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Fulltext Biosynthesis of Saxitoxin in Marine Dinoflagellates: An Omics Perspective

Akbar MA, Mohd Yusof NY, Tahir NI, Ahmad A, Usup G, Sahrani FK, et al.

Mar Drugs, 2020 Feb 05;18(2).
PMID: 32033403 DOI: 10.3390/md18020103

Saxitoxin is an alkaloid neurotoxin originally isolated from the clam Saxidomus giganteus in 1957. This group of neurotoxins is produced by several species of freshwater cyanobacteria and marine dinoflagellates. The saxitoxin biosynthesis pathway was described for the first time in the 1980s and, since then, it was studied in more than seven cyanobacterial genera, comprising 26 genes that form a cluster ranging from 25.7 kb to 35 kb in sequence length. Due to the complexity of the genomic landscape, saxitoxin biosynthesis in dinoflagellates remains unknown. In order to reveal and understand the dynamics of the activity in such impressive unicellular organisms with a complex genome, a strategy that can carefully engage them in a systems view is necessary. Advances in omics technology (the collective tools of biological sciences) facilitated high-throughput studies of the genome, transcriptome, proteome, and metabolome of dinoflagellates. The omics approach was utilized to address saxitoxin-producing dinoflagellates in response to environmental stresses to improve understanding of dinoflagellates gene-environment interactions. Therefore, in this review, the progress in understanding dinoflagellate saxitoxin biosynthesis using an omics approach is emphasized. Further potential applications of metabolomics and genomics to unravel novel insights into saxitoxin biosynthesis in dinoflagellates are also reviewed.

Matched MeSH terms: Dinoflagellida/genetics*; Dinoflagellida/metabolism*
Are Pyrodinium blooms in the Southeast Asian region recurring and spreading? A view at the end of the millennium

Azanza RV, Taylor FJ

Ambio, 2001 Sep;30(6):356-64.
PMID: 11757284

Pyrodinium bahamense (var. compressum) has been the only dinoflagellate species that has caused major public health and economic problems in the Southeast Asian region for more than 2 decades now. It produces saxitoxin, a suite of toxins that cause Paralytic Shellfish Poisoning (PSP). A serious toxicological problem affecting many countries of the world, mild cases of this poisoning can occur within 30 minutes while in extreme cases, death through respiratory paralysis may occur within 2-24 hrs of ingestion of intoxicated shellfish. Blooms of the organism have been reported in Malaysia, Brunei Darussalam, the Philippines and Indonesia. The ASEAN-Canada Red Tide Network has recorded 31 blooms of the organism in 26 areas since 1976 when it first occurred in Sabah, Malaysia. As of 1999, the most hard hit country has been the Philippines which has the greatest number of areas affected (18) and highest number of Paralytic Shellfish Poisoning (PSP) cases (about 1995). Malaysia has reported a total of 609 PSP cases and 44 deaths while Brunei has recorded 14 PSP cases and no fatalities. Indonesia, on the other hand has a record of 427 PSP cases and 17 deaths. Studies on ecological/environmental impacts of these blooms have not been done in the region. Estimates of economic impacts have shown that the loss could be up to USD 300,000 day-1. Most of the data and information useful for understanding Pyrodinium bloom dynamics have come from harmful/toxic algal monitoring and research that have developed to different degrees in the various countries in the region affected by the organism's bloom. Regional collaborative research and monitoring efforts can help harmonize local data sets and ensure their quality and availability for comparative analysis and modeling. Temporal patterns of the blooms at local and regional scales and possible signals and trends in the occurrence/recurrence and spread of Pyrodinium blooms could be investigated. Existing descriptive and simple predictive models of Pyrodinium blooms can be improved and refined to help in the management of the wild harvest and aquaculture of shellfish in a region where the people are dependent on these resources for their daily food sustainance and livelihood.

Matched MeSH terms: Dinoflagellida*
Fulltext Loktanella spp. Gb03 as an algicidal bacterium, isolated from the culture of Dinoflagellate Gambierdiscus belizeanus

Bloh AH, Usup G, Ahmad A

Vet World, 2016 Feb;9(2):142-6.
PMID: 27051199 DOI: 10.14202/vetworld.2016.142-146

AIM: Bacteria associated with harmful algal blooms can play a crucial role in regulating algal blooms in the environment. This study aimed at isolating and identifying algicidal bacteria in Dinoflagellate culture and to determine the optimum growth requirement of the algicidal bacteria, Loktanella sp. Gb-03.
MATERIALS AND METHODS: The Dinoflagellate culture used in this study was supplied by Professor Gires Usup's Laboratory, School of Environmental and Natural Resources Sciences, Faculty of Science and Technology, University Kebangsaan Malaysia, Malaysia. The culture was used for the isolation of Loktanella sp., using biochemical tests, API 20 ONE kits. The fatty acid content of the isolates and the algicidal activity were further evaluated, and the phenotype was determined through the phylogenetic tree.
RESULTS: Gram-negative, non-motile, non-spore-forming, short rod-shaped, aerobic bacteria (Gb01, Gb02, Gb03, Gb04, Gb05, and Gb06) were isolated from the Dinoflagellate culture. The colonies were pink in color, convex with a smooth surface and entire edge. The optimum growth temperature for the Loktanella sp. Gb03 isolate was determined to be 30°C, in 1% of NaCl and pH7. Phylogenetic analysis based on 16S rRNA gene sequences showed that the bacterium belonged to the genus Loktanella of the class Alphaproteobacteria and formed a tight cluster with the type strain of Loktanella pyoseonensis (97.0% sequence similarity).
CONCLUSION: On the basis of phenotypic, phylogenetic data and genetic distinctiveness, strain Gb-03, were placed in the genus Loktanella as the type strain of species. Moreover, it has algicidal activity against seven toxic Dinoflagellate. The algicidal property of the isolated Loktanella is vital, especially where biological control is needed to mitigate algal bloom or targeted Dinoflagellates.

Matched MeSH terms: Dinoflagellida
Fulltext Development of species-specific molecular marker for Pyrodinium bahamense var. compressum (Gonyaulacales, Dinophyceae) based on 18S ribosomal DNA sequence

CHIN, W. L., ANTON, A., KUMAR, S.V., TEOH, P. L.

Buletin Persatuan Genetik Malaysia, 2010;17(1):5-8.
MyJurnal

In Malaysia, harmful algal blooms often occur along the coastal waters of west Sabah, where one of the causative organisms is the toxin-producing dinoflagellate, Pyrodinium bahamense var. compressum. A total of five P. bahamense var. compressum isolates were obtained from four locations and were cultured in f/2 medium. A Polymerase Chain Reaction (PCR) based technique was developed and used to screen for the presence of the dinoflagellate, P. bahamense var. compressum. A dinoflagellate-specific primer pair was designed based on sequences of P. bahamense var. compressum to amplify the 18S small subunit ribosomal DNA (rDNA) sequences. The rDNA of the P. bahamense var. compressum isolates were obtained. A species-specific primer pair was designed to target a 600 bp rDNA sequence of the target dinoflagellate. The primer pair targeting P. bahamense var. compressum did not yield any product with the fifteen algae cultures used as negative controls, but only amplified the rDNA of P. bahamense var. compressum cultures. The PCR method for identification of P. bahamense var. compressum was also applied on twenty field samples collected with plankton net. P. bahamense var. compressum cells were detected by PCR in five field samples and were confirmed by direct sequencing. From this study, a species-specific primer pair was obtained to identify the target species, P. bahamense var. compressum, among the natural complex communities of seawater.

Matched MeSH terms: Dinoflagellida
Evidence of okadaic acid production in a cultured strain of the marine dinoflagellate Prorocentrum rhathymum from Malaysia

Caillaud A, de la Iglesia P, Campàs M, Elandaloussi L, Fernández M, Mohammad-Noor N, et al.

Toxicon, 2010 Feb-Mar;55(2-3):633-7.
PMID: 19631680 DOI: 10.1016/j.toxicon.2009.07.016

Protein phosphatase inhibition assay (PPIA), Neuroblastoma cell-based assay (Neuro-2a CBA) and LC-MS/MS analysis revealed for the first time the production of okadaic acid (OA) by a Prorocentrum rhathymum strain. Low amounts of OA were detected by LC-MS/MS analysis. Inhibition of PP2A activity and a weak toxicity to the Neuro-2a CBA were also observed.

Matched MeSH terms: Dinoflagellida/metabolism*; Dinoflagellida/ultrastructure; Dinoflagellida/chemistry
Blooms of diatom and dinoflagellate associated with nutrient imbalance driven by cycling of nitrogen and phosphorus in anaerobic sediments in Johor Strait (Malaysia)

Chai X, Li X, Hii KS, Zhang Q, Deng Q, Wan L, et al.

Mar Environ Res, 2021 Jul;169:105398.
PMID: 34171592 DOI: 10.1016/j.marenvres.2021.105398

Coastal eutrophication is one of the pivotal factors driving occurrence of harmful algal blooms (HABs), whose underlying mechanism remained unclear. To better understand the nutrient regime triggering HABs and their formation process, the phytoplankton composition and its response to varying nitrogen (N) and phosphorus (P), physio-chemical parameters in water and sediment in Johor Strait in March 2019 were analyzed. Surface and sub-surface HABs were observed with the main causative species of Skeletonema, Chaetoceros and Karlodinium. The ecophysiological responses of Skeletonema to the low ambient N/P ratio such as secreting alkaline phosphatase, regulating cell morphology (volume; surface area/volume ratio) might play an important role in dominating the community. Anaerobic sediment iron-bound P release and simultaneous N removal by denitrification and anammox, shaped the stoichiometry of N and P in water column. The decrease of N/P ratio might shift the phytoplankton community into the dominance of HABs causative diatoms and dinoflagellates.

Matched MeSH terms: Dinoflagellida*
Fulltext The genetic intractability of Symbiodinium microadriaticum to standard algal transformation methods

Chen JE, Barbrook AC, Cui G, Howe CJ, Aranda M

PLoS One, 2019;14(2):e0211936.
PMID: 30779749 DOI: 10.1371/journal.pone.0211936

Modern transformation and genome editing techniques have shown great success across a broad variety of organisms. However, no study of successfully applied genome editing has been reported in a dinoflagellate despite the first genetic transformation of Symbiodinium being published about 20 years ago. Using an array of different available transformation techniques, we attempted to transform Symbiodinium microadriaticum (CCMP2467), a dinoflagellate symbiont of reef-building corals, with the view to performing subsequent CRISPR-Cas9 mediated genome editing. Plasmid vectors designed for nuclear transformation containing the chloramphenicol resistance gene under the control of the CaMV p35S promoter as well as several putative endogenous promoters were used to test a variety of transformation techniques including biolistics, electroporation and agitation with silicon carbide whiskers. Chloroplast-targeted transformation was attempted using an engineered Symbiodinium chloroplast minicircle encoding a modified PsbA protein expected to confer atrazine resistance. We report that we have been unable to confer chloramphenicol or atrazine resistance on Symbiodinium microadriaticum strain CCMP2467.

Matched MeSH terms: Dinoflagellida/genetics*; Dinoflagellida/metabolism
Taxonomic assignment of the benthic toxigenic dinoflagellate Gambierdiscus sp. type 6 as Gambierdiscus balechii (Dinophyceae), including its distribution and ciguatoxicity

Dai X, Mak YL, Lu CK, Mei HH, Wu JJ, Lee WH, et al.

Harmful Algae, 2017 07;67:107-118.
PMID: 28755713 DOI: 10.1016/j.hal.2017.07.002

Recent molecular phylogenetic studies of Gambierdiscus species flagged several new species and genotypes, thus leading to revitalizing its systematics. The inter-relationships of clades revealed by the primary sequence information of nuclear ribosomal genes (rDNA), however, can sometimes be equivocal, and therefore, in this study, the taxonomic status of a ribotype, Gambierdiscus sp. type 6, was evaluated using specimens collected from the original locality, Marakei Island, Republic of Kiribati; and specimens found in Rawa Island, Peninsular Malaysia, were further used for comparison. Morphologically, the ribotype cells resembled G. scabrosus, G. belizeanus, G. balechii, G. cheloniae and G. lapillus in thecal ornamentation, where the thecal surfaces are reticulate-foveated, but differed from G. scabrosus by its hatchet-shaped Plate 2', and G. belizeanus by the asymmetrical Plate 3'. To identify the phylogenetic relationship of this ribotype, a large dataset of the large subunit (LSU) and small subunit (SSU) rDNAs were compiled, and performed comprehensive analyses, using Bayesian-inference, maximum-parsimony, and maximum-likelihood, for the latter two incorporating the sequence-structure information of the SSU rDNA. Both the LSU and SSU rDNA phylogenetic trees displayed an identical topology and supported the hypothesis that the relationship between Gambierdiscus sp. type 6 and G. balechii was monophyletic. As a result, the taxonomic status of Gambierdiscus sp. type 6 was revised, and assigned as Gambierdiscus balechii. Toxicity analysis using neuroblastoma N2A assay confirmed that the Central Pacific strains were toxic, ranging from 1.1 to 19.9 fg P-CTX-1 eq cell-1, but no toxicity was detected in a Western Pacific strain. This suggested that the species might be one of the species contributing to the high incidence rate of ciguatera fish poisoning in Marakei Island.

Matched MeSH terms: Dinoflagellida/classification*; Dinoflagellida/cytology; Dinoflagellida/ultrastructure
Fulltext Genome Sequence of Bacillus sp. Strain UMTAT18 Isolated from the Dinoflagellate Alexandrium tamiyavanichii Found in the Straits of Malacca

Danish-Daniel M, Ming GH, Mohd Noor ME, Sung YY, Usup G

Genome Announc, 2016 Oct 6;4(5).
PMID: 27795265 DOI: 10.1128/genomeA.01106-16

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

Matched MeSH terms: Dinoflagellida
Responses of phytoplankton community to eutrophication in Semerak Lagoon (Malaysia)

Er HH, Lee LK, Lim ZF, Teng ST, Leaw CP, Lim PT

Environ Sci Pollut Res Int, 2018 Aug;25(23):22944-22962.
PMID: 29858995 DOI: 10.1007/s11356-018-2389-0

Effects of aquaculture activities on the environmental parameters and phytoplankton community structure were investigated in a semi-enclosed lagoon located at Semerak River, Malaysia. Elevated concentrations of phosphate and ammonia were observed at the aquaculture area and the inner lagoon. Relatively low dissolved oxygen, high total chlorophyll a, and high phytoplankton abundances but low species richness were recorded. Chaetoceros, Pseudo-nitzschia brasiliana, Blixaea quinquecornis, and Skeletonema blooms were observed, and some were associated with anoxia condition. Eutrophication level assessed by UNTRIX suggests that the water quality in the lagoon is deteriorating. Dissolved inorganic phosphorus and nitrogen at the impacted area were 15 and 12 times higher than the reference sites, respectively. Such trophic status indices could provide a useful guideline for optimal aquaculture management plan to reduce the environmental impact caused by aquaculture.

Matched MeSH terms: Dinoflagellida
Unfamiliar partnerships limit cnidarian holobiont acclimation to warming

Herrera M, Klein SG, Schmidt-Roach S, Campana S, Cziesielski MJ, Chen JE, et al.

Glob Chang Biol, 2020 Jul 06.
PMID: 32627905 DOI: 10.1111/gcb.15263

Enhancing the resilience of corals to rising temperatures is now a matter of urgency, leading to growing efforts to explore the use of heat tolerant symbiont species to improve their thermal resilience. The notion that adaptive traits can be retained by transferring the symbionts alone, however, challenges the holobiont concept, a fundamental paradigm in coral research. Holobiont traits are products of a specific community (holobiont) and all its co-evolutionary and local adaptations, which might limit the retention or transference of holobiont traits by exchanging only one partner. Here, we evaluate how interchanging partners affect the short- and long-term performance of holobionts under heat stress using clonal lineages of the cnidarian model system Aiptasia (host and Symbiodiniaceae strains) originating from distinct thermal environments. Our results show that holobionts from more thermally variable environments have higher plasticity to heat stress, but this resilience could not be transferred to other host genotypes through the exchange of symbionts. Importantly, our findings highlight the role of the host in determining holobiont productivity in response to thermal stress and indicate that local adaptations of holobionts will likely limit the efficacy of interchanging unfamiliar compartments to enhance thermal tolerance.

Matched MeSH terms: Dinoflagellida
Diverse harmful microalgal community assemblages in the Johor Strait and the environmental effects on its community dynamics

Hii KS, Mohd-Din M, Luo Z, Tan SN, Lim ZF, Lee LK, et al.

Harmful Algae, 2021 07;107:102077.
PMID: 34456026 DOI: 10.1016/j.hal.2021.102077

Coastal ecosystems are often subjected to anthropogenic disturbances that lead to water quality deterioration and an increase in harmful algal bloom (HAB) events. Using the next-generation molecular tool of 18S rDNA metabarcoding, we examined the community assemblages of HAB species in the Johor Strait, Malaysia between May 2018 and September 2019, covering 19 stations across the strait. The molecular operational taxonomic units (OTUs) of HAB taxa retrieved from the dataset (n = 194) revealed a much higher number of HAB taxa (26 OTUs) than before, with 12 taxa belong to new records in the strait. As revealed in the findings of this study, the diversity and community structure of HAB taxa varied significantly over time and space. The most common and abundant HAB taxa in the strait (frequency of occurrence >70%) comprised Heterosigma akashiwo, Fibrocapsa japonica, Pseudo-nitzschia pungens, Dinophysis spp., Gymnodinium catenatum, Alexandrium leei, and A. tamiyavanichii. Also, our results demonstrated that the HAB community assemblages in the strait were dependent on the interplay of environmental variables that influence by the monsoonal effects. Different HAB taxa, constitute various functional types, occupied and prevailed in different environmental niches across space and time, leading to diverse community assemblages and population density. This study adds to the current understandings of HAB dynamics and provides a robust overview of temporal-spatial changes in HAB community assemblages along the environmental gradients in a tropical eutrophic coastal ecosystem.

Matched MeSH terms: Dinoflagellida*
Physiological and transcriptional responses to inorganic nutrition in a tropical Pacific strain of Alexandrium minutum: Implications for the saxitoxin genes and toxin production

Hii KS, Lim PT, Kon NF, Takata Y, Usup G, Leaw CP

Harmful Algae, 2016 06;56:9-21.
PMID: 28073499 DOI: 10.1016/j.hal.2016.04.005

Saxitoxins (STXs) constitute a family of potent sodium channel blocking toxins, causative agents of paralytic shellfish poisoning (PSP), and are produced by several species of marine dinoflagellates and cyanobacteria. Two STX-core genes, sxtA and sxtG, have been well elucidated in Alexandrium but the expression of these genes under various nutritional modes in tropical species remains unclear. This study investigates the physiological responses of a tropical Pacific strain of Alexandrium minutum growing with nitrate or ammonium, and with various nitrogen to phosphorus (N:P) supply ratios. The transcriptional responses of the sxt genes were observed. Likewise, a putative sxtI encoding O-carbamoyltransferase (herein designated as AmsxtI) was recovered from the transcriptomic data, and its expression was investigated. The results revealed that the cellular toxin quota (Qt) was higher in P-depleted, nitrate-grown cultures. With cultures at similar N:P (<16), cells grown with excess ammonium showed a higher Qt than those grown with nitrate. sxtA1 was not expressed under any culture conditions, suggesting that this gene might not be involved in STX biosynthesis by this strain. Conversely, sxtA4 and sxtG showed positive correlations with Qt, and were up-regulated in P-depleted, nitrate-grown cultures and with excess ambient ammonium. On the other hand, AmsxtI was expressed only when induced by P-depletion, suggesting that this gene may play an important role in P-recycling metabolism, while simultaneously enhancing toxin production.

Matched MeSH terms: Dinoflagellida/genetics; Dinoflagellida/physiology*
Transcriptional and physiological responses to inorganic nutrition in a tropical Pacific strain of Alexandrium minutum: Implications for nutrient uptakes and assimilation

Hii KS, Lim PT, Kon NF, Usup G, Gu H, Leaw CP

Gene, 2019 Aug 30;711:143950.
PMID: 31255736 DOI: 10.1016/j.gene.2019.143950

The marine dinoflagellate Alexandrium minutum is known to produce saxitoxins that cause paralytic shellfish poisoning in human worldwide through consumption of the contaminated shellfish mollusks. Despite numerous studies on the growth physiology and saxitoxin production of this species, the knowledge on the molecular basis of nutrient uptakes in relation to toxin production in this species is limited. In this study, relative expressions of the high-affinity transporter genes of nitrate, ammonium, and phosphate (AmNrt2, AmAmt1 and AmPiPT1) and the assimilation genes, nitrate reductase (AmNas), glutamine synthase (AmGSIII) and carbamoyl phosphate synthase (AmCPSII) from A. minutum were studied in batch clonal culture condition with two nitrogen sources (nitrate: NO3- or ammonium: NH4+) under different N:P ratios (high-P: N:P of 14 and 16, and low-P: N:P of 155). The expression of AmAmt1 was suppressed in excess NH4+-grown condition but was not observed in AmNrt2 and AmNas. Expressions of AmAmt1, AmNrt2, AmNas, AmGSIII, AmCPSII, and AmPiPT1 were high in P-deficient condition, showing that A. minutum is likely to take up nutrients for growth under P-stress condition. Conversely, relative expression of AmCPSII was incongruent with cell growth, but was well correlated with toxin quota, suggesting that the gene might involve in arginine metabolism and related toxin production pathway. The expression of AmGSIII is found coincided with higher toxin production and is believed to involve in mechanism to detoxify the cells from excess ammonium stress. The gene regulation observed in this study has provided better insights into the ecophysiology of A. minutum in relation to its adaptive strategies in unfavorable environments.

Matched MeSH terms: Dinoflagellida/growth & development*; Dinoflagellida/metabolism
An evaluation of the genus Amphidinium (Dinophyceae) combining evidence from morphology, phylogenetics, and toxin production, with the introduction of six novel species

Karafas S, Teng ST, Leaw CP, Alves-de-Souza C

Harmful Algae, 2017 09;68:128-151.
PMID: 28962975 DOI: 10.1016/j.hal.2017.08.001

The genus Amphidinium is an important group of athecated dinoflagellates because of its high abundance in marine habitats, its member's ability to live in a variety of environmental conditions and ability to produce toxins. Furthermore, the genus is of particular interest in the biotechnology field for its potential in the pharmaceutical arena. Taxonomically the there is a history of complication and confusion over the proper identities and placements of Amphidinium species due to high genetic variability coupled with high morphological conservation. Thirteen years has passed since the most recent review of the group, and while many issues were resolved, some remain. The present study used microscopy, phylogenetics of the 28S region of rDNA, secondary structure of the ITS2 region of rDNA, compensatory base change data, and cytotoxicity data from Amphidinium strains collected world-wide to elucidate remaining confusion. This holistic approach using multiple lines of evidence resulted in a more comprehensive understanding of the morphological, ecological, and genetic characteristics that are attributed to organisms belonging to Amphidinium, including six novel species: A. fijiensis, A. magnum, A. paucianulatum, A. pseudomassartii, A. theodori, and A. tomasii.

Matched MeSH terms: Dinoflagellida/classification*; Dinoflagellida/cytology*; Dinoflagellida/ultrastructure
Phylogenetic relationships, morphological variation, and toxin patterns in the Alexandrium ostenfeldii (Dinophyceae) complex: implications for species boundaries and identities

Kremp A, Tahvanainen P, Litaker W, Krock B, Suikkanen S, Leaw CP, et al.

J Phycol, 2014 Feb;50(1):81-100.
PMID: 26988010 DOI: 10.1111/jpy.12134

Alexandrium ostenfeldii (Paulsen) Balech and Tangen and A. peruvianum (Balech and B.R. Mendiola) Balech and Tangen are morphologically closely related dinoflagellates known to produce potent neurotoxins. Together with Gonyaulax dimorpha Biecheler, they constitute the A. ostenfeldii species complex. Due to the subtle differences in the morphological characters used to differentiate these species, unambiguous species identification has proven problematic. To better understand the species boundaries within the A. ostenfeldii complex we compared rDNA data, morphometric characters and toxin profiles of multiple cultured isolates from different geographic regions. Phylogenetic analysis of rDNA sequences from cultures characterized as A. ostenfeldii or A. peruvianum formed a monophyletic clade consisting of six distinct groups. Each group examined contained strains morphologically identified as either A. ostenfeldii or A. peruvianum. Though key morphological characters were generally found to be highly variable and not consistently distributed, selected plate features and toxin profiles differed significantly among phylogenetic clusters. Additional sequence analyses revealed a lack of compensatory base changes in ITS2 rRNA structure, low to intermediate ITS/5.8S uncorrected genetic distances, and evidence of reticulation. Together these data (criteria currently used for species delineation in dinoflagellates) imply that the A. ostenfeldii complex should be regarded a single genetically structured species until more material and alternative criteria for species delimitation are available. Consequently, we propose that A. peruvianum is a heterotypic synonym of A. ostenfeldii and this taxon name should be discontinued.

Matched MeSH terms: Dinoflagellida
Life-history stages of natural bloom populations and the bloom dynamics of a tropical Asian ribotype of Alexandrium minutum

Lau WLS, Law IK, Liow GR, Hii KS, Usup G, Lim PT, et al.

Harmful Algae, 2017 12;70:52-63.
PMID: 29169568 DOI: 10.1016/j.hal.2017.10.006

In 2015, a remarkably high density bloom of Alexandrium minutum occurred in Sungai Geting, a semi-enclosed lagoon situated in the northeast of Peninsular Malaysia, causing severe discoloration and contaminated the benthic clams (Polymesoda). Plankton and water samples were collected to investigate the mechanisms of bloom development of this toxic species. Analysis of bloom samples using flow cytometry indicated that the bloom was initiated by the process of active excystment, as planomycetes (>4C cells) were observed in the early stage of the bloom. Increase in planozygotes (2C cells) was evident during the middle stage of the bloom, coinciding with an abrupt decrease in salinity and increase of temperature. The bloom was sustained through the combination of binary division of vegetative cells, division of planozygotes, and cyst germination through continuous excystment. Nutrient depletion followed by precipitation subsequently caused the bloom to terminate. This study provides the first continuous record of in situ life-cycle stages of a natural bloom population of A. minutum through a complete bloom cycle. The event has provided a fundamental understanding of the pelagic life-cycle stages of this tropical dinoflagellate, and demonstrated a unique bloom development characteristic shared among toxic Alexandrium species in coastal embayments.

Matched MeSH terms: Dinoflagellida/growth & development*
New scenario for speciation in the benthic dinoflagellate genus Coolia (Dinophyceae)

Leaw CP, Tan TH, Lim HC, Teng ST, Yong HL, Smith KF, et al.

Harmful Algae, 2016 05;55:137-149.
PMID: 28073527 DOI: 10.1016/j.hal.2016.02.010

In this study, inter- and intraspecific genetic diversity within the marine harmful dinoflagellate genus Coolia Meunier was evaluated using isolates obtained from the tropics to subtropics in both Pacific and Atlantic Ocean basins. The aim was to assess the phylogeographic history of the genus and to clarify the validity of established species including Coolia malayensis. Phylogenetic analysis of the D1-D2 LSU rDNA sequences identified six major lineages (L1-L6) corresponding to the morphospecies Coolia malayensis (L1), C. monotis (L2), C. santacroce (L3), C. palmyrensis (L4), C. tropicalis (L5), and C. canariensis (L6). A median joining network (MJN) of C. malayensis ITS2 rDNA sequences revealed a total of 16 haplotypes; however, no spatial genetic differentiation among populations was observed. These MJN results in conjunction with CBC analysis, rDNA phylogenies and geographical distribution analyses confirm C. malayensis as a distinct species which is globally distributed in the tropical to warm-temperate regions. A molecular clock analysis using ITS2 rDNA revealed the evolutionary history of Coolia dated back to the Mesozoic, and supports the hypothesis that historical vicariant events in the early Cenozoic drove the allopatric differentiation of C. malayensis and C. monotis.

Matched MeSH terms: Dinoflagellida
Fulltext Effects of substratum and depth on benthic harmful dinoflagellate assemblages

Lee LK, Lim ZF, Gu H, Chan LL, Litaker RW, Tester PA, et al.

Sci Rep, 2020 07 09;10(1):11251.
PMID: 32647125 DOI: 10.1038/s41598-020-68136-6

Microhabitats influence the distribution and abundance of benthic harmful dinoflagellate (BHAB) species. Currently, much of the information on the relationships between BHABs and microhabitat preferences is based on non-quantitative anecdotal observations, many of which are contradictory. The goal of this study was to better quantify BHAB and microhabitat relationships using a statistically rigorous approach. Between April 2016 to May 2017, a total of 243 artificial substrate samplers were deployed at five locations in the Perhentian Islands, Malaysia while simultaneous photo-quadrat surveys were performed to characterize the benthic substrates present at each sampling site. The screen samplers were retrieved 24 h later and the abundances of five BHAB genera, Gambierdiscus, Ostreopsis, Coolia, Amphidinium, and Prorocentrum were determined. Substrate data were then analyzed using a Bray-Curtis dissimilarity matrix to statistically identify distinct microhabitat types. Although BHABs were associated with a variety of biotic and abiotic substrates, the results of this study demonstrated differing degrees of microhabitat preference. Analysis of the survey results using canonical correspondence analysis explained 70.5% (horizontal first axis) and 21.6% (vertical second axis) of the constrained variation in the distribution of various genera among microhabitat types. Prorocentrum and Coolia appear to have the greatest range being broadly distributed among a wide variety of microhabitats. Amphidinium was always found in low abundances and was widely distributed among microhabitats dominated by hard coral, turf algae, sand and silt, and fleshy algae and reached the highest abundances there. Gambierdiscus and Ostreopsis had more restricted distributions. Gambierdiscus were found preferentially associated with turf algae, hard coral and, to a lesser extent, fleshy macroalgae microhabitats. Ostreopsis, almost always more abundant than Gambierdiscus, preferred the same microhabitats as Gambierdiscus and were found in microbial mats as well. With similar habitat preferences Ostreopsis may serve as an indicator organism for the presence of Gambierdiscus. This study provides insight into how BHAB-specific microhabitat preferences can affect toxicity risks.

Matched MeSH terms: Dinoflagellida
Salinity effect on growth and toxin production of four tropical Alexandrium species (Dinophyceae)

Lim PT, Ogata T

Toxicon, 2005 May;45(6):699-710.
PMID: 15804519

Four tropical PSP toxins-producing dinoflagellates, Alexandrium minutum, Alexandrium tamiyavanichii, Alexandrium tamarense and Alexandrium peruvianum from Malaysian waters were studied to investigate the influences of salinity on growth and toxin production. Experiments were conducted on constant temperature 25 degrees C, 140 microE mol m(-2) s(-1) and under 14:10 light:dark photo-cycle with salinity ranged from 2 to 30 psu. The PSP-toxin congeners, GTX 1-6, STX, dcSTX, NEO and C1-C2 were analysed by high performance liquid chromatography. Salinity tolerance of the four species in decreasing order is A. minutum>A. peruvianum>A. tamarense>A. tamiyavanichii. Specific growth rates and maximum densities varied among these species with A. minutum recorded as the highest, 0.5 day(-1) and 6 x 10(4) cells L(-1). Toxin content decreased with elevated salinities in A. minutum, the highest toxin content was about 12 fmole cell(-1) at 5 psu. In A. tamiyavanichii, toxin content peaked at optimal growth salinity (20 and 25 psu). Toxin content of A. tamarense, somehow peaked at sub-optimal growth salinity (15 and 30 psu). Results of this study implied that salinity fluctuation not only influenced the growth physiology but also toxin production of these species.

Matched MeSH terms: Dinoflagellida/growth & development*; Dinoflagellida/metabolism*

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