Science.gov

Sample records for alexandrium catenella whedon

  1. Geographic structure evidenced in the toxic dinoflagellate Alexandrium pacificum Litaker (A. catenella - group IV (Whedon & Kofoid) Balech) along Japanese and Chinese coastal waters.

    PubMed

    Genovesi, Benjamin; Berrebi, Patrick; Nagai, Satoshi; Reynaud, Nathalie; Wang, Jinhui; Masseret, Estelle

    2015-09-15

    The intra-specific diversity and genetic structure within the Alexandrium pacificum Litaker (A. catenella - Group IV) populations along the Temperate Asian coasts, were studied among individuals isolated from Japan to China. The UPGMA dendrogram and FCA revealed the existence of 3 clusters. Assignment analysis suggested the occurrence of gene flows between the Japanese Pacific coast (cluster-1) and the Chinese Zhejiang coast (cluster-2). Human transportations are suspected to explain the lack of genetic difference between several pairs of distant Japanese samples, hardly explained by a natural dispersal mechanism. The genetic isolation of the population established in the Sea of Japan (cluster-3) suggested the existence of a strong ecological and geographical barrier. Along the Pacific coasts, the South-North current allows limited exchanges between Chinese and Japanese populations. The relationships between Temperate Asian and Mediterranean individuals suggested different scenario of large-scale dispersal mechanisms. PMID:26188429

  2. Parallel Analyses of Alexandrium catenella Cell Concentrations and Shellfish Toxicity in the Puget Sound▿

    PubMed Central

    Dyhrman, Sonya T.; Haley, Sheean T.; Borchert, Jerry A.; Lona, Bob; Kollars, Nicole; Erdner, Deana L.

    2010-01-01

    Alexandrium catenella is widespread in western North America and produces a suite of potent neurotoxins that cause paralytic shellfish poisoning (PSP) in humans and have deleterious impacts on public health and economic resources. There are seasonal PSP-related closures of recreational and commercial shellfisheries in the Puget Sound, but the factors that influence cell distribution, abundance, and relationship to paralytic shellfish toxins (PSTs) in this system are poorly described. Here, a quantitative PCR assay was used to detect A. catenella cells in parallel with state shellfish toxicity testing during the 2006 bloom season at 41 sites from April through October. Over 500,000 A. catenella cells liter−1 were detected at several stations, with two main pulses of cells driving cell distribution, one in June and the other in August. PSTs over the closure limit of 80 μg of PST 100 per g of shellfish tissue were detected at 26 of the 41 sites. Comparison of cell numbers and PST data shows that shellfish toxicity is preceded by an increase in A. catenella cells in 71% of cases. However, cells were also observed in the absence of PSTs in shellfish, highlighting the complex relationship between A. catenella and the resulting shellfish toxicity. These data provide important information on the dynamics of A. catenella cells in the Puget Sound and are a first step toward assessing the utility of plankton monitoring to augment shellfish toxicity testing in this system. PMID:20495054

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

    PubMed

    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

  4. 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

  5. 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.

  6. 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

  7. 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

  8. Enzymatic digestive activity and absorption efficiency in Tagelus dombeii upon Alexandrium catenella exposure

    NASA Astrophysics Data System (ADS)

    Fernández-Reiriz, M. J.; Navarro, J. M.; Cisternas, B. A.; Babarro, J. M. F.; Labarta, U.

    2013-12-01

    We analyzed absorption efficiency (AE) and digestive enzyme activity (amylase, cellulase complex, and laminarinase) of the infaunal bivalve Tagelus dombeii originating from two geographic sites, Corral-Valdivia and Melinka-Aysén, which have different long-term paralytic shellfish poisoning (PSP) exposure rates. We report the effects of past feeding history (origin) on T. dombeii exposed to a mixed diet containing the toxic dinoflagellate Alexandrium catenella and another dinoflagellate-free control diet over a 12-day period in the laboratory. Absorption efficiency values of T. dombeii individuals that experienced PSP exposure in their habitat (Melinka-Aysén) remained unchanged during exposure to toxic food in the laboratory. In contrast, T. dombeii from a non-PSP exposure field site (Corral-Valdivia) showed a significant reduction in AE with toxic exposure time. This study established that the amylase and cellulase complexes were the most important enzymes in the digestive glands of Tagelus from both sites. The temporal evolution of enzymatic activity under toxic diet was fitted to exponential (amylase and cellulase) and to a logarithmic (laminarinase) models. In all fits, we found significant effect of origin in the model parameters. At the beginning of the experiment, higher enzymatic activity was observed for clams from Corral-Valdivia. The amylase activity decreased with time exposure for individuals from Corral and increased for individuals from Melinka. Cellulase activity did not vary over time for clams from Corral, but increased for individuals from Melinka and laminarinase activity decreased over time for individuals from Corral and remained unchanged over time for Melinka. A feeding history of exposure to the dinoflagellate A. catenella was reflected in the digestive responses of both T. dombeii populations.

  9. Gene expression and molecular evolution of sxtA4 in a saxitoxin producing dinoflagellate Alexandrium catenella.

    PubMed

    Wiese, Maria; Murray, Shauna A; Alvin, Alfonsus; Neilan, Brett A

    2014-12-15

    Dinoflagellates of the genus Alexandrium produce the neurotoxin saxitoxin (STX), responsible for paralytic shellfish poisoning (PSP) and accumulates in marine invertebrates. The recent identification of STX biosynthesis genes allowed us to investigate the expression of sxtA4 at different growth stages in Alexandrium catenella Group IV. We found no significant differences in expression of sxtA4, despite significant differences in STX levels at different growth stages (P < 0.023). Three reference genes were tested for normalisation: actin, cytochrome b (cob), and the large subunit ribosomal RNA (LSU rDNA). cob was most stably expressed but the combination of two reference genes, actin and cob, resulted in the best stability factor. Most genomic sequences of sxtA4 from A. catenella were in a clade that included sequences from Alexandrium fundyense Group I, however, one paralogue was not related to the others, suggesting recombination or lateral transfer. A comparison of the sxtA4 cDNA sequences with genomic DNA sequences indicated the possibility of transcript editing and the preferential transcription of certain genomic DNA loci. The results show that, in dinoflagellates, post-transcriptional mechanisms play a major role in the regulation of saxitoxin biosynthesis. PMID:25301480

  10. Transcriptome de novo assembly sequencing and analysis of the toxic dinoflagellate Alexandrium catenella using the Illumina platform.

    PubMed

    Zhang, Shu; Sui, Zhenghong; Chang, Lianpeng; Kang, Kyoungho; Ma, Jinhua; Kong, Fanna; Zhou, Wei; Wang, Jinguo; Guo, Liliang; Geng, Huili; Zhong, Jie; Ma, Qingxia

    2014-03-10

    In this article, high-throughput de novo transcriptomic sequencing was performed in Alexandrium catenella, which provided the first view of the gene repertoire in this dinoflagellate based on next-generation sequencing (NGS) technologies. A total of 118,304 unigenes were identified with an average length of 673bp (base pair). Of these unigenes, 77,936 (65.9%) were annotated with known proteins based on sequence similarities, among which 24,149 and 22,956 unigenes were assigned to gene ontology categories (GO) and clusters of orthologous groups (COGs), respectively. Furthermore, 16,467 unigenes were mapped onto 322 pathways using the Kyoto Encyclopedia of Genes and Genomes Pathway database (KEGG). We also detected 1143 simple sequence repeats (SSRs), in which the tri-nucleotide repeat motif (69.3%) was the most abundant. The genetic facts and significance derived from the transcriptome dataset were suggested and discussed. All four core nucleosomal histones and linker histones were detected, in addition to the unigenes involved in histone modifications.190 unigenes were identified as being involved in the endocytosis pathway, and clathrin-dependent endocytosis was suggested to play a role in the heterotrophy of A. catenella. A conserved 22-nt spliced leader (SL) was identified in 21 unigenes which suggested the existence of trans-splicing processing of mRNA in A. catenella. PMID:24440238

  11. Identification of microRNAs in the Toxigenic Dinoflagellate Alexandrium catenella by High-Throughput Illumina Sequencing and Bioinformatic Analysis

    PubMed Central

    Geng, Huili; Sui, Zhenghong; Zhang, Shu; Du, Qingwei; Ren, Yuanyuan; Liu, Yuan; Kong, Fanna; Zhong, Jie; Ma, Qingxia

    2015-01-01

    Micro-ribonucleic acids (miRNAs) are a large group of endogenous, tiny, non-coding RNAs consisting of 19–25 nucleotides that regulate gene expression at either the transcriptional or post-transcriptional level by mediating gene silencing in eukaryotes. They are considered to be important regulators that affect growth, development, and response to various stresses in plants. Alexandrium catenella is an important marine toxic phytoplankton species that can cause harmful algal blooms (HABs). To date, identification and function analysis of miRNAs in A. catenella remain largely unexamined. In this study, high-throughput sequencing was performed on A. catenella to identify and quantitatively profile the repertoire of small RNAs from two different growth phases. A total of 38,092,056 and 32,969,156 raw reads were obtained from the two small RNA libraries, respectively. In total, 88 mature miRNAs belonging to 32 miRNA families were identified. Significant differences were found in the member number, expression level of various families, and expression abundance of each member within a family. A total of 15 potentially novel miRNAs were identified. Comparative profiling showed that 12 known miRNAs exhibited differential expression between the lag phase and the logarithmic phase. Real-time quantitative RT-PCR (qPCR) was performed to confirm the expression of two differentially expressed miRNAs that were one up-regulated novel miRNA (aca-miR-3p-456915), and one down-regulated conserved miRNA (tae-miR159a). The expression trend of the qPCR assay was generally consistent with the deep sequencing result. Target predictions of the 12 differentially expressed miRNAs resulted in 1813target genes. Gene ontology (GO) analysis and the Kyoto Encyclopedia of Genes and Genomes pathway database (KEGG) annotations revealed that some miRNAs were associated with growth and developmental processes of the alga. These results provide insights into the roles that miRNAs play in the growth of

  12. Isolation of an algicide from a marine bacterium and its effects against the toxic dinoflagellate Alexandrium catenella and other harmful algal bloom species.

    PubMed

    Kim, Yun Sook; Son, Hong-Joo; Jeong, Seong-Yun

    2015-08-01

    The aim of this study was to isolate and identify bacteria demonstrating an algicidal effect against Alexandrium catenella and to determine the activity and range of any algicide discovered. The morphological and biochemical attributes of an algicidal bacterium, isolate YS-3, and analysis of its 16S rRNA gene sequence revealed it to be a member of the genus Brachybacterium. This organism, designated Brachybacterium sp. YS-3, showed the greatest effect against A. catenella cells of all bacteria isolated, and is assumed to produce secondary metabolites. When 10% solutions of culture filtrates from this strain were applied to A. catenella cultures, over 90% of cells were killed within 9 h. Bioassay-guided isolation of the algicide involved led to the purification and identification of an active compound. Based on physicochemical and spectroscopic data, including nuclear magnetic resonance and mass analyses, this compound was identified as 1-acetyl-β-carboline. This algicide showed significant activity against A. catenella and a wide range of harmful algal bloom (HAB)-forming species. Taken together, our results suggest that Brachybacterium sp. YS-3 and its algicide represent promising candidates for use in HAB control. PMID:26224453

  13. 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

  14. 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

  15. 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

    2014-09-01

    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

  16. 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

  17. Influence of Environmental Factors on the Paralytic Shellfish Toxin Content and Profile of Alexandrium catenella (Dinophyceae) Isolated from the Mediterranean Sea

    PubMed Central

    Laabir, Mohamed; Collos, Yves; Masseret, Estelle; Grzebyk, Daniel; Abadie, Eric; Savart, Véronique; Sibat, Manoella; Amzil, Zouher

    2013-01-01

    Laboratory experiments were designed to study the toxin content and profile of the Alexandrium catenella strain ACT03 (isolated from Thau Lagoon, French Mediterranean) in response to abiotic environmental factors under nutrient-replete conditions. This dinoflagellate can produce various paralytic shellfish toxins with concentrations ranging from 2.9 to 50.3 fmol/cell. The toxin profile was characterized by carbamate toxins (GTX3, GTX4 and GTX5) and N-sulfocarbamoyl toxins (C1, C2, C3 and C4). C2 dominated at 12–18 °C, but only for salinities ranging from 10 to 25 psu, whereas GTX5 became dominant at temperatures ranging from 21 to 30 °C at almost all salinities. There was no significant variation in the cellular toxin amount from 18 °C to 27 °C for salinities ranging between 30 and 40 psu. At salinities of 10 to 25 psu, the toxin concentrations always remained below 20 fmol/cell. Toxin content was stable for irradiance ranging from 10 to 70 μmol photons/m2/s then slightly increased. Overall, the toxin profile was more stable than the toxin content (fmol/cell), except for temperature and/or salinity values different from those recorded during Alexandrium blooms in Thau Lagoon. PMID:23676417

  18. Accumulation and depuration profiles of PSP toxins in the short-necked clam Tapes japonica fed with the toxic dinoflagellate Alexandrium catenella.

    PubMed

    Samsur, Mohamad; Yamaguchi, Yasunaga; Sagara, Takefumi; Takatani, Tomohiro; Arakawa, Osamu; Noguchi, Tamao

    2006-09-01

    A toxic dinoflagellate responsible for paralytic shellfish poisoning (PSP), Alexandrium catenella (Ac) was fed to the short-necked clam Tapes japonica, and the accumulation and depuration profiles of PSP toxins were investigated by means of high-performance liquid chromatography with postcolumn fluorescence derivatization (HPLC-FLD). The short-necked clams ingested more than 99% of the Ac cells (4 x 10(7)cells) supplied once at the beginning of experiment, and accumulated a maximal amount of toxin (185 nmol/10 clams) after 12h. The rate of toxin accumulation at that time was 23%, which rapidly decreased thereafter. Composition of the PSP toxin accumulated in the clams obviously different from that of Ac even 0.5h after the cell supply, the proportion of C1+2 being much higher than in Ac, although the reason remains to be elucidated. In contrast, a higher ratio of gonyautoxin (GTX)1+4 than in Ac was detected in the toxin profiles of clam excrements. The variation in toxin composition derived presumably from the transformation of toxin analogues in clams was observed from 0.5h, such as reversal of the ratio of C1 to C2, and appearance of carbamate (saxitoxin (STX), neoSTX and GTX2, 3) and decarbamoyl (dc) derivatives (dcSTX and dcGTX2, 3), which were undetectable in Ac cells. The total amount of toxin distributed over Ac cells, clams and their excrements gradually declined, and only 1% of supplied toxin was detected at the end of experiment. PMID:16887162

  19. 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

  20. Toxin variability in cultured and natural populations of Alexandrium tamarense from southern South America - Evidences of diversity and environmental regulation.

    PubMed

    Montoya, Nora G; Fulco, Vanesa K; Carignan, Mario O; Carreto, José I

    2010-12-01

    Cell content and composition of paralytic shellfish toxins of 10 cultured strains and 6 natural populations of Alexandrium tamarense from the Argentine sea, were analyzed. These data were compiled with previously published data into a comprehensive view of the toxin composition of the complex A. tamarense/Alexandrium catenella from southern South America. The N-sulfocarbamoyl derivatives C1,2 were predominant in almost all the cultured strains. The second major derivatives were GTX1,4, although the GTX1,4/C1,2 ratio varied largely. Some strains contain relatively high amounts of GTX2,3 (up to 29%) and/or neoSTX (up to 24%). In all strains STX was a minor component (0-4.4%) whereas GTX5 was present only in Alexandrium catenella isolates. Similarity analysis based upon toxin profiles showed that cultured strains from Argentine, Brazil, Chile and Uruguay clustered together. However, whereas some strains from the same geographic area exhibited different toxin profiles, and consistently fell out in separate subgroups, strains from Chile are grouped in a unique subgroup. In contrast to cultured strains, C1,2 were minor components among field populations. The highly toxic GTX1,4 were predominant in all spring field populations (69.1-93.6%). Moreover, their toxin cell content (163.9-261.4 fmol cell(-1)) and toxicity (68.2-93.0 pg STX equiv. cell(-1)) were several times higher that showed by the cultured strains. Field populations are more closely related to one another than to the cultured strains. However a less toxic and morphologically distinctive autumn population, contained GTX2,3 as the quasi unique (88.5%) toxin derivative clustered separately. Variability in toxin content and composition of A. tamarense field populations were well correlated with in situ temperature and nitrate concentration. Whereas toxin cell content and GTX1,4 (mol %) increased following saturation functions, GTX2,3 (mol %) decrease exponentially with the increase of the in situ nitrate

  1. WITHDRAWN: Gene expression and molecular evolution of sxtA4 in a saxitoxin producing dinoflagellate Alexandrium catenella.

    PubMed

    Wiese, Maria; Murray, Shauna A; Alvin, Alfonsus; Neilan, Brett A

    2014-07-28

    This article has been withdrawn at the request of the authors and editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID:25080311

  2. [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. PMID:17465118

  3. 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

  4. Paralytic Shellfish Toxins in Protogonyaulax tamarensis and Protogonyaulax catenella in Axenic Culture 1

    PubMed Central

    Boczar, Barbara A.; Beitler, Mark K.; Liston, John; Sullivan, John J.; Cattolico, Rose Ann

    1988-01-01

    Paralytic shellfish toxin concentrations were measured and individual toxin profiles were monitored in axenic batch cultures of Protogonyaulax tamarensis and Protogonyaulax catenella. High pressure liquid chromatographic methods were used that allowed the separation of all 12 known paralytic shellfish poisons, including toxins C1, C2, and C3, from a single sample. In isolates of both Protogonyaulax species, total toxin levels were relatively low after inoculation, increased rapidly in early to mid-exponential growth to a value 100 to 300% of that at the initial time point, then decreased by 86 to 95% as the culture aged. Although the concentrations of individual toxins per cell followed the same general pattern as that seen for total moles of toxin per cell, variability in toxin profile with culture age was observed. In P. tamarensis, the mole percent of neosaxitoxin increased substantially from 8 to 44% as total toxin levels per cell decreased. A concomitant decrease in the mole percent of saxitoxin with culture age was noted. Although not as precipitous, changes in the mole percent of specific toxins from P. catenella were also observed. The mole percent of gonyautoxins I and IV increased, while that of gonyautoxins II and III decreased. These data suggest that the toxin profile in isolates of Protogonyaulax can change, sometimes significantly, with changing environmental variables. PMID:16666456

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. Phylogenetic Diversity and Specificity of Bacteria Closely Associated with Alexandrium spp. and Other Phytoplankton

    PubMed Central

    Jasti, Suresh; Sieracki, Michael E.; Poulton, Nicole J.; Giewat, Michael W.; Rooney-Varga, Juliette N.

    2005-01-01

    While several studies have suggested that bacterium-phytoplankton interactions have the potential to dramatically influence harmful algal bloom dynamics, little is known about how bacteria and phytoplankton communities interact at the species composition level. The objective of the current study was to determine whether there are specific associations between diverse phytoplankton and the bacteria that co-occur with them. We determined the phylogenetic diversity of bacterial assemblages associated with 10 Alexandrium strains and representatives of the major taxonomic groups of phytoplankton in the Gulf of Maine. For this analysis we chose xenic phytoplankton cultures that (i) represented a broad taxonomic range, (ii) represented a broad geographic range for Alexandrium spp. isolates, (iii) grew under similar cultivation conditions, (iv) had a minimal length of time since the original isolation, and (v) had been isolated from a vegetative phytoplankton cell. 16S rRNA gene fragments of most Bacteria were amplified from DNA extracted from cultures and were analyzed by denaturing gradient gel electrophoresis and sequencing. A greater number of bacterial species were shared by different Alexandrium cultures, regardless of the geographic origin, than by Alexandrium species and nontoxic phytoplankton from the Gulf of Maine. In particular, members of the Roseobacter clade showed a higher degree of association with Alexandrium than with other bacterial groups, and many sequences matched sequences reported to be associated with other toxic dinoflagellates. These results provide evidence for specificity in bacterium-phytoplankton associations. PMID:16000752

  13. Distribution of Alexandrium fundyense and A. pacificum (Dinophyceae) in the Yellow Sea and Bohai Sea.

    PubMed

    Gao, Yan; Yu, Ren-Cheng; Chen, Jian-Hua; Zhang, Qing-Chun; Kong, Fan-Zhou; Zhou, Ming-Jiang

    2015-07-15

    This study characterizes the distribution of two closely related, causative species of paralytic shellfish poisoning – Alexandrium fundyense and A. pacificum – within the Yellow Sea (YS) and Bohai Sea (BS). These two Alexandrium species are distinguished for the first time in a regional field study using species-specific, quantitative PCR (qPCR) based assays. Both qPCR assays target the large subunit ribosomal DNA gene and were used to analyze net-concentrated phytoplankton samples collected in May 2012. A. fundyense was mainly distributed in YS, while A. pacificum was confined to an area adjacent to the Changjiang River estuary. The different distribution of the two species is interpreted as evidence of their distinct bloom ecology. Expanded efforts implementing these assays offer the ability to discriminate the dynamics of A. fundyense and A. pacificum blooms and provide a more sound basis for monitoring toxic Alexandrium species in this region. PMID:26026250

  14. Effects of temperature and salinity on the growth of Alexandrium (Dinophyceae) isolates from the Salish Sea.

    PubMed

    Bill, Brian D; Moore, Stephanie K; Hay, Levi R; Anderson, Donald M; Trainer, Vera L

    2016-04-01

    Toxin-producing blooms of dinoflagellates in the genus Alexandrium have plagued the inhabitants of the Salish Sea for centuries. Yet the environmental conditions that promote accelerated growth of this organism, a producer of paralytic shellfish toxins, is lacking. This study quantitatively determined the growth response of two Alexandrium isolates to a range of temperatures and salinities, factors that will strongly respond to future climate change scenarios. An empirical equation, derived from observed growth rates describing the temperature and salinity dependence of growth, was used to hindcast bloom risk. Hindcasting was achieved by comparing predicted growth rates, calculated from in situ temperature and salinity data from Quartermaster Harbor, with corresponding Alexandrium cell counts and shellfish toxin data. The greatest bloom risk, defined at μ >0.25 d(-1) , generally occurred from April through November annually; however, growth rates rarely fell below 0.10 d(-1) . Except for a few occasions, Alexandrium cells were only observed during the periods of highest bloom risk and paralytic shellfish toxins above the regulatory limit always fell within the periods of predicted bloom occurrence. While acknowledging that Alexandrium growth rates are affected by other abiotic and biotic factors, such as grazing pressure and nutrient availability, the use of this empirical growth function to predict higher risk time frames for blooms and toxic shellfish within the Salish Sea provides the groundwork for a more comprehensive biological model of Alexandrium bloom dynamics in the region and will enhance our ability to forecast blooms in the Salish Sea under future climate change scenarios. PMID:27037588

  15. 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.

  16. Georges Bank: a leaky incubator of Alexandrium fundyense blooms

    PubMed Central

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

    2012-01-01

    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. PMID:24976691

  17. 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.

  18. 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...

  19. 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 ...

  20. 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...

  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. 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

  3. 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. PMID:26032975

  4. An Alexandrium Spp. Cyst Record from Sequim Bay, Washington State, USA, and its Relation to Past Climate Variability(1).

    PubMed

    Feifel, Kirsten M; Moore, Stephanie K; Horner, Rita A

    2012-06-01

    Since the 1970s, Puget Sound, Washington State, USA, has experienced an increase in detections of paralytic shellfish toxins (PSTs) in shellfish due to blooms of the harmful dinoflagellate Alexandrium. Natural patterns of climate variability, such as the Pacific Decadal Oscillation (PDO), and changes in local environmental factors, such as sea surface temperature (SST) and air temperature, have been linked to the observed increase in PSTs. However, the lack of observations of PSTs in shellfish prior to the 1950s has inhibited statistical assessments of longer-term trends in climate and environmental conditions on Alexandrium blooms. After a bloom, Alexandrium cells can enter a dormant cyst stage, which settles on the seafloor and then becomes entrained into the sedimentary record. In this study, we created a record of Alexandrium spp. cysts from a sediment core obtained from Sequim Bay, Puget Sound. Cyst abundances ranged from 0 to 400 cysts · cm(-3) and were detected down-core to a depth of 100 cm, indicating that Alexandrium has been present in Sequim Bay since at least the late 1800s. The cyst record allowed us to statistically examine relationships with available environmental parameters over the past century. Local air temperature and sea surface temperature were positively and significantly correlated with cyst abundances from the late 1800s to 2005; no significant relationship was found between PDO and cyst abundances. This finding suggests that local environmental variations more strongly influence Alexandrium population dynamics in Puget Sound when compared to large-scale changes. PMID:27011070

  5. Inhibition of the growth of Alexandrium tamarense by algicidal substances in Chinese fir (Cunninghamia lanceolata).

    PubMed

    Yang, Wei-Dong; Liu, Jie-Sheng; Li, Hong-Ye; Zhang, Xin-Lian; Qi, Yu-Zao

    2009-10-01

    The wood sawdust from Chinese fir (Cunninghamia lanceolata) exhibited stronger inhibition on the growth of Alexandrium tamarense than those from alder (Alnus cremastogyne), pine (Pinus massoniana), birch (Betula alnoides) and sapele (Entandrophragma cylindricum). The water extract, acetone-water extract and essential oil from fir sawdust were all shown to inhibit the growth of A. tamarense. The inhibition of fir essential oil was the strongest among all the above wood sources while the half effective concentration was only 0.65 mg/L. These results suggested that the fir essential oil may play an important role in the algicidal effect of Chinese fir. PMID:19634014

  6. 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. PMID:26841307

  7. 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

  8. Two-dimensional electrophoresis analysis of proteins extracted from Alexandrium sp. LC3

    NASA Astrophysics Data System (ADS)

    Li, Hao; Miao, Jinlai; Cui, Fengxia; Li, Guangyou

    2007-10-01

    Two-dimensional electrophoresis(2-DE) of protein extracted and purified from Alexandrium sp. LC3 was conducted. In the SDS-PAGE study, the relative molecular weights of the proteins were mainly in the range of 14kDa-31kDa and 43kDa-66kDa, and more proteins were detected between 14kDa and 31kDa. With the improved protein preparation, the two-dimensional electrophoresis patterns indicated that the relative molecular weights of the proteins were between 14kDa and 100kDa, and most of them ranged from 14kDa to 31kDa. This was consistent with the result of the SDS-PAGE analysis. The isoelectric points were found to lie between 3.0 and 8.0, and most of them were in the range of 3.0 6.0. Better separation effect was acquired with pre-prepared immobilized gradient (IPG) strip (pH3 5.6), and about 320 protein spots could be visualized on the 2-DE map by staining. Within pH3 10 and pH3 5.6 strips, the protein samples of Alexandrium sp. LC3 could be separated well.

  9. First Report of Parasitism on the Toxic Dinoflagellate Alexandrium minutum Halim

    NASA Astrophysics Data System (ADS)

    Erard-Le Denn, E.; Chrétiennot-Dinet, M.-J.; Probert, I.

    2000-01-01

    Blooms of the toxic red tide phytoplankton Alexandrium minutum Halim (Dinophyceae) have frequently occurred during recent years in the estuaries of northern Brittany, France. Some months after a bloom in the Penzé River in 1997, many Alexandrium cells in samples maintained in the dark at 14 °C were observed to be infected by the sporocysts of an unknown parasite, which, upon exposure to increased light intensity and temperature, ejected many small biflagellate zoospores. The parasite was found to infect laboratory cultures of several other dinoflagellate species, and estimates of parasite-induced mortality indicate that this parasite is capable of removing a significant fraction of dinoflagellate biomass in a short time, raising the possibility of its use as a biological control agent of toxic dinoflagellate blooms. The effect of this parasite on natural A. minutum populations remains, however, to be estimated. This paper presents video images used in a preliminary identification and life cycle elucidation of the parasite, which may be affiliated with the Apicomplexan complex.

  10. 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.

  11. 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. PMID:25431527

  12. 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

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

    PubMed Central

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

    2016-01-01

    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. PMID:27181871

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

    PubMed

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

    2012-01-01

    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

  15. 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

  16. 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. PMID:24927989

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

    PubMed

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

    2016-01-01

    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. PMID:27181871

  18. 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

  19. 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

    2015-10-01

    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

  20. [Seasonal dynamics of genus Alexandrium (potentially toxic dinoflagellate) in the lagoon of Bizerte (North of Tunisia) and controls by the abiotic factors].

    PubMed

    Bouchouicha Smida, Donia; Sahraoui, Inès; Mabrouk, Hassine Hadj; Sakka Hlaili, Asma

    2012-06-01

    Some species of the genus Alexandrium are known as potential producers of saxitoxin, a neurotoxin that causes the paralytic shellfish poisoning (PSP) syndrome. Blooming of these species, especially in shellfish farms can affect the aquaculture production and harm human health. Seasonal dynamics of Alexandrium spp. abundance in relationship to environmental factors was investigated from November 2007 to February 2009 at six stations in the Bizerte lagoon, an important shellfish farming area situated in SW Mediterranean. The sampling stations represented different hydrological and trophic conditions: one station TJ (Tinja) is affected by the river plume; two stations (Chaara [Ch] and Canal [Ca]) are influenced by marine inflow (particularly in summer), industrial and urban effluents; and the three other stations (Menzel Abdelrahmen [MA], Menzel Jemil [MJ] and Douaouda [Do]) are located close to shellfish farms. Cell abundance of Alexandrium spp. varied among stations and months. Species of this genus showed a sporadic appearance, but they reached high concentration (0.67-7 × 10(5)cells L(-1)). Maximal cell density was detected in autumn (November 2007; station MA), at salinity of 37.5, temperature of 16 °C and NH(4)(+) level of 55.45 μM. During this month, Alexandrium spp. abundance accounted for a large fraction (61%) of the harmful phytoplankton. The statistical analysis revealed that Alexandrium concentrations were positively correlated with N:P ratio and NH4+ levels. Thus, the eutrophic waters of the lagoon favour the growth of Alexandrium, which seemed to have preference for N-nutrient loading from antrophogenic activities, as ammonium. Blooms of these potential harmful algae may constitute a potential threat in this coastal lagoon of the southern Mediterranean. Consequently, it is necessary to be well vigilant and to do regular monitoring of Alexandrium species. PMID:22721562

  1. First report of goniodomin A production by the dinoflagellate Alexandrium pseudogonyaulax developing in southern Mediterranean (Bizerte Lagoon, Tunisia).

    PubMed

    Zmerli Triki, Habiba; Laabir, Mohamed; Moeller, Peter; Chomérat, Nicolas; Kéfi Daly-Yahia, Ons

    2016-03-01

    The dinoflagellate Alexandrium pseudogonyaulax is widely distributed around the world including the Mediterranean waters. The objectives of this study were to determine the morphology and phylogenic affiliation of A. pseudogonyaulax strain isolated from Bizerte Lagoon (Mediterranean waters, Tunisia) and investigate its toxicity. Molecular analyses confirmed the morphological identification of the isolated strain (APBZ12) as A. pseudogonyaulax. Moreover, it showed that it is 100% identical with strains of this species found in New Zealand, Japan, China and North Sea (Norway and Denmark) suggesting that this species is cosmopolitan. Until now, no toxin studies have been conducted on fully characterized (morphologically and molecularly) A. pseudogonyaulax. Cellular toxin production was determined using high pressure liquid chromatography coupled to mass spectrometry (HPLC/MS). Results showed for the first time that A. pseudogonyaulax contains goniodomin A (GDA), a highly toxic macrolide polyether previously shown to be produced by two other dinoflagellate species Alexandrium monilatum (Hsia et al., 2006) and Alexandrium hiranoi (erroneously identified as A. pseudogonyaulax in Murakami et al., 1988) in American and Japanese waters, respectively. This biologically active toxin has been associated over decades with fish mortality. Our study showed that the cell extracts of APBZ12 showed an important bioactivity using GH4C1 rat pituitary cytotoxicity bioassay. PMID:26748155

  2. 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. PMID:26592649

  3. 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.

  4. The sxt Gene and Paralytic Shellfish Poisoning Toxins as Markers for the Monitoring of Toxic Alexandrium Species Blooms.

    PubMed

    Penna, Antonella; Perini, Federico; Dell'Aversano, Carmela; Capellacci, Samuela; Tartaglione, Luciana; Giacobbe, Maria Grazia; Casabianca, Silvia; Fraga, Santiago; Ciminiello, Patrizia; Scardi, Michele

    2015-12-15

    Paralytic shellfish poisoning (PSP) is a serious human illness caused by the ingestion of seafood contaminated with saxitoxin and its derivatives (STXs). These toxins are produced by some species of marine dinoflagellates within the genus Alexandrium. In the Mediterranean Sea, toxic Alexandrium spp. blooms, especially of A. minutum, are frequent and intense with negative impact to coastal ecosystem, aquaculture practices and other economic activities. We conducted a large scale study on the sxt gene and toxin distribution and content in toxic dinoflagellate A. minutum of the Mediterranean Sea using both quantitative PCR (qPCR) and HILIC-HRMS techniques. We developed a new qPCR assay for the estimation of the sxtA1 gene copy number in seawater samples during a bloom event in Syracuse Bay (Mediterranean Sea) with an analytical sensitivity of 2.0 × 10° sxtA1 gene copy number per reaction. The linear correlation between sxtA1 gene copy number and microalgal abundance and between the sxtA1 gene and STX content allowed us to rapidly determine the STX-producing cell concentrations of two Alexandrium species in environmental samples. In these samples, the amount of sxtA1 gene was in the range of 1.38 × 10(5) - 2.55 × 10(8) copies/L and the STX concentrations ranged from 41-201 nmol/L. This study described a potential PSP scenario in the Mediterranean Sea. PMID:26580419

  5. 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.

  6. Isolation of bacteria from toxic dinoflagellate Alexandrium minutum and their effects on algae toxicity.

    PubMed

    Lu, Y H; Chai, T J; Hwang, D F

    2000-11-01

    Attempts were made to isolate the bacteria from toxic dinoflagellate Alexandrium minutum T1 and to study the effect of these bacteria on the growth and toxicity of A. minutum T1. It was found that intracellular bacterial species including Pasteurella haemolytica, Pseudomonas vesicularis, and Sphingomonas sp., and extracellular bacterial species including Pasteurolla pneumotropica, Morganella wisconsensis, Flavobacterium oryzihabitans, Pseudomonas pseudomallei, and Sphingomonas sp. All of them were cultured and determined to have non-PSP-producing ability. The maximum cell number of A. minutum cultured without isolated bacteria was higher than that cultured with isolated bacteria. The total toxicity of A. minutum cultured with bacteria was similar to that of A. minutum T1 cultured without bacteria from lag phase to stationary phase, but it was lower after stationary phase. The growth of A. minutum T1 cultured without antibiotics was also better than that cultured with antibiotics. The total toxicity of A. minutum cultured without antibiotics was higher than that of A. minutum cultured with antibiotics. However, the cell toxicity of A. minutum did not decrease even if the culture medium was added with antibiotics. PMID:11126517

  7. 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.

  8. Biotransformation of mycosporine like amino acids (MAAs) in the toxic dinoflagellate Alexandrium tamarense.

    PubMed

    Callone, Alan I; Carignan, Mario; Montoya, Nora G; Carreto, José I

    2006-09-01

    Changes in mycosporine-like amino acids (MAAs) induced by the increase of photosynthetically active radiation (PAR) were studied in the toxic dinoflagellate Alexandrium tamarense. Cultures of A. tamarense were maintained at exponential growth under low (25 micromol quanta m(-2)s(-1)) PAR irradiance. The cultures were nutrient enriched and one day later exposed to higher irradiance (150 micromol quanta m(-2)s(-1)). The content of MAAs was determined by means of high performance liquid chromatography (HPLC). Eleven MAAs, including some partially characterized compounds, were identified. The MAAs synthesis induction can be described as a two-stage process. The first one involves the net synthesis of the MAAs bi-substituted by amino acids. In the second stage these compounds were transformed into other secondary MAAs. The two most prominent changes were observed in the concentration of porphyra-334 and palythene. The cellular concentration of porphyra-334 increased during the first 2h of exposure to higher irradiance and then decreased rapidly. In contrast, the cellular concentration of palythene showed a continuous accumulation since the beginning of the exposure. In A. tamarense the main route of MAAs transformation has porphyra-334 as a precursor of a sequential conversion resulting in the accumulation of palythene. PMID:16697653

  9. Divinyl chlorophyll a in the marine eukaryotic protist Alexandrium ostenfeldii (Dinophyceae).

    PubMed

    Rodríguez, Francisco; Garrido, José Luis; Sobrino, Cristina; Johnsen, Geir; Riobó, Pilar; Franco, José; Aamot, Inga; Ramilo, Isabel; Sanz, Noelia; Kremp, Anke

    2016-02-01

    Here it is reported the first detection of DV-chl a together with the usual chl a in the marine dinoflagellate Alexandrium ostenfeldii from the Baltic Sea. Growth response and photosynthetic parameters were examined at two irradiances (80 and 240 μmol photons m(-2) s(-1)) and temperatures (15 °C and 19 °C) in a divinylic strain (AOTV-OS20) versus a monovinylic one (AOTV-OS16), using in vivo chl a fluorescence kinetics of PSII to characterize photosynthetic parameters by pulse amplitude modulated fluorescence, (14)C assimilation rates and toxin analyses. The divinylic isolate exhibited slower growth and stronger sensitivity to high irradiance than normal chl a strain. DV-chl a : chl a ratios decreased along time (from 11.3 to < 0.5 after 10 months) and to restore them sub-cloning and selection of strains with highest DV-chl a content was required. A mutation and/or epigenetic changes in the expression of divinyl reductase gene/s in A. ostenfeldii may explain this altered pigment composition. Despite quite severe limitations (reduced fitness and gradual loss of DV-chl a content), the DV-chl a-containing line in A. ostenfeldii could provide a model organism in photosynthetic studies related with chl biosynthesis and evolution. PMID:26337730

  10. 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

  11. 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.

  12. Diversity and dynamics of a widespread bloom of the toxic dinoflagellate Alexandrium fundyense.

    PubMed

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

    2011-01-01

    Historically, cosmopolitan phytoplankton species were presumed to represent largely unstructured populations. However, the recent development of molecular tools to examine genetic diversity have revealed differences in phytoplankton taxa across geographic scales and provided insight into the physiology and ecology of blooms. Here we describe the genetic analysis of an extensive bloom of the toxic dinoflagellate Alexandrium fundyense that occurred in the Gulf of Maine in 2005. This bloom was notable for its intensity and duration, covering hundreds of kilometers and persisting for almost two months. Genotypic analyses based on microsatellite marker data indicate that the open waters of the northeastern U.S. harbor a single regional population of A. fundyense comprising two genetically distinct sub-populations. These subpopulations were characteristic of early- and late-bloom samples and were derived from the northern and southern areas of the bloom, respectively. The temporal changes observed during this study provide clear evidence of succession during a continuous bloom and show that selection can act on the timescale of weeks to significantly alter the representation of genotypes within a population. The effects of selection on population composition and turnover would be magnified if sexual reproduction were likewise influenced by environmental conditions. We hypothesize that the combined effects of differential growth and reproduction rates serves to reduce gene flow between the sub-populations, reinforcing population structure while maintaining the diversity of the overall regional population. PMID:21829565

  13. 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

  14. 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. PMID:25170191

  15. Initial observations of the 2005 Alexandrium fundyense bloom in southern New England: General patterns and mechanisms

    NASA Astrophysics Data System (ADS)

    Anderson, Donald M.; Keafer, Bruce A.; McGillicuddy, Dennis J.; Mickelson, Michael J.; Keay, Kenneth E.; Scott Libby, P.; Manning, James P.; Mayo, Charles A.; Whittaker, David K.; Michael Hickey, J.; He, Ruoying; Lynch, Daniel R.; Smith, Keston W.

    2005-09-01

    From May to July, 2005, an extensive bloom of Alexandrium fundyense occurred along the coast of southern New England. The outbreak eventually closed shellfish beds from central Maine to Massachusetts, including Nantucket Island and portions of Martha's Vineyard, and resulted in the closure of 40,000 km 2 of offshore federal waters as well. The coastal Alexandrium bloom was exceptional in several ways: high toxin levels were measured farther south than ever before in New England; levels of toxicity in many locations were higher than previously observed at those stations; for the first time toxicity at some locations was above quarantine levels; cell concentrations far exceeded those observed in the coastal waters of southern New England in the past; and for the first time in the region the governors of Maine and Massachusetts officially declared the red tide to be a disaster, clearing the way for federal assistance. Initial observations suggest that several factors contributed to this bloom. Abundant rainfall and heavy snowmelt substantially increased the amount of fresh water entering the Gulf of Maine. Combined with other freshwater inputs, we hypothesize that this provided macro- and micro-nutrients, a stratified water column, and a transport mechanism that led to high cell abundances and broad, region-wide dispersal of the organism. Warm temperatures in western waters also would have favored A. fundyense growth. In addition, several storms with strong winds out of the northeast occurred at times when cells were abundant and in locations where the winds could advect them into Massachusetts and Cape Cod Bays and keep them there, leading to high cell concentrations and toxicity. Another contributing factor may have been the high abundance of newly deposited cysts in western Gulf of Maine sediments, as documented in a fall 2004 survey. Here, we evaluate this bloom and the patterns of toxicity in light of the conceptual models for A. fundyense dynamics developed

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

    PubMed Central

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

    2012-01-01

    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 (Ci) 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. PMID:21593032

  17. Effects of toxic dinoflagellate Alexandrium tamarense on the energy budgets and growth of two marine bivalves.

    PubMed

    Li, Siu-Chung; Wang, Wen-Xiong; Hsieh, Dennis P H

    2002-03-01

    Harmful algal blooms (HAB) may impose a serious threat to aquatic lives and human health. We determined the effects of a toxic dinoflagellate Alexandrium tamarense (clone ATCIO1, isolated from Hong Kong coastal waters) on the energy budget, quantified as scope for growth (SFG), and the growth rate of the manila clam Ruditapes philippinarum and the green-lipped mussel Perna viridis. To quantify the SFG, clams and mussels were dosed with different amounts of toxic A. tamarense for 6 days, resulting in different toxin burdens in the tissues. Clearance rate, absorption efficiency, and respiration rate were subsequently measured in these toxin containing bivalves. Clearance rate significantly declined at the highest toxin burden in the clams only, while there was no significant difference in the clearance rate among different treatments for the mussels. The respiration rate of either bivalve was not significantly affected by toxin accumulation in the tissues. In contrast, the absorption efficiency of both clams and mussels decreased, in a concentration-dependent manner for mussels but not for clams, when the tissue accumulated the toxins. With an increase in paralytic shellfish poisoning (PSP) toxin burden, SFG in both clams and mussels was significantly reduced, primarily because of the decrease of absorption efficiency. The growth rate of juvenile clam R. philippinarum, measured as an increment in tissue dry weight over a 15 d exposure period, was significantly lower during their feeding on toxic dinoflagellate than the growth rate of clams feeding on the diatom Thalassiosira pseudonana. The juvenile mussel P. viridis, however, exhibited similar growth rates after feeding on the toxic dinoflagellates and the nontoxic diatom. This study showed that SFG can provide a sensitive and integrated measure of the effect of HAB on the physiology of bivalves. Clam R. philippinarum may be more sensitive, in terms of their energy budget, to PSP toxin accumulation than the mussel P

  18. 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

  19. 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

  20. Life history, excystment features, and growth characteristics of the Mediterranean harmful dinoflagellate Alexandrium pseudogonyaulax.

    PubMed

    Zmerli Triki, Habiba; Laabir, Mohamed; Kéfi Daly-Yahia, Ons

    2015-10-01

    Studies considering the biology and ecology of the toxic bloom-forming species, Alexandrium pseudogonyaulax, are rare. Our results highlight five features not described before in A. pseudogonyaulax life cycle: (i) A. pseudogonyaulax gametes showed two modes of conjugation, anisogamy and isogamy, (ii) sexual conjugation occurs either in the dark or in the light phase by engulfment or a fusion process, (iii) the presence of planozygote and newly formed cysts in monoclonal culture suggests homothallism, (iv) newly formed cysts have very dark vesicular content and are mostly unparatabulated when observed under light microscope and (v) natural resting cysts are able to give either a planomeiocyte or two vegetative cells. Cyst viability was enhanced after 5 months of cold storage (4°C), with excystment rate reaching 97% after 3 d of incubation. Excystment rate was highest (43%-79%) in Enriched Natural Sea Water diluted culture medium, whereas few germling cells were able to survive without the culture medium (0%-13%). Salinity-irradiance experiments revealed that the highest cell concentrations occur at high irradiances for all the tested salinities. Vegetative growth rates generally increased with increasing irradiance, and were less dependent on salinity variations. The relatively low growth rate, low cell densities in the laboratory, and the notable capacity of producing cysts along growth phases of A. pseudogonyaulax could explain the occurrence of high resting cysts densities in the sediment of Bizerte lagoon and the relatively low abundances of vegetative cells in the water column. PMID:26986892

  1. Glutathione S-Transferase Regulation in Calanus finmarchicus Feeding on the Toxic Dinoflagellate Alexandrium fundyense

    PubMed Central

    Roncalli, Vittoria; Jungbluth, Michelle J.; Lenz, Petra H.

    2016-01-01

    The effect of the dinoflagellate, Alexandrium fundyense, on relative expression of glutathione S-transferase (GST) transcripts was examined in the copepod Calanus finmarchicus. Adult females were fed for 5-days on one of three experimental diets: control (100% Rhodomonas spp.), low dose of A. fundyense (25% by volume, 75% Rhodomonas spp.), and high dose (100% A. fundyense). Relative expression of three GST genes was measured using RT-qPCR on days 0.5, 1, 2 and 5 in two independent experiments. Differential regulation was found for the Delta and the Sigma GSTs between 0.5 to 2 days, but not on day 5 in both experiments. The third GST, a microsomal, was not differentially expressed in either treatment or day. RT-qPCR results from the two experiments were similar, even though experimental females were collected from the Gulf of Maine on different dates and their reproductive output differed. In the second experiment, expression of 39 GSTs was determined on days 2 and 5 using RNA-Seq. Global gene expression analyses agreed with the RT-qPCR results. Furthermore, the RNA-Seq measurements indicated that only four GSTs were differentially expressed under the experimental conditions, and the response was small in amplitude. In summary, the A. fundyense diet led to a rapid and transient response in C. finmarchicus in three cytosolic GSTs, while a fourth GST (Omega I) was significantly up-regulated on day 5. Although there was some regulation of GSTs in response the toxic dinoflagellate, the tolerance to A. fundyense by C. finmarchicus is not dependent on the long-term up-regulation of specific GSTs. PMID:27427938

  2. 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

  3. 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

    2012-01-01

    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. PMID:21593032

  4. Biogeochemical cycles during the species succession from Skeletonema costatum to Alexandrium tamarense in northern Hiroshima Bay

    NASA Astrophysics Data System (ADS)

    Yamamoto, Tamiji; Inokuchi, Yuko; Sugiyama, Teiji

    2004-12-01

    Temporal variations, budgets and cycling patterns of major biophilic elements [phosphorus (P), nitrogen (N) and silicon (Si)] in northern Hiroshima Bay during species succession from Skeletonema costatum to Alexandrium tamarense were documented using data collected on a weekly basis in 1996 and 1997. The budget calculations using a box model were made from the perspective of both particulate formation and decomposition of particulate matter. Thus, the overall circulation patterns of these three elements and their differences were examined in the present study. The turnover time of P was longer than that of N, indicating that the utilization efficiency of P by organisms is low. Along with this, absolute concentrations of dissolved inorganic phosphorus in this season and elemental ratios calculated from various forms of biophilic elements support the idea that DIP is too low for phytoplankton to utilize it. However, the utilization efficiency of P was higher in 1997 than in 1996, when A. tamarense formed a more intensive bloom, suggesting that these organisms could retain elements. When the bloom of A. tamarense formed in 1997, the residence time of water was extremely long, indicating that stagnation of water is a primary factor for bloom formation. In contrast to the large fluctuation in the residence time of water, residence times of biophilic elements were almost constant, indicating the importance of biological processes occurring in the system. On the other hand, in 1996, the residence times of biophilic elements were usually longer compared to that of water. This suggests that the particulate formation process by phytoplankton acts as a "nutrient trap" in combination with estuarine circulation.

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

    PubMed

    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

  6. Ichthyotoxicity studies of milkfish Chanos chanos fingerlings exposed to a harmful dinoflagellate Alexandrium minutum.

    PubMed

    Chen, C -Y.; Chou, H -N.

    2001-07-30

    Milkfish (Chanos chanos Forsskal) fingerlings were treated with toxic, nontoxic dinoflagellate Alexandrium minutum cells or toxic algal extract in the water medium without any aeration. Mortality of fish increased with increasing concentrations of toxic, nontoxic algal cells and water-soluble toxic algae extract. Milkfish fingerlings, which were exposed to toxic algae (1.5x10(4)-3.0x10(4) cells/ml) or algal extract [5.13x10(3)-2.05x10(4) cells/ml, 0.195 MU/10(4) cells (toxin concentration)] for 24 h, revealed by light microscopic observations a noticeable edema, hyperplasia and necrosis of secondary gill lamellae. The same toxicological symptom was observed in fish exposed to pure saxitoxin (STX) (6.475x10(-2) &mgr;g/ml) in the water medium. A higher critical oxygen pressure and oxygen consumption rate were also found in the milkfish fingerlings exposed to toxic algae extract (5.13x10(3)-2.05x10(4) cells/ml) and STX (6.475x10(-2) &mgr;g/ml). The cells of nontoxic A. minutum did not cause the gill damage to milkfish, and the extract of nontoxic algae did not cause an increase in oxygen consumption rate or critical oxygen demand of milkfish. From these results, we infer that toxic cells and its extract cause nonspecific response in gill tissues of milkfish. An instant increase in oxygen consumption rate and oxygen demand may be one of the major causes of fish death. PMID:11445088

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

    NASA Astrophysics Data System (ADS)

    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 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.

  8. Enzymatic Permeabilization of the Thecate Dinoflagellate Alexandrium minutum (Dinophyceae) Yields Detection of Intracellularly Associated Bacteria via Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization▿

    PubMed Central

    Palacios, Lucía; Marín, Irma

    2008-01-01

    The enzymatic permeabilization procedure described here allows the detection of intracellular bacteria in the thecate dinoflagellate Alexandrium minutum by using catalyzed reporter deposition-fluorescence in situ hybridization. The combined use of propidium iodide and calcofluor for confocal laser scanning microscopy, together with general and specific fluorescent bacterial probes, demonstrated the intracellular presence of bacteria, including members of the phylum Bacteroidetes. PMID:18263745

  9. 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. PMID:25621399

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

    PubMed

    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

  11. 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

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

    NASA Astrophysics Data System (ADS)

    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

  13. Development of a real-time PCR assay for rapid detection and quantification of Alexandrium minutum (a Dinoflagellate).

    PubMed

    Galluzzi, Luca; Penna, Antonella; Bertozzini, Elena; Vila, Magda; Garcés, Esther; Magnani, Mauro

    2004-02-01

    The marine dinoflagellate genus Alexandrium includes a number of species which produce neurotoxins responsible for paralytic shellfish poisoning (PSP), which in humans may cause muscular paralysis, neurological symptoms, and, in extreme cases, death. A. minutum is the most widespread toxic PSP species in the western Mediterranean basin. The monitoring of coastal waters for the presence of harmful algae also normally involves microscopic examinations of phytoplankton populations. These procedures are time consuming and require a great deal of taxonomic experience, thus limiting the number of specimens that can be analyzed. Because of the genetic diversity of different genera and species, molecular tools may also help to detect the presence of target microorganisms in marine field samples. In this study, we developed a real-time PCR-based assay for rapid detection of all toxic species of the Alexandrium genus in both fixative-preserved environmental samples and cultures. Moreover, we developed a real-time quantitative PCR assay for the quantification of A. minutum cells in seawater samples. Alexandrium genus-specific primers were designed on the 5.8S rDNA region. Primer specificity was confirmed by using BLAST and by amplification of a representative sample of the DNA of other dinoflagellates and diatoms. Using a standard curve constructed with a plasmid containing the ITS1-5.8S-ITS2 A. minutum sequence and cultured A. minutum cells, we determined the absolute number of 5.8S rDNA copies per cell. Consequently, after quantification of 5.8S rDNA copies in samples containing A. minutum cells, we were also able to estimate the number of cells. Several fixed A. minutum bloom sea samples from Arenys Harbor (Catalan Coast, Spain) were analyzed using this method, and quantification results were compared with standard microscopy counting methods. The two methods gave comparable results, confirming that real-time PCR could be a valid, fast alternative procedure for the

  14. Draft genome sequence of Mameliella alba strain UMTAT08 isolated from clonal culture of toxic dinoflagellate Alexandrium tamiyavanichii.

    PubMed

    Danish-Daniel, Muhd; Han Ming, Gan; Noor, Mohd Ezhar Mohd; Yeong, Yik Sung; Usup, Gires

    2016-12-01

    Mameliella alba strain UMTAT08 was isolated from clonal culture of paralytic shellfish toxin producing dinoflagellate, Alexandrium tamiyavanichii. Genome of the strain UMTAT08 was sequenced in order to gain insights into the dinoflagellate-bacteria interactions. The draft genome sequence of strain UMTAT08 contains 5.84Mbp with an estimated G + C content of 65%, 5717 open reading frames, 5 rRNAs and 49 tRNAs. It contains genes related to nutrients uptake, quorum sensing and environmental tolerance related genes. Gene clusters for the biosynthesis of type 1 polyketide synthase, bacteriocin, microcin, terpene and ectoine were also identified. This is suggesting that the bacterium possesses diverse adaptation strategy to survive within the dinoflagellate phycosphere. The draft genome sequence and annotation have been deposited at DDBJ/EMBL/GenBank under the accession number JSUQ00000000. PMID:27625991

  15. 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. PMID:27251197

  16. Identification of gymnodimine D and presence of gymnodimine variants in the dinoflagellate Alexandrium ostenfeldii from the Baltic Sea.

    PubMed

    Harju, Kirsi; Koskela, Harri; Kremp, Anke; Suikkanen, Sanna; de la Iglesia, Pablo; Miles, Christopher O; Krock, Bernd; Vanninen, Paula

    2016-03-15

    Gymnodimines are lipophilic toxins produced by the marine dinoflagellates Karenia selliformis and Alexandrium ostenfeldii. Currently four gymnodimine analogues are known and characterized. Here we describe a novel gymnodimine and a range of gymnodimine related compounds found in an A. ostenfeldii isolate from the northern Baltic Sea. Gymnodimine D (1) was extracted and purified from clonal cultures, and characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS), nuclear magnetic resonance (NMR) spectroscopy, and liquid chromatography-high resolution mass spectrometry (LC-HRMS) experiments. The structure of 1 is related to known gymnodimines (2-5) with a six-membered cyclic imine ring and several other fragments typical of gymnodimines. However, the carbon chain in the gymnodimine macrocyclic ring differs from the known gymnodimines in having two tetrahydrofuran rings in the macrocyclic ring. PMID:26829651

  17. Competition among Dinoflagellate Alexandrium tamarense, raphidophyte Heterosigma carterae and diatom Skeletonema costatum under combinations of two temperatures and five salinities

    NASA Astrophysics Data System (ADS)

    Yan, Tian; Zhou, Mingjiang; Qian, Peiyuan

    2003-09-01

    Competition among HAB (Harmful Algal Bloom) species Dinoflagellate Alexandrium tamarense, Raphidophyte Heterosigma carterae, and Diatom Skeletonema costatum was studied in the laboratory. Experiments with these three major HAB species under combinations of different salinities (10, 18, 25, 30, 35) and temperatures (19°C, 25°C) were carried out. The results showed that S. costatum successfully competed with the other two species at salinities of 18, 25, 30, and 35 at temperatures of 19°C and 25°C. However, H. carterae showed its advantage at low salinity of 10 and became the single dominant species at salinity 10 and 25°C. A. tamarense could not compete successfully with the other two species especially at low salinities. However, it could remain at low density in the presence of higher densities of other algae.

  18. Environmental Factors and Seasonal Occurrence of the Dinoflagellate Alexandrium minutum, a PSP Potential Producer, in a Mediterranean Lagoon

    NASA Astrophysics Data System (ADS)

    Giacobbe, M. G.; Oliva, F. D.; Maimone, G.

    1996-05-01

    Seasonal fluctuations of hydrobiological parameters were studied, throughout 1992, in a shallow Mediterranean area (Ganzirri lagoon), affected by the dinoflagellate Alexandrium minutum, a potential producer of saxitoxins. Concentrations of dissolved inorganic nitrogen (DIN) and N:P ratios were high in autumn and winter, when the waters were well mixed and free from A. minutumcells. The spring appearance of this species coincided with enhanced rainfall and freshwater runoff and with stabilization of the water column, whereas cell dispersion was observed in summer. Although A. minutumnever dominated the algal community, cell densities were maximal in May, when values of salinity did not exceed 30; during this month, there were reduced amounts of DIN, N:P atomic ratios close to 16:1 and a minor competitive pressure by other phytoplankters. Subsequent increases in values of the above parameters—in particular, of the ammonium nitrogen—were associated with a decline in cell numbers, and final disappearance of A. minutumcells.

  19. 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.

  20. Relationship of proteomic variation and toxin synthesis in the dinoflagellate Alexandrium tamarense CI01 under phosphorus and inorganic nitrogen limitation.

    PubMed

    Jiang, Xi-Wen; Wang, Jing; Gao, Yue; Chan, Leo Lai; Lam, Paul Kwan Sing; Gu, Ji-Dong

    2015-10-01

    Paralytic shellfish toxins (PSTs) are originated from cyanobacteria and dinoflagellates, including Alexandrium tamarense, the common dinoflagellate species. In this study, a toxic dinoflagellate strain of A. tamarense CI01 was selected for studying the PSTs' concentration and the related protein variation during the whole cell cycle under different nutrient conditions. High-performance liquid chromatography, 2-D DIGE and Western blotting were used collectively for protein profiling and identification. Results showed that the toxin content was suppressed under nitrogen limiting condition, but enhanced in phosphorous limiting medium. Based on the results of proteomics analysis, 7 proteins were discovered to be related to the PSTs biosynthesis of A. tamarense CI01, including S-adenosylhomocysteine hydrolase, ornithine cyclodeaminase, argininosuccinate synthase, methyluridine methyltransferase cystine ABC transporter, phosphoserine phosphatase, argininosuccinate synthase and acyl-CoA dehydrogenase, which corresponds to the metabolism of the methionine, cysteine, ornithine, arginine and proline. Moreover, some photosynthesis relating proteins also increased their expression during PST synthesis period in A. tamarense CI01, such as phosphoenolpyruvate carboxylase, chloroplast phosphoglycerate kinase, peridinin-chlorophyll α-binding protein, Mg(2+) transporter protein and chloroplast phosphoglycerate kinase. The above findings are in support of our hypothesis that these proteins are involved in toxin biosynthesis of A. tamarense CI01, but cause-and-effect mechanisms need to be investigated in further studies. PMID:26239440

  1. Ribosomal DNA organization patterns within the dinoflagellate genus Alexandrium as revealed by FISH: life cycle and evolutionary implications.

    PubMed

    Figueroa, Rosa Isabel; Cuadrado, Angeles; Stüken, Anke; Rodríguez, Francisco; Fraga, Santiago

    2014-05-01

    Dinoflagellates are a group of protists whose genome differs from that of other eukaryotes in terms of size (contains up to 250pg per haploid cell), base composition, chromosomal organization, and gene expression. But rDNA gene mapping of the active nucleolus in this unusual eukaryotic genome has not been carried out thus far. Here we used FISH in dinoflagellate species belonging to the genus Alexandrium (genome sizes ranging from 21 to 170 pg of DNA per haploid genome) to localize the sequences encoding the 18S, 5.8S, and 28S rRNA genes. The results can be summarized as follows: 1) Each dinoflagellate cell contains only one active nucleolus, with no hybridization signals outside it. However, the rDNA organization varies among species, from repetitive clusters forming discrete nuclear organizer regions (NORs) in some to specialized "ribosomal chromosomes" in other species. The latter chromosomes, never reported before in other eukaryotes, are mainly formed by rDNA genes and appeared in the species with the highest DNA content. 2) Dinoflagellate chromosomes are first characterized by several eukaryotic features, such as structural differentiation (centromere-like constrictions), size differences (dot chromosomes), and SAT (satellite) chromosomes. 3) NOR patterns prove to be useful in discriminating between cryptic species and life cycle stages in protists. PMID:24846057

  2. 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

  3. The role of dissolved organic matter (DOM) quality in the growth enhancement of Alexandrium fundyense (Dinophyceae) in laboratory culture(1).

    PubMed

    Cawley, Kaelin M; Koerfer, Verena; McKnight, Diane M

    2013-06-01

    Several algal species responsible for harmful algal blooms (HABs), such as Alexandrium fundyense, are mixotrophic under certain environmental conditions. The ability to switch between photosynthetic and heterotrophic modes of growth may play a role in the development of HABs in coastal regions. We examined the influence of humic dissolved organic matter (HDOM) derived from terrestrial (plant/soil) and microbial sources on the growth of A. fundyense. We found that a terrestrially derived HDOM, Suwannee River humic acid (SRHA), did enhance A. fundyense growth; however, a microbially derived HDOM, Pony Lake fulvic acid (PLFA) did not enhance growth. A. fundyense grows in association with bacteria in culture and we observed that bacterial cell densities were much lower in A. fundyense cultures than in bacteria-only cultures, consistent with bacterial grazing by A. fundyense in culture. In bacteria-only cultures with added algal exudates (EX), the addition of PLFA and SRHA resulted in a slight increase in bacterial cell density compared to cultures without HDOM added. Changes over time in the chemical quality of the HDOM in the A. fundyense cultures reflected contributions of microbially derived material with similar characteristics as the PLFA. Overall, these results suggest that the chemical differences between SRHA and PLFA are responsible for the greater effect of SRHA on A. fundyense growth, and that the differential effect is not a result of an effect on the growth of associated bacteria. PMID:27007043

  4. 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

  5. First Evidence of Altererythrobacter sp. LY02 with Indirect Algicidal Activity on the Toxic Dinoflagellate, Alexandrium tamarense.

    PubMed

    Li, Yi; Liu, Lei; Xu, Yanting; Guan, Chengwei; Lei, Xueqian; Zheng, Wei; Wang, Hailei; Zheng, Tianling

    2016-10-01

    Alexandrium tamarense is a toxic harmful algal blooms (HABs) causing species, which poses great threat to human health and marine economy. In this study, we isolated an algicidal bacterium Altererythrobacter sp. LY02 towards to A. tamarense and later investigated the algicidal activity, algicidal mode, characteristics of algicidal active substance and algicidal procedure. The results indicated that the cell-free filtrate of strain LY02 showed high algicidal effect on algal growth, however, bacterial cells almost lost algicidal activity. The algicidal active substance was temperature- and pH-stability, and its molecular weight was less than 1000 Da, and was a non-proteinaceous material or non-polysaccharide, mid-polar substance. Under the algicidal effect of active substance, the morphology and structure of A. tamarense cells were seriously damaged as well as organelles. Our study confirmed that the algicidal active substance could be used as an excellent bio-agent for controlling HABs caused by A. tamarense. PMID:27422436

  6. Improvement of lipid production in the marine strains Alexandrium minutum and Heterosigma akashiwo by utilizing abiotic parameters.

    PubMed

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

    2012-01-01

    Two different strains of microalgae, one raphidophyte and one dinoflagellate, were tested under different abiotic conditions with the goal of enhancing lipid production. Whereas aeration was crucial for biomass production, nitrogen deficiency and temperature were found to be the main abiotic parameters inducing the high-level cellular accumulation of neutral lipids. Net neutral lipid production and especially triacylglycerol (TAG) per cell were higher in microalgae (>200% in Alexandrium minutum, and 30% in Heterosigma akashiwo) under treatment conditions (25°C; 330 μM NaNO(3)) than under control conditions (20°C; 880 μM NaNO(3)). For both algal species, oil production (free fatty acids plus TAG fraction) was also higher under treatment conditions (57 mg L(-1) in A. minutum and 323 mg L(-1) in H. akashiwo). Despite the increased production and accumulation of lipids in microalgae, the different conditions did not significantly change the fatty acids profiles of the species analyzed. These profiles consisted of saturated fatty acids (SAFA) and polyunsaturated fatty acids (PUFA) in significant proportions. However, during the stationary phase, the concentrations per cell of some PUFAs, especially arachidonic acid (C20:4n6), were higher in treated than in control algae. These results suggest that the adjustment of abiotic parameters is a suitable and one of the cheapest alternatives to obtain sufficient quantities of microalgal biomass, with high oil content and minimal changes in the fatty acid profile of the strains under consideration. PMID:21766212

  7. Investigation of the 2006 Alexandrium fundyense bloom in the Gulf of Maine: In-situ observations and numerical modeling

    NASA Astrophysics Data System (ADS)

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

    2009-09-01

    In-situ observations and a coupled bio-physical model were used to study the germination, initiation, and development of the Gulf of Maine (GOM) Alexandrium fundyense bloom in 2006. Hydrographic measurements and comparisons with GOM climatology indicate that 2006 was a year with normal coastal water temperature, salinity, current and river runoff conditions. A. fundyense cyst abundance in bottom sediments preceding the 2006 bloom was at a moderate level compared to other recent annual cyst survey data. We used the coupled bio-physical model to hindcast coastal circulation and A. fundyense cell concentrations. Field data including water temperature, salinity, velocity time series and surface A. fundyense cell concentration maps were applied to gauge the model's fidelity. The coupled model is capable of reproducing the hydrodynamics and the temporal and spatial distributions of A. fundyense cell concentration reasonably well. Model hindcast solutions were further used to diagnose physical and biological factors controlling the bloom dynamics. Surface wind fields modulated the bloom's horizontal and vertical distribution. The initial cyst distribution was found to be the dominant factor affecting the severity and the interannual variability of the A. fundyense bloom. Initial cyst abundance for the 2006 bloom was about 50% of that prior to the 2005 bloom. As the result, the time-averaged gulf-wide cell concentration in 2006 was also only about 60% of that in 2005. In addition, weaker alongshore currents and episodic upwelling-favorable winds in 2006 reduced the spatial extent of the bloom as compared with 2005.

  8. Investigation of the Gulf of Maine Circulation and Alexandrium fundyense Bloom in summer 2006: In Situ Observations and Numerical Modeling

    NASA Astrophysics Data System (ADS)

    Li, Y.; He, R.; McGillicuddy, D. J.; Anderson, D. M.; Keafer, B. A.

    2008-12-01

    In situ observations and a coupled bio-physical model are used to study the Gulf of Maine (GOM) coastal ocean hydrodynamics, and the initiation, and development of the Alexandrium fundyense bloom in 2006. Hydrographic measurements indicate that 2006 is a year with normal coastal water temperature, salinity, current and river runoff conditions in comparison with the GOM climatology. A. fundyense cyst observations preceding the 2006 bloom show lower concentrations compared to other recent survey data. A coupled bio- physical model is used to hindcast coastal hydrodynamics and A. fundyense cell concentrations. Field data including water temperature, salinity, velocity time series and surface cell concentration maps are used to gauge the model's fidelity, indicating this coupled model is able to reproduce coastal circulation and the temporal and spatial distributions of A. fundyense cell concentration reasonably well. Model hindcast solutions are further used to diagnose physical and biological factors controlling the bloom dynamics. The initial cyst distribution is found to be the key factor controlling the severity of the bloom. Surface wind fields play an important role in modulating bloom's horizontal and vertical distribution. During the bloom's initiation phase, seasonal warming is a major factor determining the net growth rate. Later in the season, nutrient availability and cell mortality collectively control three-dimensional bloom distribution.

  9. 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

  10. 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

  11. 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

  12. 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. PMID:24858898

  13. Effect of Oxidative Stress Induced by Brevibacterium sp. BS01 on a HAB Causing Species-Alexandrium tamarense

    PubMed Central

    Zhou, Yanyan; Zhang, Bangzhou; Zhang, Su; Li, Dong; Chen, Zhangran; Li, Yi; Bai, Shijie; Lv, Jinglin; Zheng, Wei; Tian, Yun; Zheng, Tianling

    2013-01-01

    Harmful algal blooms occur all over the world, destroying aquatic ecosystems and threatening other organisms. The culture supernatant of the marine algicidal actinomycete BS01 was able to lysis dinoflagellate Alexandrium tamarense ATGD98-006. Physiological and biochemical responses to oxidative stress in A. tamarense were investigated to elucidate the mechanism involved in BS01 inhibition of algal growth. Transmission electron microscope analysis revealed that there were some chloroplast abnormalities in response to BS01 supernatant. The decrease in cellular-soluble protein content suggested that cell growth was greatly inhibited at high concentration of BS01 supernatant. The increase in the levels of reactive oxygen species (ROS) and malondialdehyde contents following exposure to BS01 supernatant indicated that algal cells suffered from oxidative damage. The content of pigment was significantly decreased after 12 h treatment, which indicated that the accumulation of ROS destroyed pigment synthesis. Moreover, the decrease of Fv/Fm ratio suggested that in the photosynthetic system, the dominant sites producing ROS were destroyed by the supernatant of the BS01 culture. The activities of the antioxidant enzymes including superoxide dismutase and peroxidase increased in a short time and decreased slightly with increasing exposure time. A real-time PCR assay showed changes in the transcript abundances of two photosynthetic genes, psbA and psbD. The results showed that BS01 supernatant reduced the expression of the psbA gene after 2 h exposure, but the expression of the psbD gene was increased at concentrations of 1.0 and 1.5%. Our results demonstrated that the expression of the psbA gene was inhibited by the BS01 supernatant, which might block the electron transport chain, significantly enhancing ROS level and excess activity of the antioxidant system. The accumulation of ROS destoryed pigment synthesis and membrane integrity, and inhibited or ultimately killed the

  14. 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.

  15. 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

  16. 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

  17. Factors determining the dynamics of toxic blooms of Alexandrium minutum during a 10-year study along the shallow southwestern Mediterranean coasts

    NASA Astrophysics Data System (ADS)

    Abdenadher, Moufida; Hamza, Asma; Fekih, Wafa; Hannachi, Imen; Zouari Bellaaj, Amel; Bradai, Mohamed Nejmeddine; Aleya, Lotfi

    2012-06-01

    Many blooms of the toxic dinoflagellate Alexandrium minutum have been recorded since 1990 in the Gulf of Gabes (southwestern Mediterranean Sea). To understand the determining factors of bloom formation, we studied the distribution of A. minutum in relation to environmental factors in samples taken at shallow sandy (<1 m) beach stations and a lagoon between 1997 and 2006. This was accompanied by laboratory experiments to identify A. minutum. The species forms harmful algal blooms (HABs) in stations subjected to anthropogenic eutrophication and in confined lagoons, living under conditions of varying salinity which gives it a unique opportunity to broaden its physiological tolerance and increase its colonisation potential. Increases in phosphorus appear to be more important than nitrogen or temperature in the control of A. minutum. The stations sampled all along the coast present specific hydrographic properties (shallowness, turbulence) suggesting that factors other than temperature and nutrients influence Alexandrium distribution, the exception being Boughrara Lagoon where the species developed in accordance with published data. Our findings and their interpretations indicate that the mechanism of the sudden A. minutum blooms along the nearshore of the Gulf of Gabes was complex and differed from that of true coastal ecosystems.

  18. Differences in the toxin profiles of Alexandrium ostenfeldii (Dinophyceae) strains isolated from different geographic origins: Evidence of paralytic toxin, spirolide, and gymnodimine.

    PubMed

    Salgado, Pablo; Riobó, Pilar; Rodríguez, Francisco; Franco, José M; Bravo, Isabel

    2015-09-01

    Among toxin-producing dinoflagellates of the genus Alexandrium, Alexandrium ostenfeldii is the only species able to produce paralytic shellfish poisoning (PSP) toxins, spirolides (SPXs) and gymnodimines (GYMs). In this study we characterized and compared three A. ostenfeldii strains isolated from the Baltic, Mediterranean, and southern Chile Seas with respect to their toxin profiles, morphology, and phylogeny. Toxin analyses by HPLC-FD and LC-HRMS revealed differences in the toxin profiles of the three strains. The PSP toxin profiles of the southern Chile and Baltic strains were largely the same and included gonyautoxin (GTX)-3, GTX-2, and saxitoxin (STX), although the total PSP toxin content of the Chilean strain (105.83 ± 72.15 pg cell(-1)) was much higher than that of the Baltic strain (4.04 ± 1.93 pg cell(-1)). However, the Baltic strain was the only strain that expressed detectable amounts of analogues of GYM-A and GYM-B/-C (48.27 ± 26.12 pg GYM-A equivalents cell(-1)). The only toxin expressed by the Mediterranean strain was 13-desmethyl SPX-C (13dMeC; 2.85 ± 4.76 pg cell(-1)). Phylogenetic analysis based on the LSU rRNA showed that the studied strains belonged to distinct molecular clades. The toxin profiles determined in this study provide further evidence of the taxonomic complexity of this species. PMID:26093028

  19. 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

  20. A novel algicide: evidence of the effect of a fatty acid compound from the marine bacterium, Vibrio sp. BS02 on the harmful dinoflagellate, Alexandrium tamarense.

    PubMed

    Li, Dong; Zhang, Huajun; Fu, Lijun; An, Xinli; Zhang, Bangzhou; Li, Yi; Chen, Zhangran; Zheng, Wei; Yi, Lin; Zheng, Tianling

    2014-01-01

    Alexandrium tamarense is a notorious bloom-forming dinoflagellate, which adversely impacts water quality and human health. In this study we present a new algicide against A. tamarense, which was isolated from the marine bacterium Vibrio sp. BS02. MALDI-TOF-MS, NMR and algicidal activity analysis reveal that this compound corresponds to palmitoleic acid, which shows algicidal activity against A. tamarense with an EC50 of 40 μg/mL. The effects of palmitoleic acid on the growth of other algal species were also studied. The results indicate that palmitoleic acid has potential for selective control of the Harmful algal blooms (HABs). Over extended periods of contact, transmission electron microscopy shows severe ultrastructural damage to the algae at 40 μg/mL concentrations of palmitoleic acid. All of these results indicate potential for controlling HABs by using the special algicidal bacterium and its active agent. PMID:24626054

  1. On an extremely dense bloom of the dinoflagellate Alexandrium tamarense in lagoons of the PO river delta: Impact on the environment

    NASA Astrophysics Data System (ADS)

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

    1996-06-01

    An extremely dense bloom of the potentially toxic dinoflagellate Alexandrium tamarense was observed in the lagoons of Cà Pisani (Veneto, Italy) in summer 1993. The lagoons were experiencing a significant eutrophication impact, receiving waste waters from intensive fish culture plants. During their bloom dinoflagellates in the lagoons reached densities of 2 to 4 × 10 6 cells·dm -3 and a biomass of over 100 g·m -3. The bloom produced drastic ecological changes in the lagoons. It caused nocturnal anoxia, mortality of macrophytes and the build-up of labile organic matter in the water column. Grazing by the tintinnid Favella sp. contributed to the termination of the bloom of the flagellates. The results show that coastal aqua culture probably stimulates dinoflagellate blooms in shallow brackish lagoons.

  2. Historic 2005 toxic bloom of Alexandrium fundyense in the west Gulf of Maine: 1. In situ observations of coastal hydrography and circulation

    NASA Astrophysics Data System (ADS)

    He, Ruoying; McGillicuddy, Dennis J.

    2008-07-01

    An extensive Alexandrium fundyense bloom occurred along the coast of the Gulf of Maine in late spring and early summer 2005. To understand the physical aspects of bloom's initiation and development, in situ observations from both a coast-wide ship survey and the coastal observing network were used to characterize coastal circulation and hydrography during that time period. Comparisons between these in situ observations and their respective long-term means revealed anomalous ocean conditions during May 2005: waters were warmer and fresher coast-wide owing to more surface heating and river runoff; coastal currents were at least 2 times stronger than their climatological means. Surface winds were also anomalous in the form of both episodic bursts of northeast winds and a downwelling-favorable mean condition. These factors may have favored more vigorous along-shore transport and nearshore aggregation of toxic A. fundyense cells (a red tide) in 2005.

  3. 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

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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

  9. 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

    PubMed Central

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

    2013-01-01

    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

  10. The toxic dinoflagellate Alexandrium minutum disrupts daily rhythmic activities at gene transcription, physiological and behavioral levels in the oyster Crassostrea gigas.

    PubMed

    Tran, Damien; Ciutat, Aurélie; Mat, Audrey; Massabuau, Jean-Charles; Hégaret, Hélène; Lambert, Christophe; Le Goic, Nelly; Soudant, Philippe

    2015-01-01

    The objective of the present work was to study the effect of the harmful alga Alexandrium minutum on the daily rhythm of the oyster Crassostrea gigas. Many metabolic and physiological functions are rhythmic in living animals. Their cycles are modeled in accordance with environmental cycles such as the day/night cycle, which are fundamental to increase the fitness of an organism in its environment. A disruption of rhythmic activities is known to possibly impact the health of an animal. This study focused in C. gigas, on a gene known to be involved in circadian rhythmicity, cryptochrome gene (CgCry), on putative clock-controlled genes involved in metabolic and physiological functions, on the length cycle of the style, a structure involved in digestion, and on the rhythmicity of valve activity involved in behavior. The results indicate that daily activity is synchronized at the gene level by light:dark cycles in C. gigas. A daily rhythm of valve activity and a difference in crystalline style length between scotophase and photophase were also demonstrated. Additionally, A. minutum exposure was shown to alter cyclic activities: in exposed oysters, gene transcription remained at a constant low level throughout a daily cycle, valve opening duration remained maximal and crystalline style length variation disappeared. The results show that a realistic bloom of A. minutum clearly can disrupt numerous and diverse molecular, physiological and behavioral functions via a loss of rhythmicity. PMID:25461744

  11. 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

  12. Historical records from dated sediment cores reveal the multidecadal dynamic of the toxic dinoflagellate Alexandrium minutum in the Bay of Brest (France).

    PubMed

    Klouch, Khadidja Z; Schmidt, Sabine; Andrieux-Loyer, Françoise; Le Gac, Mickaël; Hervio-Heath, Dominique; Qui-Minet, Zujaila N; Quéré, Julien; Bigeard, Estelle; Guillou, Laure; Siano, Raffaele

    2016-07-01

    The multiannual dynamic of the cyst-forming and toxic marine dinoflagellate Alexandrium minutum was studied over a time scale of about 150 years by a paleoecological approach based on ancient DNA (aDNA) quantification and cyst revivification data obtained from two dated sediment cores of the Bay of Brest (Brittany, France). The first genetic traces of the species presence in the study area dated back to 1873 ± 6. Specific aDNA could be quantified by a newly developed real-time PCR assay in the upper core layers, in which the germination of the species (in up to 17-19-year-old sediments) was also obtained. In both cores studied, our quantitative paleogenetic data showed a statistically significant increasing trend in the abundance of A. minutum ITS1 rDNA copies over time, corroborating three decades of local plankton data that have documented an increasing trend in the species cell abundance. By comparison, paleogenetic data of the dinoflagellate Scrippsiella donghaienis did not show a coherent trend between the cores studied, supporting the hypothesis of the existence of a species-specific dynamic of A. minutum in the study area. This work contributes to the development of paleoecological research, further showing its potential for biogeographical, ecological and evolutionary studies on marine microbes. PMID:27162179

  13. 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

  14. 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

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

    PubMed

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

    NASA Astrophysics Data System (ADS)

    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 were small compared to the inventory of cysts in the upper few centimeters of sediment but could potentially have more relevance immediately after deposition.

  17. 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., Jr.

    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.

  18. 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.

  19. 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

  20. 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.

  1. 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., Jr.; 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.

  2. 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

  3. Physiological and pathological changes in the eastern oyster Crassostrea virginica infested with the trematode Bucephalus sp. and exposed to the toxic dinoflagellate Alexandrium fundyense.

    PubMed

    Lassudrie, Malwenn; Wikfors, Gary H; Sunila, Inke; Alix, Jennifer H; Dixon, Mark S; Combot, Doriane; Soudant, Philippe; Fabioux, Caroline; Hégaret, Hélène

    2015-03-01

    Effects of experimental exposure to Alexandrium fundyense, a Paralytic Shellfish Toxin (PST) producer known to affect bivalve physiological condition, upon eastern oysters, Crassostrea virginica with a variable natural infestation of the digenetic trematode Bucephalus sp. were determined. After a three-week exposure to cultured A. fundyense or to a control algal treatment with a non-toxic dinoflagellate, adult oysters were assessed for a suite of variables: histopathological condition, hematological variables (total and differential hemocyte counts, morphology), hemocyte functions (Reactive Oxygen Species (ROS) production and mitochondrial membrane potential), and expression in gills of genes involved in immune responses and cellular protection (MnSOD, CAT, GPX, MT-IV, galectin CvGal) or suspected to be (Dominin, Segon). By comparing individual oysters infested heavily with Bucephalus sp. and uninfested individuals, we found altered gonad and digestive gland tissue and an inflammatory response (increased hemocyte concentration in circulating hemolymph and hemocyte infiltrations in tissues) associated with trematode infestation. Exposure to A. fundyense led to a higher weighted prevalence of infection by the protozoan parasite Perkinsus marinus, responsible for Dermo disease. Additionally, exposure to A. fundyense in trematode-infested oysters was associated with the highest prevalence of P. marinus infection. These observations suggest that the development of P. marinus infection was advanced by A. fundyense exposure, and that, in trematode-infested oysters, P. marinus risk of infection was higher when exposed to A. fundyense. These effects were associated with suppression of the inflammatory response to trematode infestation by A. fundyense exposure. Additionally, the combination of trematode infestation and A. fundyense exposure caused degeneration of adductor muscle fibers, suggesting alteration of valve movements and catch state, which could increase

  4. 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.

  5. 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., Jr.; 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

  6. 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

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

    PubMed Central

    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

    2013-01-01

    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 abundance

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

    PubMed Central

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

    2013-01-01

    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

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

    PubMed Central

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

    2015-01-01

    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

  10. 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

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

    PubMed

    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 also have

  12. 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

  13. 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

  14. 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. PMID:24841716

  15. sxtA-Based Quantitative Molecular Assay To Identify Saxitoxin-Producing Harmful Algal Blooms in Marine Waters ▿ †

    PubMed Central

    Murray, Shauna A.; Wiese, Maria; Stüken, Anke; Brett, Steve; Kellmann, Ralf; Hallegraeff, Gustaaf; Neilan, Brett A.

    2011-01-01

    The recent identification of genes involved in the production of the potent neurotoxin and keystone metabolite saxitoxin (STX) in marine eukaryotic phytoplankton has allowed us for the first time to develop molecular genetic methods to investigate the chemical ecology of harmful algal blooms in situ. We present a novel method for detecting and quantifying the potential for STX production in marine environmental samples. Our assay detects a domain of the gene sxtA that encodes a unique enzyme putatively involved in the sxt pathway in marine dinoflagellates, sxtA4. A product of the correct size was recovered from nine strains of four species of STX-producing Alexandrium and Gymnodinium catenatum and was not detected in the non-STX-producing Alexandrium species, other dinoflagellate cultures, or an environmental sample that did not contain known STX-producing species. However, sxtA4 was also detected in the non-STX-producing strain of Alexandrium tamarense, Tasmanian ribotype. We investigated the copy number of sxtA4 in three strains of Alexandrium catenella and found it to be relatively constant among strains. Using our novel method, we detected and quantified sxtA4 in three environmental blooms of Alexandrium catenella that led to STX uptake in oysters. We conclude that this method shows promise as an accurate, fast, and cost-effective means of quantifying the potential for STX production in marine samples and will be useful for biological oceanographic research and harmful algal bloom monitoring. PMID:21841034

  16. Profiling the fatty acids from a strain of the microalgae Alexandrium tamarense by means of high-speed counter-current chromatography and gas chromatography coupled with mass spectrometry.

    PubMed

    Hammann, Simon; Tillmann, Urban; Schröder, Markus; Vetter, Walter

    2013-10-18

    Fatty acids of microalgae have been studied as potential chemotaxonomic markers, to reveal plausible lipid phycotoxins or in the context of mass production of algal biofuels. The planctonic microalgae Alexandrium tamarense (Dinophyceae) is a common harmful algal bloom species that often proliferates in eutrophic costal waters. Alexandrium blooms are the proximal source of toxins associated with paralytic shellfish poisoning (PSP), a neurological affliction that has caused human illness for centuries via consumption of contaminated shellfish. However, data on the fatty acid composition of A. tamarense is currently limited. For this reason, we cultivated a well-defined strain of A. tamarense (Alex2, group I, North American clade) in order to study both its major and minor fatty acids. The harvested microalgae were transesterified and the fatty acid methyl esters were fractionated by means of high-speed counter-current chromatography (HSCCC). The resulting 31 HSCCC fractions were analyzed by gas chromatography with mass spectrometry (GC/MS). Unknown substances were identified by transferring assorted HSCCC fractions into picolinyl or pyrrolidide derivatives. Twenty fatty acids (range 0.2-22.9% contribution to total fatty acids) were identified in the unfractionated sample with 14:0, 16:0, 18:1n-9, 18:4n-3, 18:5n-3 and 22:6n-3 representing>80% of the total fatty acids. HSCCC fractionation enabled the identification of further 22 trace fatty acids contributing between ∼0.01 and 0.2% to total fatty acids. The fatty acids included several branched-chain fatty acids as well as scarcely reported fatty acids like 11-methyl-18:1n-6tr or 18:2Δ4,9. In order to enable a better comparability and repeatability of HSCCC fractionations, we calculated for each HSCCC fraction the total volume of mobile phase, which had passed the HSCCC. From this volume we subtracted the volume of extruded stationary phase and divided the corrected volume by the total coil volume. These elution

  17. A simple and innovative method for species identification of phytoplankton cells on minute quantities of DNA.

    PubMed

    Masseret, Estelle; Enquebecq, Marjorie; Laabir, Mohamed; Genovesi, Benjamin; Vaquer, André; Avarre, Jean-Christophe

    2010-12-01

    Dinoflagellates belonging to the genus Alexandrium are often involved in harmful algal blooms. Their ecological exploration is thus essential to increase our knowledge on these toxic events. Yet, population genetic studies, taxonomic identification and environmental monitoring are hampered by major constraints: the necessity to establish monoclonal cultures from environmental samples and the sensitivity of available molecular tools. The present work describes a very simple and sensitive method for extraction and amplification of DNA at the infra-single-cell level. Its on-slide format allows for easy visual control of both quality and quantity of the templates. Combined with a semi-multiplex PCR protocol designed on the 18S-28S rDNA-ITS region of Alexandrium catenella and Alexandrium tamarense, this procedure allowed the identification and discrimination of these species from both monoclonal cultures and natural samples. PMID:23766275

  18. 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

  19. Trophic accumulation of PSP toxins in zooplankton during Alexandrium fundyense blooms in Casco Bay, Gulf of Maine, April June 1998. I. Toxin levels in A. fundyense and zooplankton size fractions

    NASA Astrophysics Data System (ADS)

    Doucette, Gregory J.; Turner, Jefferson T.; Powell, Christine L.; Keafer, Bruce A.; Anderson, Donald M.

    2005-09-01

    The transfer of marine algal toxins involving a range of phytoplanktivorous vectors is well documented as a means of exposing organisms at higher trophic levels (including humans) to these naturally occurring yet harmful compounds. While previous studies have examined the potential for, and dynamics of, algal toxin accumulation by individual zooplankton species, few have attempted to distinguish the contribution of various grazer size classes to toxin trophic transfer in natural communities and characterize some of the factors that can influence this process. The current investigation was aimed at describing the size-fractioned (64-100, 100-200, 200-500, >500 μm) accumulation of paralytic shellfish poisoning (PSP) toxins by zooplankton in Casco Bay and the adjacent coastal waters of the Gulf of Maine during a series of cruises from April to June 1998. Several variables, including the abundance of PSP toxin-producing Alexandrium fundyense, in-water toxin concentrations associated with this dinoflagellate, and algal toxin cell quotas, were measured and their relationship to zooplankton toxin accumulation assessed. A principal finding of this work was the ability of any grazer size class examined (including grazers present in the 20-64 μm A. fundyense-containing fraction) to serve as an initial vector for introducing PSP toxins into the Casco Bay food web at various times during the sampling period, thereby providing multiple potential routes of toxin trophic transfer. In addition, trends observed in the coincident mapping of A. fundyense cells and their associated toxin were generally in agreement, yet did not remain closely coupled at all times. Therefore, although A. fundyense abundance can be a reasonable indicator of PSP toxin presence in the phytoplankton, this relationship can vary considerably and lead to situations where elevated toxin levels occur at low cell concentrations and vice versa. The uncoupling of A. fundyense cell and in-water toxin

  20. 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.

  1. 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

  2. Trophic accumulation of PSP toxins in zooplankton during Alexandrium fundyense blooms in Casco Bay, Gulf of Maine, April-June 1998. II. . Zooplankton abundance and size-fractionated community composition

    NASA Astrophysics Data System (ADS)

    Turner, Jefferson T.; Doucette, Gregory J.; Keafer, Bruce A.; Anderson, Donald M.

    2005-09-01

    During spring blooms of the toxic dinoflagellate Alexandrium fundyense in Casco Bay, Maine in 1998, we investigated vectorial intoxication of various zooplankton size fractions with PSP toxins, including zooplankton community composition from quantitative zooplankton samples (>102 μm), as well as zooplankton composition in relation to toxin levels in various size fractions (20-64, 64-100, 100-200, 200-500, >500 μm). Zooplankton abundance in 102 μm mesh samples was low (most values<10,000 animals m -3) from early April through early May, but increased to maxima in mid-June (cruise mean=121,500 animals m -3). Quantitative zooplankton samples (>102 μm) were dominated by copepod nauplii, and Oithona similis copepodites and adults at most locations except for those furthest inshore. At these inshore locations, Acartia hudsonica copepodites and adults were usually dominant. Larger copepods such as Calanus finmarchicus, Centropages typicus, and Pseudocalanus spp. were found primarily offshore, and at much lower abundances than O. similis. Rotifers, mainly present from late April to late May, were most abundant inshore. The marine cladoceran Evadne nordmani was sporadically abundant, particularly in mid-June. Microplankton in 20-64 μm size fractions was generally dominated by A. fundyense, non-toxic dinoflagellates, and tintinnids. Microplankton in 64-100 μm size fractions was generally dominated by larger non-toxic dinoflagellates, tintinnids, aloricate ciliates, and copepod nauplii, and in early May, rotifers. Some samples (23%) in the 64-100 μm size fractions contained abundant cells of A. fundyense, presumably due to sieve clogging, but most did not contain A. fundyense cells. This suggests that PSP toxin levels in those samples were due to vectorial intoxication of microzooplankters such as heterotrophic dinoflagellates, tintinnids, aloricate ciliates, rotifers, and copepod nauplii via feeding on A. fundyense cells. Dominant taxa in zooplankton fractions varied

  3. 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

  4. 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

  5. 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

  6. 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. PMID:26254582

  7. 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

  8. 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

  9. 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.

  10. 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

    2015-12-01

    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.

  11. Oligotrophication and emergence of picocyanobacteria and a toxic dinoflagellate in Thau lagoon, southern France

    NASA Astrophysics Data System (ADS)

    Collos, Yves; Bec, Béatrice; Jauzein, Cécile; Abadie, Eric; Laugier, Thierry; Lautier, Jacques; Pastoureaud, Annie; Souchu, Philippe; Vaquer, André

    2009-01-01

    Time series data have been examined in Thau lagoon (Southern France) from 1972 to 2006 for water temperature, salinity, nutrients and from 1987 to 2006 for phytoplankton. A first main trend identified is an increase in mean annual water temperature (1.5 °C over 33 years or 0.045 °C/year) that was not evenly distributed among seasons. The highest rate of increase was in the spring (+ 3.0 °C over 33 years), followed by summer (+ 2.0 °C) and fall (+ 1.7 °C). In winter, no significant increase over the 33 year period could be found. A second clear trend is a large decrease in soluble reactive phosphorus (SRP) concentration over the same 33 year period (summer values decreased from 10 µM to 1 µM, while winter values decreased from 3 µM to undetectable at present). Nitrate concentrations depended mainly on rainfall events and watershed runoff. Ammonium data were too fragmentary to be useful. N/P ratios expressed the traditional way of DIN/SRP cannot be used for phytoplankton that are not strict autotrophs. The recent and almost simultaneous appearance of both picocyanobacteria (mostly Synechococcus) and the toxic dinoflagellate Alexandrium catenella in Thau seem to be related to reduced nutrient loading and the increase in water temperature. A. catenella blooms occur either in the spring or the fall when water temperature is near 20 °C and remains so for several weeks with winds speeds below 2-3 m s - 1 . Picocyanobacterial growth is stimulated by increased summer temperatures, and lowered SRP levels provide picocyanobacteria an ecological advantage over other phytoplankton classes, in particular diatoms such as Skeletonema costatum whose cell densities have decreased over the last 8 years in summer and fall, but not in winter. An hypothesis is presented according to which A. catenella is not stimulated by increased temperatures, but is able to use picocyanobacteria for growth, and this provides this organism an additional resource over other strictly

  12. Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae

    NASA Astrophysics Data System (ADS)

    Huang, Jian; Wen, Ruobing; Bao, Zhenmin; Sui, Zhenghong; Sun, Ningbo; Kang, Kyoungho

    2012-05-01

    Over the last several decades, harmful algal blooms (HABs) have become a serious environmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H. akashiwo cells. Five HABs species ( Alexandrium catenella, Thalassiosira sp., Cryptomonas sp., Alexandrium tamarense and Symbiodinium sp.), were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%-91.6% of the cells were captured from unialgal culture by IMBs, and only 5.3%-12.5% of the four other HAB microalgae species were captured, indicating that the constructed IMBs combined specifically with the H. akashiwo cells. At the same time, flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique, 91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques, especially immuno-magnetic separation, have low associated cost, and are fast and simple processes compared with other techniques currently in use.

  13. The quantitative real-time PCR applications in the monitoring of marine harmful algal bloom (HAB) species.

    PubMed

    Penna, Antonella; Antonella, Penna; Galluzzi, Luca; Luca, Galluzzi

    2013-10-01

    In the last decade, various molecular methods (e.g., fluorescent hybridization assay, sandwich hybridization assay, automatized biosensor detection, real-time PCR assay) have been developed and implemented for accurate and specific identification and estimation of marine toxic microalgal species. This review focuses on the recent quantitative real-time PCR (qrt-PCR) technology developed for the control and monitoring of the most important taxonomic phytoplankton groups producing biotoxins with relevant negative impact on human health, the marine environment, and related economic activities. The high specificity and sensitivity of the qrt-PCR methods determined by the adequate choice of the genomic target gene, nucleic acid purification protocol, quantification through the standard curve, and type of chemical detection method make them highly efficient and therefore applicable to harmful algal bloom phenomena. Recent development of qrt-PCR-based assays using the target gene of toxins, such as saxitoxin compounds, has allowed more precise quantification of toxigenic species (i.e., Alexandrium catenella) abundance. These studies focus only on toxin-producing species in the marine environment. Therefore, qrt-PCR technology seems to offer the advantages of understanding the ecology of harmful algal bloom species and facilitating the management of their outbreaks. PMID:23247526

  14. Toxicity of benz(a)anthracene and fluoranthene to marine phytoplankton in culture: does cell size really matter?

    PubMed

    Ben Othman, Hiba; Leboulanger, Christophe; Le Floc'h, Emilie; Mabrouk, Hassine Hadj; Hlaili, Asma Sakka

    2012-12-01

    The toxicity of benz(a)anthracene and fluoranthene (polycyclic aromatic hydrocarbons, PAHs) was evaluated on seven species of marine algae in culture belonging to pico-, nano-, and microphytoplankton, exposed to increasing concentrations of up to 2 mg L(-1). The short-term (24h) toxicity was assessed using chlorophyll a fluorescence transients, linked to photosynthetic parameters. The maximum quantum yield Fv/Fm was lower at the highest concentrations tested and the toxicity thresholds were species-dependent. For acute effects, fluoranthene was more toxic than benz(a)anthracene, with LOECs of 50.6 and 186 μg L(-1), respectively. After 72 h exposure, there was a dose-dependent decrease in cell density, fluoranthene being more toxic than benz(a)anthracene. The population endpoint at 72 h was affected to a greater extent than the photosynthetic endpoint at 24h. EC50 was evaluated using the Hill model, and species sensitivity was negatively correlated to cell biovolume. The largest species tested, the dinoflagellate Alexandrium catenella, was almost insensitive to either PAH. The population endpoint EC50s for fluoranthene varied from 54 μg L(-1) for the picophytoplankton Picochlorum sp. to 418 μg L(-1) for the larger diatom Chaetoceros muelleri. The size/sensitivity relationship is proposed as a useful model when there is a lack of ecotoxicological data on hazardous chemicals, especially in marine microorganisms. PMID:23122731

  15. Boundary influences on HAB phytoplankton ecology in a stratification-enhanced upwelling shadow

    NASA Astrophysics Data System (ADS)

    Ryan, J. P.; McManus, M. A.; Kudela, R. M.; Lara Artigas, M.; Bellingham, J. G.; Chavez, F. P.; Doucette, G.; Foley, D.; Godin, M.; Harvey, J. B. J.; Marin, R.; Messié, M.; Mikulski, C.; Pennington, T.; Py, F.; Rajan, K.; Shulman, I.; Wang, Z.; Zhang, Y.

    2014-03-01

    Coastal marine ecosystems are profoundly influenced by processes that originate from their boundaries. These include fluid boundaries—with the atmosphere, oceanic boundary currents and terrestrial aquatic systems, as well as solid boundaries—with the seafloor and coast. Phytoplankton populations transfer complexly interacting boundary influences into the biosphere. In this contribution, we apply data from an ocean observing and modeling system to examine boundary influences driving phytoplankton ecology in Monterey Bay, CA, USA. The study was focused on species that may cause harmful algal blooms (HABs). During September-October 2010, autonomous molecular analytical devices were moored at two locations characterized by different degrees of stratification and exposure to upwelling dynamics. The time-series revealed multiple transitions in local HAB phytoplankton communities, involving diatoms (Pseudo-nitzschia spp.), dinoflagellates (Alexandrium catenella), and raphidophytes (Heterosigma akashiwo). Observational and model results showed that the biological transitions were closely related to environmental changes that resulted from a variety of boundary processes—responses of oceanic circulation to wind forcing, influxes of different water types that originated outside the bay, and emergence of strongly stratified nearshore water into the greater bay. Boundary processes were further implicated at patch scales. High-resolution mapping and sampling of a phytoplankton-enriched patch were conducted in a Lagrangian framework using autonomous underwater vehicles. These highly resolved measurements showed that small-scale spatial patterns in the toxicity of Pseudo-nitzschia populations were related to the coupling of resuspended sediments from the bottom boundary layer to the surface mixed layer.

  16. 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

  17. 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

  18. Drag increase and drag reduction found in phytoplankton and bacterial cultures in laminar flow: Are cell surfaces and EPS producing rheological thickening and a Lotus-leaf Effect?

    NASA Astrophysics Data System (ADS)

    Jenkinson, Ian R.; Sun, Jun

    2014-03-01

    The laminar-flow viscosity of ocean and other natural waters consists of a Newtonian aqueous component contributed by water and salts, and a non-Newtonian one contributed mainly by exopolymeric polymers (EPS) derived largely from planktonic algae and bacteria. Phytoplankton and EPS form thin layers in stratified waters, often associated with density discontinuities. A recent model (Jenkinson and Sun, 2011. J. Plankton Res., 33, 373-383) investigated possible thalassorheological control of pycnocline thickness (PT) by EPS secreted by the harmful dinoflagellate Karenia mikimotoi. The model, based on published measurements of viscosity increase by this species, found that whether it can influence PT depends on the relationship between increased viscosity, deformation rates/stresses and length scale, which the present work has investigated. To do this, flow rate vs. hydrostatic pressure (and hence wall stress) was measured in cultures (relative to that in reference water) in capillaries of 5 radii 0.35-1.5 mm, close to oceanic-turbulence Kolmogorov length. We compared cultures of the potentially harmful algae, K. mikimotoi, Alexandrium catenella, Prorocentrum donghaiense, Skeletonema costatum, Phaeodactylum tricornutum and the bacterium Escherichia coli. Drag increase, ascribed to rheological thickening by EPS, occurred in the smallest capillaries, but drag reduction (DR) occurred in the largest ones. Since this occurred at Reynolds numbers Re too small for turbulence (or turbulent DR) to occur, this was laminar-flow DR. It may have been superhydrophobic DR (SDR), associated with the surfaces of the plankton and bacteria. SDR is associated with the self-cleaning Lotus-leaf Effect, in which water and dirt are repelled from surfaces bearing nm- to µm-sized irregularities coated with hydrophobic polymers. Because DR decreased measured viscosity and EPS thickening increased it, we could not validate the model. DR, however, represents hitherto unknown phenomenon in the

  19. [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. PMID:16311702

  20. EFFECT OF FLUID SHEAR AND IRRADIANCE ON POPULATION GROWTH AND CELLULAR TOXIN CONTENT OF THE DINOFLAGELLATE ALEXANDRIUM FUNDYENSE.

    EPA Science Inventory

    The potential for in situ turbulence to inhibit dinoflagellate population growth has been demonstrated by experimentally exposing dinoflagellate cultures to quantified shear flow. However, despite interest in understanding environmental factors that affect the growth of toxic din...

  1. 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

  2. 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

  3. 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.

  4. 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 ...

  5. Berseem clover seeding rate and harvest management effects on forage yields and nutrient uptake in a swine-effluent spray field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Research on how berseem clover (Trifolium alexandrium L.) seed rate (SR) affects winter and summer forage yields in swine-effluent spray fields is lacking for the southeastern USA. This 3-yr study determined optimum berseem SR for dry matter (DM) yield and nutrient uptake in the winter cover crop an...

  6. Intraspecific facilitation by allelochemical mediated grazing protection within a toxigenic dinoflagellate population

    PubMed Central

    John, Uwe; Tillmann, Urban; Hülskötter, Jennifer; Alpermann, Tilman J.; Wohlrab, Sylke; Van de Waal, Dedmer B.

    2015-01-01

    Dinoflagellates are a major cause of harmful algal blooms (HABs), with consequences for coastal marine ecosystem functioning and services. Alexandrium fundyense (previously Alexandrium tamarense) is one of the most abundant and widespread toxigenic species in the temperate Northern and Southern Hemisphere and produces paralytic shellfish poisoning toxins as well as lytic allelochemical substances. These bioactive compounds may support the success of A. fundyense and its ability to form blooms. Here we investigate the impact of grazing on monoclonal and mixed set-ups of highly (Alex2) and moderately (Alex4) allelochemically active A. fundyense strains and a non-allelochemically active conspecific (Alex5) by the heterotrophic dinoflagellate Polykrikos kofoidii. While Alex4 and particularly Alex5 were strongly grazed by P. kofoidii when offered alone, both strains grew well in the mixed assemblages (Alex4 + Alex5 and Alex2 + Alex5). Hence, the allelochemical active strains facilitated growth of the non-active strain by protecting the population as a whole against grazing. Based on our results, we argue that facilitation among clonal lineages within a species may partly explain the high genotypic and phenotypic diversity of Alexandrium populations. Populations of Alexandrium may comprise multiple cooperative traits that act in concert with intraspecific facilitation, and hence promote the success of this notorious HAB species. PMID:25411447

  7. THE DEVELOPMENT OF A SINGLE-CELL FIELD DIAGNOSTIC FOR NITROGEN LIMITATION IN HARMFUL ALGAE

    EPA Science Inventory

    This project proposes the development of an assay to identify cell-specific acetylglucosaminidase activity using the substrate ELF-NAG (ELF 97 N'-acetylglucosaminide). Preliminary data indicates this enzyme is nitrogen-regulated in Alexandrium and that this ass...

  8. 78 FR 78783 - Magnuson-Stevens Fishery Conservation and Management Act Provisions; Fisheries of the...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-27

    ... death from PSP. On June 16, 2005, NMFS published an emergency rule (70 FR 35047) closing the area... defining when ``an emergency'' involving a fishery exists (62 FR 44421; August 21, 1997). This guidance... by the alga Alexandrium fundyense, which can form blooms commonly referred to as red tides. Red...

  9. Evolutionary Acquisition and Loss of Saxitoxin Biosynthesis in Dinoflagellates: the Second “Core” Gene, sxtG

    PubMed Central

    Orr, Russell J. S.; Stüken, Anke; Murray, Shauna A.

    2013-01-01

    Saxitoxin and its derivatives are potent neurotoxins produced by several cyanobacteria and dinoflagellate species. SxtA is the initial enzyme in the biosynthesis of saxitoxin. The dinoflagellate full mRNA and partial genomic sequences have previously been characterized, and it appears that sxtA originated in dinoflagellates through a horizontal gene transfer from a bacterium. So far, little is known about the remaining genes involved in this pathway in dinoflagellates. Here we characterize sxtG, an amidinotransferase enzyme gene that putatively encodes the second step in saxitoxin biosynthesis. In this study, the entire sxtG transcripts from Alexandrium fundyense CCMP1719 and Alexandrium minutum CCMP113 were amplified and sequenced. The transcripts contained typical dinoflagellate spliced leader sequences and eukaryotic poly(A) tails. In addition, partial sxtG transcript fragments were amplified from four additional Alexandrium species and Gymnodinium catenatum. The phylogenetic inference of dinoflagellate sxtG, congruent with sxtA, revealed a bacterial origin. However, it is not known if sxtG was acquired independently of sxtA. Amplification and sequencing of the corresponding genomic sxtG region revealed noncanonical introns. These introns show a high interspecies and low intraspecies variance, suggesting multiple independent acquisitions and losses. Unlike sxtA, sxtG was also amplified from Alexandrium species not known to synthesize saxitoxin. However, amplification was not observed for 22 non-saxitoxin-producing dinoflagellate species other than those of the genus Alexandrium or G. catenatum. This result strengthens our hypothesis that saxitoxin synthesis has been secondarily lost in conjunction with sxtA for some descendant species. PMID:23335767

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

    PubMed

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

    2013-03-01

    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 results

  11. 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

  12. Characterization of Satellite DNA from Three Marine Dinoflagellates (Dinophyceae): Glenodinium sp. and Two Members of the Toxic Genus, Protogonyaulax 1

    PubMed Central

    Boczar, Barbara A.; Liston, John; Cattolico, Rose Ann

    1991-01-01

    Using CsCl-Hoechst dye or CsCl-ethidium bromide gradients, satellite and nuclear DNAs were separated and characterized in three marine dinoflagellates: Glenodinium sp., and two toxic dinoflagellates, Protogonyaulax tamarensis and Protogonyaulax catenella. In all three dinoflagellates, the lowest density fraction, satellite DNA1, hybridized to chloroplast genes derived from terrestrial plants and/or other algae. Dinoflagellate chloroplast DNAs exhibited molecular sizes of 114 to 125 kilobase pairs, which is consistent with plastid sizes determined for other chromophytic algae (120-150 kilobase pairs). Mitochondrial DNA was not resolved from nuclear DNA in this system. Two additional satellite DNAs, satellite DNA2 and satellite DNA3, recovered from P. tamarensis and P. catenella were similar to one another, both within and between species, when characterized by restriction enzyme analysis. These satellites were 85 to 95 kilobase pairs in size, and exhibited restriction fragments that hybridized to yeast nuclear ribosomal RNA genes. Restriction enzyme analyses and DNA hybridization studies of cpDNA document that the two Protogonyaulax isolates are not evolutionarily identical. ImagesFigure 1Figure 2Figure 3Figure 4 PMID:16668443

  13. Phytoplankton invasions: comments on the validity of categorizing the non-indigenous dinoflagellates and diatoms in European seas.

    PubMed

    Gómez, Fernando

    2008-04-01

    The validity of categorizing the diatoms and dinoflagellates reported in the literature as non-indigenous phytoplankton in the European Seas was investigated. Species that are synonymous are often included as separate species (Gessnerium mochimaensis=Alexandrium monilatum, Gymnodinium nagasakiense=Karenia mikimotoi, Pleurosigma simonsenii=P. planctonicum), while other species names are synonyms of cosmopolitan taxa (Prorocentrum redfieldii=P. triestinum, Pseliodinium vaubanii=Gyrodinium falcatum, Gonyaulax grindleyi=Protoceratium reticulatum, Asterionella japonica=Asterionellopsis glacialis). Epithets of an exotic etymology (i.e. japonica, sinensis, indica) imply that a cosmopolitan species may be non-indigenous, and several taxa are even considered as non-indigenous in their type locality (Alexandrium tamarense and A. pseudogoniaulax). The records of Alexandrium monilatum, A. leei and Corethron criophilum are doubtful. Cold or warm-water species expand their geographical ranges or increase their abundances to detectable levels during cooling (Coscinodiscus wailesii) or warming periods (Chaetoceros coarctatus, Proboscia indica, Pyrodinium bahamense). These are a few examples of marginal dispersal associated with climatic events instead of species introductions from remote areas. The number of non-indigenous phytoplankton species in European Seas has thus been excessively inflated. PMID:18295804

  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. Dinomyces arenysensis gen. et sp. nov. (Rhizophydiales, Dinomycetaceae fam. nov.), a chytrid infecting marine dinoflagellates.

    PubMed

    Lepelletier, Frédéric; Karpov, Sergey A; Alacid, Elisabet; Le Panse, Sophie; Bigeard, Estelle; Garcés, Esther; Jeanthon, Christian; Guillou, Laure

    2014-03-01

    Environmental 18S rRNA gene surveys of microbial eukaryotes have recently revealed the diversity of major parasitic agents in pelagic freshwater systems, consisting primarily of chytrid fungi. To date, only a few studies have reported the presence of chydrids in the marine environment and a limited number of marine chytrids have been properly identified and characterized. Here, we report the isolation and cultivation of a marine chytrid from samples taken during a bloom of the toxic dinoflagellate Alexandrium minutum in the Arenys de Mar harbour (Mediterranean Sea, Spain). Cross-infections using cultures and natural phytoplankton communities revealed that this chytrid is only able to infect certain species of dinoflagellates, with a rather wide host range but with a relative preference for Alexandrium species. Phylogenetic analyses showed that it belongs to the order Rhizophydiales, but cannot be included in any of the existing families within this order. Several ultrastructural characters confirmed the placement of this taxon within the Rhizophydiales as well its novelty notably in terms of zoospore structure. This marine chytridial parasitoid is described as a new genus and species, Dinomyces arenysensis, within the Dinomycetaceae fam. nov. PMID:24709472

  16. Characterization of mycosporine-serine-glycine methyl ester, a major mycosporine-like amino acid from dinoflagellates: a mass spectrometry study.

    PubMed

    Carignan, Mario O; Carreto, José I

    2013-08-01

    Several unknown mycosporine-like amino acids (MAAs) have been previously isolated from some cultured species of toxic dinoflagellates of the Alexandrium genus (Dinophyceae). One of them, originally called M-333, was tentatively identified as a shinorine methyl ester, but the precise nature of this compound is still unknown. Using a high-resolution reversed-phase liquid chromatography mass spectrometry analyses (HPLC/MS), we found that natural populations of the red tide dinoflagellate Prorocentrum micans Ehrenberg showed a net dominance of M-333 together with lesser amounts of other MAAs. We also documented the isolation and characterization of this MAA from natural dinoflagellate populations and from Alexandrium tamarense (Lebour) Balech cultures. Using a comparative fragmentation study in electrospray mass spectrometry between deuterated and non-deuterated M-333 compounds and synthesized mono and dimethyl esters of shinorine, this novel compound was characterized as mycosporine-serine-glycine methyl ester, a structure confirmed by nuclear magnetic resonance. These isobaric compounds can be differentiated by their fragmentation patterns in MS(3) experiments because the extension and the specific site of the methylation changed the fragmentation pathway. PMID:27007200

  17. 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.

  18. 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

  19. Molecularly imprinted polymer as efficient sorbent of solid-phase extraction for determination of gonyautoxin 1,4 in seawater followed by high-performance liquid chromatography-fluorescence detection.

    PubMed

    Mei, Xiao-Qi; He, Xiu-Ping; Wang, Jiang-Tao

    2016-08-01

    A kind of new molecularly imprinted polymer (MIP) was synthesized by bulk polymerization using guanosine as dummy template molecule, α-methacrylic acid as functional monomer and ethylene glycol dimethyl acrylic ester as crosslinker. Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) showed that the MIP had homogenous and uniform-sized cavities. It was confirmed that the MIP had higher binding affinity and selectivity towards gonyautoxins 1,4 (GTX 1,4) than the non-imprinted polymer (NIP) according to the static equilibrium adsorption. An off-line molecularly imprinted solid-phase extraction (MISPE) method followed by high-performance liquid chromatography with fluorescence detection (HPLC-FLD) was established for the analysis of GTX 1,4. 0.1 mol/L acetic acid and 95:5 (v:v) methanol/water were optimized as the washing and elution solutions, respectively. The recoveries of spiked cultured seawater samples were satisfactory, as high as 88 %. Using this method, the concentrations of GTX 1,4 from cultured seawater samples of Alexandrium minutum and Alexandrium tamarense were detected to be 1.10 μg/L and 0.99 μg/L, respectively. Graphical Abstract The synthesis of molecularly imprinted polymer and molecularly imprinted solid-phase extraction analysis for gonyautoxin 1,4. PMID:27317255

  20. Dynamics of a mesoscale eddy off Cape Ann, Massachusetts in May 2005

    NASA Astrophysics Data System (ADS)

    Jiang, Mingshun; Zhou, Meng; Libby, Scott P.; Anderson, Donald M.

    2011-11-01

    Observations and numerical modeling indicate that a mesoscale anti-cyclonic eddy formed south of Cape Ann at the northern entrance of Massachusetts Bay (MB) during May 2005, when large river discharges in the western Gulf of Maine and two strong Nor'easters passing through the regions led to an unprecedented toxic Alexandrium fundyense bloom (red tide). Both model results and field measurements suggest that the western Maine Coastal Current separated from Cape Ann around May 7-8, and the eddy formed on around May 10. The eddy was trapped at the formation location for about a week before detaching from the coastline and moving slowly southward on May 17. Both model results and theoretical analysis suggest that the separation of the coastal current from the coast and subsequent eddy formation were initiated at the subsurface by an adverse pressure gradient between Cape Ann and MB due to the higher sea level set up by onshore Ekman transport and higher density in downstream MB. After the formation, the eddy was maintained by the input of vorticity transported by the coastal current from the north, and local vorticity generation around the cape by the horizontal gradients of wind-driven currents, bottom stress, and water density induced by the Merrimack River plume. Observations and model results indicate that the anti-cyclonic eddy significantly changed the pathway of nutrient and biota transport into the coastal areas and enhanced phytoplankton including Alexandrium abundances around the perimeter of the eddy and in the western coast of MB.

  1. Accumulation and depuration of paralytic shellfish poisoning toxins by laboratory cultured purple clam Hiatula diphos Linnaeus.

    PubMed

    Chou, Hong Nong; Huang, Chen Ping; Chen, Chih Yu

    2005-10-01

    Purple clams (Hiatula diphos Linnaeus) accumulate paralytic shellfish poisoning (PSP) toxins produced by a toxic strain of the dinoflagellate Alexandrium minutum Halim in a laboratory study. The maximal toxicity of PSP toxins attained 31.3m MU/g after 20 days exposure. The toxin profile of H. diphos was similar to that reported for A. minutum at the end of the exposure period; and GTX1 was dominant. GTX congeners were found in muscle on day 16 and day 20, these substances could be detected during the depuration period as well. GTX1 was detected in the siphon only on day 32. The results show that H. diphos accumulates PSP toxins according to the amount and toxin profile of ingested A. minutum. PMID:16137734

  2. 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.

  3. 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. PMID:25176277

  4. 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.

  5. 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.

  6. 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

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

    USGS Publications Warehouse

    McGillicuddy, D.J., Jr.; 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.

  8. 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.

  9. 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.

  10. Removal of Two Species of Harmful Algae Using Gramine Modified Montmorillonite.

    PubMed

    Liu, Su-Li; Wang, Wen-Wen; Liu, Fei; Li, Hong-Ye; Liu, Jie-Sheng; Yang, Wei-Dong

    2016-04-01

    A series of gramine modified montmorillonites (gramine-MMTs) were prepared and characterized by thermogravimetric analysis, X-ray diffraction and zeta potential measurements. These modified clays with various amounts of gramine all exhibited higher removal efficiencies of both Chattonella marina and Alexandrium tamarense than did natural clay. With the augmentation of gramine content in the modified clays, the removal efficiency increased. The 24 h LC50 values of gramine-MMT with 12.68 % of gramine on C. marina and A. tamarense were 9.16 and 10.21 mg L(-1), respectively. These data suggest gramine-MMT might be a promising material to remediate harmful algal blooms. PMID:26920698

  11. 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. PMID:27050798

  12. 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

  13. 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.

  14. The Dinoflagellate Toxin 20-Methyl Spirolide-G Potently Blocks Skeletal Muscle and Neuronal Nicotinic Acetylcholine Receptors.

    PubMed

    Couesnon, Aurélie; Aráoz, Rómulo; Iorga, Bogdan I; Benoit, Evelyne; Reynaud, Morgane; Servent, Denis; Molgó, Jordi

    2016-01-01

    The cyclic imine toxin 20-methyl spirolide G (20-meSPX-G), produced by the toxigenic dinoflagellate Alexandrium ostenfeldii/Alexandrium peruvianum, has been previously reported to contaminate shellfish in various European coastal locations, as revealed by mouse toxicity bioassay. The aim of the present study was to determine its toxicological profile and its molecular target selectivity. 20-meSPX-G blocked nerve-evoked isometric contractions in isolated mouse neuromuscular preparations, while it had no action on contractions elicited by direct electrical stimulation, and reduced reversibly nerve-evoked compound muscle action potential amplitudes in anesthetized mice. Voltage-clamp recordings in Xenopus oocytes revealed that 20-meSPX-G potently inhibited currents evoked by ACh on Torpedo muscle-type and human α7 nicotinic acetylcholine receptors (nAChR), whereas lower potency was observed in human α4β2 nAChR. Competition-binding assays showed that 20-meSPX-G fully displaced [³H]epibatidine binding to HEK-293 cells expressing the human α3β2 (Ki = 0.040 nM), whereas a 90-fold lower affinity was detected in human α4β2 nAChR. The spirolide displaced [(125)I]α-bungarotoxin binding to Torpedo membranes (Ki = 0.028 nM) and in HEK-293 cells expressing chick chimeric α7-5HT₃ nAChR (Ki = 0.11 nM). In conclusion, this is the first study to demonstrate that 20-meSPX-G is a potent antagonist of nAChRs, and its subtype selectivity is discussed on the basis of molecular docking models. PMID:27563924

  15. 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. PMID:26965334

  16. 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

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

    2015-10-01

    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

  17. Toxin Levels and Profiles in Microalgae from the North-Western Adriatic Sea—15 Years of Studies on Cultured Species

    PubMed Central

    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

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

    PubMed

    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

  19. 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

  20. 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

    2015-12-01

    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

  1. 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

  2. Feeding, respiration, and excretion of the Black Sea Noctiluca scintillans MacCartney in summer

    NASA Astrophysics Data System (ADS)

    Drits, A. V.; Nikishina, A. B.; Sergeeva, V. M.; Solov'ev, K. A.

    2013-07-01

    Studies were conducted at the end of June 2011 in the coastal region of the northeastern part of the Black Sea. The bulk of the Noctiluca scintillans population was observed in the thermocline and reached a density of 40000 ind./m3. Analysis of digestive vacuoles content showed that Noctiluca could consume cells of Neoceratium tripos and N. furca, which had been considered inedible for Black Sea zooplankton, as well as temporary cysts of dinoflagellates, presumably of the toxic genus Alexandrium. The Noctiluca population consumed in total 10-30% of the abundance of temporary cysts, 2-29% of primary production, and 2-9% of potential Calanus euxinus egg production. For the first time, the excretion rates of ammonium nitrogen and mineral phosphorus were measured for N. scintillans. Our calculations showed that in summer, excretion by Noctiluca contributed from 4 to 18% and from 15 to 53% of phytoplankton total nitrogen and phosphorus requirements, respectively. The specific growth rate of Noctiluca (0.17-0.35) in summer, estimated from data on the daily food intake and respiration rate, was close to the values obtained in spring.

  3. Results of a Saxitoxin Proficiency Test Including Characterization of Reference Material and Stability Studies

    PubMed Central

    Harju, Kirsi; Rapinoja, Marja-Leena; Avondet, Marc-André; Arnold, Werner; Schär, Martin; Luginbühl, Werner; Kremp, Anke; Suikkanen, Sanna; Kankaanpää, Harri; Burrell, Stephen; Söderström, Martin; Vanninen, Paula

    2015-01-01

    A saxitoxin (STX) proficiency test (PT) was organized as part of the Establishment of Quality Assurance for the Detection of Biological Toxins of Potential Bioterrorism Risk (EQuATox) project. The aim of this PT was to provide an evaluation of existing methods and the European laboratories’ capabilities for the analysis of STX and some of its analogues in real samples. Homogenized mussel material and algal cell materials containing paralytic shellfish poisoning (PSP) toxins were produced as reference sample matrices. The reference material was characterized using various analytical methods. Acidified algal extract samples at two concentration levels were prepared from a bulk culture of PSP toxins producing dinoflagellate Alexandrium ostenfeldii. The homogeneity and stability of the prepared PT samples were studied and found to be fit-for-purpose. Thereafter, eight STX PT samples were sent to ten participating laboratories from eight countries. The PT offered the participating laboratories the possibility to assess their performance regarding the qualitative and quantitative detection of PSP toxins. Various techniques such as official Association of Official Analytical Chemists (AOAC) methods, immunoassays, and liquid chromatography-mass spectrometry were used for sample analyses. PMID:26602927

  4. Long term phytoplankton monitoring, including harmful algal blooms, in the Bay of Fundy, eastern Canada

    NASA Astrophysics Data System (ADS)

    Martin, J. L.; Hanke, A. R.; LeGresley, M. M.

    2009-01-01

    A monitoring program was initiated in May 1987 to study phytoplankton populations in the Bay of Fundy, southwest New Brunswick, eastern Canada. Samples are collected for phytoplankton distribution and abundance at five locations in the Bay of Fundy. Other parameters measured include plant nutrients (ammonia, nitrite, nitrate, phosphate and silicate), Secchi depth, and depth profiles for fluorescence, oxygen, temperature and salinity. Alexandrium fundyense abundance from the 5 sites and between years is compared to physical and chemical properties of seawater using principle component analyses (PCA) to identify factors showing the greatest amount of variance in temporal and spatial distribution patterns. Analysis of A. fundyense abundance over the 19-year period 1987-2005 indicates that cell abundance from one year does not reflect the following year's phytoplankton concentration, and nitrate values and cell densities appear to have a negative relationship. A further comparison between the 2 years 2004 and 2005 (years with very different intensities of A. fundyense maximum cell concentrations) further supported these findings. Preliminary analyses indicate that many species abundances and intensities appear to be more climate or weather related than nutrient flux related. Examination of relationships between harmful algal bloom (HAB) cell density, nutrients and environmental variables indicates that there is no evidence that HABs are linked to eutrophication processes at the temporal and spatial scales of the study.

  5. New Insights into the Parasitoid Parvilucifera sinerae Life Cycle: The Development and Kinetics of Infection of a Bloom-forming Dinoflagellate Host.

    PubMed

    Alacid, Elisabet; Reñé, Albert; Garcés, Esther

    2015-12-01

    Parvilucifera sinerae is a parasitoid of dinoflagellates, the major phytoplankton group responsible for harmful algal bloom events. Here we provide a detailed description of both the life cycle of P. sinerae, based on optical, confocal, and transmission electron microscopy observations, and its infection kinetics and dynamics. P. sinerae completes its life cycle in 3-4 days. The zoospore encounters and penetrates the host cell within 24h after its addition to the host culture. Inside the host, the parasitoid develops a trophocyte, which constitutes the longest stage of its life cycle. The trophocyte replicates and divides by schizogony to form hundreds of new zoospores contained within a sporangium. Under laboratory conditions, P. sinerae has a short generation time, a high rate of asexual reproduction, and is highly prevalent (up to 80%) in the Alexandrium minutum population. Prevalence was shown to depend on both the parasitoid inoculum size and host density, which increase the encounter probability rate. The parasitoid infection parameters described in this study are the first reported for the genus Parvilucifera. They show that P. sinerae is well-adapted to its dinoflagellate hosts and may be an important factor in the termination of A. minutum blooms in the natural environment. PMID:26605683

  6. Effect of the Algaecide Palmitoleic Acid on the Immune Function of the Bay Scallop Argopecten irradians.

    PubMed

    Chi, Cheng; Giri, Sib Sankar; Jun, Jin Woo; Kim, Hyoun Joong; Kim, Sang Guen; Yun, Saekil; Park, Se Chang

    2016-01-01

    Palmitoleic acid (PA), an algicidal compound, is used against the toxin producing dinofagelate Alexandrium tamarense, however, its impact on the edible bay scallop (Argopecten irradians) is still unclear. Therefore, we investigated the impacts of effective algicidal concentrations (20, 40, and 80 mg/L) of PA on immune responses in A. irradians. Various immune parameters including acid phosphatase (ACP) activity, superoxide dismutase (SOD), lysozyme, phagocytic activity, total protein, malondialdehyde (MDA) level, and reactive oxygen species (ROS) production and the expression of immune-related genes (PrxV, CLT-6, MT, and BD) were measured at 3, 6, 12, 24, and 48 h post-exposure (hpe) to PA. Lysozyme activity was lower in scallops at 12-48 hpe to 80 mg/L. SOD, ACP activity, ROS production, the total protein, and MDA level was higher at 12 to 48 hpe with different concentrations of PA. Phagocytic activity increased at 6-12 hpe to 40-80 mg/L of PA, but decreased at 24-48 hpe. The expressions of genes PrxV, CLT-6, MT and BD down-regulated at 3 hpe were observed, while differential expressions from 6-48 hpe with different concentrations of PA. The present study demonstrated that immersing A. irradians in PA at effective concentrations could result in differential effects on non-specific immune responses and expressions of immune-related genes. PMID:27171074

  7. Seasonal and interannual variability in Gulf of Maine hydrodynamics: 2002-2011

    NASA Astrophysics Data System (ADS)

    Li, Yizhen; He, Ruoying; McGillicuddy, Dennis J.

    2014-05-01

    In situ observations including long-term moored meteorological and oceanographic measurements and multi-year gulf-wide ship survey data are used to quantify interannual variability of surface wind, river runoff, and hydrographic conditions in the Gulf of Maine during summers 2002-2011. The cumulative upwelling index shows that upwelling (downwelling)-favorable wind conditions were most persistent in 2010 (2005) over the 10-year study period. River discharge was highest in 2005; peak runoff occurred in early April in 2010 as opposed to late April to middle May in other years. Moored time series show that coastal water temperature was 0.5-2 °C warmer than average in summer 2010, and about 2 °C colder than average in 2004. Coastal salinity in April 2010 was the lowest in the 10-year study period. Both moored Acoustic Doppler Current Profiler (ADCP) current measurements and dynamic height/geostrophic velocity calculations based on gulf-wide ship survey data show May-June 2010 had one of the weakest alongshore transports in the western Gulf of Maine during the 10-year study period, likely associated with intrusions of warm slope water and fresher-than-usual Scotian Shelf water. Comparisons of coastal currents to the Paralytic Shellfish Poisoning (PSP) closure maps resulting from Alexandrium fundyense blooms suggest a linkage between alongshore transport and the downstream extent of toxicity.

  8. 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.

  9. 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.

  10. Paralytic shellfish poisoning (PSP) in Margarita Island, Venezuela.

    PubMed

    La Barbera-Sánchez, Amelia; Franco Soler, Jose; Rojas de Astudillo, Luisa; Chang-Yen, Ivan

    2004-09-01

    A severe outbreak of Paralytic Shellfish Poisoning (PSP) occurred in Manzanillo and Guayacán, northwestern coast of Margarita Island, Venezuela, between August and October 1991. A bloom of dinoflagellates including Prorocentrum gracile, Gymnodinium catenatum and Alexandrium tamarense seemed to be responsible for this outbreak. Levels of PSP toxins in mussels (Perna perna) exceeded the international safety limit of saxitoxin, 80 microg STX/100 microg meat. PSP toxin values varied between 2548 and 115 microg STX/100 g meat in Manzanillo, and between 1422 and 86 microg STX/100 g meat in Guayacán. At both locations, the highest levels were detected in August, when 24 patients exhibited typical symptoms of PSP toxicity after consuming cooked mussels (16 required hospitalization). A high pressure liquid chromatographic (HPLC) procedure was recently used on the 1991 samples. The major toxin detected in samples of both locations was decarbamoyl saxitoxin (dcSTX), but low concentrations of saxitoxin were also found in Manzanillo samples. Gonyautoxins GTX1, GTX2 and GTX3 were detected only at Guayacán, while in both locations, decarbamoylgonyatouxin (dcGTX2,3) toxins were detected. These findings represent the first time that causative toxins of PSP in Venezuela have been chemically identified, and confirm the presence of dcSTX and dcGTX in mussels from the Caribbean Sea. The presence of dcSTX and dcGTX in shellfish is indicative that Gymnodinium catenatum was a causative organism for outbreak of PSP. PMID:17465121

  11. 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

  12. Prominent human health impacts from several marine microbes: history, ecology, and public health implications.

    PubMed

    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

  13. Australian dust storm associated with extensive Aspergillus sydowii fungal "bloom" in coastal waters.

    PubMed

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

    2014-06-01

    A massive central Australian dust storm in September 2009 was associated with abundant fungal spores (150,000/m(3)) 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

  14. 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. PMID:25918403

  15. Neurotoxic Alkaloids: Saxitoxin and Its Analogs

    PubMed Central

    Wiese, Maria; D’Agostino, Paul M.; Mihali, Troco K.; Moffitt, Michelle C.; Neilan, Brett A.

    2010-01-01

    Saxitoxin (STX) and its 57 analogs are a broad group of natural neurotoxic alkaloids, commonly known as the paralytic shellfish toxins (PSTs). PSTs are the causative agents of paralytic shellfish poisoning (PSP) and are mostly associated with marine dinoflagellates (eukaryotes) and freshwater cyanobacteria (prokaryotes), which form extensive blooms around the world. PST producing dinoflagellates belong to the genera Alexandrium, Gymnodinium and Pyrodinium whilst production has been identified in several cyanobacterial genera including Anabaena, Cylindrospermopsis, Aphanizomenon Planktothrix and Lyngbya. STX and its analogs can be structurally classified into several classes such as non-sulfated, mono-sulfated, di-sulfated, decarbamoylated and the recently discovered hydrophobic analogs—each with varying levels of toxicity. Biotransformation of the PSTs into other PST analogs has been identified within marine invertebrates, humans and bacteria. An improved understanding of PST transformation into less toxic analogs and degradation, both chemically or enzymatically, will be important for the development of methods for the detoxification of contaminated water supplies and of shellfish destined for consumption. Some PSTs also have demonstrated pharmaceutical potential as a long-term anesthetic in the treatment of anal fissures and for chronic tension-type headache. The recent elucidation of the saxitoxin biosynthetic gene cluster in cyanobacteria and the identification of new PST analogs will present opportunities to further explore the pharmaceutical potential of these intriguing alkaloids. PMID:20714432

  16. 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. PMID:26913820

  17. Cell bioassay for paralytic shellfish poisoning (PSP): comparison with postcolumn derivatization liquid chromatographic analysis and application to the monitoring of PSP in shellfish.

    PubMed

    Hayashi, Rumiko; Saito, Hiroshi; Okumura, Masanao; Kondo, Fumio

    2006-01-25

    We performed a neuroblastoma cell (Neuro2a) culture assay modified slightly from a method reported previously to provide a simple and sensitive evaluation of paralytic shellfish poisoning (PSP) toxicity in shellfish. The cell bioassay was just as sensitive for C-toxins as for gonyautoxins. The sensitivity of our cell bioassay was 4 times that of the current standard mouse bioassay. Using the cell bioassay, we evaluated PSP toxicity in 361 shellfish samples collected from Mikawa Bay and Ise Bay, Aichi Prefecture, Japan, from April 1999-March 2002. The results were compared with those obtained in a postcolumn derivatization liquid chromatographic analysis. PSP toxins were detected in 236/361 samples by both assays, and there was a fairly good correlation (r = 0.9001, n = 236, p < 0.001) between the results from the two assays. We applied this cell bioassay when short-necked clams in the bay turned poisonous in 2001. The chronological changes in PSP toxicity in the short-necked clams were analyzed and compared with those of the cell density of poisonous plankton (Alexandrium tamarense) occurring in the bay. The PSP toxicity in shellfish peaked 2 weeks after the cell density reached a maximum. We recommend using the cell bioassay for routine monitoring of PSP toxicity in shellfish living in natural marine environments. PMID:16417278

  18. Phylogenetic analysis of microalgae based on highly abundant proteins using mass spectrometry.

    PubMed

    Lee, Hae-Won; Roh, Seong Woon; Cho, Kichul; Kim, Kil-Nam; Cha, In-Tae; Yim, Kyung June; Song, Hye Seon; Nam, Young-Do; Oda, Tatsuya; Chung, Young-Ho; Kim, Soo Jung; Choi, Jong-Soon; Kim, Daekyung

    2015-01-01

    The blooms of toxic phototrophic microorganisms, such as microalgae and cyanobacteria, which are typically found in freshwater and marine environments, are becoming more frequent and problematic in aquatic systems. Due to accumulation of toxic algae, harmful algal blooms (HABs) exert negative effects on aquatic systems. Therefore, rapid detection of harmful microalgae is important for monitoring the occurrence of HABs. Mass spectrometry-based methods have become sensitive, specific techniques for the identification and characterization of microorganisms. Matrix-assisted laser desorption/ionization (MALDI) with time-of-flight (TOF) mass spectrometry (MS) allows us to measure a unique molecular fingerprint of highly abundant proteins in a microorganism and has been used for the rapid, accurate identification of bacteria and fungi in clinical microbiology. Here, we tested the specificity of MALDI-TOF MS using microalgal strains (Heterocapsa, Alexandrium, Nannochloropsis, Chaetoceros, Chlorella, and Dunaliella spp.). Our research suggested that this method was comparable in terms of the rapid identification of microalgea to conventional methods based on genetic information and morphology. Thus, this efficient mass spectrometry-based technique may have applications in the rapid identification of harmful microorganisms from aquatic environmental samples. PMID:25476355

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

    PubMed

    Paranjpye, Rohinee N; Nilsson, William B; Liermann, Martin; Hilborn, Elizabeth D; George, Barbara J; Li, Quanlin; Bill, Brian D; Trainer, Vera L; Strom, Mark S; Sandifer, Paul A

    2015-12-01

    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 Dinophysis), the abundance of diatoms and dinoflagellates, chlorophyll-a and temperature, salinity and macronutrients at five sites in Washington State from 2008-2009. The variability in V. parahaemolyticus density was explained predominantly by strong seasonal trends where maximum densities occurred in June, 2 months prior to the highest seasonal water temperature. In spite of large geographic differences in temperature, salinity and nutrients, there was little evidence of corresponding differences in V. parahaemolyticus density. In addition, there was no evident relationship between V. parahaemolyticus and indices of HAB genera, perhaps due to a lack of significant HAB events during the sampling period. The only nutrient significantly associated with V. parahaemolyticus density after accounting for the seasonal trend was silicate. This negative relationship may be caused by a shift in cell wall structure for some diatom species to a chitinous substrate preferred by V. parahaemolyticus. Results from our study differ from those in other regions corroborating previous findings that environmental factors that trigger vibrio and HAB events may differ depending on geographic locations. Therefore caution should be used when applying results from one region to another. PMID:26454066

  20. Application of a laser fluorometer for discriminating phytoplankton species

    NASA Astrophysics Data System (ADS)

    Chen, Peng; Pan, Delu; Mao, Zhihua

    2015-04-01

    A portable laser-induced fluorescence system for discriminating phytoplankton species has been developed. It consists of a high pulsed repetition frequency (10-kHz) microchip laser at 405 nm, a reflective fluorescent probe and a broadband micro spectrometer. The measured fluorescent spectra were overlapped by various fluorescent components, and were then decomposed by a bi-Gaussian mixture model. A spectral shape description index was designed to characterize fluorescent spectral shapes for descriminating the phytoplankton species cultured in our laboratory. Using clustering analysis, the samples of eight phytoplankton species belonging to two divisions of Bacillariophyta and Dinophyta were divided into six categories: 1) Chaetoceros debilis, Thalassiosira rotula; 2) Prorocentrum donghaiense, Prorocentrum dentatum; 3) Gymnodinium simplex; 4) Alexandrium tamarense; 5) Karenia mikimotoi; and 6) Akashiwo sanguinea. The phytoplankton species belonging to Bacillariophyta were well separated from those belonging to Dinophyta. In addition, the phytoplankton species belonging to Dinophyta were successfully distinguished from each other at genus level. The portable system is expected to be used both in vivo and in the field.

  1. Discrimination of phytoplankton classes using characteristic spectra of 3D fluorescence spectra

    NASA Astrophysics Data System (ADS)

    Zhang, Qian-Qian; Lei, Shu-He; Wang, Xiu-Lin; Wang, Lei; Zhu, Chen-Jian

    2006-02-01

    The discrimination of phytoplankton classes using the characteristic fluorescence spectra extracted from three-dimensional fluorescence spectra was investigated. Single species cultures of 11 phytoplankton species, representing 5 major phytoplankton divisions, were used. The 3D fluorescence spectra of the cultures grown at different temperatures (20 and 15 °C) and illumination intensities (140, 80 and 30 μM m -2 s -1) were measured and their feature extraction methods were explored. Ordering Rayleigh and Raman scattering data as zero, the obtained excitation-emission matrices were processed by both singular value decomposition (SVD) and trilinear decomposition methods. The resulting first principal component can be regarded as the characteristic spectrum of the original 3D fluorescence spectrum. The analysis shows that such characteristic spectra have a discriminatory capability. At different temperatures, the characteristic spectra of Isochrysis galbana, Platymonas helgolanidica and Skeletonema costatuma have high degrees of similarity to their own species samples, while the spectra similarities of Alexandrium tamarense, Prorocentrum dentatum, Pseudo-nitzschia pungens, Chaetoceros curvisetus, Ch. Debilis, Ch. Didymus and Synechococcus sp. are not as significant as the other three species. C. curvisetus, Ch. Debilis and Ch. Didymus, belonging to genus Chaetoceros, have identical spectra and cannot be discriminated at all. Regarding all six diatom species as one class, the average discriminant error rate is below 9%. It is worth mentioning that the diatom class can be distinguished from A. tamarense and P. dentatum, which belong to Dinophyta.

  2. Formation of a volunteer harmful algal bloom network in British Columbia, Canada, following an outbreak of diarrhetic shellfish poisoning.

    PubMed

    McIntyre, Lorraine; Cassis, David; Haigh, Nicola

    2013-11-01

    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

  3. Marine dinoflagellates show induced life-history shifts to escape parasite infection in response to water-borne signals.

    PubMed

    Toth, Gunilla B; Norén, Fredrik; Selander, Erik; Pavia, Henrik

    2004-04-01

    Many dinoflagellate species form dormant resting cysts as a part of their life cycle, and in some freshwater species, hatching of these cysts can be delayed by the presence of water-borne signals from grazing zooplankton. Some marine dinoflagellates can form temporary cysts, which may function to resist unfavourable short-term environmental conditions. We investigated whether the marine dinoflagellate Alexandrium ostenfeldii is able to induce an increased resistance to the parasitic flagellate Parvilucifera infectans by forming temporary cysts. We performed several laboratory experiments where dinoflagellates were exposed either to direct contact with parasites or to filtered water from cultures of parasite-infected conspecifics (parasite-derived signals). Infection by P. infectans is lethal to motile A. ostenfeldii cells, but temporary cysts were more resistant to parasite infection. Furthermore, A. ostenfeldii induced a shift in life-history stage (from motile cells to temporary cysts) when exposed to parasite-derived water-borne signals. The response was relaxed within a couple of hours, indicating that A. ostenfeldii may use this behaviour as a short-term escape mechanism to avoid parasite infection. The results suggest that intraspecies chemical communication evoked by biotic interactions can be an important mechanism controlling life-history shifts in marine dinoflagellates, which may have implications for the development of toxic algal blooms. PMID:15209107

  4. Green alga Ulva pertusa--a new source of bioactive compounds with antialgal activity.

    PubMed

    Ying-ying, Sun; Hui, Wang; Gan-lin, Guo; Yin-fang, Pu; Bin-lun, Yan; Chang-hai, Wang

    2015-07-01

    We tested the effects of solvent fractions (FA, FB, FC, and FD), which partitioned by liquid-liquid extraction from the methanol extract of Ulva pertusa, on the growth of red tide microalgae (Karenia mikimitoi, Skeletonema costatum, Alexandrium tamarense, Heterosigma akashiwo, Prorocentrum donghaiense), and FA, FB, and FC exhibited significantly antialgal activity. The chemical constituent analysis showed the existence of bioactive compounds such as phenols and alkaloids. Further, four solvent fractions were applied to silica gel column and repeated preparative TLC to produce 13 samples and their purity qualified as thin-layer chromatographic grade. Among these purified samples, FA111, FB411, FC411, FD111, and FD211 exhibited stronger antialgal activity. Furthermore, their functional groups were analyzed by colorimetric methods and UV spectra data. FD111 and FD211 were temptatively identified as alkaloids; the others were initially identified as phenolic acids. This is a preliminary study and the structure identification of these purified samples requires further investigation. While concentration of these purified samples in this algae was very small, they showed excellent effects against red tide microalgae. PMID:25724801

  5. 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

  6. Bioluminescent response of individual dinoflagellate cells to hydrodynamic stress measured with millisecond resolution in a microfluidic device.

    PubMed

    Latz, Michael I; Bovard, Michelle; VanDelinder, Virginia; Segre, Enrico; Rohr, Jim; Groisman, Alex

    2008-09-01

    Dinoflagellate bioluminescence serves as a model system for examining mechanosensing by suspended motile unicellular organisms. The response latency, i.e. the delay time between the mechanical stimulus and luminescent response, provides information about the mechanotransduction and signaling process, and must be accurately known for dinoflagellate bioluminescence to be used as a flow visualization tool. This study used a novel microfluidic device to measure the response latency of a large number of individual dinoflagellates with a resolution of a few milliseconds. Suspended cells of several dinoflagellate species approximately 35 microm in diameter were directed through a 200 microm deep channel to a barrier with a 15 microm clearance impassable to the cells. Bioluminescence was stimulated when cells encountered the barrier and experienced an abrupt increase in hydrodynamic drag, and was imaged using high numerical aperture optics and a high-speed low-light video system. The average response latency for Lingulodinium polyedrum strain HJ was 15 ms (N>300 cells) at the three highest flow rates tested, with a minimum latency of 12 ms. Cells produced multiple flashes with an interval as short as 5 ms between individual flashes, suggesting that repeat stimulation involved a subset of the entire intracellular signaling pathway. The mean response latency for the dinoflagellates Pyrodinium bahamense, Alexandrium monilatum and older and newer isolates of L. polyedrum ranged from 15 to 22 ms, similar to the latencies previously determined for larger dinoflagellates with different morphologies, possibly reflecting optimization of dinoflagellate bioluminescence as a rapid anti-predation behavior. PMID:18723546

  7. 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

  8. 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. PMID:23300940

  9. 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

  10. 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.

  11. First direct fluorescence polarization assay for the detection and quantification of spirolides in mussel samples.

    PubMed

    Otero, Paz; Alfonso, Amparo; Alfonso, Carmen; Aráoz, Rómulo; Molgó, Jordi; Vieytes, Mercedes R; Botana, Luis M

    2011-09-01

    In 2009, we achieve the first inhibition FP assay to detect imine cyclic toxins. In the present paper we propose a new FP assay for direct quantify spirolides. This new method has resulted in significant improvement of sensitivity, rapidity and accessibility. In the method design, nicotinic acetylcholine receptor from Torpedo marmorata membranes labelled with a derivative of fluorescein was used. Spirolides, 13-desmethyl spirolide C (13-desMeC) and 13,19-didesmethyl spirolide C (13,19-didesMeC) were extracted and purified from cultures of the Alexandrium ostenfeldii dinoflagellate. Data showed the decrease of FP when toxin concentration was increased. Thus, a relationship between the FP units and the spirolides amount present in a sample was obtained. This direct assay is a reproducible, simple and very sensitive method with a detection limit about 25 nM for 13-desMeC and 150 nM for 13,19-didesMeC. The procedure was used to measure spirolides in mussel samples using an extraction and clean up protocol suitable for the FP assay. Results obtained show that this method is able to quantify 13-desMeC in the range of 50-350 μg kg(-1) meat. Other liposoluble toxins did not interfere with the assay, proving a specific method. Moreover, the matrix do not affect in the range of toxin concentrations that involving risk of spirolides intoxication. PMID:21801889

  12. Patterns of Post-Glacial Genetic Differentiation in Marginal Populations of a Marine Microalga

    PubMed Central

    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 (FST) 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. PMID:23300940

  13. Results of a Saxitoxin Proficiency Test Including Characterization of Reference Material and Stability Studies.

    PubMed

    Harju, Kirsi; Rapinoja, Marja-Leena; Avondet, Marc-André; Arnold, Werner; Schär, Martin; Luginbühl, Werner; Kremp, Anke; Suikkanen, Sanna; Kankaanpää, Harri; Burrell, Stephen; Söderström, Martin; Vanninen, Paula

    2015-12-01

    A saxitoxin (STX) proficiency test (PT) was organized as part of the Establishment of Quality Assurance for the Detection of Biological Toxins of Potential Bioterrorism Risk (EQuATox) project. The aim of this PT was to provide an evaluation of existing methods and the European laboratories' capabilities for the analysis of STX and some of its analogues in real samples. Homogenized mussel material and algal cell materials containing paralytic shellfish poisoning (PSP) toxins were produced as reference sample matrices. The reference material was characterized using various analytical methods. Acidified algal extract samples at two concentration levels were prepared from a bulk culture of PSP toxins producing dinoflagellate Alexandrium ostenfeldii. The homogeneity and stability of the prepared PT samples were studied and found to be fit-for-purpose. Thereafter, eight STX PT samples were sent to ten participating laboratories from eight countries. The PT offered the participating laboratories the possibility to assess their performance regarding the qualitative and quantitative detection of PSP toxins. Various techniques such as official Association of Official Analytical Chemists (AOAC) methods, immunoassays, and liquid chromatography-mass spectrometry were used for sample analyses. PMID:26602927

  14. Comparison of paralytic shellfish poisoning toxin between carnivorous crabs (Telmessus acutidens and Charybdis japonica) and their prey mussel (Mytilus galloprovincialis) in an inshore food chain.

    PubMed

    Oikawa, Hiroshi; Fujita, Tsuneo; Saito, Ken; Watabe, Shugo; Satomi, Masataka; Yano, Yutaka

    2004-05-01

    Paralytic shellfish poisoning toxin in two shore crab species, Telmessus acutidens and Charybdis japonica, were compared with the toxin in the prey mussel Mytilus galloprovincialis and causative dinoflagellates Alexandrium tamarense, all having been collected at Onahama, Fukushima Prefecture, in the northern part of Japan. When the toxicities were detected in mussels by mouse bioassays, 73.7% of the sampled T. acutidens were toxic in the hepatopancreas. T. acutidens has been found to become toxic for three years, therefore, it can be concluded that the crab commonly and repeatedly accumulate the toxins via the food chain at Onahama. C. japonica was also expected to be a possible vector species, because small quantities of the toxins were detected in eight specimens of the crab by HPLC analysis. By the comparison of the toxin profiles in the dinoflagellates, mussels and the crab T. acutidens, reductive conversions of GTX1 and GTX4 were observed when the toxins passed through the three species in the food chain. But increases of STX and neoSTX by further reductive process were not observed in the crab. The absence of the STX group toxins in the crab suggests that the crab eliminates the toxin before such reductive process occur. PMID:15109892

  15. Optofluidic characterization of marine algae using a microflow cytometer.

    PubMed

    Hashemi, Nastaran; Erickson, Jeffrey S; Golden, Joel P; Ligler, Frances S

    2011-09-01

    The effects of global warming, pollution in river effluents, and changing ocean currents can be studied by characterizing variations in phytoplankton populations. We demonstrate the design and fabrication of a Microflow Cytometer for characterization of phytoplankton. Guided by chevron-shaped grooves on the top and bottom of a microfluidic channel, two symmetric sheath streams wrap around a central sample stream and hydrodynamically focus it in the center of the channel. The lasers are carefully chosen to provide excitation light close to the maximum absorbance wavelengths for the intrinsic fluorophores chlorophyll and phycoerythrin, and the excitation light is coupled to the flow cytometer through the use of an optical fiber. Fluorescence and light scatter are collected using two multimode optical fibers placed at 90-degree angles with respect to the excitation fiber. Light emerging from these collection fibers is directed through optical bandpass filters into photomultiplier tubes. The cytometer measured the optical and side scatter properties of Karenia b., Synechococcus sp., Pseudo-Nitzchia, and Alexandrium. The effect of the sheath-to-sample flow-rate ratio on the light scatter and fluorescence of these marine microorganisms was investigated. Reducing the sample flow rate from 200 μL/min to 10 μL/min produced a more tightly focused sample stream and less heterogeneous signals. PMID:22662031

  16. 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. PMID:16677346

  17. Homology-Driven Proteomics of Dinoflagellates with Unsequenced Genomes Using MALDI-TOF/TOF and Automated De Novo Sequencing

    PubMed Central

    Wang, Da-Zhi; Li, Cheng; Xie, Zhang-Xian; Dong, Hong-Po; Lin, Lin; Hong, Hua-Sheng

    2011-01-01

    This study developed a multilayered, gel-based, and underivatized strategy for de novo protein sequence analysis of unsequenced dinoflagellates using a MALDI-TOF/TOF mass spectrometer with the assistance of DeNovo Explorer software. MASCOT was applied as the first layer screen to identify either known or unknown proteins sharing identical peptides presented in a database. Once the confident identifications were removed after searching against the NCBInr database, the remainder was searched against the dinoflagellate expressed sequence tag database. In the last layer, those borderline and nonconfident hits were further subjected to de novo interpretation using DeNovo Explorer software. The de novo sequences passing a reliability filter were subsequently submitted to nonredundant MS-BLAST search. Using this layer identification method, 216 protein spots representing 158 unique proteins out of 220 selected protein spots from Alexandrium tamarense, a dinoflagellate with unsequenced genome, were confidently or tentatively identified by database searching. These proteins were involved in various intracellular physiological activities. This study is the first effort to develop a completely automated approach to identify proteins from unsequenced dinoflagellate databases and establishes a preliminary protein database for various physiological studies of dinoflagellates in the future. PMID:21977052

  18. 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

  19. 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 . PMID:24320746

  20. 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. PMID:23922056

  1. 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. PMID:25076815

  2. 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. PMID:25682049

  3. 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.

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

    PubMed

    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

  5. 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

  6. Pharmacokinetic and toxicological data of spirolides after oral and intraperitoneal administration.

    PubMed

    Otero, Paz; Alfonso, Amparo; Rodríguez, Paula; Rubiolo, Juan A; Cifuentes, José Manuel; Bermúdez, Roberto; Vieytes, Mercedes R; Botana, Luis M

    2012-02-01

    Spirolides are a kind of marine toxins included in the cyclic imine toxin group and produced by the dinoflagellate Alexandrium ostenfeldii. This study shows for the first time a complete and detailed description about the symptoms observed in mice when these toxins were intraperitoneal (i.p.) administered. It is also compared the i.p. toxicity of 13-desmethyl spirolide C (13-desMeC), 13,19-didesMeC (13,19-didesMeC) and 20-methyl spirolide G (20-Me-G) in experiments performed with highly purified toxins. The bioassay indicates that 13-desMeC and 13,19-didesMeC are extremely toxic compounds which have a LD(50) of 27.9μg/kg and 32.2μg/kg, respectively. However, when 20-MeG was i.p administrated with dose up 63.5μg/kg, no deaths were recorded. In order to evaluate the oral toxicity, spirolides were administered by gastric intubation into mice. Then, samples of blood, urine and faeces were collected and analyzed by liquid chromatography-mass spectrometry tandem (LC-MS/MS) technique. Spirolides appear in blood at 15min and in urine after 1h of being toxin administered. In summary, in this paper, it is provided new data about the toxicity, absorption, and excretion of spirolides in mouse. So far, little information is available on this item but necessary for spirolide regulation in the European Union (EU). PMID:22100396

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

    PubMed

    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. 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. PMID:25765159

  9. Nutrient and Phytoplankton Analysis of a Mediterranean Coastal Area

    NASA Astrophysics Data System (ADS)

    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. 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

  11. 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

  12. 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.

  13. Allelopathic interactions between the macroalga Ulva pertusa and eight microalgal species

    NASA Astrophysics Data System (ADS)

    Nan, Chunrong; Zhang, Haizhi; Zhao, Guangqiang

    2004-11-01

    Growth of Ulva pertusa and eight microalgal species, Heterosigma akashiwo, Skeletonema costatum, Tetraselmis subcordiformis, Nitzschia closterium, Chaetoceros gracile, Chroomonas placoidea 1967, Isochrysis galbana 8701, and Alexandrium tamarense, was examined in a series of batch, semi-continuous and isolated co-cultures ( U. pertusa and one microalgal species). The results of the experiments with co-cultures confirmed the secretion of allelopathic substances by U. pertusa. Growth was significantly ( p<0.05) suppressed in each of the macroalgal species in batch co-cultures, nutrient replete semi-continuous co-cultures and isolated co-cultures. The percentage growth reduction varied between 42 and 100% in batch co-cultures, between 28 and 100% in semi-continuous co-cultures, and between 21 and 100% in isolated co-cultures. In addition, we examined the potential allelopathic effect of U. pertusa culture filtrate. The Ulva culture filtrate significantly ( p<0.01) inhibited the growth of C. placoidea from 2 days after incubation until the end of the experiment, and it exhibited no inhibitory effect on the growth of the other microalgal species. This may suggest that the allelochemicals released from U. pertusa are rapidly degradable. The microalgae tested exhibited different (stimulatory, inhibitory or no) effects on the growth of U. pertusa. U. pertusa grew faster with H. akashiwo (+16%) and S. costatum (+9%), less with T. subcordiformis (-20%), N. closterium (-23%) and C. gracile (-30%), but was not significantly affected by I. galbana, A. tamarense and C. placoidea. The microalgae tested exhibited no clear allelopathic effects on U. pertusa.

  14. 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. PMID:19999976

  15. Competition of bloom-forming marine phytoplankton at low nutrient concentrations.

    PubMed

    Hu, Hanhua; Zhang, Jun; Chen, Weidong

    2011-01-01

    Competition of three bloom-forming marine phytoplankton (diatom Skeletonema costatum, and dinoflagellates Prorocentrum minimum and Alexandrium tamarense) was studied through a series of multispecies cultures with different nitrate (NaNO3) and phosphate (NaH2PO4) levels and excess silicate to interpret red tide algae succession. S. costatum outgrew the other two dinoflagellates in nitrate and phosphate replete cultures with 10 micromol/L Na2SiO3. Under nitrate limited (8.82 micromol/L NaNO3) conditions, the growth of S. costatum was also dominant when phosphate concentrations were from 3.6 to 108 micromol/L. Cell density of the two dinoflagellates only increased slightly, to less than 400 and 600 cells/mL, respectively. Cell density of S. costatum decreased with time before day 12, and then increased to 4000 cells/mL (1.5 mg/L dry biomass) at NaNO3 concentrations between 88.2 and 882 micromol/L with limited phosphate (0.36 micromol/L NaH2PO4) levels. In addition, P. minimum grew well with a maximal cell density of 1690-2100 cells/mL (0.5-0.6 mg/L dry biomass). Although S. costatum initially grew fast, its cell density decreased quickly with time later in the growth phase and the two dinoflagellates were dominant under the nitrate-limited and high nitrate conditions with limited phosphate. These results indicated that the diatom was a poor competitor compared to the two dinoflagellates under limited phosphate; however, it grew well under limited nitrate when growth of the dinoflagellates was near detection limits. PMID:21793409

  16. 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.

  17. 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

  18. 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

  19. 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

  20. Comparison of three protein extraction procedures from toxic and non-toxic dinoflagellates for proteomics analysis.

    PubMed

    Jiang, Xi-Wen; Wang, Jing; Chan, Leo Lai; Lam, Paul Kwan Sing; Gu, Ji-Dong

    2015-08-01

    Three methods for extraction and preparation of high-quality proteins from both toxic and non-toxic dinoflagellates for proteomics analysis, including Trizol method, Lysis method and Tris method, were compared with the subsequent protein separation profiles using 2-D differential gel electrophoresis (2-D DIGE), Coomassie Blue and silver staining. These methods showed suitability for proteins with different pIs and molecular weights. Tris method was better for low molecular weight and low pI protein isolation; whereas both Lysis and Trizol method were better for high-molecular weight and high pI protein purification. Trizol method showed good results with Alexandrium species and Gynodinium species, and the background in gel was much clearer than the other two methods. At the same time, only Lysis method caused breaking down of the target proteins. On the other hand, Trizol method obtained higher concentration of ribulose-1,5-bisphosphate carboxylase/oxygenase proteins by Western-blotting, while Tris method was the best for peridinin-chlorophyll-protein complexes protein and T1 protein preparation. DIGE was better than Coomassie Blue and silver staining, except for some limitations, such as the high cost of the dyes, relatively short shelf life and the requirements for extensive and special image capturing equipment. Some proteins related to PSTs synthesis in dinoflagellates are hydrophobic with high molecular weight or binding on membranes and Trizol method performed better than Tris method for these proteins. The Trizol method and 2-D DIGE were effective combination for proteomics investigations of dinoflagellates. This procedure allows reliable and high recovery efficiency of proteins from dinoflagellates for better understanding on their occurrence and toxin-production for physiological and biochemical information. PMID:26197730

  1. 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

  2. Seasonal dynamics of harmful algae in outer Oslofjorden monitored by microarray, qPCR, and microscopy.

    PubMed

    Dittami, Simon M; Hostyeva, Vladyslava; Egge, Elianne Sirnæs; Kegel, Jessica U; Eikrem, Wenche; Edvardsen, Bente

    2013-10-01

    Monitoring of marine microalgae is important to predict and manage harmful algal blooms. Microarray Detection of Toxic ALgae (MIDTAL) is an FP7-funded EU project aiming to establish a multi-species microarray as a tool to aid monitoring agencies. We tested the suitability of different prototype versions of the MIDTAL microarray for the monthly monitoring of a sampling station in outer Oslofjorden during a 1-year period. Microarray data from two different versions of the MIDTAL chip were compared to results from cell counts (several species) and quantitative real-time PCR (qPCR; only Pseudochattonella spp.). While results from generation 2.5 microarrays exhibited a high number of false positive signals, generation 3.3 microarray data generally correlated with microscopy and qPCR data, with three important limitations: (1) Pseudo-nitzschia cells were not reliably detected, possibly because cells were not sufficiently retained during filtration or lysed during the extraction, and because of low sensitivity of the probes; (2) in the case of samples with high concentrations of non-target species, the sensitivity of the arrays was decreased; (3) one occurrence of Alexandrium pseudogonyaulax was not detected due to a 1-bp mismatch with the genus probe represented on the microarray. In spite of these shortcomings our data demonstrate the overall progress made and the potential of the MIDTAL array. The case of Pseudochattonella - where two morphologically similar species impossible to separate by light microscopy were distinguished - in particular, underlines the added value of molecular methods such as microarrays in routine phytoplankton monitoring. PMID:23325054

  3. 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. PMID:26986890

  4. 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

  5. Discovery of Nuclear-Encoded Genes for the Neurotoxin Saxitoxin in Dinoflagellates

    PubMed Central

    Stüken, Anke; Orr, Russell J. S.; Kellmann, Ralf; Murray, Shauna A.; Neilan, Brett A.; Jakobsen, Kjetill S.

    2011-01-01

    Saxitoxin is a potent neurotoxin that occurs in aquatic environments worldwide. Ingestion of vector species can lead to paralytic shellfish poisoning, a severe human illness that may lead to paralysis and death. In freshwaters, the toxin is produced by prokaryotic cyanobacteria; in marine waters, it is associated with eukaryotic dinoflagellates. However, several studies suggest that saxitoxin is not produced by dinoflagellates themselves, but by co-cultured bacteria. Here, we show that genes required for saxitoxin synthesis are encoded in the nuclear genomes of dinoflagellates. We sequenced >1.2×106 mRNA transcripts from the two saxitoxin-producing dinoflagellate strains Alexandrium fundyense CCMP1719 and A. minutum CCMP113 using high-throughput sequencing technology. In addition, we used in silico transcriptome analyses, RACE, qPCR and conventional PCR coupled with Sanger sequencing. These approaches successfully identified genes required for saxitoxin-synthesis in the two transcriptomes. We focused on sxtA, the unique starting gene of saxitoxin synthesis, and show that the dinoflagellate transcripts of sxtA have the same domain structure as the cyanobacterial sxtA genes. But, in contrast to the bacterial homologs, the dinoflagellate transcripts are monocistronic, have a higher GC content, occur in multiple copies, contain typical dinoflagellate spliced-leader sequences and eukaryotic polyA-tails. Further, we investigated 28 saxitoxin-producing and non-producing dinoflagellate strains from six different genera for the presence of genomic sxtA homologs. Our results show very good agreement between the presence of sxtA and saxitoxin-synthesis, except in three strains of A. tamarense, for which we amplified sxtA, but did not detect the toxin. Our work opens for possibilities to develop molecular tools to detect saxitoxin-producing dinoflagellates in the environment. PMID:21625593

  6. Dinoflagellate cysts in recent marine sediments from Thermaikos Gulf, Greece: Effects of resuspension events on vertical cyst distribution

    NASA Astrophysics Data System (ADS)

    Giannakourou, A.; Orlova, T. Y.; Assimakopoulou, G.; Pagou, K.

    2005-12-01

    A qualitative and semi-quantitative study of recent dinoflagellate cysts has been undertaken in the NW part of Aegean Sea, Thermaikos Gulf (Eastern Mediteranean), before (September 2001), during (October 2001) and after 120 days (February 2002) of intensive trawling activities. This is the first survey of recent dinoflagellate cysts from Greek marine coastal environments. Sediment samples were collected with a corer and the vertical distribution of the cysts was studied at five different layers, from 0 to 10 cm. Dinoflagellate cysts were both abundant and diverse. Cysts were found over the whole sampling area and periods, with concentrations ranging between 247-3202 cysts cm -3. Thirty-six cyst types were encountered, of which 32 were identified to species level, representing 12 genera. It seems that significant local resuspension, related to the onset of the trawling period and stirring up of the sediment, contributed to mixing of the upper layers, resulting to more homogenous cyst profiles in the sediment. Viable cysts constituted 16-60% of the total cyst abundance. The abundance peaks of viable cysts within the subsurface sediment layers, observed during the undisturbed period, disappeared during October. In February, the reduction of cyst concentration was associated to a loss of viable cysts, whilst the ratio of viable/empty cysts ranged between 0.30 and 0.67. The abundance of the different dinoflagellate species, in their active form, was monitored in order to detect any relationship between the concentration of cysts in the top 10 cm of sediment and blooms of algae in the water column. Cysts of potentially toxic species, causing Paralitic Shellfish Poisoning (PSP), such as Alexandrium cf. tamarense, A. cf. affine, A. cf. minutum, as well as Gymnodinium catenatum, were detected in the cyst survey.

  7. Microarray testing for the presence of toxic algae monitoring programme in Galicia (NW Spain).

    PubMed

    Dittami, Simon M; Pazos, Yolanda; Laspra, Melchor; Medlin, Linda K

    2013-10-01

    Rapid and reliable detection of harmful algae in coastal areas and shellfish farms is an important requirement of monitoring programmes. Monitoring of toxic algae by means of traditional methods, i.e., light microscopy, can be time consuming when many samples have to be routinely analysed. Reliable species identification requires expensive equipment and trained personnel to carry out the analyses. However, all techniques for the monitoring of harmful algae usually require transportation of samples to specialised laboratories. In many monitoring laboratories, results are usually obtained within five working days after receiving the sample and therefore preventative measures are not always possible. Molecular technologies are rapidly improving the detection of phytoplankton and their toxins and the speed at which the results can be obtained. Assays are based on the discrimination of the genetic differences of the different species and species-specific probes can be designed. Such probes have been adapted to a microarray or phylochip format and assessed in several EU monitoring sites. Microarray results are presented for 1 year of field samples validated with cell counts from concentrated samples taken during toxic events from the weekly sampling of the Galician Monitoring Programme done by INTECMAR. The Galician monitoring laboratory does their own counting and their results are posted on their web site within 24 h. There was good correlation between cells present and microarray signals. In the few cases of false negatives, these can be attributed to poor RNA extraction of the target species, viz. Prorocentrum or Dinophysis. Where potential false positives were encountered, the smaller volume taken for cell counts as compared to the upto 300 times more volume taken for RNA extraction for the microarray is likely the cause for these differences, making the microarray more sensitive. The microarray was able to provide better species resolution in Alexandrium and Pseudo

  8. 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. PMID:25893798

  9. Detection of marine toxins, brevetoxin-3 and saxitoxin, in seawater using neuronal networks.

    PubMed

    Kulagina, Nadezhda V; Mikulski, Christina M; Gray, Samuel; Ma, Wu; Doucette, Gregory J; Ramsdell, John S; Pancrazio, Joseph J

    2006-01-15

    There is a need for assay systems that can detect known and unanticipated neurotoxins associated with harmful algal blooms. The present work describes our attempt to monitor the presence of brevetoxin-3 (PbTx-3) and saxitoxin (STX) in a seawater matrix using the neuronal network biosensor (NNB). The NNB relies on cultured mammalian neurons grown over microelectrode arrays, where the inherent bioelectrical activity of the network manifested as extracellular action potentials can be monitored noninvasively. Spinal cord neuronal networks were prepared from embryonic mice and the mean spike rate across the network was analyzed before and during exposure to the toxins. Extracellular action potentials from the network are highly sensitive not only to purified STX and PbTx-3, but also when in combination with matrixes such as natural seawater and algal growth medium. Detection limits for STX and PbTx-3, respectively, are 0.031 and 0.33 nM in recording buffer and 0.076 and 0.48 nM in the presence of 25-fold-diluted seawater. Our results demonstrated that neuronal networks could be used for analysis of Alexandrium fundyense (STX-producer) and Karenia brevis (PbTx-producer) algal samples lysed directly in the seawater-based growth medium and appropriately diluted with HEPES-buffered recording medium. The cultured network responded by changes in mean spike rate to the presence of STX-or PbTx-producing algae but not to the samples of two non-STX and non-PbTx isolates of the same algal genera. This work provides evidence that the NNB has the capacity to rapidly detect toxins associated with cells of toxic algal species or as dissolved forms present in seawater and hasthe potential for monitoring toxin levels during harmful algal blooms. PMID:16468405

  10. Observations of exchange between eastern Casco Bay and the western Gulf of Maine

    NASA Astrophysics Data System (ADS)

    Janzen, C. D.; Churchill, J. H.; Pettigrew, N. R.

    2005-09-01

    Exchange of water between eastern Casco Bay and the adjacent Gulf of Maine shelf is examined to assess the circulation processes that impact the distribution and occurrence of a toxic dinoflagellate, Alexandrium fundyense, in eastern Casco Bay. Over the inner shelf adjacent to the bay, tidal variance is weak, and the across-shelf current is highly coherent and in phase with the along-shelf wind stress. Although tidal current variance increases as one advances into the bay, non-tidal currents account for 30-40% of the across-shelf current variance at the bay entrance. Between the shelf and the bay interior is a transition region, where the circulation response to wind forcing changes as the wind adjusts to the changing orientation of the shoreline. Far from shore, the overall large-scale coastline orientation dominates the wind-driven response, but within a few internal Rossby radii, the local coastline clearly dominates the flow patterns and across-shelf wind becomes locally shore-parallel inside the bay. Within the bay interior, the across-shelf wind is highly coherent and in phase with the near-surface subtidal across-shelf current. The Kennebec River north of the study area supplies freshwater to eastern Casco Bay in all seasons. A pool of low-density, relatively fresh water at the entrance to the bay sets up an across-shelf density gradient that is reversed from a typical estuary, and likely contributes to the mean surface on-shelf transport in this region. Surface-drifter trajectories observed over the course of the study suggest that both the across-shelf wind and the across-shelf density gradient are important in driving surface up-bay transport and in the retention of surface-dwelling organisms in eastern Casco Bay.

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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

  17. 13C fractionation of dinoflagellates - a new proxy for past CO2 levels?

    NASA Astrophysics Data System (ADS)

    Hoins, M.; Van de Waal, D. B.; Eberlein, T.; Reichart, G.; Sluijs, A.; Rost, B.

    2012-12-01

    Reliable reconstructions of atmospheric CO2 levels prior to ~1 million years ago are required to quantify climate sensitivity as well as ocean acidification in response to past carbon cycle perturbations. Despite recent progress, however, uncertainties in reconstructed values especially from the Paleogene and older, are still very large. We aim to develop a new proxy for CO2 concentrations based on the carbon isotopic fractionation of autotrophic marine dinoflagellates. Dinoflagellates feature RubisCO (type II) with the lowest CO2 affinity of all eukaryote phytoplankton, which makes this group inherently sensitive to changes in carbonate chemistry. Along with growth and carbon production also the 13C versus 12C incorporation, i.e., the 13C fractionation (ɛp), will likely be affected. Hence, the carbon isotopic composition of dinoflagellates may ultimately reflect the prevailing atmospheric CO2 concentrations. Crucially, microfossils of dinoflagellates, i.e. organic dinoflagellate cysts, have been recovered from ocean sediments as old as the Triassic (i.e. ~215 Ma BP). We performed dilute batch experiments with four dinoflagellate species: Alexandrium tamarense, Scrippsiella trochoidea, Gonyaulax spinifera and Protoceratium reticulatum. Cells were grown at various CO2 concentrations representing the Last Glacial Maximum (180 ppm), present day (380 ppm), and future projections (and estimates for distant past levels; 950 ppm and 1400 ppm). Consistent with expectations, ɛp increased with CO2 concentration in all four species (with a slope of up to 0.19 ‰ μM-1), while growth and carbon production (μc) had little effect. ɛp in relation to CO2 varied between species and strains, but the relation of ɛp to μc/CO2 was more consistent. First results of underlying processes affecting ɛp, including carbon acquisition and leakage, will be discussed. Considering that the cysts of P. reticulatum (Operculodinium centrocarpum) and G. spinifera (Spiniferites sp.) are

  18. 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

  19. Seasonal variations in phytoplankton growth and microzooplankton grazing in a temperate coastal embayment, Korea

    NASA Astrophysics Data System (ADS)

    Kim, Sunju; Park, Myung Gil; Moon, Changho; Shin, Kyoungsoon; Chang, Man

    2007-01-01

    Microzooplankton grazing on coastal phytoplankton was determined by the dilution method from May 2002 to April 2003 at a fixed site located in the Jinhae Bay, Korea. During the dilution experiments, our study site exhibited a wide range of chlorophyll a concentrations (0.29-127.42 μg l -1), and the species composition of the phytoplankton community changed dramatically over a year, shifting from the predominance of chain-forming diatoms, particularly Chaetoceros spp., Leptocylindrus danicus, Pseudonitzschia pungens, and Skeletonema costatum, between May and September 2002, to a massive bloom of the dinoflagellates, Alexandrium spp. in October 2002, to a dominance of cryptophytes ( Chroomonas sp.) between November 2002 and March 2003, and then again to a prevalence of diatoms toward the end of the experiment. Both nutrients enriched ( μ n) and in situ phytoplankton growth rates ( μ0) showed pronounced seasonal variations, ranging from 0.11 to 2.87 d -1 and from -0.63 to 2.08 d -1, respectively. With regard to both variables, the lowest values were obtained during the fall and winter seasons. The average ratio of μ0/ μ n was 0.96 (SE = 0.08), thereby indicating that phytoplankton growth in the study site was not nutrient-limited. Microzooplankton grazing rates showed the large fluctuations (0-3.86 d -1) over an annual cycle, with non-significant and/or negative grazing frequently (62% of 29 measurements) detected. Relatively high grazing rates did occur frequently at the times during which a large phytoplankton biomass and/or large-sized phytoplankton dominance were observed. Our results contribute to the growing body of evidence suggesting that microzooplankton are important phytoplankton consumers in communities dominated by large phytoplankton, and also bolster the notion that size-based models of food web relationships may be of limited predictive value. The observed large fluctuations in grazing rates over a year, coupled with frequent non-significant and

  20. Feeding by the newly described mixotrophic dinoflagellate Paragymnodinium shiwhaense: feeding mechanism, prey species, and effect of prey concentration.

    PubMed

    Yoo, Yeong Du; Jeong, Hae Jin; Kang, Nam Seon; Song, Jae Yoon; Kim, Kwang Young; Lee, Gitack; Kim, Juhyoung

    2010-01-01

    To investigate the feeding by the newly described mixotrophic dinoflagellate Paragymnodinium shiwhaense (GenBank accession number=AM408889), we explored the feeding process and the kinds of prey species that P. shiwhaense is able to feed on using several different types of microscopes, including a transmission electron microscope and high-resolution video-microscopy. In addition, we measured the growth and ingestion rates of P. shiwhaense on its optimal algal prey Amphidinium carterae as a function of prey concentration. We also measured these parameters for edible prey at a single concentration at which the growth and ingestion rates of P. shiwhaense on A. carterae were saturated. Paragymnodinium shiwhaense feed on algal prey using a peduncle after anchoring the prey by a tow filament. Among the algal prey offered, P. shiwhaense ingested small algal species that had equivalent spherical diameters (ESDs) < or =11 microm (e.g. the prymnesiophyte Isochrysis galbana, the cryptophytes Teleaulax sp. and Rhodomonas salina, the raphidophyte Heterosigma akashiwo, and the dinoflagellates Heterocapsa rotundata and A. carterae). However, it did not feed on larger algal species that had ESDs > or =12 microm (e.g. the dinoflagellates Prorocentrum minimum, Heterocapsa triquetra, Scrippsiella trochoidea, Alexandrium tamarense, Prorocentrum micans, Gymnodinium catenatum, Akashiwo sanguinea, and Lingulodinium polyedrum) or the small diatom Skeletonema costatum. The specific growth rates for P. shiwhaense feeding upon A. carterae increased rapidly with increasing mean prey concentration before saturating at concentrations of ca. 350 ng C/ml (5,000 cells/ml). The maximum specific growth rate (i.e. mixotrophic growth) of P. shiwhaense on A. carterae was 1.097/d at 20 degrees C under a 14:10 h light-dark cycle of 20 microE/m(2)/s, while its growth rate (i.e. phototrophic growth) under the same light conditions without added prey was -0.224/d. The maximum ingestion and clearance rates

  1. [Growth inhibition of the four species of red tide microalgae by extracts from Enteromorpha prolifera extracted with the five solvents].

    PubMed

    Sun, Ying-Ying; Liu, Xiao-Xiao; Wang, Chang-Hai

    2010-06-01

    To study the effects of extracts of Enteromorpha prolifera on the growth of the four species of red tide microalgae (Amphidinium hoefleri, Karenia mikimitoi, Alexandrium tamarense and Skeletonema costatum), the extracts were extracted with five solvents (methanol, acetone, ethyl acetate, chloroform and petroleum ether), respectively. Based on the observation of algal morphology and the measurement of algal density, cell size and the contents of physiological indicators (chlorophyll, protein and polysaccharide), the results showed methanol extracts of E. prolifera had the strongest action. The inhibitory effects of A. hoefleri, K. mikimitoi, A. tamarense and S. costatum by the methanol extracts were 54.0%, 48.1%, 44.0% and 37.5% in day 10, respectively. The extracts of E. prolifera extracted with methanol, acetone and ethyl acetate caused cavities, pieces and pigment reduction in cells, and those with chloroform and petroleum ether caused goffers on cells. The extracts of E. prolifera extracted with all the five solvents decreased athletic ability of the cells, among which those extracted with ethyl acetate, chloroform and petroleum ether decreased cell size of test microalgae. The further investigation found that the methanol extracts significantly decreased contents of chlorophyll, protein and polysaccharide in the cells of those microalgae. The inhibitory effect of chlorophyll, protein and polysaccharide contents of four species of microalgae by the methanol extracts was about 51%. On the basis of the above experiments, dry powder of E. prolifera were extracts with methanol, and extracts were obtained. The methanol extracts were partitioned to petroleum ether phase, ethyl acetate phase, n-butanol phase and distilled water phase by liquid-liquid fractionation, and those with petroleum ether and ethyl acetate significantly inhibited the growth of all test microalgae, and the inhibitory effect of four species of microalgae by those two extracts was above 25% in day

  2. 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

  3. 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

  4. Transformation of paralytic shellfish poisoning toxins in UK surf clams (Spisula solida) for targeted production of reference materials.

    PubMed

    Turner, Andrew D; Lewis, Adam M; O'Neil, Alison; Hatfield, Robert G

    2013-04-01

    The periodic occurrence of Paralytic Shellfish Poisoning (PSP) toxins in UK surf clams and the recent move away from biological assays for PSP testing resulted in the need to determine method performance characteristics for the replacement analytical method in this species. With the requirement for laboratory reference materials to aid this validation together with known issues relating to toxin transformation in live clams and homogenised tissue, there was the need to assess the toxin transformation characteristics of PSP toxins in surf clam tissue. Initial work examined the rates of toxin transformation in UK surf clam tissue incubated with toxin standards, showing rapid transformation of N-sulfocarbamoyl toxins with slower transformation of carbamate toxins. Full transformational pathways were determined using a combination of three different analytical methods and confirmed the major expected transformations involving decarbamoylation, with some evidence for additional reaction pathways. Results obtained from the analysis of surf clam and oyster tissues incubated with varying concentrations of toxic Alexandrium algae highlighted expected transformation reactions, although significant differences were observed in the extent of the transformations amongst the range of toxins studied, with less efficient transformation of N-hydroxylated toxins as compared with other carbamate and N-sulfocarbamoyl toxins. Analysis of PSP-toxic incurred oyster, scallop and mussel tissues incubated with variable proportions of surf clam tissue showed large differences in the extent of the transformations. Total conversion of N-sulfocarbamoyl toxins was confirmed at low relative proportions of surf clam tissue in all three species, whereas transformation of carbamate toxins was found to occur only in the presence of higher proportions of surf clam tissue in oysters and mussels in comparison with scallops. Results enabled the production of three laboratory reference materials prepared

  5. 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

  6. Phycotoxins.

    PubMed

    Quilliam, M A

    1999-01-01

    The 1997-1998 period brought many new developments to the phycotoxin field. There were several reviews on phycotoxins in general, on their toxicological evaluation, and on their analysis. The ecophysiology, biosynthesis, and metabolism of polyether toxins and paralytic shellfish poisoning (PSP) toxins were also reviewed. The proceedings of the Eighth International Conference on Harmful Algae (Vigo, Spain, June 25-29, 1997) have been published and provide an excellent source of information on phycotoxins and toxic plankton bloom research. In addition, the much anticipated proceedings of the IX International IUPAC Symposium on Mycotoxins and Phycotoxins (Rome, Italy, May 27-31, 1996) have been published. Further evidence was provided to support the theory that Prorocentrum lima is the source organism for diarrhetic shellfish poisoning (DSP) toxins in Nova Scotian shellfish. In another study, different Prorocentrum species and isolates were analyzed for DSP toxins. In addition to detecting some new compounds, such as a DTX1 isomer, it was found that toxins were produced by both axenic and nonaxenic batch cultures, indicating that bacteria are probably not involved in the biosynthesis. The source organism for the spirolides, a family of fast-acting toxins reported from Nova Scotia, Canada, was determined to be Alexandrium ostenfeldii, a species that is found worldwide. The biogenetic origin of yessotoxin was reported to be Protoceratium reticulatum, another widely occurring organism. A great deal of attention and research funding has been directed at the serious problems associated with Pfiesteria piscicida. Analysts are eagerly awaiting publication of toxin structures, which will then allow the development of analytical methods. An incident of the mass mortality of California sea lions was reported in the Monterey area in May 1998. Analyses of tissue and urine samples revealed the presence of domoic acid. High levels of domoic acid were also found in anchovies and