PSP toxin levels and plankton community composition and abundance in size-fractionated vertical profiles during spring/summer blooms of the toxic dinoflagellate Alexandrium fundyense in the Gulf of Maine and on Georges Bank, 2007, 2008, and 2010: 2. Plankton community composition and abundance

    PubMed Central

    Petitpas, Christian M.; Turner, Jefferson T.; Deeds, Jonathan R.; Keafer, Bruce A.; McGillicuddy, Dennis J.; Milligan, Peter J.; Shue, Vangie; White, Kevin D.; Anderson, Donald M.

    2015-01-01

    As part of the Gulf of Maine Toxicity (GOMTOX1) project, we determined Alexandrium fundyense abundance, paralytic shellfish poisoning (PSP) toxin levels in various plankton size fractions, and the community composition of potential grazers of A. fundyense in plankton size fractions during blooms of this toxic dinoflagellate in the coastal Gulf of Maine and on Georges Bank in spring and summer of 2007, 2008, and 2010. PSP toxins and A. fundyense cells were found throughout the sampled water column (down to 50 m) in the 20–64 μm size fractions. While PSP toxins were widespread throughout all size classes of the zooplankton grazing community, the majority of the toxin was measured in the 20–64 μm size fraction. A. fundyense cellular toxin content estimated from field samples was significantly higher in the coastal Gulf of Maine than on Georges Bank. Most samples containing PSP toxins in the present study had diverse assemblages of grazers. However, some samples clearly suggested PSP toxin accumulation in several different grazer taxa including tintinnids, heterotrophic dinoflagellates of the genus Protoperidinium, barnacle nauplii, the harpacticoid copepod Microsetella norvegica, the calanoid copepods Calanus finmarchicus and Pseudocalanus spp., the marine cladoceran Evadne nordmanni, and hydroids of the genus Clytia. Thus, a diverse assemblage of zooplankton grazers accumulated PSP toxins through food-web interactions. This raises the question of whether PSP toxins pose a potential human health risk not only from nearshore bivalve shellfish, but also potentially from fish and other upper-level consumers in zooplankton-based pelagic food webs. PMID:26236112

  5. PSP toxin levels and plankton community composition and abundance in size-fractionated vertical profiles during spring/summer blooms of the toxic dinoflagellate Alexandrium fundyense in the Gulf of Maine and on Georges Bank, 2007, 2008, and 2010: 2. Plankton community composition and abundance.

    PubMed

    Petitpas, Christian M; Turner, Jefferson T; Deeds, Jonathan R; Keafer, Bruce A; McGillicuddy, Dennis J; Milligan, Peter J; Shue, Vangie; White, Kevin D; Anderson, Donald M

    2014-05-01

    As part of the Gulf of Maine Toxicity (GOMTOX) project, we determined Alexandrium fundyense abundance, paralytic shellfish poisoning (PSP) toxin levels in various plankton size fractions, and the community composition of potential grazers of A. fundyense in plankton size fractions during blooms of this toxic dinoflagellate in the coastal Gulf of Maine and on Georges Bank in spring and summer of 2007, 2008, and 2010. PSP toxins and A. fundyense cells were found throughout the sampled water column (down to 50 m) in the 20-64 μm size fractions. While PSP toxins were widespread throughout all size classes of the zooplankton grazing community, the majority of the toxin was measured in the 20-64 μm size fraction. A. fundyense cellular toxin content estimated from field samples was significantly higher in the coastal Gulf of Maine than on Georges Bank. Most samples containing PSP toxins in the present study had diverse assemblages of grazers. However, some samples clearly suggested PSP toxin accumulation in several different grazer taxa including tintinnids, heterotrophic dinoflagellates of the genus Protoperidinium, barnacle nauplii, the harpacticoid copepod Microsetella norvegica, the calanoid copepods Calanus finmarchicus and Pseudocalanus spp., the marine cladoceran Evadne nordmanni, and hydroids of the genus Clytia. Thus, a diverse assemblage of zooplankton grazers accumulated PSP toxins through food-web interactions. This raises the question of whether PSP toxins pose a potential human health risk not only from nearshore bivalve shellfish, but also potentially from fish and other upper-level consumers in zooplankton-based pelagic food webs.

  6. PSP toxin levels and plankton community composition and abundance in size-fractionated vertical profiles during spring/summer blooms of the toxic dinoflagellate Alexandrium fundyense in the Gulf of Maine and on Georges Bank, 2007, 2008, and 2010: 1. Toxin levels

    PubMed Central

    Deeds, Jonathan R.; Petitpas, Christian M.; Shue, Vangie; White, Kevin D.; Keafer, Bruce A.; McGillicuddy, Dennis J.; Milligan, Peter J.; Anderson, Donald M.; Turner, Jefferson T.

    2014-01-01

    As part of the NOAA ECOHAB funded Gulf of Maine Toxicity (GOMTOX)1 project, we determined Alexandrium fundyense abundance, paralytic shellfish poisoning (PSP) toxin composition, and concentration in quantitatively-sampled size-fractionated (20–64, 64–100, 100–200, 200–500, and > 500 μm) particulate water samples, and the community composition of potential grazers of A. fundyense in these size fractions, at multiple depths (typically 1, 10, 20 m, and near-bottom) during 10 large-scale sampling cruises during the A. fundyense bloom season (May–August) in the coastal Gulf of Maine and on Georges Bank in 2007, 2008, and 2010. Our findings were as follows: (1) when all sampling stations and all depths were summed by year, the majority (94% ± 4%) of total PSP toxicity was contained in the 20–64 μm size fraction; (2) when further analyzed by depth, the 20–64 μm size fraction was the primary source of toxin for 97% of the stations and depths samples over three years; (3) overall PSP toxin profiles were fairly consistent during the three seasons of sampling with gonyautoxins (1, 2, 3, and 4) dominating (90.7% ± 5.5%), followed by the carbamate toxins saxitoxin (STX) and neosaxitoxin (NEO) (7.7% ± 4.5%), followed by n-sulfocarbamoyl toxins (C1 and 2, GTX5) (1.3% ± 0.6%), followed by all decarbamoyl toxins (dcSTX, dcNEO, dcGTX2&3) (< 1%), although differences were noted between PSP toxin compositions for nearshore coastal Gulf of Maine sampling stations compared to offshore Georges Bank sampling stations for 2 out of 3 years; (4) surface cell counts of A. fundyense were a fairly reliable predictor of the presence of toxins throughout the water column; and (5) nearshore surface cell counts of A. fundyense in the coastal Gulf of Maine were not a reliable predictor of A. fundyense populations offshore on Georges Bank for 2 out of the 3 years sampled. PMID:25076816

  7. PSP toxin levels and plankton community composition and abundance in size-fractionated vertical profiles during spring/summer blooms of the toxic dinoflagellate Alexandrium fundyense in the Gulf of Maine and on Georges Bank, 2007, 2008, and 2010: 1. Toxin levels.

    PubMed

    Deeds, Jonathan R; Petitpas, Christian M; Shue, Vangie; White, Kevin D; Keafer, Bruce A; McGillicuddy, Dennis J; Milligan, Peter J; Anderson, Donald M; Turner, Jefferson T

    2014-05-01

    As part of the NOAA ECOHAB funded Gulf of Maine Toxicity (GOMTOX) project, we determined Alexandrium fundyense abundance, paralytic shellfish poisoning (PSP) toxin composition, and concentration in quantitatively-sampled size-fractionated (20-64, 64-100, 100-200, 200-500, and > 500 μm) particulate water samples, and the community composition of potential grazers of A. fundyense in these size fractions, at multiple depths (typically 1, 10, 20 m, and near-bottom) during 10 large-scale sampling cruises during the A. fundyense bloom season (May-August) in the coastal Gulf of Maine and on Georges Bank in 2007, 2008, and 2010. Our findings were as follows: (1) when all sampling stations and all depths were summed by year, the majority (94% ± 4%) of total PSP toxicity was contained in the 20-64 μm size fraction; (2) when further analyzed by depth, the 20-64 μm size fraction was the primary source of toxin for 97% of the stations and depths samples over three years; (3) overall PSP toxin profiles were fairly consistent during the three seasons of sampling with gonyautoxins (1, 2, 3, and 4) dominating (90.7% ± 5.5%), followed by the carbamate toxins saxitoxin (STX) and neosaxitoxin (NEO) (7.7% ± 4.5%), followed by n-sulfocarbamoyl toxins (C1 and 2, GTX5) (1.3% ± 0.6%), followed by all decarbamoyl toxins (dcSTX, dcNEO, dcGTX2&3) (< 1%), although differences were noted between PSP toxin compositions for nearshore coastal Gulf of Maine sampling stations compared to offshore Georges Bank sampling stations for 2 out of 3 years; (4) surface cell counts of A. fundyense were a fairly reliable predictor of the presence of toxins throughout the water column; and (5) nearshore surface cell counts of A. fundyense in the coastal Gulf of Maine were not a reliable predictor of A. fundyense populations offshore on Georges Bank for 2 out of the 3 years sampled.

  8. Progress in Understanding Algal Bloom-Mediated Fish Kills: The Role of Superoxide Radicals, Phycotoxins and Fatty Acids

    PubMed Central

    Dorantes-Aranda, Juan José; Seger, Andreas; Mardones, Jorge I.; Nichols, Peter D.; Hallegraeff, Gustaaf M.

    2015-01-01

    Quantification of the role of reactive oxygen species, phycotoxins and fatty acids in fish toxicity by harmful marine microalgae remains inconclusive. An in vitro fish gill (from rainbow trout Oncorhynchus mykiss) assay was used to simultaneously assess the effect in superoxide dismutase, catalase and lactate dehydrogenase enzymatic activities caused by seven species of ichthyotoxic microalgae (Chattonella marina, Fibrocapsa japonica, Heterosigma akashiwo, Karenia mikimotoi, Alexandrium catenella, Karlodinium veneficum, Prymnesium parvum). Quantification of superoxide production by these algae was also performed. The effect of purified phycotoxins and crude extracts was compared, and the effect of fatty acids is discussed. The raphidophyte Chattonella was the most ichthyotoxic (gill cell viability down to 35%) and also the major producer of superoxide radicals (14 pmol cell-1 hr-1) especially after cell lysis. The raphidophyte Heterosigma and dinoflagellate Alexandrium were the least toxic and had low superoxide production, except when A. catenella was lysed (5.6 pmol cell-1 hr-1). Catalase showed no changes in activity in all the treatments. Superoxide dismutase (SOD) and lactate dehydrogenase exhibited significant activity increases of ≤23% and 51.2% TCC (total cellular content), respectively, after exposure to C. marina, but SOD showed insignificant changes with remaining algal species. A strong relationship between gill cell viability and superoxide production or superoxide dismutase was not observed. Purified brevetoxins PbTx-2 and -3 (from Karenia brevis, LC50 of 22.1 versus 35.2 μg mL-1) and karlotoxin KmTx-2 (from Karlodinium; LC50 = 380 ng mL-1) could almost entirely account for the fish killing activity by those two dinoflagellates. However, the paralytic shellfish toxins (PST) GTX1&4, C1&C2, and STX did not account for Alexandrium ichthyotoxicity. Only aqueous extracts of Alexandrium were cytotoxic (≤65% decrease of viability), whereas crude

  9. Progress in Understanding Algal Bloom-Mediated Fish Kills: The Role of Superoxide Radicals, Phycotoxins and Fatty Acids.

    PubMed

    Dorantes-Aranda, Juan José; Seger, Andreas; Mardones, Jorge I; Nichols, Peter D; Hallegraeff, Gustaaf M

    2015-01-01

    Quantification of the role of reactive oxygen species, phycotoxins and fatty acids in fish toxicity by harmful marine microalgae remains inconclusive. An in vitro fish gill (from rainbow trout Oncorhynchus mykiss) assay was used to simultaneously assess the effect in superoxide dismutase, catalase and lactate dehydrogenase enzymatic activities caused by seven species of ichthyotoxic microalgae (Chattonella marina, Fibrocapsa japonica, Heterosigma akashiwo, Karenia mikimotoi, Alexandrium catenella, Karlodinium veneficum, Prymnesium parvum). Quantification of superoxide production by these algae was also performed. The effect of purified phycotoxins and crude extracts was compared, and the effect of fatty acids is discussed. The raphidophyte Chattonella was the most ichthyotoxic (gill cell viability down to 35%) and also the major producer of superoxide radicals (14 pmol cell-1 hr-1) especially after cell lysis. The raphidophyte Heterosigma and dinoflagellate Alexandrium were the least toxic and had low superoxide production, except when A. catenella was lysed (5.6 pmol cell-1 hr-1). Catalase showed no changes in activity in all the treatments. Superoxide dismutase (SOD) and lactate dehydrogenase exhibited significant activity increases of ≤23% and 51.2% TCC (total cellular content), respectively, after exposure to C. marina, but SOD showed insignificant changes with remaining algal species. A strong relationship between gill cell viability and superoxide production or superoxide dismutase was not observed. Purified brevetoxins PbTx-2 and -3 (from Karenia brevis, LC50 of 22.1 versus 35.2 μg mL-1) and karlotoxin KmTx-2 (from Karlodinium; LC50 = 380 ng mL-1) could almost entirely account for the fish killing activity by those two dinoflagellates. However, the paralytic shellfish toxins (PST) GTX1&4, C1&C2, and STX did not account for Alexandrium ichthyotoxicity. Only aqueous extracts of Alexandrium were cytotoxic (≤65% decrease of viability), whereas crude

  10. β-N-methylamino-l-alanine (BMAA) and isomers: Distribution in different food web compartments of Thau lagoon, French Mediterranean Sea.

    PubMed

    Réveillon, Damien; Abadie, Eric; Séchet, Véronique; Masseret, Estelle; Hess, Philipp; Amzil, Zouher

    2015-09-01

    The neurotoxin BMAA (β-N-methylamino-l-alanine) and its isomer DAB (2,4-diaminobutyric acid) have been detected in seafood worldwide, including in Thau lagoon (French Mediterranean Sea). A cluster of amyotrophic lateral sclerosis (ALS), a neurodegenerative disease associated with BMAA, has also been observed in this region. Mussels, periphyton (i.e. biofilms attached to mussels) and plankton were sampled between July 2013 and October 2014, and analyzed using HILIC-MS/MS. BMAA, DAB and AEG (N-(2-aminoethyl)glycine) were found in almost all the samples of the lagoon. BMAA and DAB were present at 0.58 and 0.83, 2.6 and 3.3, 4.0 and 7.2 μg g(-1) dry weight in plankton collected with nets, periphyton and mussels, respectively. Synechococcus sp., Ostreococcus tauri, Alexandrium catenella and eight species of diatoms were cultured and screened for BMAA and analogs. While Synechococcus sp., O. tauri and A. catenella did not produce BMAA under our culture conditions, four diatoms species contained both BMAA and DAB. Hence, diatoms may be a source of BMAA for mussels. Unlike other toxins produced by microalgae, BMAA and DAB were detected in significant amounts in tissues other than digestive glands in mussels.

  11. Functional Genomics of a Non-Toxic Alexandrium Lusitanicum Culture

    DTIC Science & Technology

    2007-02-01

    electrophoresis. Proteomics 2:1169-1186. Chang, P. 2004. Lack of interaction between AFLR and AFLJ contributes to nonaflatoxigenicity of Aspergillus sojae ...nonaflatoxigenicity of Aspergillus sojae . Journal of Biotechnology 107: 245-253. Doyle, J. J., and J. L. Doyle. 1990. Isolation of plant DNA from fresh tissue. Focus

  12. A shift in the dominant toxin-producing algal species in central California alters phycotoxins in food webs

    USGS Publications Warehouse

    Jester, R.; Lefebvre, K.; Langlois, G.; Vigilant, V.; Baugh, K.; Silver, M.W.

    2009-01-01

    In California, the toxic algal species of primary concern are the dinoflagellate Alexandrium catenella and members of the pennate diatom genus Pseudo-nitzschia, both producers of potent neurotoxins that are capable of sickening and killing marine life and humans. During the summer of 2004 in Monterey Bay, we observed a change in the taxonomic structure of the phytoplankton community-the typically diatom-dominated community shifted to a red tide, dinoflagellate-dominated community. Here we use a 6-year time series (2000-2006) to show how the abundance of the dominant harmful algal bloom (HAB) species in the Bay up to that point, Pseudo-nitzschia, significantly declined during the dinoflagellate-dominated interval, while two genera of toxic dinoflagellates, Alexandrium and Dinophysis, became the predominant toxin producers. This change represents a shift from a genus of toxin producers that typically dominates the community during a toxic bloom, to HAB taxa that are generally only minor components of the community in a toxic event. This change in the local HAB species was also reflected in the toxins present in higher trophic levels. Despite the small contribution of A. catenella to the overall phytoplankton community, the increase in the presence of this species in Monterey Bay was associated with an increase in the presence of paralytic shellfish poisoning (PSP) toxins in sentinel shellfish and clupeoid fish. This report provides the first evidence that PSP toxins are present in California's pelagic food web, as PSP toxins were detected in both northern anchovies (Engraulis mordax) and Pacific sardines (Sardinops sagax). Another interesting observation from our data is the co-occurrence of DA and PSP toxins in both planktivorous fish and sentinel shellfish. We also provide evidence, based on the statewide biotoxin monitoring program, that this increase in the frequency and abundance of PSP events related to A. catenella occurred not just in Monterey Bay, but also

  13. Algicidal Effects of a Novel Marine Pseudoalteromonas Isolate (Class Proteobacteria, Gamma Subdivision) on Harmful Algal Bloom Species of the Genera Chattonella, Gymnodinium, and Heterosigma

    PubMed Central

    Lovejoy, Connie; Bowman, John P.; Hallegraeff, Gustaaf M.

    1998-01-01

    During a bacterial survey of the Huon Estuary in southern Tasmania, Australia, we isolated a yellow-pigmented Pseudoalteromonas strain (class Proteobacteria, gamma subdivision), designated strain Y, that had potent algicidal effects on harmful algal bloom species. This organism was identified by 16S rRNA sequencing as a strain with close affinities to Pseudoalteromonas peptidysin. This bacterium caused rapid cell lysis and death (within 3 h) of gymnodinoids (including Gymnodinium catenatum) and raphidophytes (Chattonella marina and Heterosigma akashiwo). It caused ecdysis of armored dinoflagellates (e.g., Alexandrium catenella, Alexandrium minutum, and Prorocentrum mexicanum), but the algal cultures then recovered over the subsequent 24 h. Strain Y had no effect on a cryptomonad (Chroomonas sp.), a diatom (Skeletonema sp.), a cyanobacterium (Oscillatoria sp.), and two aplastidic protozoans. The algicidal principle of strain Y was excreted into the seawater medium and lost its efficacy after heating. Another common bacterial species, Pseudoalteromonas carrageenovora, was isolated at the same time and did not have these algicidal effects. The minimum concentrations of strain Y required to kill G. catenatum were higher than the mean concentrations found in nature under nonbloom conditions. However, the new bacterium showed a chemotactic, swarming behavior that resulted in localized high concentrations around target organisms. These observations imply that certain bacteria could play an important role in regulating the onset and development of harmful algal blooms. PMID:9687434

  14. Occurrence of four species of algae in the marine water of Hong Kong.

    PubMed

    Chai, Yemao; Deng, Wen-Jing; Qin, Xing; Xu, Xiangrong

    2017-01-05

    Harmful algal blooms (HABs) have broken out frequently throughout the world in recent decades; they are caused by the rapid multiplication of algal cells in near-coastal waters polluted with nitrogen and phosphorus and greatly affect the quality of marine water and human health. Over the past several decades, climate change and increasing environmental degradation have provided favourable growth conditions for certain phytoplankton species. Therefore, it is essential to rapidly identify and enumerate harmful marine algae to control these species. In this study, quantitative PCR (qPCR) was used to detect four representative species of HABs that are widespread in the marine water of Hong Kong, namely, Alexandrium catenella, Pseudo-nitzschia spp., Karenia mikimotoi and Heterosigma akashiwo. We applied qPCR with the dye SYBR Green to detect Alexandrium spp. and Pseudo-nitzschia spp. and used TaqMan probe for the enumeration of Karenia mikimotoi and Heterosigma akashiwo. The total genomic DNA of these algae from Hong Kong marine water was extracted successfully using the CTAB method, and for each kind of alga, we constructed a ten-fold series of recombinant plasmid solutions containing certain gene fragments of 18S rDNA and ITS1-5.8S-ITS2 as standard samples. Ten-fold dilutions of the DNA of known numbers of the extracted algal cells were also used to create an additional standard curve. In this way, the relationship between the cell number and the related plasmid copy number was established. The qPCR assay displayed high sensitivity in monitoring marine water samples in which the low concentrations of harmful algae were not detected accurately by traditional methods. The results showed that the cell numbers of the four species were all in low abundance. For Alexandrium catenella, the cell abundances at 12 sites ranged from 3.8×10(2) to 4.3×10(3)cellsL(-1), while H. akashiwo, K. mikimotoi and Pseudo-nitzschia ranged from 1.1×10(2) to 1.3×10(3), from 23 to 6.5×10

  15. Impact of several harmful algal bloom (HAB) causing species, on life history characteristics of rotifer Brachionus plicatilis Müller

    NASA Astrophysics Data System (ADS)

    Lin, Jianing; Yan, Tian; Zhang, Qingchun; Zhou, Mingjiang

    2016-07-01

    In recent years, harmful algal blooms (HABs) have occurred frequently along the coast of China, and have been exhibiting succession from diatom- to dinoflagellate-dominated blooms. To examine the effects of different diatom and dinoflagellate HABs, the life history parameters of rotifers ( Brachionus plicatilis Müller) were measured after exposure to different concentrations of HAB species. The HAB species examined included a diatom ( Skeletonema costatum) and four dinoflagellates ( Prorocentrum donghaiense, Alexandrium catenella, Prorocentrum lima and Karlodinium veneficum). Compared with the control treatment (CT), the diatom S. costatum showed no adverse impacts on rotifers. Exposure to dinoflagellates at densities equivalent to those measured in the field resulted in a reduction in all the life history parameters measured. This included a reduction in: lifetime egg production (CT: 20.34 eggs/ind.) reduced to 10.11, 3.22, 4.17, 7.16 eggs/ind., life span (CT: 394.53 h) reduced to 261.11, 162.90, 203.67, 196 h, net reproductive rate (CT: 19.51/ind.) reduced to 3.01, 1.26, 3.53, 5.96/ind., finite rate of increase (CT: 1.47/d) reduced to 1.16, 1.03, 1.33, 1.38/d, and intrinsic rate of population increase (CT: 0.39/d) reduced to 0.15, 0.03, 0.28, 0.32/d, for the dinoflagellates P. donghaiense, A. catenella, P. lima and K. veneficum, respectively. The results showed that the diatom S. costatum had no detrimental consequences on the reproduction and growth of B. plicatilis, however, the four dinoflagellates tested did show adverse effects. This suggests that dinoflagellate HABs may suppress microzooplankton, resulting in an increase in algal numbers.

  16. Uncovering the Complex Transcriptome Response of Mytilus chilensis against Saxitoxin: Implications of Harmful Algal Blooms on Mussel Populations

    PubMed Central

    Detree, Camille; Núñez-Acuña, Gustavo; Roberts, Steven; Gallardo-Escárate, Cristian

    2016-01-01

    Saxitoxin (STX), a principal phycotoxin contributing to paralytic shellfish poisoning, is largely produced by marine microalgae of the genus Alexandrium. This toxin affects a wide range of species, inducing massive deaths in fish and other marine species. However, marine bivalves can resist and accumulate paralytic shellfish poisons. Despite numerous studies on the impact of STX in marine bivalves, knowledge regarding STX recognition at molecular level by benthic species remains scarce. Therefore, the aim of this study was to identify novel genes that interact with STX in the Chilean mussel Mytilus chilensis. For this, RNA-seq and RT-qPCR approaches were used to evaluate the transcriptomic response of M. chilensis to a purified STX as well as in vivo Alexandrium catenella exposure. Approximately 800 million reads were assembled, generating 138,883 contigs that were blasted against the UniProt Mollusca database. Pattern Recognition Receptors (PRRs) involved in mussel immunity, such as Toll-like receptors, tumor necrosis factor receptors, and scavenger-like receptors were found to be strongly upregulated at 8 and 16 h post-STX injection. These results suggest an involvement of PRRs in the response to STX, as well as identifying potential, novel STX-interacting receptors in this Chilean mussel. This study is the first transcriptomic overview of the STX-response in the edible species M. chilensis. However, the most significant contribution of this work is the identification of immune receptors and pathways potentially involved in the recognition and defense against STX’s toxicity and its impact of harmful algae blooms on wild and cultivated mussel populations. PMID:27764234

  17. Seasonal phytoplankton dynamics in extreme southern South America (Beagle Channel, Argentina)

    NASA Astrophysics Data System (ADS)

    Almandoz, Gastón O.; Hernando, Marcelo P.; Ferreyra, Gustavo A.; Schloss, Irene R.; Ferrario, Martha E.

    2011-08-01

    The phytoplankton biomass and composition dynamics in relation with environmental factors were explored during an annual cycle (July 2006-2007) for the first time in the Beagle Channel. Samples were examined by light and scanning electron microscopy and carbon biomass was estimated using measured cell biovolumes. The annual cycle was characterized by a sharp contrast between low density and biomass during the autumn-winter period and a significant increase during spring and summer. The maximum biomass peak was observed during early spring (141 μg C l - 1 and 8.7 μg Chl a l - 1 ) and was associated with a bloom of the diatom genus Chaetoceros. In spite of a strong nutrient depletion that lead to the subsequent bloom decline, a second peak of biomass was observed after 2 weeks, mainly represented by the diatoms T. mendiolana and T. anguste-lineata. Towards the end of spring and during summer diatom density decreased and assemblages were enriched by unidentified tiny phytoflagellates and dinoflagellates. A total of 138 taxa were identified during the study period. From these, 10 are known as potentially toxic species: Alexandrium catenella, A. ostenfeldii, A. tamarense, Dinophysis acuminata, D. rotundata, Prorocentrum cordatum, Pseudo-nitzschia australis, P. calliantha, P. fraudulenta and P. cf. seriata. Their presence is noteworthy since the area sustains a growing aquiculture production.

  18. Analysis of Toxic and Non-Toxic Alexandrium (Dinophyceae) Species Using Ribosomal RNA Gene Sequences

    DTIC Science & Technology

    1993-02-01

    1990). Late Mesozoic and Cenozoic paleoceanography of the northern polar oceans. In: Grantz, A., Johnson, L., Sweeny, U.. (eds.). The Geology of...Cultures represent isolates from North America, Western Europe, Thailand, Japan, Australia and the ballast water of several cargo vessels, and include... eastern North America. The B gene is considered to be a pseudogene. A restriction fragment L-ngth polymorphism (RFLP) assay developed to detect the A

  19. Differential Growth Responses of Marine Phytoplankton to Herbicide Glyphosate

    PubMed Central

    Wang, Cong; Lin, Xin; Li, Ling; Lin, Senjie

    2016-01-01

    Glyphosate is a globally popular herbicide to kill weeds and its wide applications may lead to accumulation in coastal oceans as a source of phosphorus (P) nutrient or growth inhibitor of phytoplankton. We studied the physiological effects of glyphosate on fourteen species representing five major coastal phytoplankton phyla (haptophyta, bacillariophyta, dinoflagellata, raphidophyta, and chlorophyta). Based on growth responses to different concentrations of glyphosate under contrasting dissolved inorganic phosphorus (DIP) conditions, we found that phytoplankton species could be classified into five groups. Group I (Emiliania huxleyi, Skeletonema costatum, Phaeodactylum tricornutum) could utilize glyphosate as sole P-source to support growth in axenic culture, but in the presence of DIP, they were inhibited by both 36-μM and 360-μM glyphosate. Group II (Karenia mikimotoi, Prorocentrum minimum, Dunaliella tertiolecta, Symbiodinium sp., Heterosigma akashiwo and Alexandrium catenella) could not utilize glyphosate as sole P-source to support growth, and in the presence of DIP growth was not affected by 36-μM but inhibited by 360-μM glyphosate. Glyphosate consistently enhanced growth of Group III (Isochrysis galbana) and inhibited Group IV (Thalassiosira weissflogii, Thalassiosira pseudonana and Chattonella marina) regardless of DIP condition. Group V (Amphidinium carterae) exhibited no measurable response to glyphosate regardless of DIP condition. This grouping is not congruent with the phylogenetic relationships of the phytoplankton species suggesting functional differentiation driven by environmental pressure. We conclude that glyphosate could be used as P-source by some species while is toxic to some other species and yet has no effects on others. The observed differential effects suggest that the continued use of glyphosate and increasing concentration of this herbicide in the coastal waters will likely exert significant impact on coastal marine phytoplankton

  20. [Vibrio parahaemolyticus infections and algal intoxications as emergent public health problems in Chile].

    PubMed

    Hernández, Cristina; Ulloa, Juanita; Vergara, José Antonio; Espejo, Romilio; Cabello, Felipe

    2005-09-01

    There is interest in the paradigm that relates environmental sea changes to the emergence of diseases that affect both aquatic organisms in the sea and human beings. The emergence of Vibrio parahaemolyticus as an important cause of epidemic summer diarrhea in 2004 and 2005, confined mainly to the tenth region in Chile, could be a manifestation of this trend. This and other areas of the country have also experienced several outbreaks of paralytic shellfish poisoning (PSP), diarrheal shellfish poisoning (DSP) and amnesic shellfish poisoning (ASP) caused by harmful algal blooms (HAB) of Alexandrium catenella, Dinophysis acuta and Pseudonitzchia species, respectively. The short historical record of these pathological phenomena in Chile suggests that they are increasing in frequency and expanding their geographical range. The V parahaemolyticus isolates responsible for the Chilean outbreaks correspond mainly to the pandemic strain O3:K6. HAB found in Chile and the intoxications caused by them have similar biological characteristics to those described in other areas of the world. The tenth region, the area where these problems are emerging, produces approximately 80-90% of the shellfish consumed in Chile and a large proportion of the shellfish that is exported. Prevention of these public health problems can be attained by developing policies that increase environmental surveillance for Vibrios and toxic algae, improve the epidemiological surveillance of acute diarrhea and algal intoxications after the ingestion of raw bivalves, and educate the population on the mode of transmission of these diseases. Scientific capacity and laboratories need to be developed to widen the limited knowledge of the biology of Vibrio and toxic algae and the environmental factors that favor their emergence as public health and economic problems in Chile.

  1. Differential Growth Responses of Marine Phytoplankton to Herbicide Glyphosate.

    PubMed

    Wang, Cong; Lin, Xin; Li, Ling; Lin, Senjie

    2016-01-01

    Glyphosate is a globally popular herbicide to kill weeds and its wide applications may lead to accumulation in coastal oceans as a source of phosphorus (P) nutrient or growth inhibitor of phytoplankton. We studied the physiological effects of glyphosate on fourteen species representing five major coastal phytoplankton phyla (haptophyta, bacillariophyta, dinoflagellata, raphidophyta, and chlorophyta). Based on growth responses to different concentrations of glyphosate under contrasting dissolved inorganic phosphorus (DIP) conditions, we found that phytoplankton species could be classified into five groups. Group I (Emiliania huxleyi, Skeletonema costatum, Phaeodactylum tricornutum) could utilize glyphosate as sole P-source to support growth in axenic culture, but in the presence of DIP, they were inhibited by both 36-μM and 360-μM glyphosate. Group II (Karenia mikimotoi, Prorocentrum minimum, Dunaliella tertiolecta, Symbiodinium sp., Heterosigma akashiwo and Alexandrium catenella) could not utilize glyphosate as sole P-source to support growth, and in the presence of DIP growth was not affected by 36-μM but inhibited by 360-μM glyphosate. Glyphosate consistently enhanced growth of Group III (Isochrysis galbana) and inhibited Group IV (Thalassiosira weissflogii, Thalassiosira pseudonana and Chattonella marina) regardless of DIP condition. Group V (Amphidinium carterae) exhibited no measurable response to glyphosate regardless of DIP condition. This grouping is not congruent with the phylogenetic relationships of the phytoplankton species suggesting functional differentiation driven by environmental pressure. We conclude that glyphosate could be used as P-source by some species while is toxic to some other species and yet has no effects on others. The observed differential effects suggest that the continued use of glyphosate and increasing concentration of this herbicide in the coastal waters will likely exert significant impact on coastal marine phytoplankton

  2. Blooms of the Toxic Dinoflagellate Alexandrium fundyense in the Gulf of Maine: Investigations Using a Physical-Biological Model

    DTIC Science & Technology

    2005-02-01

    B. A., Bronzino , A. C., Matrai, P., Thompson, B., Keller, M., McGillicuddy, D. J., Hyatt, J., submitted. Experimental and modeling observations of...and Oceanography (submitted). Anderson, D. M., Stock, C. A., Keafer, B. A., Bronzino , A. C., Matrai, P., Thompson, B., Keller, M., McGillicuddy, D. J...submitted). Anderson, D. M., Stock, C. A., Keafer, B. A., Bronzino , A. C., Matrai, P., Thompson, B., Keller, M., McGillicuddy, D. J., Hyatt, J

  3. Adaptations and selection of harmful and other dinoflagellate species in upwelling systems. 2. Motility and migratory behaviour

    NASA Astrophysics Data System (ADS)

    Smayda, T. J.

    2010-04-01

    The motility and migrational behaviour of upwelling dinoflagellates as adaptations for growth in upwelling systems is evaluated. Traits considered include hydrodynamic streamlining; chain formation; motility rates of single cells and chains; adaptations to turbulence; turbulence sensing; and migrational scattering to avoid turbulence, including its role in the maintenance of indigenous populations. Motility rates are compared to vertical mixing and upwelling rates. Diverse combinations of cell shape, size and motility rates characterize the dinoflagellate species selected for growth in physically energetic upwelling systems. Specific or unique combinations of cell shape, size, propulsion system and swimming rate are not evident. The traits are shared with dinoflagellates generally, and probably reflect their swim-based ecology. Experimental evidence - primarily from Alexandrium catenella - suggests upwelling dinoflagellates can sense turbulence leading to three distinct, but coherent, adaptive responses: chain formation (in such species); increased swimming speed (including non-chain-forming species); and the capacity to re-orient swimming trajectory in response to changes in turbulence, and at time-scales appropriate to survival and growth in the turbulence field being experienced. The added swimming power that dinoflagellates gain through chain formation does not appear to be a major requirement for their selection or success in upwelling systems. Only three of the 42 most prominent dinoflagellates that bloom in eastern boundary upwelling systems form chains, a representation far below expectations. Most chain-forming dinoflagellates are excluded from those upwelling systems. The role of temperature in this exclusion is evaluated. Field and experimental evidence suggests that strong turbulence would be required to overwhelm the swimming-based ecology of the upwelling dinoflagellates and deter their blooms. The Yamazaki-Kamykowski model demonstrating that the

  4. Coastal thin layer dynamics: Consequences to biology and optics

    NASA Astrophysics Data System (ADS)

    Sullivan, James M.; Donaghay, Percy L.; Rines, Jan E. B.

    2010-01-01

    Thin layers are fine-scale structures with high concentrations of organisms or particles occurring over very small vertical scales (a few meters or less), but with large horizontal scales, often extending for many kilometers. Because of their small vertical scales, thin layers are traditionally under sampled, but when proper measurement techniques are used, thin layers have been found to be ubiquitous in stratified oceans. A multi-investigator, interdisciplinary study of thin layers was sponsored by the US Office of Naval Research under a research initiative termed: Layered Organization in the Coastal Ocean (LOCO). The goal of this program was to understand the properties of coastal thin layers and the interacting physical, chemical, biological and optical processes responsible for their formation, maintenance and dissipation. As part of this program, fine-scale vertical profiles (cm resolution) of biological, physical and chemical properties were made hourly over periods spanning 1-3 weeks during three summers in Monterey Bay, California USA. The vertical profiles were made using arrays of moored autonomous profilers. In total, these profilers made ~2000 individual vertical profiles and provided a unique view of phytoplankton thin layer spatial-temporal dynamics. The autonomous profiler data were supplemented with high-resolution ship-based profiling and discrete water sampling for identifications of organisms. Persistent phytoplankton thin layers were observed during each year in Monterey Bay; however, each year had very different biological and physical dynamics. During 2002, thin layers were dominated by the non-motile and potentially toxic diatom genus Pseudo-nitzschia; during 2005, thin layers were dominated by the highly motile dinoflagellate species Akashiwo sanguinea; and during 2006, a more complex phytoplankton assemblage was present, but thin layers of the toxic dinoflagellate species Alexandrium catenella frequently occurred. The variability in the

  5. Dissolved saxitoxin causes transient inhibition of sensorimotor function in larval Pacific herring (Clupea harengus pallasi) Kathi A. Lefebvre , N

    USGS Publications Warehouse

    Lefebvre, Kathi A.; Elder, Nancy E.; Hershberger, Paul K.; Trainer, Vera L.; Stehr, Carla M.; Scholz, Nathaniel L.

    2005-01-01

    Herring (Clupea harengus pallasi) spawning sites in Puget Sound, Washington overlap spatially and temporally with blooms of Alexandrium catenella, a toxic dinoflagellate species responsible for paralytic shellfish poisoning. Consequently, newly hatched herring larvae may be regularly exposed to the suite of dissolved paralytic shellfish toxins that are released into the water column from toxic cells during blooms. To date, virtually nothing is known about the impacts of these neurotoxins on early developmental stages of marine fish. In the present study, herring larvae at three ages, 0 days post hatch (dph), 4 dph, and 11 dph, were exposed to dissolved saxitoxin (STX) in 24-h and multi-day exposures. All larvae were examined for sensorimotor function (i.e. spontaneous swimming behavior and touch response). Significant reductions in spontaneous and touch-activated swimming behavior occurred within 1 h of exposure. EC50s at 1 h of exposure were 1,500, 840, and 700 μg STX equiv. l−1 for larvae introduced to STX at 0, 4, and 11 dph, respectively. This progressive age-specific increase in STX-induced paralysis suggests that older larvae were more sensitive to the toxin than younger larvae. Interestingly, herring larvae at all ages exhibited a significant degree of neurobehavioral recovery within 4–24 h of continuous exposure relative to the 1-h time point. This recovery of normal motor behaviors was not observed in previous studies with freshwater zebrafish (Danio rerio) larvae under the same continuous exposure conditions, suggesting that an adaptive detoxification or toxin sequestration mechanism may have evolved in some species of marine fish larvae. Our data reveal that (1) dissolved STX is bioavailable to marine finfish larvae, (2) the toxin is a paralytic agent with potencies that differ between developmental stages, and (3) STX-induced sensorimotor inhibition occurs rapidly but is transient in marine larvae. Collectively, these results suggest that

  6. 77 FR 72994 - Magnuson-Stevens Fishery Conservation and Management Act Provisions; Fisheries of the...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-07

    ... that oceanographic conditions and alga sampling data suggest that the northern section of the Temporary... by the alga Alexandrium fundyense, which can form blooms commonly referred to as red tides. Red...

  7. 76 FR 72125 - Magnuson-Stevens Fishery Conservation and Management Act Provisions; Fisheries of the...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-22

    ... that oceanographic conditions and alga sampling data suggest that the northern section of the Temporary... cause PSP. These toxins are produced by the alga Alexandrium fundyense, ] which can form blooms...

  8. Environmental influences on the seasonal distribution of Vibrio parahaemolyticus in the Pacific Northwest of the USA

    EPA Science Inventory

    Populations of Vibrio parahaemolyticus in the environment can be influenced by numerous factors. We assessed the correlation of total (tl+) and potentially virulent (tdh+) V. parahaemolyticus in water with three harmful algal bloom (HAB) genera (Pseudo-nitzschia, Alexandrium and ...

  9. Case study: The Avengers 3D: cinematic techniques and digitally created 3D

    NASA Astrophysics Data System (ADS)

    Clark, Graham D.

    2013-03-01

    Marvel's THE AVENGERS was the third film Stereo D collaborated on with Marvel; it was a summation of our artistic development of what Digitally Created 3D and Stereo D's artists and toolsets affords Marvel's filmmakers; the ability to shape stereographic space to support the film and story, in a way that balances human perception and live photography. We took our artistic lead from the cinematic intentions of Marvel, the Director Joss Whedon, and Director of Photography Seamus McGarvey. In the digital creation of a 3D film from a 2D image capture, recommendations to the filmmakers cinematic techniques are offered by Stereo D at each step from pre-production onwards, through set, into post. As the footage arrives at our facility we respond in depth to the cinematic qualities of the imagery in context of the edit and story, with the guidance of the Directors and Studio, creating stereoscopic imagery. Our involvement in The Avengers was early in production, after reading the script we had the opportunity and honor to meet and work with the Director Joss Whedon, and DP Seamus McGarvey on set, and into post. We presented what is obvious to such great filmmakers in the ways of cinematic techniques as they related to the standard depth cues and story points we would use to evaluate depth for their film. Our hope was any cinematic habits that supported better 3D would be emphasized. In searching for a 3D statement for the studio and filmmakers we arrived at a stereographic style that allowed for comfort and maximum visual engagement to the viewer.

  10. Effect of different solvents extracts and mode of action of Loktanella spp. Gb03 on toxic dinoflagellate

    NASA Astrophysics Data System (ADS)

    Hameed, Anmar; Usup, Gires; Ahmad, Asmat

    2016-11-01

    This study was aimed to evaluate the algicidal activity of Loktanella sp. Gb-03 bacterial extracts against toxic dinoflagellate, using various polar and non-polar solvents. For this purpose, six different solvent extracts were prepared (i.e. methanol, ethyl acetate, hexane, chloroform, acetonitrile and water). Ratio of 1:100 (v:v) (extract to dinoflagellate culture) of each extract was used for preliminary algicidal activity screening against toxic dinoflagellate Coolia malaynesis. Dinoflagellate cells at the stationary phase (1.0 × 103 cells/ mL) were treated with 1% (v/v) of each extract by using 24-well microplate. The plates were then incubated for 24 hours at dinoflagellate culture condition (under a light intensity of 140 µmol m-2s-1 and 12:12 hours light:dark photoperiod). The result of algicidal activity screening showed that all 6 extracts from Loktanella sp. Gb-03 had different ranges of algicidal activity against the toxic dinoflagellates. Ethyl acetate extract showed the highest activity against C. malaynesis and also other harmful dinoflagellate (Alexandrium sp. Alexandrium leei, Alexandrium affine, Alexandrium tamiyavanichi, Alexandrium tamarense, Gambierdiscus belizeanus, and Ostreopsis). This study was the first to explore the algicidal activity of Loktanella sp. Gb-03 extracts against toxic dinoflagellate with ethyl acetate as the best solvent to extract algicidal active compounds.

  11. 77 FR 53164 - Magnuson-Stevens Fishery Conservation and Management Act Provisions; Fisheries of the...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-31

    ... of surfclams and ocean quahogs since 1990 due to red tide blooms that cause paralytic shellfish... alga Alexandrium fundyense, which can form blooms commonly referred to as red tides, or harmful algal... as requested by the Mid-Atlantic Fishery Management Council, NMFS also recognizes that...

  12. Interannual variability in the timing of New England shellfish toxicity and relationships to environmental forcing

    PubMed Central

    Nair, Apurva; Thomas, Andrew C.; Borsuk, Mark E.

    2013-01-01

    Routine monitoring along the coast of the Gulf of Maine (GoM) reveals shellfish toxicity nearly every summer, but at varying times, locations, and magnitudes. The responsible toxin is known to be produced by the dinoflagellate Alexandrium fundyense, yet there is little apparent association between Alexandrium abundance and shellfish toxicity. One possibility is that toxic cells are persistent in offshore areas and variability in shellfish toxicity is caused not by changes in overall abundance, but rather by variability in transport processes. Measurements of offshore Alexandrium biomass are scarce, so we bypass cell abundance as an explanatory variable and focus instead on the relations between shellfish toxicity and concurrent metrics of GoM meteorology, hydrology, and oceanography. While this yields over two decades (1985–2005) of data representing a variety of interannual conditions, the toxicity data are gappy in spatial and temporal coverage. We address this through a combination of parametric curve fitting and hierarchical cluster analysis to reveal eight archetypical modes of seasonal toxicity timing. Groups of locations are then formed that have similar interannual patterns in these archetypes. Finally, the interannual patterns within each group are related to available environmental metrics using classification trees. Results indicate that a weak cross-shore sea surface temperature (SST) gradient in the summer is the strongest correlate of shellfish toxicity, likely by signifying a hydrological connection between offshore Alexandrium populations and near-shore shellfish beds. High cumulative downwelling wind strength early in the season is revealed as a precursor consistent with this mechanism. Although previous studies suggest that alongshore transport is important in moving Alexandrium from the eastern to western GoM, alongshore SST gradient is not an important correlate of toxicity in our study. We conclude by discussing the implications of our

  13. Morphology and phylogeny of Triadinium polyedricum (Pouchet) Dodge (Dinophyceae) from Korean coastal waters

    NASA Astrophysics Data System (ADS)

    Shin, Hyeon Ho; Li, Zhun; Kim, Eun Song; Youn, Joo Yeon; Jeon, Seul Gi; Oh, Seok Jin; Lim, Weol-Ae

    2016-12-01

    To identify features that can be used to differentiate Triadinium polyedricum from other related species, such as Fukuyoa paulensis and Alexandrium species, the detailed morphology and phylogeny of T. polyedricum collected from Korean coastal waters were investigated. The cells had a plate formula of Po, 3', 7″, 5‴, 1p and 2″″, which is consistent with morphological descriptions in previous reports. Large subunit ribosomal DNA sequences also revealed that T. polyedricum from Korean coastal waters is identical to previously recorded isolates. T. polyedricum is morphologically characterized by a ventral pore in the 1″ plate that is comparable to F. paulensis and Alexandrium species. This result indicates that the location and presence of this ventral pore seems suitable for differentiating T. polyedricum from other related species.

  14. Gene duplication, loss and selection in the evolution of saxitoxin biosynthesis in alveolates.

    PubMed

    Murray, Shauna A; Diwan, Rutuja; Orr, Russell J S; Kohli, Gurjeet S; John, Uwe

    2015-11-01

    A group of marine dinoflagellates (Alveolata, Eukaryota), consisting of ∼10 species of the genus Alexandrium, Gymnodinium catenatum and Pyrodinium bahamense, produce the toxin saxitoxin and its analogues (STX), which can accumulate in shellfish, leading to ecosystem and human health impacts. The genes, sxt, putatively involved in STX biosynthesis, have recently been identified, however, the evolution of these genes within dinoflagellates is not clear. There are two reasons for this: uncertainty over the phylogeny of dinoflagellates; and that the sxt genes of many species of Alexandrium and other dinoflagellate genera are not known. Here, we determined the phylogeny of STX-producing and other dinoflagellates based on a concatenated eight-gene alignment. We determined the presence, diversity and phylogeny of sxtA, domains A1 and A4 and sxtG in 52 strains of Alexandrium, and a further 43 species of dinoflagellates and thirteen other alveolates. We confirmed the presence and high sequence conservation of sxtA, domain A4, in 40 strains (35 Alexandrium, 1 Pyrodinium, 4 Gymnodinium) of 8 species of STX-producing dinoflagellates, and absence from non-producing species. We found three paralogs of sxtA, domain A1, and a widespread distribution of sxtA1 in non-STX producing dinoflagellates, indicating duplication events in the evolution of this gene. One paralog, clade 2, of sxtA1 may be particularly related to STX biosynthesis. Similarly, sxtG appears to be generally restricted to STX-producing species, while three amidinotransferase gene paralogs were found in dinoflagellates. We investigated the role of positive (diversifying) selection following duplication in sxtA1 and sxtG, and found negative selection in clades of sxtG and sxtA1, clade 2, suggesting they were functionally constrained. Significant episodic diversifying selection was found in some strains in clade 3 of sxtA1, a clade that may not be involved in STX biosynthesis, indicating pressure for diversification

  15. Grazer cues induce stealth behavior in marine dinoflagellates

    PubMed Central

    Selander, Erik; Jakobsen, Hans H.; Lombard, Fabien; Kiørboe, Thomas

    2011-01-01

    Chain formation is common among phytoplankton organisms but the underlying reasons and consequences are poorly understood. Here we show that chain formation is strongly impaired by waterborne cues from copepod grazers in the dinoflagellate Alexandrium tamarense. Chains of Alexandrium cells exposed to copepod cues responded by splitting into single cells or shorter chains. Motion analysis revealed significantly lower swimming velocities for single cells compared with chains, with two- to fivefold higher simulated predator encounter rates for two- and four-cell chains, respectively. In addition, the few remaining two-cell chains in grazed treatments were swimming at approximately half the speed of two-cell chains in treatments without grazers, which reduced encounter rates with grazers to values similar to that of single cells. Chain length plasticity and swimming behavior constitute unique mechanisms to reduce encounters with grazers. We argue that dinoflagellates can regulate the balance between motility and predator avoidance by adjusting chain length. The high predator encounter rate for motile chains may have contributed to the low prevalence of chain formation in motile phytoplankton compared with in nonmotile phytoplankton where chain formation is more common. PMID:21368128

  16. Repercussions of salinity changes and osmotic stress in marine phytoplankton species

    NASA Astrophysics Data System (ADS)

    D'ors, A.; Bartolomé, M. C.; Sánchez-Fortún, S.

    2016-06-01

    The short-term effect of low salinity was studied using laboratory protocols on some coastal phytoplankton species such as chlorophycea Tetraselmis suecica, among diatom the strain Nitzschia N1c1 and dinoflagellates Alexandrium minutum and Prorocentrum lima. All of cultures were exposed to low salinities, and cell growth rate, photosynthetic quantum yield (ΦPSII), and gross photosynthesis (Pg) were analyzed. Growth rate inhibition was similar in all species, and all of them also tolerate short-term exposures to salinities in the range 5-35. There were no significant differences between ΦPSII and Pg endpoints from Tetraselmis suecica and Nitzschia sp., while Alexandrium minutum and Prorocentrum lima displayed a higher affectation rate on Pg than on ΦPSII activity. The influence of low salinity was higher on respiration in T. suecica, while both dinoflagellates had higher net photosynthesis. Nitzschia sp. exhibited similar involvement of the two photosynthetic parameters. Therefore, although the four phytoplankton monocultures studied are able to survive in internal areas of estuaries under low salinity conditions, the photosynthetic activity is more affected than the growth rate in all phytoplankton communities studied except in chlorophycea T. suecica, which has increased tolerance for this salinity decrease.

  17. When Naked Became Armored: An Eight-Gene Phylogeny Reveals Monophyletic Origin of Theca in Dinoflagellates

    PubMed Central

    Orr, Russell J. S.; Murray, Shauna A.; Stüken, Anke; Rhodes, Lesley; Jakobsen, Kjetill S.

    2012-01-01

    The dinoflagellates are a diverse lineage of microbial eukaryotes. Dinoflagellate monophyly and their position within the group Alveolata are well established. However, phylogenetic relationships between dinoflagellate orders remain unresolved. To date, only a limited number of dinoflagellate studies have used a broad taxon sample with more than two concatenated markers. This lack of resolution makes it difficult to determine the evolution of major phenotypic characters such as morphological features or toxin production e.g. saxitoxin. Here we present an improved dinoflagellate phylogeny, based on eight genes, with the broadest taxon sampling to date. Fifty-five sequences for eight phylogenetic markers from nuclear and mitochondrial regions were amplified from 13 species, four orders, and concatenated phylogenetic inferences were conducted with orthologous sequences. Phylogenetic resolution is increased with addition of support for the deepest branches, though can be improved yet further. We show for the first time that the characteristic dinoflagellate thecal plates, cellulosic material that is present within the sub-cuticular alveoli, appears to have had a single origin. In addition, the monophyly of most dinoflagellate orders is confirmed: the Dinophysiales, the Gonyaulacales, the Prorocentrales, the Suessiales, and the Syndiniales. Our improved phylogeny, along with results of PCR to detect the sxtA gene in various lineages, allows us to suggest that this gene was probably acquired separately in Gymnodinium and the common ancestor of Alexandrium and Pyrodinium and subsequently lost in some descendent species of Alexandrium. PMID:23185516

  18. A mechanism for offshore initiation of harmful algal blooms in the coastal Gulf of Maine

    USGS Publications Warehouse

    McGillicuddy, D.J.; Signell, R.P.; Stock, C.A.; Keafer, B.A.; Keller, M.D.; Hetland, R.D.; Anderson, D.M.

    2003-01-01

    A combination of observations and model results suggest a mechanism by which coastal blooms of the toxic dinoflagellate Alexandrium fundyense can be initiated from dormant cysts located in offshore sediments. The mechanism arises from the joint effects of organism behavior and the wind-driven response of a surface-trapped plume of fresh water originating from riverine sources. During upwelling-favorable winds, the plume thins vertically and extends offshore; downwelling winds thicken the plume and confine it to the nearshore region. In the western Gulf of Maine, the offshore extent of the river plume during upwelling conditions is suffcient to entrain upward-swimming A. fundyense cells germinated from offshore cyst beds. Subsequent downwelling conditions then transport those populations towards the coast.

  19. The influence of extracellular compounds produced by selected Baltic cyanobacteria, diatoms and dinoflagellates on growth of green algae Chlorella vulgaris

    NASA Astrophysics Data System (ADS)

    Żak, Adam; Kosakowska, Alicja

    2015-12-01

    Secondary metabolites produced by bacteria, fungi, algae and plants could affect the growth and development of biological and agricultural systems. This natural process that occurs worldwide is known as allelopathy. The main goal of this work was to investigate the influence of metabolites obtained from phytoplankton monocultures on the growth of green algae Chlorella vulgaris. We selected 6 species occurring in the Baltic Sea from 3 different taxonomic groups: cyanobacteria (Aphanizomenon flos-aquae; Planktothrix agardhii), diatoms (Thalassiosira pseudonana; Chaetoceros wighamii) and dinoflagellates (Alexandrium ostenfeldii; Prorocentrum minimum). In this study we have demonstrated that some of selected organisms caused allelopathic effects against microalgae. Both the negative and positive effects of collected cell-free filtrates on C. vulgaris growth, chlorophyll a concentration and fluorescence parameters (OJIP, QY, NPQ) have been observed. No evidence has been found for the impact on morphology and viability of C. vulgaris cells.

  20. Evidence of paralytic shellfish poisoning toxin in milkfish in South Taiwan.

    PubMed

    Chou, H-N; Chung, Y-C; Cho, W-C; Chen, C-Y

    2003-06-01

    Natural phytoplankton blooms of the dinoflagellate Alexandrium minutum, milkfish (Chanos chanos) exposed to natural blooms, sediment and mangrove crab (Scylla serrata) were analysed for paralytic shellfish poisoning toxins by high-performance liquid chromatography. The toxin profiles of milkfish and mangrove crab were similar to that of A. minutum collected from blooming fishponds. In a laboratory A. minutum-blooming environment, the stomach and intestine of milkfish accumulated paralytic shellfish poisoning toxins during the exposure period. The non-visceral tissues were non-toxic. However, milkfish lost their entire body burden of toxin on the first day of transferring to a toxic algae-free environment. The result shows that milkfish concentrate paralytic shellfish poisoning toxins in digestive organs and did not retain toxins.

  1. [Effects of marine bacteria on the growth and toxin production of red-tide algae under different pH and salinities].

    PubMed

    Su, Jianqiang; Zheng, Tianling; Hu, Zhong; Xu, Jingsen; Yu, Zhiming; Song, Xiuxian

    2003-07-01

    The effects of strain S10 isolated from sediments of Xiamen Western Sea Area on the growth and paralytic shellfish poison (PSP) production of Alexandrium tamarense at different pH and salinities were studied. The results showed that the alga grew well at pH 6-8 and at salinity of 20-34. The toxicity of A. tamarense varied markedly at different pH and salinities: it decreased with increasing pH, while increased with salinity and reached its peak value at the salinity of 30, and then declined. The strain S10 inhibited the growth and the PSP production of A. tamarense at different pH and salinities. It had the best inhibitory function on the growth of A. tamarense at pH 7 and salinity of 34. The best inhibitory function on the PSP production of A. tamarense was at pH 7, but this inhibitory function was not related to salinity.

  2. Paralytic shellfish poisoning toxin profile of mussels Perna perna from southern Atlantic coasts of Morocco.

    PubMed

    Abouabdellah, Rachid; Taleb, Hamid; Bennouna, Asmae; Erler, Katrin; Chafik, Abdeghani; Moukrim, Abdelatif

    2008-04-01

    During the monitoring programme of harmful algal blooms established along the south Atlantic coast of Morocco, a bimonthly determination of harmful algae and phycotoxins analysis in Perna perna was carried out from May 2003 to December 2004. Results of mouse bioassay (in organs and whole flesh) showed a seasonal evolution of paralytic shellfish poisoning (PSP) toxin. The mussel's contamination was associated with the occurrence in water of Alexandrium minutum. The PSP toxin profile obtained with high-performance liquid chromatography (HPLC/FD) revealed the dominance of gonyautoxins GTX2 and GTX3 and a minority of GTX1, GTX4 and saxitoxin (STX). This profile explains that the toxicity was mainly associated with A. minutum.

  3. Isolation, purification and spectrometric analysis of PSP toxins from moraxella sp., a bacterium associated with a toxic dinoflagellate

    SciTech Connect

    Boyce, S.D.; Doucette, G.J.

    1994-12-31

    Paralytic shellfish poisoning (PSP) is a seafood intoxication syndrome caused by the injestion of shellfish contaminated with toxins produced by algae known as dinoflagellates. The PSP toxins, saxitoxin and its derivatives, act to block voltage-dependent sodium channels and can cause paralysis and even death at higher doses. It is well documented that bacteria coexist with many harmful or toxic algal species, though the exact nature of the association in relation to toxin production is unknown. Recently, the bacterium Moraxella sp. was isolated from the PSP toxin producing dinoflagellate Alexandrium tamarense. Through HPLC analysis and saxitoxin receptor binding assays performed on crude bacterial extracts, it appears that Moraxella sp. is capable of producing saxitoxin and several of its derivatives. However, physical confirmation (e.g. mass spectrometry) of these results is still needed.

  4. Genome of the R-body producing marine alphaproteobacterium Labrenzia alexandrii type strain (DFL-11T)

    PubMed Central

    Fiebig, Anne; Pradella, Silke; Petersen, Jörn; Päuker, Orsola; Michael, Victoria; Lünsdorf, Heinrich; Göker, Markus; Klenk, Hans-Peter; Wagner-Döbler, Irene

    2013-01-01

    Labrenzia alexandrii Biebl et al. 2007 is a marine member of the family Rhodobacteraceae in the order Rhodobacterales, which has thus far only partially been characterized at the genome level. The bacterium is of interest because it lives in close association with the toxic dinoflagellate Alexandrium lusitanicum. Ultrastructural analysis reveals R-bodies within the bacterial cells, which are primarily known from obligate endosymbionts that trigger “killing traits” in ciliates (Paramecium spp.). Genomic traits of L. alexandrii DFL-11T are in accordance with these findings, as they include the reb genes putatively involved in R-body synthesis. Analysis of the two extrachromosomal elements suggests a role in heavy-metal resistance and exopolysaccharide formation, respectively. The 5,461,856 bp long genome with its 5,071 protein-coding and 73 RNA genes consists of one chromosome and two plasmids, and has been sequenced in the context of the Marine Microbial Initiative. PMID:24019989

  5. Isolation and characterization of pigmented algicidal bacteria from seawater

    NASA Astrophysics Data System (ADS)

    Shaima, A.; Gires, U.; Asmat, A.

    2014-09-01

    Some dinoflagellate species are toxic and widely distributed in Malaysian marines ecosystems. They can cause many problems to aquatic life due to the production of various potential and natural toxins that accumulate in filter feeding shellfish and cause food poisoning to human. In recent decades, bacteria have been widely used as a biological control against these harmful algae. In the present study, pigmented bacteria isolated from marine water of Port Dickson beach was studied for their anti-algal activity towards toxic dinoflagellate Alexandrium minutum. Four isolates were studied and only one was capable of inhibiting algal growth when treated with bacterial culture. The algilytic effect on dinoflagellate was evaluated based on direct cell count under the microscope. Results showed that only isolate Sdpd-310 with orange colour has an inhibitory effect on A. minutum growth. This study demonstrated the rapid algicidal activity of a marine pigmented bacteria against the toxic dinoflagellate A. minutum.

  6. Exploring the erodibility of sediments and harmful algal blooms in the Gulf of Maine

    USGS Publications Warehouse

    Butman, Bradford; Dickhudt, Patrick J.; Keafer, Bruce A.

    2012-01-01

    Investigators at the U.S. Geological Survey (USGS) are cooperating with scientists at Woods Hole Oceanographic Institution (WHOI) to investigate harmful algal blooms along the New England coast in the Gulf of Maine. These blooms are caused by cysts of the dinoflagellate Alexandrium fundyense that overwinter in the bottom sediments and germinate in spring. Depending on conditions such as temperature, light, nutrient levels, and currents, these single-celled organismscan create a bloom along the coast, called ‘red tides.’Shellfish that have ingested these cells in sufficient concentration can become toxic to humans and require that the shellfisheries be closed. After the spring bloom, the organisms form cysts that sink to the sea floor and are sequestered in the bottom sediments over the winter.

  7. Copepods induce paralytic shellfish toxin production in marine dinoflagellates

    PubMed Central

    Selander, Erik; Thor, Peter; Toth, Gunilla; Pavia, Henrik

    2006-01-01

    Among the thousands of unicellular phytoplankton species described in the sea, some frequently occurring and bloom-forming marine dinoflagellates are known to produce the potent neurotoxins causing paralytic shellfish poisoning. The natural function of these toxins is not clear, although they have been hypothesized to act as a chemical defence towards grazers. Here, we show that waterborne cues from the copepod Acartia tonsa induce paralytic shellfish toxin (PST) production in the harmful algal bloom-forming dinoflagellate Alexandrium minutum. Induced A. minutum contained up to 2.5 times more toxins than controls and was more resistant to further copepod grazing. Ingestion of non-toxic alternative prey was not affected by the presence of induced A. minutum. The ability of A. minutum to sense and respond to the presence of grazers by increased PST production and increased resistance to grazing may facilitate the formation of harmful algal blooms in the sea. PMID:16769640

  8. Fatal paralytic shellfish poisoning in Kittlitz's Murrelet (Brachyramphus brevirostris) nestlings, Alaska, USA

    USGS Publications Warehouse

    Shearn-Bochsler, Valerie I.; Lance, Ellen W.; Corcoran, Robin; Piatt, John; Bodenstein, Barbara; Frame, Elizabeth; Lawonn, James

    2014-01-01

    Paralytic shellfish poisoning (PSP) is an acute toxic illness in humans resulting from ingestion of shellfish contaminated with a suite of neurotoxins (saxitoxins) produced by marine dinoflagellates, most commonly in the genus Alexandrium. Poisoning also has been sporadically suspected and, less often, documented in marine wildlife, often in association with an outbreak in humans. Kittlitz's Murrelet (Brachyramphus brevirostris) is a small, rare seabird of the Northern Pacific with a declining population. From 2008 to 2012, as part of a breeding ecology study, multiple Kittlitz's Murrelet nests on Kodiak Island, Alaska, were monitored by remote cameras. During the 2011 and 2012 breeding seasons, nestlings from several sites died during mild weather conditions. Remote camera observations revealed that the nestlings died shortly after consuming sand lance (Ammodytes hexapterus), a fish species known to biomagnify saxitoxin. High levels of saxitoxin were subsequently documented in crop content in 87% of nestling carcasses. Marine bird deaths from PSP may be underreported.

  9. All aboard! A biological survey of ballast water onboard vessels spanning the North Atlantic Ocean.

    PubMed

    Steichen, Jamie L; Schulze, Anja; Brinkmeyer, Robin; Quigg, Antonietta

    2014-10-15

    Global movement of nonindigenous species, within ballast water tanks across natural barriers, threatens coastal and estuarine ecosystem biodiversity. In 2012, the Port of Houston ranked 10th largest in the world and 2nd in the US (waterborne tonnage). Ballast water was collected from 13 vessels to genetically examine the eukaryotic microorganism diversity being discharged into the Port of Houston, Texas (USA). Vessels took ballast water onboard in North Atlantic Ocean between the Port of Malabo, Africa and Port of New Orleans, Louisiana, (USA). Twenty genera of Protists, Fungi and Animalia were identified from at least 10 phyla. Dinoflagellates were the most diverse and dominant identified (Alexandrium, Exuviaella, Gyrodinium, Heterocapsa, Karlodinium, Pfiesteria and Scrippsiella). We are reporting the first detection of Picobiliphytes, Apusozoa (Amastigomonas) and Sarcinomyces within ballast water. This study supports that global commerce by shipping contributes to long-distance transportation of eukaryotic microorganisms, increasing propagule pressure and invasion supply on ecosystems.

  10. High Specificity of a Quantitative PCR Assay Targeting a Saxitoxin Gene for Monitoring Toxic Algae Associated with Paralytic Shellfish Toxins in the Yellow Sea

    PubMed Central

    Gao, Yan; Murray, Shauna A.; Chen, Jian-Hua; Kang, Zhen-Jun; Zhang, Qing-Chun; Kong, Fan-Zhou; Zhou, Ming-Jiang

    2015-01-01

    The identification of core genes involved in the biosynthesis of saxitoxin (STX) offers a great opportunity to detect toxic algae associated with paralytic shellfish toxins (PST). In the Yellow Sea (YS) in China, both toxic and nontoxic Alexandrium species are present, which makes it a difficult issue to specifically monitor PST-producing toxic algae. In this study, a quantitative PCR (qPCR) assay targeting sxtA4, a domain in the sxt gene cluster that encodes a unique enzyme involved in STX biosynthesis, was applied to analyze samples collected from the YS in spring of 2012. The abundance of two toxic species within the Alexandrium tamarense species complex, i.e., A. fundyense and A. pacificum, was also determined with TaqMan-based qPCR assays, and PSTs in net-concentrated phytoplankton samples were analyzed with high-performance liquid chromatography coupled with a fluorescence detector. It was found that the distribution of the sxtA4 gene in the YS was consistent with the toxic algae and PSTs, and the quantitation results of sxtA4 correlated well with the abundance of the two toxic species (r = 0.857). These results suggested that the two toxic species were major PST producers during the sampling season and that sxtA-based qPCR is a promising method to detect toxic algae associated with PSTs in the YS. The correlation between PST levels and sxtA-based qPCR results, however, was less significant (r = 0.552), implying that sxtA-based qPCR is not accurate enough to reflect the toxicity of PST-producing toxic algae. The combination of an sxtA-based qPCR assay and chemical means might be a promising method for monitoring toxic algal blooms. PMID:26231652

  11. Simultaneous screening for lipophilic and hydrophilic toxins in marine harmful algae using a serially coupled reversed-phase and hydrophilic interaction liquid chromatography separation system with high-resolution mass spectrometry.

    PubMed

    Chen, Junhui; Gao, Liyuan; Li, Zhaoyong; Wang, Shuai; Li, Jingxi; Cao, Wei; Sun, Chengjun; Zheng, Li; Wang, Xiaoru

    2016-03-31

    The presence of toxins in harmful algal blooms (HABs) poses considerable concerns because of their potential adverse effects on ecological environments and human health. When marine HABs occur, efficient screening and identification of toxins in different kinds of HAB algae remains a challenge. In this study, the applicability of serial coupling of reversed-phase liquid chromatography (RPLC) and hydrophilic interaction chromatography (HILIC) combined with high resolution mass spectrometry (HR-MS) for the simultaneous screening and identification of various kinds of known lipophilic and hydrophilic toxins in HAB algae was investigated for the first time. Ultrasound-assisted extraction (UAE) was explored to extract both lipophilic and hydrophilic toxins in algae simultaneously. As in most cases, toxin standards were not available; therefore, an identification procedure based on accurate mass data and chromatographic behavior was proposed. According to this procedure, eight known lipophilic toxins and 11 hydrophilic toxins were successfully detected in a single injection, and the proposed method was validated. Satisfactory sensitivity, repeatability (RSD <14.87%) and recovery (89.4-105.8%) of the method were achieved. A major advantage of the proposed method is that it can almost detect members of all eight groups of marine algal toxins in a single run. Using this method, several known toxins in different marine toxigenic algae including Alexandrium tamarense, Alexandrium minutum and Prorocentrum lima were successfully observed and identified. This work demonstrates that RPLC/HILIC-HR-MS combined with an accurate mass list of known marine algal toxins may be used as a powerful tool for screening of different classes of known toxins in marine harmful algae.

  12. Fractal dimensions of flocs between clay particles and HAB organisms

    NASA Astrophysics Data System (ADS)

    Wang, Hongliang; Yu, Zhiming; Cao, Xihua; Song, Xiuxian

    2011-05-01

    The impact of harmful algal blooms (HABs) on public health and related economics have been increasing in many coastal regions of the world. Sedimentation of algal cells through flocculation with clay particles is a promising strategy for controlling HABs. Previous studies found that removal efficiency (RE) was influenced by many factors, including clay type and concentration, algal growth stage, and physiological aspects of HAB cells. To estimate the effect of morphological characteristics of the aggregates on HAB cell removal, fractal dimensions were measured and the RE of three species of HAB organism, Heterosigma akashiwo, Alexandrium tamarense, and Skeletonema costatum, by original clay and modified clay, was determined. For all HAB species, the modified clay had a higher RE than original clay. For the original clay, the two-dimensional fractal dimension ( D 2) was 1.92 and three-dimensional fractal dimension ( D 3) 2.81, while for the modified clay, D 2 was 1.84 and D 3 was 2.50. The addition of polyaluminum chloride (PACl) lead to a decrease of the repulsive barrier between clay particles, and resulted in lower D 2 and D 3. Due to the decrease of D 3, and the increase of the effective sticking coefficient, the flocculation rate between modified clay particles and HAB organisms increased, and thus resulted in a high RE. The fractal dimensions of flocs differed in HAB species with different cell morphologies. For example, Alexandrium tamarense cells are ellipsoidal, and the D 3 and D 2 of flocs were the highest, while for Skeletonema costatum, which has filamentous cells, the D 3 and D 2 of flocs were the lowest.

  13. Cytotoxicity of goniodomin A and B in non contractile cells.

    PubMed

    Espiña, Begoña; Cagide, Eva; Louzao, M Carmen; Vilariño, Natalia; Vieytes, Mercedes R; Takeda, Yoshiyuki; Sasaki, Makoto; Botana, Luis M

    2016-05-27

    Goniodomin A is a phycotoxin produced by the dinoflagellates Alexandrium hiranoi (formerly Goniodoma pseudogoniaulax) and Alexandrium monilatum. This polyether macrolide exerts a potent antifungal effect and disturbs the actomyosin ATPase activity and the F-actin meshwork in diverse cell types. Goniodomin B is a fused acetal isomer isolated with goniodomin A with unknown activity. Histopathological changes induced by goniodomin A postulated hepatocytes as target cells. In this study both compounds induce a time and concentration dependent fall in the viability of Clone 9 rat hepatocytes. Furthermore, for both compounds, primary rat hepatocytes are almost 10 folds less sensitive than Clone 9 cells. Goniodomin A is highly effective in the nanomolar range while micromolar concentrations of goniodomin B are necessary to observe cytoxicity. Additionally, goniodomin A induced a significant increase in the F-actin and decrease in the G-actin content of Clone 9 cells but did not change the actin of primary cultured hepatocytes. However, goniodomin B could not exert significant alterations in the cytoskeleton of neither cell type. Futhermore goniodomin A as well as goniodomin B are cytotoxic to excitable cells. Both analogues triggered a time dependent decrease on viability in BE(2)-M17 human neuroblastoma cells. In this cell model goniodomin A increased the intracellular calcium and depolarized cells. We conclude that goniodomins A and B are biologically active molecules in hepatocytes and also in excitable cells BE(2)-M17. However, the analogue goniodomin B, whose activity is described in this work for the first time, is a much less potent compound.

  14. Zooplankton interactions with toxic phytoplankton: Some implications for food web studies and algal defence strategies of feeding selectivity behaviour, toxin dilution and phytoplankton population diversity

    NASA Astrophysics Data System (ADS)

    Barreiro, A.; Guisande, C.; Maneiro, I.; Vergara, A. R.; Riveiro, I.; Iglesias, P.

    2007-11-01

    This study focuses on the interactions between toxic phytoplankton and zooplankton grazers. The experimental conditions used are an attempt to simulate situations that have, so far, received little attention. We presume the phytoplankton community to be a set of species where a population of a toxic species is intrinsically diverse by the presence of coexisting strains with different toxic properties. The other species in the community may not always be high-quality food for herbivorous zooplankton. Zooplankton populations may have developed adaptive responses to sympatric toxic phytoplankton species. Zooplankton grazers may perform a specific feeding behaviour and its consequences on fitness will depend on the species ingested, the effect of toxins, and the presence of mechanisms of toxin dilution and compensatory feeding. Our target species are a strain of the dinoflagellate Alexandrium minutum and a sympatric population of the copepod Acartia clausi. Mixed diets were used with two kinds of A. minutum cells: non-toxic and toxic. The flagellate Rhodomonas baltica and the non-toxic dinoflagellate Alexandrium tamarense were added as accompanying species. The effect of each alga was studied in separate diets. The toxic A. minutum cells were shown to have negative effects on egg production, hatching success and total reproductive output, while, in terms of its effect on fitness, the non-toxic A. minutum was the best quality food offered. R. baltica and A. tamarense were in intermediate positions. In the mixed diets, copepods showed a strong preference for toxic A. minutum cells and a weaker one for A. tamarense cells, while non-toxic A. minutum was slightly negatively selected and R. baltica strongly negatively selected. Although the level of toxins accumulated by copepods was very similar, in both the diet with only toxic A. minutum cells and in the mixed diet, the negative effects on fitness in the mixed diet could be offset by toxin dilution mechanisms. The

  15. Toxin levels and profiles in microalgae from the north-Western Adriatic Sea--15 years of studies on cultured species.

    PubMed

    Pistocchi, Rossella; Guerrini, Franca; Pezzolesi, Laura; Riccardi, Manuela; Vanucci, Silvana; Ciminiello, Patrizia; Dell'Aversano, Carmela; Forino, Martino; Fattorusso, Ernesto; Tartaglione, Luciana; Milandri, Anna; Pompei, Marinella; Cangini, Monica; Pigozzi, Silvia; Riccardi, Elena

    2012-01-01

    The Northern Adriatic Sea is the area of the Mediterranean Sea where eutrophication and episodes related to harmful algae have occurred most frequently since the 1970s. In this area, which is highly exploited for mollusk farming, the first occurrence of human intoxication due to shellfish consumption occurred in 1989, nearly 10 years later than other countries in Europe and worldwide that had faced similar problems. Until 1997, Adriatic mollusks had been found to be contaminated mostly by diarrhetic shellfish poisoning toxins (i.e., okadaic acid and dinophysistoxins) that, along with paralytic shellfish poisoning toxins (i.e., saxitoxins), constitute the most common marine biotoxins. Only once, in 1994, a toxic outbreak was related to the occurrence of paralytic shellfish poisoning toxins in the Adriatic coastal waters. Moreover, in the past 15 years, the Adriatic Sea has been characterized by the presence of toxic or potentially toxic algae, not highly widespread outside Europe, such as species producing yessotoxins (i.e., Protoceratium reticulatum, Gonyaulax spinifera and Lingulodinium polyedrum), recurrent blooms of the potentially ichthyotoxic species Fibrocapsa japonica and, recently, by blooms of palytoxin-like producing species of the Ostreopsis genus. This review is aimed at integrating monitoring data on toxin spectra and levels in mussels farmed along the coast of the Emilia-Romagna region with laboratory studies performed on the species involved in the production of those toxins; toxicity studies on toxic or potentially toxic species that have recently appeared in this area are also reviewed. Overall, reviewed data are related to: (i) the yessotoxins producing species P. reticulatum, G. spinifera and L. polyedrum, highlighting genetic and toxic characteristics; (ii) Adriatic strains of Alexandrium minutum, Alexandrium ostenfeldii and Prorocentrum lima whose toxic profiles are compared with those of strains of different geographic origins; (iii) F

  16. Prominent Human Health Impacts from Several Marine Microbes: History, Ecology, and Public Health Implications

    PubMed Central

    Bienfang, P. K.; DeFelice, S. V.; Laws, E. A.; Brand, L. E.; Bidigare, R. R.; Christensen, S.; Trapido-Rosenthal, H.; Hemscheidt, T. K.; McGillicuddy, D. J.; Anderson, D. M.; Solo-Gabriele, H. M.; Boehm, A. B.; Backer, L. C.

    2011-01-01

    This paper overviews several examples of important public health impacts by marine microbes and directs readers to the extensive literature germane to these maladies. These examples include three types of dinoflagellates (Gambierdiscus spp., Karenia brevis, and Alexandrium fundyense), BMAA-producing cyanobacteria, and infectious microbes. The dinoflagellates are responsible for ciguatera fish poisoning, neurotoxic shellfish poisoning, and paralytic shellfish poisoning, respectively, that have plagued coastal populations over time. Research interest on the potential for marine cyanobacteria to contribute BMAA into human food supplies has been derived by BMAA's discovery in cycad seeds and subsequent implication as the putative cause of amyotrophic lateral sclerosis/parkinsonism dementia complex among the Chamorro people of Guam. Recent UPLC/MS analyses indicate that recent reports that BMAA is prolifically distributed among marine cyanobacteria at high concentrations may be due to analyte misidentification in the analytical protocols being applied for BMAA. Common infectious microbes (including enterovirus, norovirus, Salmonella, Campylobacter, Shigella, Staphylococcus aureus, Cryptosporidium, and Giardia) cause gastrointestinal and skin-related illness. These microbes can be introduced from external human and animal sources, or they can be indigenous to the marine environment. PMID:20976073

  17. Analysis of Lagoonal Ecosystems in the Po River Delta Associated with Intensive Aquaculture

    NASA Astrophysics Data System (ADS)

    Sorokin, Yu. I.; Sorokin, P. Yu.; Ravagnan, G.

    1999-03-01

    Observations on structure and functioning of coastal lagoon ecosystems experiencing a high level of eutrophication impact were accomplished in three lagoons of Ca'Pisani integrated within an experimental aquaculture enterprise variously fertilized by waste effluents discharged from and intensive fish culture plant. During August and early September an extremely dense bloom of dinoflagellate Alexandrium tamarensewas recorded in these lagoons with the density of phytoplankton up to 190 g m -3of wet biomass, and primary production 2 to 6 mg Cl -1 day -1. The diel dissolved oxygen fluctuations in water column during the bloom reached 15-20 mg O 2 l -1. The wet biomass of bacterioplankton in the lagoons attained 5-9 g m -3. The microzooplankton was dominated by ciliates with biomass 1 to 19 g m -3. The daytime mesozooplankton was dominated by calanoid copepods with a biomass 0·05-0·25 g m -3, while the biomass of the demersal zooplankton at night attained 2 to 14 g m -3. In the lagoon of Ocaro, the phototrophic plankton was dominated by the symbiotic ciliate Mesodinium.The labile sulphides content in the upper layer of the bottom attained over 1 g S dm -3of wet silt. The rate of microbial sulphate reduction was 5-10 mg S dm -3day -1. The data are generalized within the energy balance in these specific anthropogenically transformed pelagic communities.

  18. Metabolites of saxitoxin analogues in bivalves contaminated by Gymnodinium catenatum.

    PubMed

    Vale, Paulo

    2010-01-01

    Bivalve metabolites of saxitoxin analogues, not present in microalgae, were recently described as an important toxin fraction in mussels contaminated by Alexandrium tamarense. These possess very low fluorescence, and require mass spectrometry detection. HILIC-MS was implemented to look for these metabolites in bivalves contaminated during Gymnodinium catenatum blooms at the Portuguese coast. The presence of M1 was tentatively identified in several bivalves, ranging from estuarine (Mytilus galloprovinciallis, Cerastoderma edule and Ruditapes decussatus) to oceanic habitat (Donax trunculus and Ensis spp.). It was hypothesized that M1 could contribute to an important fraction of the profile of STX analogues. M1 was more abundant in estuarine bivalves that retain longer PSP toxins, in the following order: mussels>cockles>clams. These data highlight that the study by fluorimetry alone of the carbamoyl, N-sulfocarbamoyl, and decarbamoyl families is manifestly insufficient to fully understand toxin dynamics in bivalves feeding on G. catenatum without a proper study of hydroxybenzoate and hydroxylated M-toxins.

  19. Performance of a sand filter in removal of micro-algae from seawater in aquaculture production systems.

    PubMed

    Sabiri, N E; Castaing, J B; Massé, A; Jaouen, P

    2012-01-01

    In this study, a sand filter was used to remove micro-algae from seawater feeding aquaculture ponds. A lab-scale sand filter was used to filter 30,000 cells/mL of Heterocapsa triquetra suspension, a non-toxic micro-alga that has morphological and dimensional (15-20 microm) similarities with Alexandrium sp., one of the smallest toxic micro-algae in seawater. Removal efficiency and capture mechanisms for a fixed superficial velocity (3.5 m/h) were evaluated in relation to size distribution and mean diameter of the sand. Various sands (average diameter ranging between 200 microm and 600 microm) were characterized and used as porous media. The structural parameters of the fixed beds were evaluated for each medium using experimental measurements of pressure drop as a function of superficial velocity over a range of Reynolds numbers covering Darcy's regime and the inertial regime. For a filtration cycle of six hours, the best efficiency (E = 90%) was obtained with the following sand characteristics: sieved sand with a range of grain diameter of 100 and 300 microm and a mean grain diameter equal to 256 microm. Results obtained show the influence of the size distribution of sand on the quality of retention of the micro-algae studied.

  20. Predator/prey interaction between Pfiesteria piscicida and Rhodomonas mediated by a marine alpha proteobacterium.

    PubMed

    Alavi, M R

    2004-01-01

    The dinoflagellate Pfiesteria piscicida coexists with bacteria in aquatic environments and as such, may interact with them at the physiological level. This study was designed to investigate the influence of bacteria, present in a clonal culture of Pfiesteria piscicida, on the predator/prey relationship of this dinoflagellate with the alga Rhodomonas. A series of replenishment experiments with bacteria isolated from P. piscicida clonal culture and the bacteria-free P. piscicida derived from the same culture were carried out. In the presence of bacteria, the number of P. piscicida increased significantly when incubated with alga Rhodomonas. This enhanced growth was almost entirely due to the increased consumption rate of Rhodomonas by P. piscicida since in bacteria-free (axenic) cultures Rhodomonas were consumed at significantly reduced rates relative to cultures with bacteria. Subsequent replenishment experiments with individual bacterial isolates showed that a single isolate was responsible for the increased predation rate of P. piscicida. The presence or absence of this specific bacterium determined the outcome of the interaction between P. piscicida and Rhodomonas. Partial sequence analysis of the 16S rDNA of this isolate indicated that it was a novel marine alpha proteobacterium with sequence similarities to a Roseobacter sp. and a bacterium recently isolated from a toxic dinoflagellate Alexandrium sp.

  1. Formation of a Volunteer Harmful Algal Bloom Network in British Columbia, Canada, Following an Outbreak of Diarrhetic Shellfish Poisoning

    PubMed Central

    McIntyre, Lorraine; Cassis, David; Haigh, Nicola

    2013-01-01

    Evidence for shellfish toxin illness in British Columbia (BC) on the west coast of Canada can be traced back to 1793. For over two hundred years, domestically acquired bivalve shellfish toxin illnesses in BC were solely ascribed to paralytic shellfish poisonings caused by algal blooms of Alexandrium. This changed in 2011, when BC experienced its first outbreak of diarrhetic shellfish poisoning (DSP). As a result of this outbreak, Canada’s first DSP symposium was held in November, 2012, in North Vancouver, BC. Three of the objectives of the symposium were to provide a forum to educate key stakeholders on this emerging issue, to identify research and surveillance priorities and to create a DSP network. The purpose of this paper is to review what is known about shellfish poisoning in BC and to describe a novel volunteer network that arose following the symposium. The newly formed network was designed for industry shellfish growers to identify harmful algae bloom events, so that they may take actions to mitigate the effects of harmful blooms on shellfish morbidity. The network will also inform public health and regulatory stakeholders of potentially emerging issues in shellfish growing areas. PMID:24172211

  2. Patterns of post-glacial genetic differentiation in marginal populations of a marine microalga.

    PubMed

    Tahvanainen, Pia; Alpermann, Tilman J; Figueroa, Rosa Isabel; John, Uwe; Hakanen, Päivi; Nagai, Satoshi; Blomster, Jaanika; Kremp, Anke

    2012-01-01

    This study investigates the genetic structure of an eukaryotic microorganism, the toxic dinoflagellate Alexandrium ostenfeldii, from the Baltic Sea, a geologically young and ecologically marginal brackish water estuary which is predicted to support evolution of distinct, genetically impoverished lineages of marine macroorganisms. Analyses of the internal transcribed spacer (ITS) sequences and Amplified Fragment Length Polymorphism (AFLP) of 84 A. ostenfeldii isolates from five different Baltic locations and multiple external sites revealed that Baltic A. ostenfeldii is phylogenetically differentiated from other lineages of the species and micro-geographically fragmented within the Baltic Sea. Significant genetic differentiation (F(ST)) between northern and southern locations was correlated to geographical distance. However, instead of discrete genetic units or continuous genetic differentiation, the analysis of population structure suggests a complex and partially hierarchic pattern of genetic differentiation. The observed pattern suggests that initial colonization was followed by local differentiation and varying degrees of dispersal, most likely depending on local habitat conditions and prevailing current systems separating the Baltic Sea populations. Local subpopulations generally exhibited low levels of overall gene diversity. Association analysis suggests predominately asexual reproduction most likely accompanied by frequency shifts of clonal lineages during planktonic growth. Our results indicate that the general pattern of genetic differentiation and reduced genetic diversity of Baltic populations found in large organisms also applies to microscopic eukaryotic organisms.

  3. Quantitative determination of paralytic shellfish toxins in cultured toxic algae by LC-MS/MS.

    PubMed

    Watanabe, Ryuichi; Matsushima, Ryoji; Harada, Tomoko; Oikawa, Hiroshi; Murata, Masakazu; Suzuki, Toshiyuki

    2013-01-01

    We developed a sample preparation and LC-MS/MS method for the determination of saxitoxins in toxic algae. Paralytic shellfish toxins (PSTs) were successfully separated by gradient elution on an amide column with the hydrophilic interaction mode and quantified with multiple reaction monitoring (MRM) detection in the positive ion mode. This method showed good performance in the summed LODs and LOQs for all 12 toxins, 25 and 84 nM, respectively. Next, extracts of cultured strains of a toxic dinoflagellate Alexandrium tamarense and a freshwater cyanobacteria Anabaena circinalis were treated in a short column of basic alumina and the toxic fractions were analysed by our LC-MS/MS method and by HPLC with fluorescence detection. Comparison of the results obtained by the two methods demonstrated that approximately equivalent results were obtained for both the dinoflagellate and the cyanobacteria. In addition, the retention time of the toxins showed acceptable shifts. Therefore, the clean-up of the toxic algal extracts by using the basic alumina column controlled unwanted chromatographic behaviour and variable ionisation efficiency during MS detection. LC-MS/MS for saxitoxins has great potential as a rapid analytical method for determining all primary saxitoxins in cultured algae.

  4. Trophic transfer of paralytic shellfish toxins from the cladoceran (Moina mongolica) to larvae of the fish (Sciaenops ocellatus).

    PubMed

    Jiang, Tian-Jiu; Wang, Da-Zhi; Niu, Tao; Xu, Yi-Xiao

    2007-10-01

    The dynamic transmission and transformation of paralytic shellfish toxins (PSTs) from the toxic dinoflagellate Alexandrium tamarense to the cladoceran Moina mongolica and subsequently to the larvae of the fish Sciaenops ocellatus were investigated under laboratory conditions. The results showed that PSTs could be transferred to S. ocellatus when they preyed on PST-containing M. mongolica. During the experimental period, A. tamarense, M. mongolica and the digestive glands of the fish larvae contained C(1/2) toxins, and the viscera of S. ocellatus contained neoSTX. The proportion of beta toxin (C2) in C(1+2) toxins increased when PSTs were transferred from A. tamarense to M. mongolica, but in the subsequent transfer from M. mongolica to S. ocellatus the proportion of alpha toxin (C1) increased. During depuration, the contents of C1 and C2 toxins in fish larvae decreased with the duration of depuration, but neoSTX remained relatively constant. The present results indicated that, using a cladoceran as the vector, PSTs can be transferred from toxic algae to a high trophic level fish and metabolized in the fish. Future work should address the metabolic characteristics of PSTs in cladocerans and the end result when they are transferred to fishes.

  5. Development and evaluation of a DNA microarray assay for the simultaneous detection of nine harmful algal species in ship ballast and seaport waters

    NASA Astrophysics Data System (ADS)

    Chen, Xianfeng; Zhou, Qianjin; Duan, Weijun; Zhou, Chengxu; Duan, Lijun; Zhang, Huili; Sun, Aili; Yan, Xiaojun; Chen, Jiong

    2016-01-01

    Rapid, high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae, which are responsible for algal blooms, such as red and green tides. In this study, we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously, namely Alexandrium tamarense, Gyrodinium instriatum, Heterosigma akashiwo, Karenia mikimotoi, Prorocentrum donghaiense, Prorocentrum minimum, Ulva compressa, Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection (LOD) of 0.5 ng of genomic DNA (orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether, 230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%, respectively, relative to conventional morphological methods. This indicated that this high-throughput, automatic, and specific method is well suited for the detection of algae in water samples.

  6. Biosynthetic route towards saxitoxin and shunt pathway

    PubMed Central

    Tsuchiya, Shigeki; Cho, Yuko; Konoki, Keiichi; Nagasawa, Kazuo; Oshima, Yasukatsu; Yotsu-Yamashita, Mari

    2016-01-01

    Saxitoxin, the most potent voltage-gated sodium channel blocker, is one of the paralytic shellfish toxins (PSTs) produced by cyanobacteria and dinoflagellates. Recently, putative biosynthetic genes of PSTs were reported in these microorganisms. We previously synthesized genetically predicted biosynthetic intermediates, Int-A’ and Int-C’2, and also Cyclic-C’ which was not predicted based on gene, and identified them all in the toxin-producing cyanobacterium Anabaena circinalis (TA04) and the dinoflagellate Alexandrium tamarense (Axat-2). This study examined the incorporation of 15N-labeled intermediates into PSTs (C1 and C2) in A. circinalis (TA04). Conversions from Int-A’ to Int-C’2, from Int-C’2 to Cyclic-C’, and from Int-A’ and Int-C’2 to C1 and C2 were indicated using high resolution-LC/MS. However, Cyclic-C’ was not converted to C1 and C2 and was detected primarily in the extracellular medium. These results suggest that Int-A’ and Int-C’2 are genuine precursors of PSTs, but Int-C’2 converts partially to Cyclic-C’ which is a shunt product excreted to outside the cells. This paper provides the first direct demonstration of the biosynthetic route towards saxitoxin and a shunt pathway. PMID:26842222

  7. Evidence for parasite-mediated selection during short-lasting toxic algal blooms

    PubMed Central

    Valero, Myriam; Alves-de-Souza, Catharina; Dia, Aliou; Lepelletier, Frédéric; Bigeard, Estelle; Jeanthon, Christian; Destombe, Christophe; Guillou, Laure

    2016-01-01

    Parasites play a role in the control of transient algal blooms, but it is not known whether parasite-mediated selection results in coevolution of the host and the parasites over this short time span. We investigated the presence of coevolution between the toxic dinoflagellate Alexandrium minutum and two naturally occurring endoparasites during blooms lasting a month in two river estuaries, using cross-inoculation experiments across time and space. Higher parasite abundance was associated with a large daily reduction in relative A. minutum abundances, demonstrating strong parasite-mediated selection. There was genetic variability in infectivity in both parasite species, and in resistance in the host. We found no evidence for coevolution in one estuary; however, in the other estuary, we found high genetic diversity in the two parasite species, fluctuations in infectivity and suggestion that the two parasites are well adapted to their host, as in ‘Red Queen’ dynamics. Thus, coevolution is possible over the short time span of a bloom, but geographically variable, and may feedback on community dynamics. PMID:27798309

  8. Combined heat shock protein 90 and ribosomal RNA sequence phylogeny supports multiple replacements of dinoflagellate plastids.

    PubMed

    Shalchian-Tabrizi, Kamran; Minge, Marianne A; Cavalier-Smith, Tom; Nedreklepp, Joachim M; Klaveness, Dag; Jakobsen, Kjetill S

    2006-01-01

    Dinoflagellates harbour diverse plastids obtained from several algal groups, including haptophytes, diatoms, cryptophytes, and prasinophytes. Their major plastid type with the accessory pigment peridinin is found in the vast majority of photosynthetic species. Some species of dinoflagellates have other aberrantly pigmented plastids. We sequenced the nuclear small subunit (SSU) ribosomal RNA (rRNA) gene of the "green" dinoflagellate Gymnodinium chlorophorum and show that it is sister to Lepidodinium viride, indicating that their common ancestor obtained the prasinophyte (or other green alga) plastid in one event. As the placement of dinoflagellate species that acquired green algal or haptophyte plastids is unclear from small and large subunit (LSU) rRNA trees, we tested the usefulness of the heat shock protein (Hsp) 90 gene for dinoflagellate phylogeny by sequencing it from four species with aberrant plastids (G. chlorophorum, Karlodinium micrum, Karenia brevis, and Karenia mikimotoi) plus Alexandrium tamarense, and constructing phylogenetic trees for Hsp90 and rRNAs, separately and together. Analyses of the Hsp90 and concatenated data suggest an ancestral origin of the peridinin-containing plastid, and two independent replacements of the peridinin plastid soon after the early radiation of the dinoflagellates. Thus, the Hsp90 gene seems to be a promising phylogenetic marker for dinoflagellate phylogeny.

  9. Potential Threats Posed by New or Emerging Marine Biotoxins in UK Waters and Examination of Detection Methodologies Used for Their Control: Cyclic Imines.

    PubMed

    Davidson, Keith; Baker, Clothilde; Higgins, Cowan; Higman, Wendy; Swan, Sarah; Veszelovszki, Andrea; Turner, Andrew D

    2015-11-26

    Cyclic imines (CIs) are a group of phytoplankton produced toxins related to shellfish food products, some of which are already present in UK and European waters. Their risk to shellfish consumers is poorly understood, as while no human intoxication has been definitively related to this group, their fast acting toxicity following intraperitoneal injection in mice has led to concern over their human health implications. A request was therefore made by UK food safety authorities to examine these toxins more closely to aid possible management strategies. Of the CI producers only the spirolide producer Alexandrium ostenfeldii is known to exist in UK waters at present but trends in climate change may lead to increased risk from other organisms/CI toxins currently present elsewhere in Europe and in similar environments worldwide. This paper reviews evidence concerning the prevalence of CIs and CI-producing phytoplankton, together with testing methodologies. Chemical, biological and biomolecular methods are reviewed, including recommendations for further work to enable effective testing. Although the focus here is on the UK, from a strategic standpoint many of the topics discussed will also be of interest in other parts of the world since new and emerging marine biotoxins are of global concern.

  10. Horizontal Gene Transfer is a Significant Driver of Gene Innovation in Dinoflagellates

    PubMed Central

    Wisecaver, Jennifer H.; Brosnahan, Michael L.; Hackett, Jeremiah D.

    2013-01-01

    The dinoflagellates are an evolutionarily and ecologically important group of microbial eukaryotes. Previous work suggests that horizontal gene transfer (HGT) is an important source of gene innovation in these organisms. However, dinoflagellate genomes are notoriously large and complex, making genomic investigation of this phenomenon impractical with currently available sequencing technology. Fortunately, de novo transcriptome sequencing and assembly provides an alternative approach for investigating HGT. We sequenced the transcriptome of the dinoflagellate Alexandrium tamarense Group IV to investigate how HGT has contributed to gene innovation in this group. Our comprehensive A. tamarense Group IV gene set was compared with those of 16 other eukaryotic genomes. Ancestral gene content reconstruction of ortholog groups shows that A. tamarense Group IV has the largest number of gene families gained (314–1,563 depending on inference method) relative to all other organisms in the analysis (0–782). Phylogenomic analysis indicates that genes horizontally acquired from bacteria are a significant proportion of this gene influx, as are genes transferred from other eukaryotes either through HGT or endosymbiosis. The dinoflagellates also display curious cases of gene loss associated with mitochondrial metabolism including the entire Complex I of oxidative phosphorylation. Some of these missing genes have been functionally replaced by bacterial and eukaryotic xenologs. The transcriptome of A. tamarense Group IV lends strong support to a growing body of evidence that dinoflagellate genomes are extraordinarily impacted by HGT. PMID:24259313

  11. Temperature dependence of an estuarine harmful algal bloom: Resolving interannual variability in bloom dynamics using a degree day approach.

    PubMed

    Ralston, David K; Keafer, Bruce A; Brosnahan, Michael L; Anderson, Donald M

    2014-01-01

    Observations of harmful algal blooms (HABs) of the dinoflagellate Alexandrium fundyense in an estuary over multiple years were used to assess drivers of their spatial and temporal variability. Nauset Estuary on Cape Cod, Massachusetts has a recurrent, self-seeding A. fundyense population that produces paralytic shellfish poisoning toxins and leads to nearly annual closure to shellfishing. Weekly surveys of the entire estuary were made in 3 of 4 consecutive years, with surveys of a subembayment during the intervening year. Major A. fundyense blooms were observed all 4 years, with maximum concentrations >10(6) cells L(-1). Concentrations were greatest in three salt ponds at the distal edges of the estuary. The bloom timing varied among the salt ponds and among years, although the blooms had similar durations and maximum cell concentrations. Nutrient concentrations did not correlate with the growth of the bloom, but differences in water temperature among years and ponds were significant. Net growth rates inferred from the surveys were similar to those from laboratory experiments, and increased linearly with temperature. A growing degree day calculation was used to account for effects of interannual variability and spatial gradients in water temperature on population development. The approach collapsed variability in the timing of bloom onset, development, and termination across years and among ponds, suggesting that this relatively simple metric could be used as an early-warning indicator for HABs in Nauset and similar areas with localized, self-seeding blooms.

  12. Diverse seed banks favour adaptation of microalgal populations to future climate conditions.

    PubMed

    Kremp, Anke; Oja, Johanna; LeTortorec, Anniina H; Hakanen, Päivi; Tahvanainen, Pia; Tuimala, Jarno; Suikkanen, Sanna

    2016-02-01

    Selection of suitable genotypes from diverse seed banks may help phytoplankton populations to cope with environmental changes. This study examines whether the high genotypic diversity found in the Baltic cyst pool of the toxic dinoflagellate Alexandrium ostenfeldii is coupled to phenotypic variability that could aid short-term adaptation. Growth rates, cellular toxicities and bioluminescence of 34 genetically different clones isolated from cyst beds of four Baltic bloom sites were determined in batch culture experiments along temperature and salinity gradients covering present and future conditions in the Baltic Sea. For all parameters a significant effect of genotype on the response to temperature and salinity changes was identified. General or site-specific effects of the two factors remained minor. Clones thriving at future conditions were different from the best performing at present conditions, suggesting that genotypic shifts may be expected in the future. Increased proportions of highly potent saxitoxin were observed as a plastic response to temperature increase, indicating a potential for higher toxicity of future blooms. The observed standing variation in Baltic seed banks of A. ostenfeldii suggests that the population is likely to persist under environmental change.

  13. Temperature dependence of an estuarine harmful algal bloom: Resolving interannual variability in bloom dynamics using a degree day approach

    PubMed Central

    Ralston, David K.; Keafer, Bruce A.; Brosnahan, Michael L.; Anderson, Donald M.

    2014-01-01

    Observations of harmful algal blooms (HABs) of the dinoflagellate Alexandrium fundyense in an estuary over multiple years were used to assess drivers of their spatial and temporal variability. Nauset Estuary on Cape Cod, Massachusetts has a recurrent, self-seeding A. fundyense population that produces paralytic shellfish poisoning toxins and leads to nearly annual closure to shellfishing. Weekly surveys of the entire estuary were made in 3 of 4 consecutive years, with surveys of a subembayment during the intervening year. Major A. fundyense blooms were observed all 4 years, with maximum concentrations >106 cells L−1. Concentrations were greatest in three salt ponds at the distal edges of the estuary. The bloom timing varied among the salt ponds and among years, although the blooms had similar durations and maximum cell concentrations. Nutrient concentrations did not correlate with the growth of the bloom, but differences in water temperature among years and ponds were significant. Net growth rates inferred from the surveys were similar to those from laboratory experiments, and increased linearly with temperature. A growing degree day calculation was used to account for effects of interannual variability and spatial gradients in water temperature on population development. The approach collapsed variability in the timing of bloom onset, development, and termination across years and among ponds, suggesting that this relatively simple metric could be used as an early-warning indicator for HABs in Nauset and similar areas with localized, self-seeding blooms. PMID:25419003

  14. Potential Threats Posed by New or Emerging Marine Biotoxins in UK Waters and Examination of Detection Methodologies Used for Their Control: Cyclic Imines

    PubMed Central

    Davidson, Keith; Baker, Clothilde; Higgins, Cowan; Higman, Wendy; Swan, Sarah; Veszelovszki, Andrea; Turner, Andrew D.

    2015-01-01

    Cyclic imines (CIs) are a group of phytoplankton produced toxins related to shellfish food products, some of which are already present in UK and European waters. Their risk to shellfish consumers is poorly understood, as while no human intoxication has been definitively related to this group, their fast acting toxicity following intraperitoneal injection in mice has led to concern over their human health implications. A request was therefore made by UK food safety authorities to examine these toxins more closely to aid possible management strategies. Of the CI producers only the spirolide producer Alexandrium ostenfeldii is known to exist in UK waters at present but trends in climate change may lead to increased risk from other organisms/CI toxins currently present elsewhere in Europe and in similar environments worldwide. This paper reviews evidence concerning the prevalence of CIs and CI-producing phytoplankton, together with testing methodologies. Chemical, biological and biomolecular methods are reviewed, including recommendations for further work to enable effective testing. Although the focus here is on the UK, from a strategic standpoint many of the topics discussed will also be of interest in other parts of the world since new and emerging marine biotoxins are of global concern. PMID:26703628

  15. Biomass and lipid production of dinoflagellates and raphidophytes in indoor and outdoor photobioreactors.

    PubMed

    Fuentes-Grünewald, C; Garcés, E; Alacid, E; Rossi, S; Camp, J

    2013-02-01

    The principal fatty acids from the lipid profiles of two autochthonous dinoflagellates (Alexandrium minutum and Karlodinium veneficum) and one raphidophyte (Heterosigma akashiwo) maintained in bubble column photobioreactors under outdoor culture conditions are described for the first time. The biomass production, lipid content and lipid productivity of these three species were determined and the results compared to those obtained when the strains were cultured indoors. Under the latter condition, the biotic values did not significantly differ among species, whereas under outdoor conditions, differences in both duplication time and fatty acids content were observed. Specifically, A. minutum had higher biomass productivity (0.35 g·L⁻¹ day⁻¹), lipid productivity (80.7 mg lipid·L⁻¹ day⁻¹) and lipid concentration (252 mg lipid·L⁻¹) at harvest time (stationary phase) in outdoor conditions. In all three strains, the growth rate and physiological response to the light and temperature fluctuations of outdoor conditions greatly impacted the production parameters. Nonetheless, the species could be successfully grown in an outdoor photobioreactor and were of sufficient robustness to enable the establishment of long-term cultures yielding consistent biomass and lipid production.

  16. Predator lipids induce paralytic shellfish toxins in bloom-forming algae

    PubMed Central

    Selander, Erik; Kubanek, Julia; Hamberg, Mats; Andersson, Mats X.; Cervin, Gunnar; Pavia, Henrik

    2015-01-01

    Interactions among microscopic planktonic organisms underpin the functioning of open ocean ecosystems. With few exceptions, these organisms lack advanced eyes and thus rely largely on chemical sensing to perceive their surroundings. However, few of the signaling molecules involved in interactions among marine plankton have been identified. We report a group of eight small molecules released by copepods, the most abundant zooplankton in the sea, which play a central role in food webs and biogeochemical cycles. The compounds, named copepodamides, are polar lipids connecting taurine via an amide to isoprenoid fatty acid conjugate of varying composition. The bloom-forming dinoflagellate Alexandrium minutum responds to pico- to nanomolar concentrations of copepodamides with up to a 20-fold increase in production of paralytic shellfish toxins. Different copepod species exude distinct copepodamide blends that contribute to the species-specific defensive responses observed in phytoplankton. The signaling system described here has far reaching implications for marine ecosystems by redirecting grazing pressure and facilitating the formation of large scale harmful algal blooms. PMID:25918403

  17. Occurrence of Harmful Algal Species and Shellfish Toxicity in Sardinia (Italy)

    PubMed Central

    Bazzoni, Anna Maria; Mudadu, Alessandro Graziano; Lorenzoni, Giuseppa; Arras, Igor; Lugliè, Antonella; Vivaldi, Barbara; Cicotelli, Valentina; Sanna, Giovanna; Tedde, Giuseppe; Ledda, Salvatore; Alesso, Enrico; Marongiu, Edoardo; Virgilio, Sebastiano

    2016-01-01

    Sardinia (Italy, north-western Mediterranean) is a commercially important producer of edible bivalve molluscs. Since the early 2000s, it was subjected to recurring cases of mussel farm closures due to toxic algal poison. Here, we present the studies on toxin concentrations and the associated potentially toxic phytoplankton distribution and abundances carried out by a regular monitoring programme in Sardinian shellfish areas, from January to May 2015. Diarrheic shellfish poisoning (DSP) toxins were detected in several bivalve molluscs samples, while paralytic shellfish poisoning (PSP) and paralytic shellfish poisoning toxins were present just once, without exceeding the legal limits. Potentially toxic algal species have been constantly present. Pseudo-nitzschia species were present during the entire study often with high abundances, while Dinophysis species reached high densities sporadically. Among PSP phytoplankton, only Alexandrium minutum Halim was found. The data obtained in this study showed an increase in the DSP toxicity in mussels in Sardinia. No clear relation between the occurrence of toxins in shellfish and the presence of potentially toxic algal species was found, although a slight correlation between DSP toxins and Dinophysis species could be supported. PMID:28058244

  18. Australian Dust Storm Associated with Extensive Aspergillus sydowii Fungal “Bloom” in Coastal Waters

    PubMed Central

    Coman, Frank; Davies, Claire; Hayashi, Aiko; McLeod, David; Slotwinski, Anita; Whittock, Lucy; Richardson, Anthony J.

    2014-01-01

    A massive central Australian dust storm in September 2009 was associated with abundant fungal spores (150,000/m3) and hyphae in coastal waters between Brisbane (27°S) and Sydney (34°S). These spores were successfully germinated from formalin-preserved samples, and using molecular sequencing of three different genes (the large subunit rRNA gene [LSU], internal transcribed spacer [ITS[, and beta-tubulin gene), they were conclusively identified as Aspergillus sydowii, an organism circumstantially associated with gorgonian coral fan disease in the Caribbean. Surprisingly, no human health or marine ecosystem impacts were associated with this Australian dust storm event. Australian fungal cultures were nontoxic to fish gills and caused a minor reduction in the motility of Alexandrium or Chattonella algal cultures but had their greatest impacts on Symbiodinium dinoflagellate coral symbiont motility, with hyphae being more detrimental than spores. While we have not yet seen any soft coral disease outbreaks on the Australian Great Barrier Reef similar to those observed in the Caribbean and while this particular fungal population was non- or weakly pathogenic, our observations raise the possibility of future marine ecosystem pathogen impacts from similar dust storms harboring more pathogenic strains. PMID:24657868

  19. Screening of surfactants for harmful algal blooms mitigation.

    PubMed

    Sun, Xiao-Xia; Han, Kyung-Nam; Choi, Joong-Ki; Kim, Eun-Ki

    2004-05-01

    Screening experiments were conducted in order to find promising synthetic surfactants for harmful algal blooms (HABs) mitigation. The chemically synthesized surfactant cocamidopropyl betaine (CAPB) showed characteristics of relatively high inhibition efficiency, high biodegradability and low cost. The motility inhibition ratios of 10 mg/L CAPB on Cochlodinium polykrikoides and Alexandrium tamarense were about 60% after 5 min. The biodegradation test indicated that the half-life of CAPB in seawater was shorter than one day and 90% was biodegraded after five days under the initial concentration of 100 mg/L at 25 degrees C. Further cell lysis experiments revealed the selective lysis effect of CAPB on different HAB organisms. More than 90% of C. polykrikoides lysed at the concentration of 10 mg/L CAPB after 24 h and at 15 mg/L CAPB after 4 h, whereas the lysis effect of CAPB on A. tamarense was slight, no more than 10% after 2 h interaction with 50 mg/L CAPB. This research provided preliminary data for CAPB as a candidate in harmful algal blooms mitigation and pointed out unresolved problems for its practical application in the meantime.

  20. Combined Effects of Ocean Acidification and Light or Nitrogen Availabilities on 13C Fractionation in Marine Dinoflagellates

    PubMed Central

    Hoins, Mirja; Eberlein, Tim; Groβmann, Christian H.; Brandenburg, Karen; Reichart, Gert-Jan; Rost, Björn; Sluijs, Appy; Van de Waal, Dedmer B.

    2016-01-01

    Along with increasing oceanic CO2 concentrations, enhanced stratification constrains phytoplankton to shallower upper mixed layers with altered light regimes and nutrient concentrations. Here, we investigate the effects of elevated pCO2 in combination with light or nitrogen-limitation on 13C fractionation (εp) in four dinoflagellate species. We cultured Gonyaulax spinifera and Protoceratium reticulatum in dilute batches under low-light (‘LL’) and high-light (‘HL’) conditions, and grew Alexandrium fundyense and Scrippsiella trochoidea in nitrogen-limited continuous cultures (‘LN’) and nitrogen-replete batches (‘HN’). The observed CO2-dependency of εp remained unaffected by the availability of light for both G. spinifera and P. reticulatum, though at HL εp was consistently lower by about 2.7‰ over the tested CO2 range for P. reticulatum. This may reflect increased uptake of (13C-enriched) bicarbonate fueled by increased ATP production under HL conditions. The observed CO2-dependency of εp disappeared under LN conditions in both A. fundyense and S. trochoidea. The generally higher εp under LN may be associated with lower organic carbon production rates and/or higher ATP:NADPH ratios. CO2-dependent εp under non-limiting conditions has been observed in several dinoflagellate species, showing potential for a new CO2-proxy. Our results however demonstrate that light- and nitrogen-limitation also affect εp, thereby illustrating the need to carefully consider prevailing environmental conditions. PMID:27153107

  1. Predator lipids induce paralytic shellfish toxins in bloom-forming algae.

    PubMed

    Selander, Erik; Kubanek, Julia; Hamberg, Mats; Andersson, Mats X; Cervin, Gunnar; Pavia, Henrik

    2015-05-19

    Interactions among microscopic planktonic organisms underpin the functioning of open ocean ecosystems. With few exceptions, these organisms lack advanced eyes and thus rely largely on chemical sensing to perceive their surroundings. However, few of the signaling molecules involved in interactions among marine plankton have been identified. We report a group of eight small molecules released by copepods, the most abundant zooplankton in the sea, which play a central role in food webs and biogeochemical cycles. The compounds, named copepodamides, are polar lipids connecting taurine via an amide to isoprenoid fatty acid conjugate of varying composition. The bloom-forming dinoflagellate Alexandrium minutum responds to pico- to nanomolar concentrations of copepodamides with up to a 20-fold increase in production of paralytic shellfish toxins. Different copepod species exude distinct copepodamide blends that contribute to the species-specific defensive responses observed in phytoplankton. The signaling system described here has far reaching implications for marine ecosystems by redirecting grazing pressure and facilitating the formation of large scale harmful algal blooms.

  2. Thermal effects on the growth and fatty acid composition of four harmful algal bloom species: Possible implications for ichthyotoxicity

    NASA Astrophysics Data System (ADS)

    Hyun, Bonggil; Ju, Se-Jong; Ko, Ah-Ra; Choi, Keun-Hyung; Jung, Seung Won; Jang, Pung-Guk; Jang, Min-Chul; Moon, Chang Ho; Shin, Kyoungsoon

    2016-09-01

    Little is known regarding how harmful algal bloom species respond to different temperatures in terms of fatty acid production. This study examined the effects of temperature on the growth rates, cell volumes, and fatty acid concentrations and compositions of four harmful algal bloom species (HABs), Akashiwo sanguinea, Alexandrium tamarense, Chattonella ovata, and Prorocentrum minimum. The HABs species were cultured at 15, 20, 25, and 30°C in a nutrient-enriched medium. Three of the species maintained optimal growth rates over a wide range of temperatures, but A. tamarense did not. The cell volumes of each species showed little change over the temperature range. The total fatty acid concentrations in A. sanguinea, A. tamarense and C. ovata decreased as the temperature increased, but P. minimum showed no trend in this respect. Polyunsaturated fatty acids (PUFAs), the key biochemical components that maintain cell membrane fluidity and which are associated with toxicity, decreased in both concentration and proportion of total fatty acids as temperature increased, except in A. sanguinea, in which the proportion of PUFAs to the total fatty acids increased. These reductions in PUFA concentration and proportion could reduce cell membrane fluidity and toxicity in HABs; however, enhanced growth and/or ruptured cells, which are considered more toxic than intact cells, could compensate for the reduced per-cell toxicity. This phenomenon might impact on the marine ecosystem and aquaculture industry.

  3. Analysis of the hydrographic conditions and cyst beds in the San Jorge Gulf, Argentina, that favor dinoflagellate population development including toxigenic species and their toxins

    NASA Astrophysics Data System (ADS)

    Krock, Bernd; Borel, C. Marcela; Barrera, Facundo; Tillmann, Urban; Fabro, Elena; Almandoz, Gastón O.; Ferrario, Martha; Garzón Cardona, John E.; Koch, Boris P.; Alonso, Cecilia; Lara, Rubén

    2015-08-01

    The overlay of cooler nutrient enriched Beagle-Magellan water with warmer nutrient depleted shelf water and a strong stratification of the water column in the San Jorge Gulf region, Argentina, coincided with relatively high dinoflagellate abundances in April 2012, up to 34,000 cells L- 1. This dinoflagellate proliferation was dominated by Ceratium spp., but environmental conditions also favored to a lesser amount the occurrence of toxigenic dinoflagellates, such as Alexandrium tamarense and Protoceratium reticulatum, whose toxins were hardly detected in any other areas along the expedition transect of the R/V Puerto Deseado between 38 and 56°S (Ushuaia-Mar del Plata) in March/April 2012. Generally vegetative cells of A. tamarense and P. reticulatum co-occurred with their respective phycotoxins in the water column and their cysts in the upper sediment layers. Two strains of A. tamarense were isolated from the bloom sample and morphologically characterized. Their PSP toxin profiles consisted of C1/2, gonyautoxins 1/4 and to a lesser amount of neosaxitoxin and confirmed earlier data from this region. The ratios between autotrophic picoplankton and heterotrophic bacteria were higher in shelf waters in the north than in Beagle-Magellan waters in the south of San Jorge Gulf.

  4. Categorizing the severity of paralytic shellfish poisoning outbreaks in the Gulf of Maine for forecasting and management

    NASA Astrophysics Data System (ADS)

    Kleindinst, Judith L.; Anderson, Donald M.; McGillicuddy, Dennis J.; Stumpf, Richard P.; Fisher, Kathleen M.; Couture, Darcie A.; Michael Hickey, J.; Nash, Christopher

    2014-05-01

    Development of forecasting systems for harmful algal blooms (HABs) has been a long-standing research and management goal. Significant progress has been made in the Gulf of Maine, where seasonal bloom forecasts are now being issued annually using Alexandrium fundyense cyst abundance maps and a population dynamics model developed for that organism. Thus far, these forecasts have used terms such as “significant”, “moderately large” or “moderate” to convey the extent of forecasted paralytic shellfish poisoning (PSP) outbreaks. In this study, historical shellfish harvesting closure data along the coast of the Gulf of Maine were used to derive a series of bloom severity levels that are analogous to those used to define major storms like hurricanes or tornados. Thirty-four years of PSP-related shellfish closure data for Maine, Massachusetts and New Hampshire were collected and mapped to depict the extent of coastline closure in each year. Due to fractal considerations, different methods were explored for measuring length of coastline closed. Ultimately, a simple procedure was developed using arbitrary straight-line segments to represent specific sections of the coastline. This method was consistently applied to each year’s PSP toxicity closure map to calculate the total length of coastline closed. Maps were then clustered together statistically to yield distinct groups of years with similar characteristics. A series of categories or levels was defined (“Level 1: Limited”, “Level 2: Moderate”, and “Level 3: Extensive”) each with an associated range of expected coastline closed, which can now be used instead of vague descriptors in future forecasts. This will provide scientifically consistent and simply defined information to the public as well as resource managers who make decisions on the basis of the forecasts.

  5. Categorizing the severity of paralytic shellfish poisoning outbreaks in the Gulf of Maine for forecasting and management.

    PubMed

    Kleindinst, Judith L; Anderson, Donald M; McGillicuddy, Dennis J; Stumpf, Richard P; Fisher, Kathleen M; Couture, Darcie A; Hickey, J Michael; Nash, Christopher

    2014-05-01

    Development of forecasting systems for harmful algal blooms (HABs) has been a long-standing research and management goal. Significant progress has been made in the Gulf of Maine, where seasonal bloom forecasts are now being issued annually using Alexandrium fundyense cyst abundance maps and a population dynamics model developed for that organism. Thus far, these forecasts have used terms such as "significant", "moderately large" or "moderate" to convey the extent of forecasted paralytic shellfish poisoning (PSP) outbreaks. In this study, historical shellfish harvesting closure data along the coast of the Gulf of Maine were used to derive a series of bloom severity levels that are analogous to those used to define major storms like hurricanes or tornados. Thirty-four years of PSP-related shellfish closure data for Maine, Massachusetts and New Hampshire were collected and mapped to depict the extent of coastline closure in each year. Due to fractal considerations, different methods were explored for measuring length of coastline closed. Ultimately, a simple procedure was developed using arbitrary straight-line segments to represent specific sections of the coastline. This method was consistently applied to each year's PSP toxicity closure map to calculate the total length of coastline closed. Maps were then clustered together statistically to yield distinct groups of years with similar characteristics. A series of categories or levels was defined ("Level 1: Limited", "Level 2: Moderate", and "Level 3: Extensive") each with an associated range of expected coastline closed, which can now be used instead of vague descriptors in future forecasts. This will provide scientifically consistent and simply defined information to the public as well as resource managers who make decisions on the basis of the forecasts.

  6. A feasibility study into the production of a freeze-dried oyster reference material for paralytic shellfish poisoning toxins.

    PubMed

    Turner, Andrew D; Lewis, Adam M; Hatfield, Robert G; Higman, Wendy A; Burrell, Stephen

    2013-10-01

    Matrix reference materials are an essential component for the validation and quality control of analytical methodologies for the quantitation of marine biotoxins in shellfish. Given the potential advantages of reference materials in powder form, a study was conducted to assess the feasibility for the production of a freeze-dried oyster tissue reference material containing a range of important paralytic shellfish poisoning toxins. One bulk sample of a wet oyster tissue homogenate was generated following mass culturing of toxic Alexandrium and oyster feeding experiments. The bulk tissue was used to prepare untreated wet frozen aliquots with the remainder being freeze-dried and processed into appropriately-sized powder samples. A pre-column oxidation LC-FLD analysis was used to confirm the absence of any chromatographic artefacts resulting from the processing and to confirm acceptable homogeneity of the tissues. Excellent stability over both the short-term (1 month) and long-term (1 year) of the freeze-dried material was demonstrated as compared with the stability of the untreated wet tissue. A post-column oxidation LC-FLD method was used to confirm the absence of toxin epimerisation in freeze-dried tissues which were observed in the wet tissues. Overall the work showed the feasibility of an approach to produce a homogenous freeze-dried oyster matrix material with enhanced stability in comparison to the untreated wet tissue. The potential for use of the process for preparation of large scale production batches of a freeze-dried CRM for paralytic shellfish poisoning toxins has therefore been demonstrated.

  7. Saxitoxin Modulates Immunological Parameters and Gene Transcription in Mytilus chilensis Hemocytes

    PubMed Central

    Astuya, Allisson; Carrera, Crisleri; Ulloa, Viviana; Aballay, Ambbar; Núñez-Acuña, Gustavo; Hégaret, Hélène; Gallardo-Escárate, Cristian

    2015-01-01

    Saxitoxin (STX) is a neurotoxin produced by dinoflagellates in diverse species, such as Alexandrium spp., and it causes paralytic shellfish poisoning (PSP) in humans after the ingestion of contaminated shellfish. Recent studies have suggested that the immune functions of bivalves could be affected by harmful algae and/or by their toxins. Herein, hemocytes are the main effector cells of the immune cellular response. In this study, we evaluated the response of hemocytes from the mussel Mytilus chilensis to STX exposure in a primary culture. Cell cultures were characterized according to size and complexity, while reactive oxygen species (ROS) production was evaluated using a dichlorofluorescein diacetate (DCFH-DA) assay. Finally, phagocytic activity was measured using both flow cytometry and fluorescence microscopy assays. Additionally, gene transcription of candidate genes was evaluated by qPCR assays. The results evidenced that exposures to different concentrations of STX (1–100 nM) for 24 h did not affect cell viability, as determined by an MTT assay. However, when hemocytes were exposed for 4 or 16 h to STX (1–100 nM), there was a modulation of phagocytic activity and ROS production. Moreover, hemocytes exposed to 100 nM of STX for 4 or 16 h showed a significant increase in transcript levels of genes encoding for antioxidant enzymes (SOD, CAT), mitochondrial enzymes (COI, COIII, CYTB, ATP6, ND1) and ion channels (K+, Ca2+). Meanwhile, C-type lectin and toll-like receptor genes revealed a bi-phase transcriptional response after 16 and 24–48 h of exposure to STX. These results suggest that STX can negatively affect the immunocompetence of M. chilensis hemocytes, which were capable of responding to STX exposure in vitro by increasing the mRNA levels of antioxidant enzymes. PMID:26154765

  8. Determining the Advantages, Costs, and Trade-Offs of a Novel Sodium Channel Mutation in the Copepod Acartia hudsonica to Paralytic Shellfish Toxins (PST)

    PubMed Central

    Finiguerra, Michael; Avery, David E.; Dam, Hans G.

    2015-01-01

    The marine copepod Acartia hudsonica was shown to be adapted to dinoflagellate prey, Alexandrium fundyense, which produce paralytic shellfish toxins (PST). Adaptation to PSTs in other organisms is caused by a mutation in the sodium channel. Recently, a mutation in the sodium channel in A. hudsonica was found. In this study, we rigorously tested for advantages, costs, and trade-offs associated with the mutant isoform of A. hudsonica under toxic and non-toxic conditions. We combined fitness with wild-type: mutant isoform ratio measurements on the same individual copepod to test our hypotheses. All A. hudsonica copepods express both the wild-type and mutant sodium channel isoforms, but in different proportions; some individuals express predominantly mutant (PMI) or wild-type isoforms (PWI), while most individuals express relatively equal amounts of each (EI). There was no consistent pattern of improved performance as a function of toxin dose for egg production rate (EPR), ingestion rate (I), and gross growth efficiency (GGE) for individuals in the PMI group relative to individuals in the PWI expression group. Neither was there any evidence to indicate a fitness benefit to the mutant isoform at intermediate toxin doses. No clear advantage under toxic conditions was associated with the mutation. Using a mixed-diet approach, there was also no observed relationship between individual wild-type: mutant isoform ratios and among expression groups, on both toxic and non-toxic diets, for eggs produced over three days. Lastly, expression of the mutant isoform did not mitigate the negative effects of the toxin. That is, the reductions in EPR from a toxic to non-toxic diet for copepods were independent of expression groups. Overall, the results did not support our hypotheses; the mutant sodium channel isoform does not appear to be related to adaptation to PST in A. hudsonica. Other potential mechanisms responsible for the adaptation are discussed. PMID:26075900

  9. Categorizing the severity of paralytic shellfish poisoning outbreaks in the Gulf of Maine for forecasting and management

    PubMed Central

    Kleindinst, Judith L.; Anderson, Donald M.; McGillicuddy, Dennis J.; Stumpf, Richard P.; Fisher, Kathleen M.; Couture, Darcie A.; Hickey, J. Michael; Nash, Christopher

    2014-01-01

    Development of forecasting systems for harmful algal blooms (HABs) has been a long-standing research and management goal. Significant progress has been made in the Gulf of Maine, where seasonal bloom forecasts are now being issued annually using Alexandrium fundyense cyst abundance maps and a population dynamics model developed for that organism. Thus far, these forecasts have used terms such as “significant”, “moderately large” or “moderate” to convey the extent of forecasted paralytic shellfish poisoning (PSP) outbreaks. In this study, historical shellfish harvesting closure data along the coast of the Gulf of Maine were used to derive a series of bloom severity levels that are analogous to those used to define major storms like hurricanes or tornados. Thirty-four years of PSP-related shellfish closure data for Maine, Massachusetts and New Hampshire were collected and mapped to depict the extent of coastline closure in each year. Due to fractal considerations, different methods were explored for measuring length of coastline closed. Ultimately, a simple procedure was developed using arbitrary straight-line segments to represent specific sections of the coastline. This method was consistently applied to each year’s PSP toxicity closure map to calculate the total length of coastline closed. Maps were then clustered together statistically to yield distinct groups of years with similar characteristics. A series of categories or levels was defined (“Level 1: Limited”, “Level 2: Moderate”, and “Level 3: Extensive”) each with an associated range of expected coastline closed, which can now be used instead of vague descriptors in future forecasts. This will provide scientifically consistent and simply defined information to the public as well as resource managers who make decisions on the basis of the forecasts. PMID:25076815

  10. Quantification and profiling of lipophilic marine toxins in microalgae by UHPLC coupled to high-resolution orbitrap mass spectrometry.

    PubMed

    Orellana, Gabriel; Van Meulebroek, Lieven; Van Vooren, Sarah; De Rijcke, Maarten; Vandegehuchte, Michiel; Janssen, Colin R; Vanhaecke, Lynn

    2015-08-01

    During the last decade, a significant increase in the occurrence of harmful algal blooms (HABs), linked to repetitive cases of shellfish contamination has become a public health concern and therefore, accurate methods to detect marine toxins in different matrices are required. In this study, we developed a method for profiling lipophilic marine microalgal toxins based on ultra-high-performance liquid chromatography coupled to high-resolution Orbitrap mass spectrometry (UHPLC-HR-Orbitrap MS). Extraction of selected toxins (okadaic acid (OA), dinophysistoxin-1 (DTX-1), pectenotoxin-2 (PTX-2), azaspiracid-1 (AZA-1), yessotoxin (YTX) and 13-desmethyl spirolide C (SPX-1)) was optimized using a Plackett-Burman design. Three key algal species, i.e., Prorocentrum lima, Protoceratium reticulatum and Alexandrium ostenfeldii were used to test the extraction efficiency of OA, YTXs and SPXs, respectively. Prorocentrum micans, fortified with certified reference solutions, was used for recovery studies. The quantitative and confirmatory performance of the method was evaluated according to CD 2002/657/EC. Limits of detection and quantification ranged between 0.006 and 0.050 ng mL(-1) and 0.018 to 0.227 ng mL(-1), respectively. The intra-laboratory reproducibility ranged from 6.8 to 11.7 %, repeatability from 6.41 to 11.5 % and mean corrected recoveries from 81.9 to 119.6 %. In addition, algae cultures were retrospectively screened for analogues and metabolites through a homemade database. Using the ToxID software programme, 18 toxin derivates were detected in the extract of three toxin producing microalgae species. In conclusion, the generic extraction and full-scan HRMS approach offers an excellent quantitative performance and simultaneously allows to profile analogues and metabolites of marine toxins in microalgae. Graphical Abstract Optimization of extraction, detection and quantification of lipophilic marine toxins in microalgae by UHPLC-HR Orbitrap MS.

  11. The diversity of algal phospholipase D homologs revealed by biocomputational analysis.

    PubMed

    Beligni, María Verónica; Bagnato, Carolina; Prados, María Belén; Bondino, Hernán; Laxalt, Ana María; Munnik, Teun; Ten Have, Arjen

    2015-10-01

    Phospholipase D (PLD) participates in the formation of phosphatidic acid, a precursor in glycerolipid biosynthesis and a second messenger. PLDs are part of a superfamily of proteins that hydrolyze phosphodiesters and share a catalytic motif, HxKxxxxD, and hence a mechanism of action. Although HKD-PLDs have been thoroughly characterized in plants, animals and bacteria, very little is known about these enzymes in algae. To fill this gap in knowledge, we performed a biocomputational analysis by means of HMMER iterative profiling, using most eukaryotic algae genomes available. Phylogenetic analysis revealed that algae exhibit very few eukaryotic-type PLDs but possess, instead, many bacteria-like PLDs. Among algae eukaryotic-type PLDs, we identified C2-PLDs and PXPH-like PLDs. In addition, the dinoflagellate Alexandrium tamarense features several proteins phylogenetically related to oomycete PLDs. Our phylogenetic analysis also showed that algae bacteria-like PLDs (proteins with putative PLD activity) fall into five clades, three of which are novel lineages in eukaryotes, composed almost entirely of algae. Specifically, Clade II is almost exclusive to diatoms, whereas Clade I and IV are mainly represented by proteins from prasinophytes. The other two clades are composed of mitochondrial PLDs (Clade V or Mito-PLDs), previously found in mammals, and a subfamily of potentially secreted proteins (Clade III or SP-PLDs), which includes a homolog formerly characterized in rice. In addition, our phylogenetic analysis shows that algae have non-PLD members within the bacteria-like HKD superfamily with putative cardiolipin synthase and phosphatidylserine/phosphatidylglycerophosphate synthase activities. Altogether, our results show that eukaryotic algae possess a moderate number of PLDs that belong to very diverse phylogenetic groups.

  12. Phytoplankton community structure and environmental parameters in aquaculture areas of Daya Bay, South China Sea.

    PubMed

    Wang, Zhaohui; Zhao, Jiangang; Zhang, Yujuan; Cao, Yu

    2009-01-01

    Environmental characteristics and phytoplankton community structure were investigated in two aquaculture areas in Dapeng Cove of Daya Bay, South China Sea, between April 2005 and June 2006. Phytoplankton abundance ranged between 5.0 and 8877.5 cells/mL, with an average of 751.8 cells/mL. The seasonal cycle of phytoplankton were demonstrated by frequent oscillations, with recurrent high abundances from late spring to autumn and a peak stage in late winter. Diatoms were the predominant phytoplankton group, accounting for 93.21% of the total abundance. The next most abundant group was the dinoflagellates, which made up only 1.24% of total abundance. High concentrations of Alexandrium tamarense (Lebour) Balech with a maximum of 603.0 cells/mL were firstly recorded in this area known for high rates of paralytic shellfish poisoning (PSP) contamination. Temperatures and salinities were within the suitable values for the growth of phytoplankton, and were important in phytoplankton seasonal fluctuations. The operation of the Daya Bay Nuclear Power Station (DNPS) exerts influences on the phytoplankton community and resulted in the high abundances of toxic dinoflagellate species during the winter months. Dissolved inorganic nitrogen (DIN) and dissolved silicate (DSi) were sufficient, and rarely limited for the growth of phytoplankton. Dissolved inorganic phosphorus (DIP) was the most necessary element for phytoplankton growth. The enriched environments accelerated the growth of small diatoms, and made for the shift in predominant species from large diatom Rhizosolenia spp. to chain-forming diatoms such as Skeletonema costatum, Pseudo-nitzschia spp. and Thalassiosira subtilis.

  13. Quorum sensing of microalgae associated marine Ponticoccus sp. PD-2 and its algicidal function regulation.

    PubMed

    Chi, Wendan; Zheng, Li; He, Changfei; Han, Bin; Zheng, Minggang; Gao, Wei; Sun, Chengjun; Zhou, Gefei; Gao, Xiangxing

    2017-12-01

    Quorum sensing (QS) systems play important roles in regulating many physiological functions of microorganisms, such as biofilm formation, bioluminescence, and antibiotic production. One marine algicidal bacterium, Ponticoccus sp. PD-2, was isolated from the microalga Prorocentrum donghaiense, and its N-acyl-homoserine lactone (AHL)-mediated QS system was verified. In this study, we analyzed the AHLs profile of strain PD-2. Two AHLs, 3-oxo-C8-HSL and 3-oxo-C10-HSL, were detected using a biosensor overlay assay and GC-MS methods. Two complete AHL-QS systems (designated zlaI/R and zlbI/R) were identified in the genome of strain PD-2. When expressed in Escherichia coli, both zlaI and zlbI genes could each produce 3-oxo-C8-HSL and 3-oxo-C10-HSL. Algicidal activity was investigated by evaluating the inhibitory rate (IR) of microalgae growth by measuring the fluorescence of viable cells. We found that the metabolites of strain PD-2 had algicidal activity against its host P. donghaiense (IR 84.81%) and two other red tide microalgae, Phaeocystis globosa (IR 78.91%) and Alexandrium tamarense (IR 67.14%). β-cyclodextrin which binds to AHLs and inhibits the QS system reduced the algicidal activity more than 50%. This indicates that inhibiting the QS system may affect the algicidal metabolites production of strain PD-2. Our study indicated that a QS-regulated algicidal system may play a potential role in the process of red tides disintegration. QS might be a potential way to control red tides.

  14. Optimization of culture conditions and medium composition for the marine algicidal bacterium Alteromonas sp. DH46 by uniform design

    NASA Astrophysics Data System (ADS)

    Lin, Jing; Zheng, Wei; Tian, Yun; Wang, Guizhong; Zheng, Tianling

    2013-09-01

    Harmful algal blooms (HABs) have led to extensive ecological and environmental issues and huge economic losses. Various HAB control techniques have been developed, and biological methods have been paid more attention. Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner, and kill or damage the algal cells. A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp. The culture conditions were optimized using a single-factor test method. Factors including carbon source, nitrogen source, temperature, initial pH value, rotational speed and salinity were studied. The results showed that the cultivation of the bacteria at 28°C and 180 r min-1 with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46. The optimal medium composition for strain DH46 was determined by means of uniform design experimentation, and the most important components influencing the cell density were tryptone, yeast extract, soluble starch, NaNO3 and MgSO4. When the following culture medium was used (tryptone 14.0g, yeast extract 1.63g, soluble starch 5.0 g, NaNO3 1.6 g, MgSO4 2.3 g in 1L), the largest bacterial dry weight (7.36 g L-1) was obtained, which was an enhancement of 107% compared to the initial medium; and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.

  15. Technical Note: Effects of tethering on herbage selection, intake and digestibility, grazing behavior, and energy expenditure by Boer x Spanish goats grazing high-quality herbage.

    PubMed

    Patra, A K; Puchala, R; Detweiler, G; Dawson, L J; Sahlu, T; Goetsch, A L

    2008-05-01

    Twenty-four yearling Boer x Spanish goats were used in a crossover experiment to determine the effects of tethering on herbage selection, intake and digestibility, grazing behavior, and energy expenditure (EE) with high-quality herbage. Four 0.72-ha paddocks of wheat (Triticum aestivum) and berseem clover (Trifolium alexandrium) were grazed in the spring. Each paddock hosted 6 animals, 3 with free movement and 3 attached to a 3-m tether that was moved daily and provided access to an area of 28.3 m(2). One animal of each treatment and paddock was used to determine herbage selection, fecal output, or grazing behavior and EE. Herbage DM mass in tethered areas before grazing averaged 2,649 and 2,981 kg/ha in periods 1 and 2, respectively. The CP concentration in ingesta was greater (P < 0.05; 23.1 and 20.3 +/- 0.82%) for free vs. tethered animals, although in vitro true DM digestion (75.7 and 76.5 +/- 1.20%, respectively) did not differ (P > 0.05) between treatments. Intake of ME based on in vitro true DM digestion and fecal output was greater (P < 0.05) for free vs. tethered animals (12.7 and 10.4 +/- 0.89 MJ/d). No treatment effects were observed (P > 0.05) for time spent ruminating or grazing (405 and 366 +/- 42.5 min/d, respectively), although mean EE was greater (P < 0.05) for free vs. tethered animals (633 and 512 +/- 27.4 kJ/kg of BW(0.75) for free and tethered, respectively), with differences (P < 0.05) between treatments at each hour of the day. Tethering animals may be acceptable to model those with free movement for some measures such as ingesta composition but appears inappropriate for others, such as energy metabolism.

  16. Transcriptome Analysis of Scrippsiella trochoidea CCMP 3099 Reveals Physiological Changes Related to Nitrate Depletion

    PubMed Central

    Cooper, Joshua T.; Sinclair, Geoffrey A.; Wawrik, Boris

    2016-01-01

    Dinoflagellates are a major component of marine phytoplankton and many species are recognized for their ability to produce harmful algal blooms (HABs). Scrippsiella trochoidea is a non-toxic, marine dinoflagellate that can be found in both cold and tropic waters where it is known to produce “red tide” events. Little is known about the genomic makeup of S. trochoidea and a transcriptome study was conducted to shed light on the biochemical and physiological adaptations related to nutrient depletion. Cultures were grown under N and P limiting conditions and transcriptomes were generated via RNAseq technology. De novo assembly reconstructed 107,415 putative transcripts of which only 41% could be annotated. No significant transcriptomic response was observed in response to initial P depletion, however, a strong transcriptional response to N depletion was detected. Among the down-regulated pathways were those for glutamine/glutamate metabolism as well as urea and nitrate/nitrite transporters. Transcripts for ammonia transporters displayed both up- and down-regulation, perhaps related to a shift to higher affinity transporters. Genes for the utilization of DON compounds were up-regulated. These included transcripts for amino acids transporters, polyamine oxidase, and extracellular proteinase and peptidases. N depletion also triggered down regulation of transcripts related to the production of Photosystems I & II and related proteins. These data are consistent with a metabolic strategy that conserves N while maximizing sustained metabolism by emphasizing the relative contribution of organic N sources. Surprisingly, the transcriptome also contained transcripts potentially related to secondary metabolite production, including a homolog to the Short Isoform Saxitoxin gene (sxtA) from Alexandrium fundyense, which was significantly up-regulated under N-depletion. A total of 113 unique hits to Sxt genes, covering 17 of the 34 genes found in C. raciborskii were detected

  17. Characterisation of algal organic matter produced by bloom-forming marine and freshwater algae.

    PubMed

    Villacorte, L O; Ekowati, Y; Neu, T R; Kleijn, J M; Winters, H; Amy, G; Schippers, J C; Kennedy, M D

    2015-04-15

    Algal blooms can seriously affect the operation of water treatment processes including low pressure (micro- and ultra-filtration) and high pressure (nanofiltration and reverse osmosis) membranes mainly due to accumulation of algal-derived organic matter (AOM). In this study, the different components of AOM extracted from three common species of bloom-forming algae (Alexandrium tamarense, Chaetoceros affinis and Microcystis sp.) were characterised employing various analytical techniques, such as liquid chromatography - organic carbon detection, fluorescence spectroscopy, fourier transform infrared spectroscopy, alcian blue staining and lectin staining coupled with laser scanning microscopy to indentify its composition and force measurement using atomic force microscopy to measure its stickiness. Batch culture monitoring of the three algal species illustrated varying characteristics in terms of growth pattern, cell concentration and AOM release. The AOM produced by the three algal species comprised mainly biopolymers (e.g., polysaccharides and proteins) but some refractory compounds (e.g., humic-like substances) and other low molecular weight acid and neutral compounds were also found. Biopolymers containing fucose and sulphated functional groups were found in all AOM samples while the presence of other functional groups varied between different species. A large majority (>80%) of the acidic polysaccharide components (in terms of transparent exopolymer particles) were found in the colloidal size range (<0.4 μm). The relative stickiness of AOM substantially varied between algal species and that the cohesion between AOM-coated surfaces was much stronger than the adhesion of AOM on AOM-free surfaces. Overall, the composition as well as the physico-chemical characteristics (e.g., stickiness) of AOM will likely dictate the severity of fouling in membrane systems during algal blooms.

  18. Phytochip: development of a DNA-microarray for rapid and accurate identification of Pseudo-nitzschia spp and other harmful algal species.

    PubMed

    Noyer, Charlotte; Abot, Anne; Trouilh, Lidwine; Leberre, Véronique Anton; Dreanno, Catherine

    2015-05-01

    Detection of harmful algal blooms has become a challenging concern because of the direct impacts on public health and economy. The identification of toxic dinoflagellates and diatoms in monitoring programs requires an extensive taxonomic expertise and is time consuming. Advances in molecular biology have allowed the development of new approaches, more rapid, accurate and cost-effective for detecting these microorganisms. In this context, we developed a new DNA microarray (called, Phytochip) for the simultaneous detection of multiple HAB species with a particular emphasis on Pseudo-nitzschia species. Oligonucleotide probes were designed along the rRNA operon. After DNA extraction, the target rDNA genes were amplified and labeled using an asymmetric PCR; then, the amplicons were hybridized to the oligonucleotide probes present on the chips. The total assay from seawater sampling to data acquisition can be performed within a working day. Specificity and sensitivity were assessed by using monoclonal cultures, mixtures of species and field samples spiked with a known amount of cultured cells. The Phytochip with its 81 validated oligonucleotide probes was able to detect 12 species of Pseudo-nitzschia and 11 species of dinoflagellates among which were 3 species of Karenia and 3 species of Alexandrium. The Phytochip was applied to environmental samples already characterized by light microscopy and cloned into DNA libraries. The hybridizations on the Phytochip were in good agreement with the sequences retrieved from the clone libraries and the microscopic observations. The Phytochip enables a reliable multiplex detection of phytoplankton and can assist a water quality monitoring program as well as more general ecological research.

  19. RNA Sequencing Revealed Numerous Polyketide Synthase Genes in the Harmful Dinoflagellate Karenia mikimotoi

    PubMed Central

    Kimura, Kei; Okuda, Shujiro; Nakayama, Kei; Shikata, Tomoyuki; Takahashi, Fumio; Yamaguchi, Haruo; Skamoto, Setsuko; Yamaguchi, Mineo; Tomaru, Yuji

    2015-01-01

    The dinoflagellate Karenia mikimotoi forms blooms in the coastal waters of temperate regions and occasionally causes massive fish and invertebrate mortality. This study aimed to elucidate the toxic effect of K. mikimotoi on marine organisms by using the genomics approach; RNA-sequence libraries were constructed, and data were analyzed to identify toxin-related genes. Next-generation sequencing produced 153,406 transcript contigs from the axenic culture of K. mikimotoi. BLASTX analysis against all assembled contigs revealed that 208 contigs were polyketide synthase (PKS) sequences. Thus, K. mikimotoi was thought to have several genes encoding PKS metabolites and to likely produce toxin-like polyketide molecules. Of all the sequences, approximately 30 encoded eight PKS genes, which were remarkably similar to those of Karenia brevis. Our phylogenetic analyses showed that these genes belonged to a new group of PKS type-I genes. Phylogenetic and active domain analyses showed that the amino acid sequence of four among eight Karenia PKS genes was not similar to any of the reported PKS genes. These PKS genes might possibly be associated with the synthesis of polyketide toxins produced by Karenia species. Further, a homology search revealed 10 contigs that were similar to a toxin gene responsible for the synthesis of saxitoxin (sxtA) in the toxic dinoflagellate Alexandrium fundyense. These contigs encoded A1–A3 domains of sxtA genes. Thus, this study identified some transcripts in K. mikimotoi that might be associated with several putative toxin-related genes. The findings of this study might help understand the mechanism of toxicity of K. mikimotoi and other dinoflagellates. PMID:26561394

  20. Differential effects of ocean acidification on carbon acquisition in two bloom-forming dinoflagellate species.

    PubMed

    Eberlein, Tim; Van de Waal, Dedmer B; Rost, Björn

    2014-08-01

    Dinoflagellates represent a cosmopolitan group of phytoplankton with the ability to form harmful algal blooms. Featuring a Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) with very low CO2 affinities, photosynthesis of this group may be particularly prone to carbon limitation and thus benefit from rising atmospheric CO2 partial pressure (pCO2) under ocean acidification (OA). Here, we investigated the consequences of OA on two bloom-forming dinoflagellate species, the calcareous Scrippsiella trochoidea and the toxic Alexandrium tamarense. Using dilute batch incubations, we assessed growth characteristics over a range of pCO2 (i.e. 180-1200 µatm). To understand the underlying physiology, several aspects of inorganic carbon acquisition were investigated by membrane-inlet mass spectrometry. Our results show that both species kept growth rates constant over the tested pCO2 range, but we observed a number of species-specific responses. For instance, biomass production and cell size decreased in S. trochoidea, while A. tamarense was not responsive to OA in these measures. In terms of oxygen fluxes, rates of photosynthesis and respiration remained unaltered in S. trochoidea whereas respiration increased in A. tamarense under OA. Both species featured efficient carbon concentrating mechanisms (CCMs) with a CO2-dependent contribution of HCO3(-) uptake. In S. trochoidea, the CCM was further facilitated by exceptionally high and CO2-independent carbonic anhydrase activity. Comparing both species, a general trade-off between maximum rates of photosynthesis and respective affinities is indicated. In conclusion, our results demonstrate effective CCMs in both species, yet very different strategies to adjust their carbon acquisition. This regulation in CCMs enables both species to maintain growth over a wide range of ecologically relevant pCO2 .

  1. Insights into a dinoflagellate genome through expressed sequence tag analysis

    PubMed Central

    Hackett, Jeremiah D; Scheetz, Todd E; Yoon, Hwan Su; Soares, Marcelo B; Bonaldo, Maria F; Casavant, Thomas L; Bhattacharya, Debashish

    2005-01-01

    Background Dinoflagellates are important marine primary producers and grazers and cause toxic "red tides". These taxa are characterized by many unique features such as immense genomes, the absence of nucleosomes, and photosynthetic organelles (plastids) that have been gained and lost multiple times. We generated EST sequences from non-normalized and normalized cDNA libraries from a culture of the toxic species Alexandrium tamarense to elucidate dinoflagellate evolution. Previous analyses of these data have clarified plastid origin and here we study the gene content, annotate the ESTs, and analyze the genes that are putatively involved in DNA packaging. Results Approximately 20% of the 6,723 unique (11,171 total 3'-reads) ESTs data could be annotated using Blast searches against GenBank. Several putative dinoflagellate-specific mRNAs were identified, including one novel plastid protein. Dinoflagellate genes, similar to other eukaryotes, have a high GC-content that is reflected in the amino acid codon usage. Highly represented transcripts include histone-like (HLP) and luciferin binding proteins and several genes occur in families that encode nearly identical proteins. We also identified rare transcripts encoding a predicted protein highly similar to histone H2A.X. We speculate this histone may be retained for its role in DNA double-strand break repair. Conclusion This is the most extensive collection to date of ESTs from a toxic dinoflagellate. These data will be instrumental to future research to understand the unique and complex cell biology of these organisms and for potentially identifying the genes involved in toxin production. PMID:15921535

  2. Stable carbon isotope fractionation of organic cyst-forming dinoflagellates: Evaluating the potential for a CO2 proxy

    NASA Astrophysics Data System (ADS)

    Hoins, Mirja; Van de Waal, Dedmer B.; Eberlein, Tim; Reichart, Gert-Jan; Rost, Björn; Sluijs, Appy

    2015-07-01

    Over the past decades, significant progress has been made regarding the quantification and mechanistic understanding of stable carbon isotope fractionation (13C fractionation) in photosynthetic unicellular organisms in response to changes in the partial pressure of atmospheric CO2 (pCO2). However, hardly any data is available for organic cyst-forming dinoflagellates while this is an ecologically important group with a unique fossil record. We performed dilute batch experiments with four harmful dinoflagellate species known for their ability to form organic cysts: Alexandrium tamarense, Scrippsiella trochoidea, Gonyaulax spinifera and Protoceratium reticulatum. Cells were grown at a range of dissolved CO2 concentrations characterizing past, modern and projected future values (∼5-50 μmol L-1), representing atmospheric pCO2 of 180, 380, 800 and 1200 μatm. In all tested species, 13C fractionation depends on CO2 with a slope of up to 0.17‰ (μmol L)-1. Even more consistent correlations were found between 13C fractionation and the combined effects of particulate organic carbon quota (POC quota; pg C cell-1) and CO2. Carbon isotope fractionation as well as its response to CO2 is species-specific. These results may be interpreted as a first step towards a proxy for past pCO2 based on carbon isotope ratios of fossil organic dinoflagellate cysts. However, additional culture experiments focusing on environmental variables other than pCO2, physiological underpinning of the recorded response, testing for possible offsets in 13C values between cells and cysts, as well as field calibration studies are required to establish a reliable proxy.

  3. Planktonic Microbes in the Gulf of Maine Area

    PubMed Central

    Li, William K. W.; Andersen, Robert A.; Gifford, Dian J.; Incze, Lewis S.; Martin, Jennifer L.; Pilskaln, Cynthia H.; Rooney-Varga, Juliette N.; Sieracki, Michael E.; Wilson, William H.; Wolff, Nicholas H.

    2011-01-01

    In the Gulf of Maine area (GoMA), as elsewhere in the ocean, the organisms of greatest numerical abundance are microbes. Viruses in GoMA are largely cyanophages and bacteriophages, including podoviruses which lack tails. There is also evidence of Mimivirus and Chlorovirus in the metagenome. Bacteria in GoMA comprise the dominant SAR11 phylotype cluster, and other abundant phylotypes such as SAR86-like cluster, SAR116-like cluster, Roseobacter, Rhodospirillaceae, Acidomicrobidae, Flavobacteriales, Cytophaga, and unclassified Alphaproteobacteria and Gammaproteobacteria clusters. Bacterial epibionts of the dinoflagellate Alexandrium fundyense include Rhodobacteraceae, Flavobacteriaceae, Cytophaga spp., Sulfitobacter spp., Sphingomonas spp., and unclassified Bacteroidetes. Phototrophic prokaryotes in GoMA include cyanobacteria that contain chlorophyll (mainly Synechococcus), aerobic anoxygenic phototrophs that contain bacteriochlorophyll, and bacteria that contain proteorhodopsin. Eukaryotic microalgae in GoMA include Bacillariophyceae, Dinophyceae, Prymnesiophyceae, Prasinophyceae, Trebouxiophyceae, Cryptophyceae, Dictyochophyceae, Chrysophyceae, Eustigmatophyceae, Pelagophyceae, Synurophyceae, and Xanthophyceae. There are no records of Bolidophyceae, Aurearenophyceae, Raphidophyceae, and Synchromophyceae in GoMA. In total, there are records for 665 names and 229 genera of microalgae. Heterotrophic eukaryotic protists in GoMA include Dinophyceae, Alveolata, Apicomplexa, amoeboid organisms, Labrynthulida, and heterotrophic marine stramenopiles (MAST). Ciliates include Strombidium, Lohmaniella, Tontonia, Strobilidium, Strombidinopsis and the mixotrophs Laboea strobila and Myrionecta rubrum (ex Mesodinium rubra). An inventory of selected microbial groups in each of 14 physiographic regions in GoMA is made by combining information on the depth-dependent variation of cell density and the depth-dependent variation of water volume. Across the entire GoMA, an estimate for the

  4. Molecular diversity patterns among various phytoplankton size-fractions in West Greenland in late summer

    NASA Astrophysics Data System (ADS)

    Elferink, Stephanie; Neuhaus, Stefan; Wohlrab, Sylke; Toebe, Kerstin; Voß, Daniela; Gottschling, Marc; Lundholm, Nina; Krock, Bernd; Koch, Boris P.; Zielinski, Oliver; Cembella, Allan; John, Uwe

    2017-03-01

    as Alexandrium fundyense and A. ostenfeldii) in the area points out the potential risk for this vulnerable ecosystem in a changing world.

  5. Determining the Advantages, Costs, and Trade-Offs of a Novel Sodium Channel Mutation in the Copepod Acartia hudsonica to Paralytic Shellfish Toxins (PST).

    PubMed

    Finiguerra, Michael; Avery, David E; Dam, Hans G

    2015-01-01

    The marine copepod Acartia hudsonica was shown to be adapted to dinoflagellate prey, Alexandrium fundyense, which produce paralytic shellfish toxins (PST). Adaptation to PSTs in other organisms is caused by a mutation in the sodium channel. Recently, a mutation in the sodium channel in A. hudsonica was found. In this study, we rigorously tested for advantages, costs, and trade-offs associated with the mutant isoform of A. hudsonica under toxic and non-toxic conditions. We combined fitness with wild-type: mutant isoform ratio measurements on the same individual copepod to test our hypotheses. All A. hudsonica copepods express both the wild-type and mutant sodium channel isoforms, but in different proportions; some individuals express predominantly mutant (PMI) or wild-type isoforms (PWI), while most individuals express relatively equal amounts of each (EI). There was no consistent pattern of improved performance as a function of toxin dose for egg production rate (EPR), ingestion rate (I), and gross growth efficiency (GGE) for individuals in the PMI group relative to individuals in the PWI expression group. Neither was there any evidence to indicate a fitness benefit to the mutant isoform at intermediate toxin doses. No clear advantage under toxic conditions was associated with the mutation. Using a mixed-diet approach, there was also no observed relationship between individual wild-type: mutant isoform ratios and among expression groups, on both toxic and non-toxic diets, for eggs produced over three days. Lastly, expression of the mutant isoform did not mitigate the negative effects of the toxin. That is, the reductions in EPR from a toxic to non-toxic diet for copepods were independent of expression groups. Overall, the results did not support our hypotheses; the mutant sodium channel isoform does not appear to be related to adaptation to PST in A. hudsonica. Other potential mechanisms responsible for the adaptation are discussed.

  6. Environmental Barcoding Reveals Massive Dinoflagellate Diversity in Marine Environments

    PubMed Central

    Stern, Rowena F.; Horak, Ales; Andrew, Rose L.; Coffroth, Mary-Alice; Andersen, Robert A.; Küpper, Frithjof C.; Jameson, Ian; Hoppenrath, Mona; Véron, Benoît; Kasai, Fumai; Brand, Jerry; James, Erick R.; Keeling, Patrick J.

    2010-01-01

    Background Dinoflagellates are an ecologically important group of protists with important functions as primary producers, coral symbionts and in toxic red tides. Although widely studied, the natural diversity of dinoflagellates is not well known. DNA barcoding has been utilized successfully for many protist groups. We used this approach to systematically sample known “species”, as a reference to measure the natural diversity in three marine environments. Methodology/Principal Findings In this study, we assembled a large cytochrome c oxidase 1 (COI) barcode database from 8 public algal culture collections plus 3 private collections worldwide resulting in 336 individual barcodes linked to specific cultures. We demonstrate that COI can identify to the species level in 15 dinoflagellate genera, generally in agreement with existing species names. Exceptions were found in species belonging to genera that were generally already known to be taxonomically challenging, such as Alexandrium or Symbiodinium. Using this barcode database as a baseline for cultured dinoflagellate diversity, we investigated the natural diversity in three diverse marine environments (Northeast Pacific, Northwest Atlantic, and Caribbean), including an evaluation of single-cell barcoding to identify uncultivated groups. From all three environments, the great majority of barcodes were not represented by any known cultured dinoflagellate, and we also observed an explosion in the diversity of genera that previously contained a modest number of known species, belonging to Kareniaceae. In total, 91.5% of non-identical environmental barcodes represent distinct species, but only 51 out of 603 unique environmental barcodes could be linked to cultured species using a conservative cut-off based on distances between cultured species. Conclusions/Significance COI barcoding was successful in identifying species from 70% of cultured genera. When applied to environmental samples, it revealed a massive amount of

  7. High Sequence Variability, Diverse Subcellular Localizations, and Ecological Implications of Alkaline Phosphatase in Dinoflagellates and Other Eukaryotic Phytoplankton

    PubMed Central

    Lin, Xin; Zhang, Huan; Cui, Yudong; Lin, Senjie

    2012-01-01

    Alkaline phosphatase (AP) is a key enzyme for phytoplankton to utilize dissolved organic phosphorus (DOP) when dissolved inorganic phosphorus is limited. While three major types of AP and their correspondingly diverse subcellular localization have been recognized in bacteria, little is known about AP in eukaryotic phytoplankton such as dinoflagellates. Here, we isolated a full-length AP cDNA from a latest-diverging dinoflagellate genus Alexandrium, and conducted comparative analyses with homologs from a relatively basal (Amphidinium carterae) and late-diverging (Karenia brevis) lineage of dinoflagellates as well as other eukaryotic algae. New data and previous studies indicate that AP is common in dinoflagellates and most other major eukaryotic groups of phytoplankton. AP sequences are more variable than many other genes studied in dinoflagellates, and are divergent among different eukaryotic phytoplankton lineages. Sequence comparison to the other characterized APs suggests that dinoflagellates and some other eukaryotic phytoplankton possess the putative AP as phoA type, but some other eukaryotic phytoplankton seem to have other types. Phylogenetic analyses based on AP amino acid sequences indicated that the “red-type” eukaryotic lineages formed a monophyletic group, suggesting a common origin of their APs. As different amino acid sequences have been found to predictably determine different spatial distribution in the cells, which may facilitate access to different pools of DOP, existing computational models were adopted to predict the subcellular localizations of putative AP in the three dinoflagellates and other eukaryotic phytoplankton. Results showed different subcellular localizations of APs in different dinoflagellates and other lineages. The linkage between AP sequence divergence, subcellular localization, and ecological niche differentiation requires rigorous experimental verification, and this study now provides a framework for such a future effort

  8. CO2-dependent carbon isotope fractionation in dinoflagellates relates to their inorganic carbon fluxes.

    PubMed

    Hoins, Mirja; Eberlein, Tim; Van de Waal, Dedmer B; Sluijs, Appy; Reichart, Gert-Jan; Rost, Björn

    2016-08-01

    Carbon isotope fractionation (εp) between the inorganic carbon source and organic matter has been proposed to be a function of pCO2. To understand the CO2-dependency of εp and species-specific differences therein, inorganic carbon fluxes in the four dinoflagellate species Alexandrium fundyense, Scrippsiella trochoidea, Gonyaulax spinifera and Protoceratium reticulatum have been measured by means of membrane-inlet mass spectrometry. In-vivo assays were carried out at different CO2 concentrations, representing a range of pCO2 from 180 to 1200 μatm. The relative bicarbonate contribution (i.e. the ratio of bicarbonate uptake to total inorganic carbon uptake) and leakage (i.e. the ratio of CO2 efflux to total inorganic carbon uptake) varied from 0.2 to 0.5 and 0.4 to 0.7, respectively, and differed significantly between species. These ratios were fed into a single-compartment model, and εp values were calculated and compared to carbon isotope fractionation measured under the same conditions. For all investigated species, modeled and measured εp values were comparable (A. fundyense, S. trochoidea, P. reticulatum) and/or showed similar trends with pCO2 (A. fundyense, G. spinifera, P. reticulatum). Offsets are attributed to biases in inorganic flux measurements, an overestimated fractionation factor for the CO2-fixing enzyme RubisCO, or the fact that intracellular inorganic carbon fluxes were not taken into account in the model. This study demonstrates that CO2-dependency in εp can largely be explained by the inorganic carbon fluxes of the individual dinoflagellates.

  9. Nutrient and phytoplankton analysis of a Mediterranean coastal area.

    PubMed

    Sebastiá, M T; Rodilla, M

    2013-01-01

    Identifying and quantifying the key anthropogenic nutrient input sources are essential to adopting management measures that can target input for maximum effect in controlling the phytoplankton biomass. In this study, three systems characterized by distinctive main nutrient sources were sampled along a Mediterranean coast transect. These sources were groundwater discharge in the Ahuir area, the Serpis river discharge in the Venecia area, and a submarine wastewater outfall 1,900 m from the coast. The study area includes factors considered important in determining a coastal area as a sensitive area: it has significant nutrient sources, tourism is a major source of income in the region, and it includes an area of high water residence time (Venecia area) which is affected by the harbor facilities and by wastewater discharges. We found that in the Ahuir and the submarine wastewater outfall areas, the effects of freshwater inputs were reduced because of a greater water exchange with the oligotrophic Mediterranean waters. On the other hand, in the Venecia area, the highest levels of nutrient concentration and phytoplankton biomass were attributed to the greatest water residence time. In this enclosed area, harmful dinoflagellates were detected (Alexandrium sp. and Dinophysis caudata). If the planned enlargement of the Gandia Harbor proceeds, it may increase the vulnerability of this system and provide the proper conditions of confinement for the dinoflagellate blooms' development. Management measures should first target phosphorus inputs as this is the most potential-limiting nutrient in the Venecia area and comes from a point source that is easier to control. Finally, we recommend that harbor environmental management plans include regular monitoring of water quality in adjacent waters to identify adverse phytoplankton community changes.

  10. Application of the standard addition method for the absolute quantification of neutral lipids in microalgae using Nile red.

    PubMed

    Bertozzini, Elena; Galluzzi, Luca; Penna, Antonella; Magnani, Mauro

    2011-10-01

    Microalgae are considered one of the best candidates for biofuel production due to their high content in neutral lipids, therefore, an accurate quantification of these lipids in microalgae is fundamental for the identification of the better candidates as biodiesel source. Nile red is a fluorescent dye widely employed for the quantification of neutral lipids in microalgae. Usually, the fluorescence intensity of the stained samples is correlated to the neutral lipid content determined with standard methods, in order to draw a standard curve and deduce the neutral lipids concentration of the unknown samples positioning their fluorescence intensity values on the curve. Standard methods used for the neutral lipids determination are laborious and often implying solvent extraction and/or other transformation (i.e. saponification or transesterification) of the sample. These methods are also time consuming and may give rise to an underestimation of the lipid content due to variable extraction yields. The approach described in this paper combines the standard addition method and the fluorometric staining using Nile red, avoiding the association of traditional neutral lipids quantification methods to the fluorometric determination. After optimization of instrument parameters and staining conditions, a linear correlation between the fluorescence intensity of each sample stained with the Nile red and its neutral lipids content deduced with the standard addition method was identified. The obtained curve allowed the direct determination of neutral lipids content maintaining a linearity range from 0.12 to 12 μg of neutral lipids per ml of sample, without need of pre-concentration. This curve was then used in the quantification of the neutral lipids content in culture of Skeletonema marinoi (Bacillariophyceae) at different days from the inoculum. This method was also successfully applied on Chaetoceros socialis (Bacillariophyceae) and Alexandrium minutum (Dinophyceae).

  11. Phosphorus cycling in the red tide incubator region of monterey bay in response to upwelling.

    PubMed

    Mackey, Katherine R M; Mioni, Cécile E; Ryan, John P; Paytan, Adina

    2012-01-01

    This study explores the cycling of phosphorus (P) in the euphotic zone following upwelling in northeastern Monterey Bay (the Red Tide Incubator region) of coastal California, with particular emphasis on how bacteria and phytoplankton that form harmful algal blooms mediate and respond to changes in P availability. In situ measurements of nutrient concentrations, phytoplankton community composition, and cell-specific alkaline phosphatase (AP) activity (determined via enzyme-labeled fluorescence assay) were measured during three cruises. Upwelling led to a 10-fold increase in dissolved inorganic (DIP) in surface waters, reaching ∼0.5 μmol L(-1). This DIP was drawn down rapidly as upwelling relaxed over a period of 1 week. Ratios of nitrate to DIP drawdown (∼5:1, calculated as the change in nitrate divided by the change in DIP) were lower than the Redfield ratio of 16:1, suggesting that luxury P uptake was occurring as phytoplankton bloomed. Dissolved organic (DOP) remained relatively constant (∼0.3 μmol L(-1)) before and immediately following upwelling, but doubled as upwelling relaxed, likely due to phytoplankton excretion and release during grazing. This transition from a relatively high DIP:DOP ratio to lower DIP:DOP ratio was accompanied by a decline in the abundance of diatoms, which had low AP activity, toward localized, spatially heterogeneous blooms of dinoflagellates in the genera Prorocentrum, Ceratium, Dinophysis, Alexandrium, and Scrippsiella that showed high AP activity regardless of ambient DIP levels. A nutrient addition incubation experiment showed that phytoplankton growth was primarily limited by nitrate, followed by DIP and DOP, suggesting that P regulates phytoplankton physiology and competition, but is not a limiting nutrient in this region. AP activity was observed in bacteria associated with lysed cell debris and aggregates of particulate organic material, where it may serve to facilitate P regeneration, as well as affixed to

  12. De Novo Assembly of a Transcriptome for Calanus finmarchicus (Crustacea, Copepoda) – The Dominant Zooplankter of the North Atlantic Ocean

    PubMed Central

    Lenz, Petra H.; Roncalli, Vittoria; Hassett, R. Patrick; Wu, Le-Shin; Cieslak, Matthew C.; Hartline, Daniel K.; Christie, Andrew E.

    2014-01-01

    Assessing the impact of global warming on the food web of the North Atlantic will require difficult-to-obtain physiological data on a key copepod crustacean, Calanus finmarchicus. The de novo transcriptome presented here represents a new resource for acquiring such data. It was produced from multiplexed gene libraries using RNA collected from six developmental stages: embryo, early nauplius (NI-II), late nauplius (NV-VI), early copepodite (CI-II), late copepodite (CV) and adult (CVI) female. Over 400,000,000 paired-end reads (100 base-pairs long) were sequenced on an Illumina instrument, and assembled into 206,041 contigs using Trinity software. Coverage was estimated to be at least 65%. A reference transcriptome comprising 96,090 unique components (“comps”) was annotated using Blast2GO. 40% of the comps had significant blast hits. 11% of the comps were successfully annotated with gene ontology (GO) terms. Expression of many comps was found to be near zero in one or more developmental stages suggesting that 35 to 48% of the transcriptome is “silent” at any given life stage. Transcripts involved in lipid biosynthesis pathways, critical for the C. finmarchicus life cycle, were identified and their expression pattern during development was examined. Relative expression of three transcripts suggests wax ester biosynthesis in late copepodites, but triacylglyceride biosynthesis in adult females. Two of these transcripts may be involved in the preparatory phase of diapause. A key environmental challenge for C. finmarchicus is the seasonal exposure to the dinoflagellate Alexandrium fundyense with high concentrations of saxitoxins, neurotoxins that block voltage-gated sodium channels. Multiple contigs encoding putative voltage-gated sodium channels were identified. They appeared to be the result of both alternate splicing and gene duplication. This is the first report of multiple NaV1 genes in a protostome. These data provide new insights into the transcriptome and

  13. Annual contribution of different plankton size classes to particulate dimethylsulfoniopropionate in a marine perturbed ecosystem

    NASA Astrophysics Data System (ADS)

    Jean, Natacha; Bogé, Gérard; Jamet, Jean-Louis; Richard, Simone; Jamet, Dominique

    2005-01-01

    Dimethylsulfoniopropionate (DMSP) concentrations were observed from October 1999 to September 2000 in a Mediterranean ecosystem (Little Bay of Toulon) submitted to eutrophication. DMSP was measured in the particulate material (DMSP p), and more specifically in the >90, 5-90 and 0.2-5 μm fractions. DMSP was first converted into dimethylsulfide (DMS) by a cold alkali treatment. DMS was then analysed by gas chromatography equipped with a flame photometric detector (FPD). DMSP p concentrations were relatively high, showing a strong temporal variability with maxima in February-March (58.8 nM). The most elevated values were recorded in the 5-90 μm size class, and represented between 60% and 100% of the total DMSP p. This fraction was mostly composed of Dinoflagellates whose biomass was significantly correlated with DMSP p concentrations. These concentrations showed better correlations with Ceratium furca, Dinophysis acuminata, Prorocentrum arcuatum and also Alexandrium minutum. The intracellular contents of DMSP were much higher in Dinoflagellates (124.9±5.7 mM) than in Diatoms (25.1±1.1 mM). A. minutum produced the strongest intracellular concentrations (3387.6±121.9 mM). High DMSP contents were also found in D. acuminata (477.4±64.3 mM) and P. arcuatum (442.2±22.9 mM). The >90 μm size class had a minor importance in DMSP production, generally below 20% of the total DMSP p. However, DMSP in the >90 μm fraction was well correlated with cladoceran abundance. Plankton cells between 0.2 and 5 μm contained a low part of the DMSP p pool, lower than 20%. Besides, no correlation was found between their algal abundances and DMSP in the 0.2-5 μm size class. Temperature and photoperiod could have influenced the Dinoflagellate development, and consequently, the DMSP p concentrations. The strong autumnal rains affected the composition of the phytoplankton community and the production of sulfur compounds. These particular climatic conditions have induced an increase in

  14. Evaluation of the MIDTAL microarray chip for monitoring toxic microalgae in the Orkney Islands, U.K.

    PubMed

    Taylor, Joe D; Berzano, Marco; Percy, Linda; Lewis, Jane

    2013-10-01

    Harmful or nuisance algal blooms can cause economic damage to fisheries and tourism. Additionally, toxins produced by harmful algae and ingested via contaminated shellfish can be potentially fatal to humans. The seas around the Orkney Islands, UK currently hold a number of toxic algal species which cause shellfishery closures in most years. Extensive and costly monitoring programs are carried out to detect harmful microalgae before they reach action levels. However, the ability to distinguish between toxic and non-toxic strains of some algae is not possible using these methods. The microarrays for the detection of toxic algae (MIDTAL) microarray contains rRNA probes for toxic algal species/strains which have been adapted and optimized for microarray use. In order to investigate the use of the chip for monitoring in the Orkney Islands, samples were collected between 2009 and 2011 from Brings Deep, Scapa Flow, Orkney Islands, UK; RNA was extracted and hybridized with generation 2 and 3.1 of the chip. The data were then compared to cell counts performed under light microscopy and in the case of Alexandrium tamarense to qPCR data targeting the saxitoxin gene and the LSU-rRNA gene. A good agreement between cell numbers and microarray signal was found for A. tamarense, Pseudo-nitzschia sp., Dinophysis sp. (r<0.5, for all) in addition to this there the chip successfully detected a large bloom of Karenia mikimotoi (r<0.70) in August and September 2011. Overall, there was good improvement in probe signal between generation 2 and generation 3.1 of the chip with much less variability and more consistent results and better correlation between the probes. The chip performed well for A. tamarense group I signal to cell numbers in calibrations (r>0.9). However, in field samples, this correlation was slightly lower suggesting interactions between all species in the sample may affect signal. Overall, the chip showed it could identify the presence of target species in field samples

  15. Microbial modulation in the biomass and toxin production of a red-tide causing alga.

    PubMed

    Zheng, T L; Su, J Q; Maskaoui, K; Yu, Z M; Hu, Z; Xu, J S; Hong, H S

    2005-01-01

    The effect of S10, a strain of marine bacteria isolated from sediment in the Western Xiamen Sea, on the growth and paralytic shellfish poison (PSP) production in the alga Alexandrium tamarense (A. tamarense) was studied under controlled experimental conditions. The results of these experiments have shown that the growth of A. tamarense is obviously inhibited by S10 at high concentrations, however no evident effect on its growth was observed at low concentrations. Its PSP production was also inhibited by S10 at different concentrations, especially at low concentrations. The toxicity of this strain of A. tamarense is about (0.95-12.14) x 10(-6) MU/cell, a peak toxicity value of 12.14 x 10(-6) MU/cell appeared on the 14th day, after which levels decreased gradually. The alga grew well in conditions of pH 6-8 and salinities of 20-34 per thousand. The toxicity of the alga varied markedly at different pH and salinity levels. Toxicity decreased as pH increased, while it increased with salinity and reached a peak value at a salinity of 30 per thousand, after which it declined gradually. S10 at a concentration of 1.02 x 10(9) cells/ml inhibited growth and the PSP production of A. tamarense at different pH and salinity levels. S10 had the strongest inhibitory function on the growth of A. tamarense under conditions of pH 7 and a salinity of 34 per thousand. The best inhibitory effect on PSP production by A. tamarense was at pH 7, this inhibitory effect on PSP production did not relate to salinity. Interactions between marine bacteria and A. tamarense were also investigated using the flow cytometer technique (FCM) as well as direct microscope counting. S10 was identified as being a member of the genus Bacillus, the difference in 16S rDNA between S10 and Bacillus halmapalus was only 2%. The mechanism involved in the inhibition of growth and PSP production of A. tamarense by this strain of marine bacteria, and the prospect of using it and other marine bacteria in the bio

  16. Understanding interannual, decadal level variability in paralytic shellfish poisoning toxicity in the Gulf of Maine: the HAB Index

    PubMed Central

    Anderson, Donald M.; Couture, Darcie A.; Kleindinst, Judith L.; Keafer, Bruce A.; McGillicuddy, Dennis J.; Martin, Jennifer L.; Richlen, Mindy L.; Hickey, J. Michael; Solow, Andrew R.

    2013-01-01

    A major goal in harmful algal bloom (HAB) research has been to identify mechanisms underlying interannual variability in bloom magnitude and impact. Here the focus is on variability in Alexandrium fundyense blooms and paralytic shellfish poisoning (PSP) toxicity in Maine, USA, over 34 years (1978 – 2011). The Maine coastline was divided into two regions -eastern and western Maine, and within those two regions, three measures of PSP toxicity (the percent of stations showing detectable toxicity over the year, the cumulative amount of toxicity per station measured in all shellfish (mussel) samples during that year, and the duration of measurable toxicity) were examined for each year in the time series. These metrics were combined into a simple HAB Index that provides a single measure of annual toxin severity across each region. The three toxin metrics, as well as the HAB Index that integrates them, reveal significant variability in overall toxicity between individual years as well as long-term, decadal patterns or regimes. Based on different conceptual models of the system, we considered three trend formulations to characterize the long-term patterns in the Index – a three-phase (mean-shift) model, a linear two-phase model, and a pulse-decline model. The first represents a “regime shift” or multiple equilibria formulation as might occur with alternating periods of sustained high and low cyst abundance or favorable and unfavorable growth conditions, the second depicts a scenario of more gradual transitions in cyst abundance or growth conditions of vegetative cells, and the third characterizes a ”sawtooth” pattern in which upward shifts in toxicity are associated with major cyst recruitment events, followed by a gradual but continuous decline until the next pulse. The fitted models were compared using both residual sum of squares and Akaike's Information Criterion. There were some differences between model fits, but none consistently gave a better fit than

  17. Trophic cascades and future harmful algal blooms within ice-free Arctic Seas north of Bering Strait: A simulation analysis

    NASA Astrophysics Data System (ADS)

    Walsh, John J.; Dieterle, Dwight A.; Chen, F. Robert; Lenes, Jason M.; Maslowski, Wieslaw; Cassano, John J.; Whitledge, Terry E.; Stockwell, Dean; Flint, Mikhail; Sukhanova, Irina N.; Christensen, John

    2011-11-01

    Within larger ice-free regions of the western Arctic Seas, subject to ongoing trophic cascades induced by past overfishing, as well as to possible future eutrophication of the drainage basins of the Yukon and Mackenzie Rivers, prior very toxic harmful algal blooms (HABs) - first associated with ∼100 human deaths near Sitka, Alaska in 1799 - may soon expand. Blooms of calcareous coccolithophores in the Bering Sea during 1997-1998 were non-toxic harbingers of the subsequent increments of other non-siliceous phytoplankton. But, now saxitoxic dinoflagellates, e.g. Alexandrium tamarense, were instead found by us within the adjacent downstream Chukchi Sea during SBI cruises of 2002 and 2003. A previous complex, coupled biophysical model had been validated earlier by ship-board observations from the Chukchi/Beaufort Seas during the summer of 2002. With inclusion of phosphorus as another chemical state variable to modulate additional competition by recently observed nitrogen-fixers, we now explore here the possible consequences of altered composition of dominant phytoplankton functional groups [diatoms, microflagellates, prymnesiophyte Phaeocystis colonies, coccolithophores, diazotrophs, and dinoflagellates] in relation to increases of the toxic A. tamarense, responding to relaxation of grazing pressure by herbivores north of Bering Strait as part of a continuing trophic cascade. Model formulation was guided by validation observations obtained during 2002-2004 from: cruises of the SBI, CHINARE, and CASES programs; moored arrays in Bering Strait; other RUSALCA cruises around Wrangel Island; and SBI helicopter surveys of the shelf-break regions of the Arctic basin. Our year-long model scenarios during 2002-2003 indicate that post bloom silica-limitation of diatoms, after smaller simulated spring grazing losses, led to subsequent competitive advantages in summer for the coccolithophores, dinoflagellates, and diazotrophs. Immediate top-down control is exerted by imposed

  18. Field-deployable colorimetric biosensor system for the rapid detection of pathogenic organisms

    NASA Astrophysics Data System (ADS)

    Duy, Janice

    The rapid identification of pathogenic organisms is necessary for recognizing and managing human and environmental health risks. Numerous detection schemes are available, but most are difficult to employ in non-laboratory settings due to their need for bulky, specialized equipment, multiple reagents, or highly trained personnel. To address this problem, a rapid, field-compatible biosensor system based on the colorimetric detection of nucleic acid hybrids was developed. Peptide nucleic acid (PNA) probes were used to capture ribosomal RNA sequences from environmental samples. Non-target nucleic acids, including single-base mismatches flanked by adenines and uracils, were removed with a micrococcal nuclease digestion step. Matched PNA-RNA hybrids remained intact and were indicated by the cyanine dye DiSC2(5). PNA-containing duplexes function as templates for the aggregation of DiSC2(5), visualized as a change in solution color from blue to purple. This transition can be measured as an increase in the solution absorbance at 540 nm (dye aggregate) at the expense of the dye monomer peak at 650 nm. These concomitant spectral changes were used to calculate a "hybridization signal" using the ratio A aggregate/Amonomer ≈ A540/A650. Testing with pathogenic environmental samples was accomplished using two model organisms: the harmful algal bloom-causing dinoflagellate Alexandrium species, and the potato wart disease-causing fungus Synchytrium endobioticum. In both cases, the colorimetric approach was able to distinguish the targets with sensitivities rivaling those of established techniques, but with the advantages of decreased hands-on time and cost. Assay fieldability was tested with a portable colorimeter designed to quantify the dye-indicated hybridization signal and assembled from commercially available components. Side-by-side testing revealed no difference in the sensing performance of the colorimeter compared to a laboratory spectrophotometer (Pearson's r=0

  19. An Oceanographic Buoy for Multidisciplinary Education and Research in a Coastal Embayment Prone to Harmful Algal Blooms

    NASA Astrophysics Data System (ADS)

    Laine, E. P.; Roesler, C.; Teegarden, G.

    2005-12-01

    In the spring of 2006 a consortium of Bowdoin College, Bigelow Laboratory for Ocean Sciences, and Saint Joseph's College of Maine will begin the operation of an oceanographic buoy in Harpswell Sound, part of the Casco Bay region of coastal Maine. Funding for acquisition of the buoy has been provided by NSF's MRI program. The sensing buoy will measure physical climatic and oceanographic variables, as well as a suite of biogeochemical indicators (nutrients, chlorophyll, light absorption, etc.). The data collected will be publicly available in real time and will contribute to the overall Gulf of Maine Ocean Observing System (GoMOOS) monitoring program, a premier and ground-breaking effort in assessing the physical and biogeochemical characteristics of the Gulf of Maine. Harpswell Sound is known as an indicator region for harmful algal blooms (HABs) of toxic Alexandrium spp. microalgae, and is an ideal location to employ long-term, comprehensive, remote and real-time monitoring to characterize model systems that promote HABs, as well as system response to changing watershed use patterns and evolving cultural eutrophication. Data acquired with the buoy's sensors, both streaming in real-time and archived in larger sets, will be used in course work at Bowdoin College and Saint Joseph's College, and will be available for use by other post-secondary institutions. Immediate applications include use of data in course work to understand the influence of physical oceanographic processes on biological processes in three dimensions and through time from an Eulerian perspective. The influence of climatic events and the geological characteristics of the surrounding watershed will also be recorded and analyzed through earth science course work. Bowdoin College has a marine research station immediately adjacent on the shore of Harpswell Sound, facilitating complementary traditional monitoring opportunities, e.g. targeted and detailed sampling of interesting features indicated by the

  20. A comparative study for PSP toxins quantification by using MBA and HPLC official methods in shellfish.

    PubMed

    Ben-Gigirey, B; Rodríguez-Velasco, M L; Otero, A; Vieites, J M; Cabado, A G

    2012-10-01

    Commission Regulation (EC) N° 2074/2005 recognises the biological method as the reference method for Paralytic Shellfish Poisoning (PSP) toxins detection in molluscs. It was amended by Commission Regulation (EC) N° 1664/2006 that accepted the so-called Lawrence method as an alternative to the reference method. The goal of this study was to compare AOAC Official Methods of Analysis 959.08 (Biological method) and 2005.06 (Prechromatographic Oxidation and Liquid Chromatography with fluorescence detection) in samples with different toxin profiles. The influence of extraction solvent in the total samples toxicity was also evaluated. A total of 40 samples including mussels, clams, scallops, razor-clams, cockles, oysters and barnacles were analysed by both official methods. Samples were selected with Alexandrium and Gymnodinium toxic profiles, from different origin and including several presentations: fresh, frozen, canned and boiled. Acetic and hydrochloric acid extractions were performed in all samples and the extracts were simultaneously analysed by both methods. Most samples were naturally contaminated and two samples were spiked. Comparison of both official methods, mouse bioassay (MBA) with HCl extraction and Liquid Chromatography with fluorescence detection (HPLC-FLD) with acetic acid extraction, led to an 85% of consistent results regarding compliance with legal limit, including samples below and above it. The linear correlation coefficient was r² = 0.69 and the paired t test (two tails, α = 0.05) indicated that there were not significant differences among both sets of data. Nevertheless, toxicity differences were found in several samples. In 15 out of 18 shellfish with a Gymnodinium toxic profile, higher toxicity levels were obtained by MBA. This fact was more evident in 7 samples, partially related to the lack of standards and the impossibility of analysing dc-NEO, C1, 2 and GTX6 at the beginning of the study. However, other factors concerning the extraction

  1. Understanding interannual, decadal level variability in paralytic shellfish poisoning toxicity in the Gulf of Maine: The HAB Index

    NASA Astrophysics Data System (ADS)

    Anderson, Donald M.; Couture, Darcie A.; Kleindinst, Judith L.; Keafer, Bruce A.; McGillicuddy, Dennis J., Jr.; Martin, Jennifer L.; Richlen, Mindy L.; Hickey, J. Michael; Solow, Andrew R.

    2014-05-01

    A major goal in harmful algal bloom (HAB) research has been to identify mechanisms underlying interannual variability in bloom magnitude and impact. Here the focus is on variability in Alexandrium fundyense blooms and paralytic shellfish poisoning (PSP) toxicity in Maine, USA, over 34 years (1978-2011). The Maine coastline was divided into two regions - eastern and western Maine, and within those two regions, three measures of PSP toxicity (the percent of stations showing detectable toxicity over the year, the cumulative amount of toxicity per station measured in all shellfish (mussel) samples during that year, and the duration of measurable toxicity) were examined for each year in the time series. These metrics were combined into a simple HAB Index that provides a single measure of annual toxin severity across each region. The three toxin metrics, as well as the HAB Index that integrates them, reveal significant variability in overall toxicity between individual years as well as long-term, decadal patterns or regimes. Based on different conceptual models of the system, we considered three trend formulations to characterize the long-term patterns in the Index - a three-phase (mean-shift) model, a linear two-phase model, and a pulse-decline model. The first represents a “regime shift” or multiple equilibria formulation as might occur with alternating periods of sustained high and low cyst abundance or favorable and unfavorable growth conditions, the second depicts a scenario of more gradual transitions in cyst abundance or growth conditions of vegetative cells, and the third characterizes a ”sawtooth” pattern in which upward shifts in toxicity are associated with major cyst recruitment events, followed by a gradual but continuous decline until the next pulse. The fitted models were compared using both residual sum of squares and Akaike's Information Criterion. There were some differences between model fits, but none consistently gave a better fit than the

  2. Temperature effects on kinetics of paralytic shellfish toxin elimination in Atlantic surfclams, Spisula solidissima

    NASA Astrophysics Data System (ADS)

    Monica Bricelj, V.; Cembella, Allan D.; Laby, David

    2014-05-01

    Surfclams, Spisula solidissima, pose a particular health risk for human consumption as they are characterized by accumulation of extremely high levels of toxins associated with paralytic shellfish poisoning (PSP), slow toxin elimination and an extremely high post-ingestive capacity for toxin bioconversion. Surfclam populations experience a wide range of temperatures along the NW Atlantic continental shelf, and are undergoing range contraction that has been attributed to global warming. In this study the influence of temperature (5, 12 and 21 °C) on detoxification kinetics of individual PSP toxins in two tissue compartments of juvenile surfclams (∼35 mm shell length) was determined under controlled laboratory conditions, over prolonged (2.4 months) depuration. Clams were toxified with a representative regional Gulf of Maine isolate of the dinoflagellate Alexandrium fundyense of known toxin profile, allowing tracking of changes in toxin composition and calculated toxicity in surfclam tissues. The visceral mass detoxified at all temperatures, although toxin loss rate increased with increasing temperature. In contrast, total toxin content and calculated toxicities in other tissues remained constant or even increased during depuration, suggesting a physiological or biochemical toxin-retention mechanism in this tissue pool and temperature-independent detoxification. In vivo toxin compositional changes in surfclam tissues found in this study provide evidence of specific toxin conversion pathways, involving both reductive and decarbamoylation pathways. We conclude that such toxin biotransformations, especially in non-visceral tissues, may introduce a discrepancy in describing kinetics of total toxicity (in saxitoxin equivalents [STXeq]) of S. solidissima over prolonged detoxification. Nevertheless, use of total toxicity values generated by routine regulatory monitoring based upon mouse bioassays or calculated from chemical analytical determination of molar toxin

  3. Phosphorus Cycling in the Red Tide Incubator Region of Monterey Bay in Response to Upwelling

    PubMed Central

    Mackey, Katherine R. M.; Mioni, Cécile E.; Ryan, John P.; Paytan, Adina

    2012-01-01

    This study explores the cycling of phosphorus (P) in the euphotic zone following upwelling in northeastern Monterey Bay (the Red Tide Incubator region) of coastal California, with particular emphasis on how bacteria and phytoplankton that form harmful algal blooms mediate and respond to changes in P availability. In situ measurements of nutrient concentrations, phytoplankton community composition, and cell-specific alkaline phosphatase (AP) activity (determined via enzyme-labeled fluorescence assay) were measured during three cruises. Upwelling led to a 10-fold increase in dissolved inorganic (DIP) in surface waters, reaching ∼0.5 μmol L−1. This DIP was drawn down rapidly as upwelling relaxed over a period of 1 week. Ratios of nitrate to DIP drawdown (∼5:1, calculated as the change in nitrate divided by the change in DIP) were lower than the Redfield ratio of 16:1, suggesting that luxury P uptake was occurring as phytoplankton bloomed. Dissolved organic (DOP) remained relatively constant (∼0.3 μmol L−1) before and immediately following upwelling, but doubled as upwelling relaxed, likely due to phytoplankton excretion and release during grazing. This transition from a relatively high DIP:DOP ratio to lower DIP:DOP ratio was accompanied by a decline in the abundance of diatoms, which had low AP activity, toward localized, spatially heterogeneous blooms of dinoflagellates in the genera Prorocentrum, Ceratium, Dinophysis, Alexandrium, and Scrippsiella that showed high AP activity regardless of ambient DIP levels. A nutrient addition incubation experiment showed that phytoplankton growth was primarily limited by nitrate, followed by DIP and DOP, suggesting that P regulates phytoplankton physiology and competition, but is not a limiting nutrient in this region. AP activity was observed in bacteria associated with lysed cell debris and aggregates of particulate organic material, where it may serve to facilitate P regeneration, as well as affixed to

  4. Impact of floral sources and processing on the antimicrobial activities of different unifloral honeys

    PubMed Central

    Elbanna, Khaled; Attalla, Khaled; Elbadry, Medhat; Abdeltawab, Awad; Gamal-Eldin, Hosny; Ramadan, Mohamed Fawzy

    2014-01-01

    Objective To study in vitro antibacterial activity and physicochemical properties of three unifloral honeys (citrus, clover and cotton honeys), and to study the impacts of storage, dilution with water (33%, w/v) and autoclaving (121 °C for 15 min) on honeys characteristics. Methods Honey samples from monofloral sources including citrus (Citrus spp.), Egyptian clover (Trifolium alexandrium) and cotton (Gossypium vitifolium) were obtained during three successive seasons (2010-2012). Physicochemical properties and antimicrobial activities of different honey samples were studies. Results In honey samples stored for 12 or 24 month, colour, hydroxymethyl furfural and acidity increased, while refractive index, water activity, total soluble solids, electrical conductivity and pH remained relatively unaffected, but H2O2 values decreased. Types of honey exhibited various degrees of antibacterial activity against different indicator bacteria, wherein the highest antibacterial activity was recorded for clover honey followed by citrus and cotton honeys, respectively. Different species of bacteria were differed in their sensitivity to honey, wherein Salmonella enteritidis was the most sensitive followed by Staphylococcus aureus, Listeria monocytogenes and Escherichia coli, respectively. Storage up to 24 months at room temperature slightly reduced the antibacterial activity. The reduction levels were about 2.6% and 4.6% after 12 and 24 months, respectively. Diluting honeys with water increased the antibacterial activity by ca. 8.3%, while autoclaving decreased the antibacterial activity by ca. 13.5%. The relative contribution of the peroxide and non-peroxide components in the total antibacterial activity of fresh honeys was investigated. The antibacterial activity of honeys was mainly attributed to non-peroxide antibacterial agents, wherein their contribution was ca. 88%, while the contribution of H2O2 was only 12%. The contribution of the thermostable antibacterial components

  5. Centers for Oceans and Human Health: a unified approach to the challenge of harmful algal blooms

    PubMed Central

    Erdner, Deana L; Dyble, Julianne; Parsons, Michael L; Stevens, Richard C; Hubbard, Katherine A; Wrabel, Michele L; Moore, Stephanie K; Lefebvre, Kathi A; Anderson, Donald M; Bienfang, Paul; Bidigare, Robert R; Parker, Micaela S; Moeller, Peter; Brand, Larry E; Trainer, Vera L

    2008-01-01

    Background Harmful algal blooms (HABs) are one focus of the national research initiatives on Oceans and Human Health (OHH) at NIEHS, NOAA and NSF. All of the OHH Centers, from the east coast to Hawaii, include one or more research projects devoted to studying HAB problems and their relationship to human health. The research shares common goals for understanding, monitoring and predicting HAB events to protect and improve human health: understanding the basic biology of the organisms; identifying how chemistry, hydrography and genetic diversity influence blooms; developing analytical methods and sensors for cells and toxins; understanding health effects of toxin exposure; and developing conceptual, empirical and numerical models of bloom dynamics. Results In the past several years, there has been significant progress toward all of the common goals. Several studies have elucidated the effects of environmental conditions and genetic heterogeneity on bloom dynamics. New methods have been developed or implemented for the detection of HAB cells and toxins, including genetic assays for Pseudo-nitzschia and Microcystis, and a biosensor for domoic acid. There have been advances in predictive models of blooms, most notably for the toxic dinoflagellates Alexandrium and Karenia. Other work is focused on the future, studying the ways in which climate change may affect HAB incidence, and assessing the threat from emerging HABs and toxins, such as the cyanobacterial neurotoxin β-N-methylamino-L-alanine. Conclusion Along the way, many challenges have been encountered that are common to the OHH Centers and also echo those of the wider HAB community. Long-term field data and basic biological information are needed to develop accurate models. Sensor development is hindered by the lack of simple and rapid assays for algal cells and especially toxins. It is also critical to adequately understand the human health effects of HAB toxins. Currently, we understand best the effects of acute