Amphidinium Operculatum Research Articles

Temporal changes in microalgal biomass and species composition on different plastic polymers in nutrient-enriched microcosm experiments

Marine plastic debris (MPD) is a potential threat to marine ecosystems, but its function as a vector for the transportation of harmful microalgae and its impact on the habitats of other marine organisms are uncertain. To address this gap in knowledge, we performed month-long experiments in 30 L microcosms that contained plates made of six different plastic polymers (polypropylene [PP], low-density polyethylene [LDPE], high-density polyethylene [HDPE], polyvinyl chloride [PVC], polyethylene terephthalate [PET], and polystyrene [PS]), and examined the time course of changes in planktonic and periphytic microalgae. There were no significant differences in the composition of periphytic microalgae or biomass among the different plastic polymers (p > 0.05). Nutrient depletion decreased the abundance of planktonic microalgae, but increased the biomass of attached periphytic microalgae (p < 0.05). In particular, analysis of the plastic plates showed that the abundance of benthic species that are responsible for harmful algal blooms (HABs), such as Amphidinium operculatum and Coolia monotis, significantly increased over time (days 21–28; p < 0.05). Our findings demonstrated that periphyton species, including benthic microalgae that cause HABs, can easily attach to different types of plastic and potentially spread to different regions and negatively impact these ecosystems. These observations have important implications for understanding the potential role of MPD in the spread of microalgae, including HABs, which pose a significant threat to marine ecosystems.

Sterols of Amphidinium species in the Operculatum Clade: Predominance of cholesterol instead of Δ8 (14) sterols previously considered Amphidinium-specific biomarkers.

The dinoflagellates Amphidinium carterae and Amphidinium corpulentum have been previously characterized as having Δ8(14) -nuclear unsaturated 4α-methyl-5α-cholest-8(14)-en-3β-ol (C28:1 ) and 4α-methyl-5α-ergosta-8(14),24(28)-dien-3β-ol (amphisterol; C29:2 ) as predominant sterols, where they comprise approximately 80% of the total sterol composition. These two sterols have hence been considered as possible major sterol biomarkers for the genus. Here, we have examined the sterols of four recently identified species of Amphidinium (Amphidinium fijiense, Amphidinium magnum, Amphidinium theodori, and Amphidinium tomasii) that are closely related to Amphidinium operculatum as part of what is termed the Operculatum Clade to show that each species has its sterol composition dominated by the common dinoflagellate sterol cholesterol (cholest-5-en-3β-ol; C27:1 ), which is found in many other dinoflagellate genera, rather than Δ8(14) sterols. While the Δ8(14) sterols 4α-methyl-5α-cholest-8(14)-en-3β-ol and 4α,23,24-trimethyl-5α-cholest-8(14),22E-dien-3β-ol (C30:2 ) were present as minor sterols along with another common dinoflagellate sterol, 4α,23,24-trimethyl-5α-cholest-22E-en-3β-ol (dinosterol; C30:1 ), in some of these four species, amphisterol was not conclusively observed. From a chemotaxonomic perspective, while this does reinforce the genus Amphidinium's ability to produce Δ8(14) sterols, albeit here as minor sterols, these results demonstrate that caution should be used when considering Δ8(14) sterols, especially amphisterol, as Amphidinium-specific biomarkers within these species where cholesterol is the predominant sterol.

Toxicity Bioassay and Cytotoxic Effects of the Benthic Marine Dinoflagellate Amphidinium operculatum.

Benthic dinoflagellates produce a wide array of bioactive compounds, primarily polyketides, that cause toxic effects on human consumers of seafood and perhaps mediate species interactions in the benthic microenvironment. This study assesses toxic and other bioactive effects of the benthic dinoflagellate Amphidinium operculatum (strain AA60) in two targeted bioassays. The brine shrimp (Artemia salina) bioassay revealed lethal effects of direct exposure to live dinoflagellate cells (Treatment A) and even higher potency with ethanolic extracts of lysed cells (Treatment D). There were no inimical bioactive effects of components released to the aqueous growth medium (Treatment B) or from aqueous cell lysates (Treatment C). The hypothesis that released bioactive compounds provide a chemical defense against metazoan grazers is therefore not supported by these results. The cytotoxic effect of ethanolic crude extracts of this dinoflagellate exhibited mild to high growth reduction effects on six human cancer cell lines. In particular, crude cell-free extracts proved highly growth-inhibitory activity towards breast and lung cancer cell lines MCF-7 and SKLU-1, respectively. Preliminary anti-cancer results indicate that natural bioactive compounds from Amphidinium are worthy of structural characterization and further toxicological investigation as potential therapeutants.

Morphology, growth, toxin production, and toxicity of cultured marine benthic dinoflagellates from Brazil and Cuba

The present study investigated selected benthic dinoflagellates isolated from different regions from the Western Atlantic, with respect to their morphology, growth, toxicity, and toxin production in culture. A total of nine strains of benthic dinoflagellates belonging to three genera were cultivated: Amphidinium massartii, Amphidinium operculatum (2 strains), Coolia malayensis (2 strains), Prorocentrum hoffmannianum (2 strains), and Prorocentrum mexicanum, whose morphological and genetic characterizations matched previous descriptions, and Prorocentrum sp., morphologically related to P. cf. norrissianum. The two strains of C. malayensis from Brazil attained the highest growth rates (0.42–0.47 day−1), but the lowest cell densities (2.2–2.9 × 104 cells mL−1) in culture. The highest cell densities were recorded for A. massartii from Cuba (3.8 × 105 cells mL−1). All species/strains investigated exhibited moderate toxicity to larvae of the brine shrimp Artemia salina; A. massartii being the most toxic species and Prorocentrum sp. the least one. Additionally, extracts of Prorocentrum species (P. hoffmannianum and Prorocentrum sp.) tested positive in mouse bioassays following intraperitoneal injection. Moderate to high concentrations of okadaic acid (OA), but no dinophysistoxins (DTXs), were found in both P. hoffmannianum strains from Cuba; but no diarrheic toxins were detected in either P. mexicanum from Cuba or Prorocentrum sp. from southern Brazil. Finally, five novel amphidinols were detected in cultures of both A. massartii (Cuba) and A. operculatum (Brazil) by LC-MS/MS, with molecular weights of 1440.8 (two isomers), 1360.8, 1287.7, and 984.6. These findings clearly indicate the need to include benthic species among the harmful microalgae surveyed in regional monitoring programs of phytoplankton.

English

Eleven strains of benthic dinoflagellates (Amphidinium carterae (D1), Prorocentrum rhathymum (D2), Symbiodinium sp. (D3), Coolia malayensis 1 (D4), Ostreopsis ovata 1 (D5), Ostreopsis ovata 2 (D6), Coolia malayensis 2 (D7), Amphidinium operculatum 1 (D8), Heterocapsa psammophila (D9), Coolia malayensis 3 (D10) and Amphidinium operculatum 2 (D11)) were collected in Jeju Island, Korea and cultured in 20 L carboys after establishing unialgal cultures. Their growth potential and biomass productivity were evaluated using two different culture media (IMK and f/2 medium); it was found that IMK medium has the potential to culture benthic dinoflagellates compared to commonly used f/2 medium. Among the benthic dinoflagellates, A. carterae (D1) had the maximum cell density (148.6 × 103 cells mL-1), growth rate (0.317 ± 0.01 divisions day-1) and biomass (0.260 ± 0.03 g L-1 dry weight) in IMK medium at 20 days of culture. Also, screened bioactivities among the methanolic extracts of cultured dinoflagellates showed A. carterae (D1) to have the highest antioxidant and anti-inflammatory effect and O. ovata 1 (D5) had the highest anticancer activity compared to the other strains. Taken together, this is the first report on the growth potential and biomass production of benthic dinoflagellate strains isolated from Jeju Island in appropriate culture medium as well as their importance in potential pharmacological applications. Key words: Amphidinium carterae, benthic dinoflagellates, biomass, bioactivities, culture conditions, Jeju Island.

Microphytobenthic dynamics in a Wadden Sea intertidal flat – Part II: Seasonal and spatial variability of non-diatom community components in relation to abiotic parameters

The microphytobenthos colonizing the intertidal flats forms an important component of the Wadden Sea. Ten sampling points along a 1-km transect were studied in a fringe area of the Solthörn tidal flat, southern North Sea, in order to determine seasonal differences in the microphytobenthos. An accompanying paper deals with the major component of the flora, the diatoms; here we, focus on the minor taxonomic groups. From May 2008 to May 2009 surface sediments were collected during low tide. Variation of environmental factors as well as microphytobenthic density (abundance and chlorophyll a) were monitored. The area investigated was a mixed-sediment mudflat, with a gradient from coarse to fine. Highest biomass was obtained in summer 2008 with 215.9 ± 12.6 mg chlorophyll a m–2. In late autumn the chlorophyll a concentration decreased continuously at all investigated stations. Lowest values were detected in December 2008. Species abundances varied considerably both along the transect and seasonally, depending on species-specific requirements as well as hydrodynamic conditions (tidal currents). Higher densities of benthic pro- and eukaryotic microalgae were observed in sites characterized by fine sediments. Apart from the diatoms, the most abundant microphytobenthic group was the cyanophytes. Coccoid cyanophytes, mainly Merismopedia sp., were most abundant during summer, with cell numbers up to 5.72 × 106 cells cm−2, while diatoms dominated in winter, spring and autumn. Filamentous cyanophytes, particularly Microcoleus chthonoplastes, were most abundant during autumn, while coccoid chlorophytes (spring: Chlorococcum submarinum, Crucigenia tetrapedia, Tetraselmis suecica), euglenophytes (summer: Euglena obtusa), dinophytes (autumn: Amphidinium operculatum, A. herdmanii) and cryptophytes (autumn: Hillea marina, Hemiselmis virescens) contributed to the microphytobenthos during warmer seasons. The statistical analysis confirmed that the composition of the microphytobenthos was related to sediment features and to characteristics of particular seasons.

Comparative analysis of dinoflagellate chloroplast genomes reveals rRNA and tRNA genes

BackgroundPeridinin-containing dinoflagellates have a highly reduced chloroplast genome, which is unlike that found in other chloroplast containing organisms. Genome reduction appears to be the result of extensive transfer of genes to the nuclear genome. Unusually the genes believed to be remaining in the chloroplast genome are found on small DNA 'minicircles'. In this study we present a comparison of sets of minicircle sequences from three dinoflagellate species.ResultsPCR was used to amplify several minicircles from Amphidinium carterae so that a homologous set of gene-containing minicircles was available for Amphidinium carterae and Amphidinium operculatum, two apparently closely related peridinin-containing dinoflagellates. We compared the sequences of these minicircles to determine the content and characteristics of their chloroplast genomes. We also made comparisons with minicircles which had been obtained from Heterocapsa triquetra, another peridinin-containing dinoflagellate. These in silico comparisons have revealed several genetic features which were not apparent in single species analyses. The features include further protein coding genes, unusual rRNA genes, which we show are transcribed, and the first examples of tRNA genes from peridinin-containing dinoflagellate chloroplast genomes.ConclusionComparative analysis of minicircle sequences has allowed us to identify previously unrecognised features of dinoflagellate chloroplast genomes, including additional protein and RNA genes. The chloroplast rRNA gene sequences are radically different from those in other organisms, and in many ways resemble the rRNA genes found in some highly reduced mitochondrial genomes. The retention of certain tRNA genes in the dinoflagellate chloroplast genome has important implications for models of chloroplast-mitochondrion interaction.

The Copy Number of Chloroplast Gene Minicircles Changes Dramatically with Growth Phase in the Dinoflagellate Amphidinium operculatum

The chloroplast genome of algae and plants typically comprises a circular DNA molecule of 100-200kb, which harbours approximately 120 genes, and is present in 50-100 copies per chloroplast. However, in peridinin dinoflagellates, an ecologically important group of unicellular algae, the chloroplast genome is fragmented into plasmid-like 'minicircles', each of 2-3kb. Furthermore, the chloroplast gene content of dinoflagellates is dramatically reduced. Only 14 genes have been found on dinoflagellate minicircles, and recent evidence from EST studies suggests that most of the genes typically located in the chloroplast in other algae and plants are located in the nucleus. In this study, Southern blot analysis was used to estimate the copy number per cell of a variety of minicircles during different growth stages in the dinoflagellate Amphidinium operculatum. It was found that minicircle copy number is low during the exponential growth stage but increases during the later growth phase to resemble the situation seen in other plants and algae. The control of minicircle replication is discussed in the light of these findings.

Novel plastid gene minicircles in the dinoflagellate Amphidinium operculatum.

Seven new minicircles, forming part of the fragmented plastid genome of the dinoflagellate Amphidinium operculatum, have been identified by PCR. Three minicircles are full-length, one of 2.6 kb encoding the 23S rRNA gene, one of 2.4 kb containing the psaB gene, and a third of 2.5 kb containing the psbD, psbE and psbI genes. This is the first report of a three-gene minicircle. All three genes are conventional in length, with the same codon bias found in other minicircle genes. The psbI gene sequence is very divergent. One empty minicircle, of 1.7 kb, and three 'microcircles', between 400 and 600 bp, have also been identified by PCR. They appear to be formed from full-length minicircles by homologous recombination and internal deletion. All three microcircles contain the core region common to minicircles, but are missing a coding region, providing further evidence that the core region is necessary for minicircle replication and maintenance.

AMPHIDINIUM REVISITED. II. RESOLVING SPECIES BOUNDARIES IN THE AMPHIDINIUM OPERCULATUM SPECIES COMPLEX (DINOPHYCEAE), INCLUDING THE DESCRIPTIONS OF AMPHIDINIUM TRULLA SP. NOV. AND AMPHIDINIUM GIBBOSUM. COMB. NOV.1

Amphidinium operculatum Claparède et Lachmann, the type species of the dinoflagellate genus Amphidinium, has long had an uncertain identity. It has been considered to be either difficult to distinguish from other similar species or a morphologically variable species itself. This has led to the hypothesis that A. operculatum represents a “species complex.” Recently, the problem of distinguishing A. operculatum from similar species has become particularly acute, because several morphologically similar species have been found to produce bioactive compounds of potential interest to the pharmaceutical industry. In this study, we cultured and examined existing cultures of several species of Amphidinium, most of which have been previously identified as A. operculatum or as species considered by some to be synonyms or varieties of A. operculatum. Thirty strains were examined using comparative LM, SEM, and partial large subunit (LSU) rDNA sequence data. Through morphological and molecular phylogenetic analyses, six distinct species were identified, including Amphidinium trulla sp. nov. and Amphidinium gibbosum comb. nov. Amphidinium operculatum was redescribed based on four cultures. Genetic variability within the examined Amphidinium species varied greatly. There was little difference among strains in partial LSU rDNA for most species, but strains of A. carterae and A. massartii Biencheler differed by as much as 4%. In both A. carterae and A. massartii, three distinct genotypes based on partial LSU rDNA were found, but no morphological differences among strains could be observed using LM or SEM. In the case of A. carterae, no biogeographically related molecular differences were found.

Physiological factors contributing to the species-specific sensitivity of four estuarine microalgal species exposed to the herbicide atrazine

Algal species vary considerably in sensitivity to the commonly used herbicide atrazine, and it is hypothesized that several factors may contribute to species-specific sensitivity. In this study four estuarine microalgal species, a planktonic chlorophyte (Dunaliella tertiolecta), a benthic chlorophyte (Ankistrodesmus sp.), a cryptophyte (Storeatula major) and a dinoflagellate (Amphidinium operculatum), were exposed to atrazine to determine toxicity and identify factors that might influence species sensitivity. Atrazine effects were examined at the population (cell density, primary productivity and biomass), cellular (biovolume), and subcellular (pigment composition, protein concentration and lipid concentration) levels. Atrazine significantly decreased cell density, productivity rate, biomass and biovolume in all the algal populations tested at atrazine concentrations ≥ 12.5 μ g l−1. Toxicity values (96 h EC50 values) for the four species ranged from 11.87 to 146.71 μ g l−1. Species sensitivity rankings varied with endpoint measured. Overall, Ankistrodesmus sp. was the most sensitive species followed by Storeatula, Dunaliella and Amphidinium. Cellular biovolume was a significantly more sensitive test endpoint for Amphidinium, chlorophyll a was a significantly more sensitive test endpoint for Ankistrodesmus sp., and phototrophic carbon assimilation was a significantly more sensitive test endpoint for both Storeatula and Amphidinium. Algal subcellular responses to atrazine were also species dependent. Chlorophyll a concentration per cell decreased in the green algae, increased in Amphidinium and did not change in Storeatula. Total lipids per cell increased in Storeatula, decreased in Amphidinium and did not change in the green algae. Ankistrodesmus sp. pigments were not significantly altered after atrazine exposure; however selected Amphidinium pigments increased per cell, and selected Dunaliella and Storeatula pigments decreased per cell in the atrazine treatments. Atrazine significantly reduced cellular biovolume in all test species. Species with smaller biovolumes and less chlorophyll a per cell tended to be more sensitive to atrazine exposure based on population growth rate.

Highly organized structure in the non-coding region of the psbA minicircle from clade C Symbiodinium.

The chloroplast genes of dinoflagellates are distributed among small, circular dsDNA molecules termed minicircles. In this paper, we describe the structure of the non-coding region of the psbA minicircle from Symbiodinium: DNA sequence was obtained from five Symbiodinium strains obtained from four different coral host species (Goniopora tenuidens, Heliofungia actiniformis, Leptastrea purpurea and Pocillopora damicornis), which had previously been determined to be closely related using LSU rDNA region D1/D2 sequence analysis. Eight distinct sequence blocks, consisting of four conserved cores interspersed with two metastable regions and flanked by two variable regions, occurred at similar positions in all strains. Inverted repeats (IRs) occurred in tandem or "twin" formation within two of the four cores. The metastable regions also consisted of twin IRs and had modular behaviour, being either fully present or completely absent in the different strains. These twin IRs are similar in sequence to double-hairpin elements (DHEs) found in the mitochondrial genomes of some fungi, and may be mobile elements or may serve a functional role in recombination or replication. Within the central unit (consisting of the cores plus the metastable regions), all IRs contained perfect sequence inverses, implying they are highly evolved. IRs were also present outside the central unit but these were imperfect and possessed by individual strains only. A central adenine-rich sequence most closely resembled one in the centre of the non-coding part of Amphidinium operculatum minicircles, and is a potential origin of replication. Sequence polymorphism was extremely high in the variable regions, suggesting that these regions may be useful for distinguishing strains that cannot be differentiated using molecular markers currently available for Symbiodinium.

Organisation and expression of the plastid genome of the dinoflagellate Amphidinium operculatum.

We show using PCR that psbC, atpA and petB genes are present in the plastid DNA minicircles from the dinoflagellate Amphidinium operculatum, extending the set of plastid genes identified from this organism. Unusually, the petBand atpA genes are located on the same minicircle. PCR using primers based on the "core" region found on all coding minicircles revealed the existence of a number of DNA minicircles with no apparent coding function. Northern analysis of total RNA from A. operculatum showed that the petB and atpA genes are represented on separate transcripts, despite being encoded in close proximity on the same minicircle. The possibility of transcript editing was investigated by RT-PCR, but psaA, psbA, psbB and atpB transcripts showed no evidence of editing, indicating that GUA can be used as an initiation codon in A. operculatum.

Cell cycle regulation in a dinoflagellate, Amphidinium operculatum: identification of the diel entraining cue and a possible role for cyclic AMP

This research describes the diel phasing of the cell cycle in the dinoflagellate, Amphidinium operculatum Claparéde and Lachmann, and investigates the mechanisms that serve to link the cell cycle to the diel cycle. Unlike many dinoflagellates, A. operculatum has a naturally high division rate of approximately 1 division day −1, which yields a nearly synchronous population, making it useful for population studies. When grown on a 16:8 h light/dark cycle, S-phase begins 10 h and mitosis 14–16 h after the onset of light, as determined by flow cytometry. Alterations in the timing of the dark/light and light/dark transitions showed that the cell cycle is entrained by the dark/light transition, with the light/dark cue being uninvolved. Cells in logarithmic phase growth also undergo diel changes in cell size (9–14 μm), reaching a maximum size late in the light phase, concurrent with mitosis. Stationary phase cells or cells blocked in G1 of the cell cycle with a cell cycle inhibitor, olomoucine, showed no size changes or reduced size changes over the diel cycle, suggesting a coupling of cell size to the cell division cycle. In Euglena, cAMP-dependent signaling appears to mediate diel phasing of the cell cycle. Therefore, the role of cAMP in cell cycle control in A. operculatum was investigated. Measurement of intracellular cAMP by radioimmunoassay (RIA) revealed that cAMP concentrations varied on a diel basis, but increases observed appeared to correlate with cell size increases, and did not correlate with light cues at the dark/light or light/dark transition. However, when cells were treated with the cAMP phosphodiesterase inhibitor, IBMX, cell cycle progression was inhibited at both the G1/S and the G2/M phase transitions. This supports a role for cAMP-dependent signaling in the dinoflagellate cell cycle and is in agreement with the documented role of cAMP in the cell cycle control of higher eukaryotes.

Unusual organisation of the plastid genome of the dinoflagellate Amphidinium operculatum

We have previously reported an anomalous organization of several plastid genes in the dinoflagellate Amphidinium operculatum. These genes are located on individual minicircles of around 2.4 kbp, with a single gene on each minicircle. Each minicircle also has a `core? region, which is similar but not identical between minicircles. This gene organization contrasts markedly with that in other plastid genomes, which typically have 100 or so genes located on one molecule. Using dinoflagellate DNA and PCR with primers based on conserved regions of several genes, we have identified additional genes for plastid proteins on minicircles. We have identified one minicircle containing two genes. PCR using primers based on the core region has allowed us to identify a number of `empty? minicircles that do not appear to contain any genes (although one empty minicircle has short regions of similarity to other genes). These empty minicircles have a recognisable core region, however. We have carried out transcript analysis of a number of minicircles, including the one carrying two genes. We have also tested the hypothesis that dinoflagellate plastid transcripts undergo editing.

IDENTIFICATION OF A CYCLIC AMP‐DEPENDENT PROTEIN KINASE IN THE DINOFLAGELLATE AMPHIDINIUM OPERCULATUM

Cyclic AMP is an important signaling molecule in both prokaryotes and eukaryotes. In eukaryotes, the action of cyclic AMP is mediated by cyclic AMP‐dependent protein kinase (PKA). Efforts to identify PKA in dinoflagellates have met with mixed success in previous studies. In Amphidinium operculatum, we have identified the presence of cAMP, and observed cell cycle inhibition by the cAMP pathway inhibitor, isobutyl methylxanthine, suggesting the presence of a eukaryotic cAMP pathway. However, crude lysates of A. operculatum failed to show in vitro kinase activity toward kemptide, a specific substrate of mammalian PKA. This may reflect the presence of unidentified low molecular weight inhibitors, because when cell lysates were fractionated by gel filtration chromatography, cAMP dependent kinase activity was identified. The molecular weight of this PKA was 66 kDa, much smaller than bovine PKA (250 kDa), but similar to that reported in the diatom Cylindrotheca (78 kDa). Current work is focusing on characterization of subunit structure of the PKA of A. operculatum, as compared with bovine PKA.

MEASUREMENT OF MICROALGAL CELL VOLUME BY FLOW CYTOMETRY

Single cell analysis by flow cytometry is a powerful tool that has been employed to identify many different characteristics of phytoplankton populations. Cell volume is an important physiological component of many cellular processes. We have used a Coulter EPICS XL flow cytometer to measure cell volume in the spheroid dinoflagellate Amphidinium operculatum as a function of forward scatter. Cell volume measurements of this alga were quantified as equivalent spherical diameters from a standard curve obtained with latex beads of known diameter. This parameter was used to monitor cell diameter throughout the cell division cycle. In log phase cultures, A. operculatum showed increasing cell volumes throughout the light phase and a maximum cell volume concurrent with the onset of cell division late in the light phase. The maximum equivalent spherical diameter measured 14 μm, while the minimum equivalent spherical diameter was 10 μm that occurred late in the dark phase. Stationary phase cultures of A. operculatum did not exhibit oscillating cell volumes throughout the diel cycle. Chemical inhibition of the cell cycle using 100 μM olomoucine diminished cell volume changes during the light phase. These results suggest a coupling of size control to the cell division cycle.

AMPHIDINIUM OPERCULATUM VAR. NOV. GIBBOSUM (DINOPHYCEAE), A FREE‐SWIMMING MARINE SPECIES PRODUCING CYTOTOXIC METABOLITES1

ABSTRACTA single‐cell isolate of Amphidinium from the northern Caribbean Sea was identified as A. operculatum Clap. et Lack. var. nov. gibbosum Maranda et Shimizu based on a morphological and ultrastructural study. This free‐swimming dinoflagellate, which was found to produce potent antitumor metabolites, is elongate and asymmetrical. It is compared with two close relatives A. klebsii Kofoid et Swezy and A. carterae Hulburt. Amphidinium operculatum var. gibbosum can be distinguished from A. klebsii on the basis of shape while it differs from A. carterae on the basis of size, shape, cell surface, and chloroplast arrangement. The shape and size of the nucleus and the presence of mucocysts also differentiate the gibbosum variety from its two relatives. The architecture of the pusule may ultimately provide the best ultrastructural discriminating character. Otherwise, the general ultra‐structure of the Caribbean isolate is typical of many unarmored photo synthetic dinoflagellates with amphiesma, condensed chromosomes, trichocysts, mitochondria with tubular cristae, and trilamellar chloroplasts.

Distribution of Amphidinium

BIOLOGICAL readers of NATURE will perhaps recollect the record of the finding of the dinoflagellate, Amphidinium operculatum (previously unknown in Britain), on the beach at Port Erin a couple of years ago. Since then it has been present in great abundance at Port Erin on many occasions; Mr. R. D. Laurie has found it at Hoylake, near Liverpool, two of our young Liverpool zoologists (R. J. Daniel and J. E. Hamilton) now at the Belmullet Whaling Station, co. Mayo, inform me that they have noticed it on the shores of Blacksod Bay, and now I have to-day found it here in abundance, staining slightly in patches and streaks the beautiful white shell-sands of Iona.

On the Occurrence of Amphidinium operculatum, Clap. &amp; Lach., in vast Quantity, at Port Erin (Isle of Man).

On the Occurrence of Amphidinium operculatum, Clap. & Lach., in vast Quantity, at Port Erin (Isle of Man) Get access W. A. Herdman, F.R.S., F.L.S. W. A. Herdman, F.R.S., F.L.S. Search for other works by this author on: Oxford Academic Google Scholar Botanical Journal of the Linnean Society, Volume 40, Issue 277, May 1912, Pages 389–393, https://doi.org/10.1111/j.1095-8339.1912.tb00876.x Published: 23 December 2008 Article history Accepted: 13 May 2008 Published: 23 December 2008