Marine Dinoflagellate Amphidinium Carterae Research Articles

The influences of dissolved inorganic and organic phosphorus on arsenate toxicity in marine diatom Skeletonema costatum and dinoflagellate Amphidinium carterae

In this study, arsenate (As(V)) uptake, bioaccumulation, subcellular distribution and biotransformation were assessed in the marine diatom Skeletonema costatum and dinoflagellate Amphidinium carterae cultured in dissolved inorganic phosphorus (DIP) and dissolved organic phosphorus (DOP). The results of 3-days As(V) exposure showed that As(V) was more toxic in DOP cultures than in DIP counterparts. The higher As accumulation contributed to more severe As(V) toxicity. The 4-h As(V) uptake kinetics followed Michaelis–Menten kinetics. The maximum uptake rates were higher in DOP cultures than those in DIP counterparts. After P addition, the half-saturation constants remained constant in S. costatum (2.42–3.07 μM) but decreased in A. carterae (from 10.9 to 3.8 μM) compared with that in the respective P-depleted counterparts. During long-term As(V) exposure, A. carterae accumulated more As than S. costatum. Simultaneously, As(V) was reduced and transformed into organic As species in DIP-cultured S. costatum, which was severely inhibited in their DOP counterparts. Only As(V) reduction occurred in A. carterae. Overall, this study demonstrated species-specific effects of DOP on As(V) toxicity, and thus provide a new insight into the relationship between As contamination and eutrophication on the basis of marine microalgae.

Antibacterial Activity and Amphidinol Profiling of the Marine Dinoflagellate Amphidinium carterae (Subclade III).

Microalgae have received growing interest for their capacity to produce bioactive metabolites. This study aimed at characterising the antimicrobial potential of the marine dinoflagellate Amphidinium carterae strain LACW11, isolated from the west of Ireland. Amphidinolides have been identified as cytotoxic polyoxygenated polyketides produced by several Amphidinium species. Phylogenetic inference assigned our strain to Amphidinium carterae subclade III, along with isolates interspersed in different geographic regions. A two-stage extraction and fractionation process of the biomass was carried out. Extracts obtained after stage-1 were tested for bioactivity against bacterial ATCC strains of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa. The stage-2 solid phase extraction provided 16 fractions, which were tested against S. aureus and E. faecalis. Fractions I, J and K yielded minimum inhibitory concentrations between 16 μg/mL and 256 μg/mL for both Gram-positive. A targeted metabolomic approach using UHPLC-HRMS/MS analysis applied on fractions G to J evidenced the presence of amphidinol type compounds AM-A, AM-B, AM-22 and a new derivative dehydroAM-A, with characteristic masses of m/z 1361, 1463, 1667 and 1343, respectively. Combining the results of the biological assays with the targeted metabolomic approach, we could conclude that AM-A and the new derivative dehydroAM-A are responsible for the detected antimicrobial bioactivity.

Antialgal and antilarval activities of bioactive compounds extracted from the marine dinoflagellate Amphidinium carterae

With the global ban on the application of organotin-based marine coatings by the International Maritime Organization, the development of environmentally friendly, low-toxic and nontoxic antifouling compounds for marine industries has become an urgent need. Marine microorganisms have been considered as a potential source of natural antifoulants. In this study, the antifouling potential of marine dinoflagellate Amphidinium carterae, the toxic and red-tide microalgae, was investigated. We performed a series of operations to extract the bioactive substances from Amphidinium carterae and tested their antialgal and antilarval activities. The crude extract of Amphidinium carterae showed significant antialgal activity and the EC50 value against Skeletonema costatum was 55.4 μg mL−1. After purification, the isolated bioactive substances (the organic extract C) exhibited much higher antialgal and antilarval activities with EC50 of 12.9 μg mL−1 against Skeletonema costatum and LC50 of 15.1 μg mL−1 against Amphibalanus amphitrite larvae. Subsequently, IR, Q-TOFMS, and GC-MS were utilized for the structural elucidation of the bioactive compounds, and a series of unsaturated and saturated 16- to 22-carbon fatty acids were detected. The data suggested the bioactive compounds isolated from Amphidinium carterae exhibited a significant inhibiting effect against the diatom Skeletonema costatum and Amphibalanus amphitrite larvae, and could be substitutes for persistent, toxic antifouling compounds.

Carteraol E, a potent polyhydroxyl ichthyotoxin from the dinoflagellate Amphidinium carterae

Carteraol E ( 1), C 74H 126O 24, a polyhydroxyl ichthyotoxin, was isolated from the lab-cultured marine dinoflagellate Amphidinium carterae. Carteraol E possessed three tetrahydropyrans, and 19 hydroxyl groups on a C69-linear aliphatic chain with a ketone moiety, an exo-methylene, and three methyl branches. The structure was elucidated by extensive analyses of 2D NMR spectra. Cateraol E exhibited potent ichthyotoxicity with LD 50 value of 0.28 μM, and antifungal activity against Aspergillus niger.

Development of an efficient method for the preparative isolation and purification of chlorophyll a from a marine dinoflagellate Amphidinium carterae by high-speed counter-current chromatography coupled with reversed-phase high-performance liquid chromatography

In our research into chlorophylls of marine dinoflagellates, chlorophyll a was separated rapidly from the hexane extract of Amphidinium carterae in three steps. The first step was silica gel column chromatography, where elution was performed with 0-50% ethyl acetate in n-hexane. The second was high-speed counter-current chromatography using a two-phase solvent system consisting of n-hexane-ethyl acetate-methanol-water (5:5:5:1, v/v), and the third step was preparative reversed-phase high-performance liquid chromatography using a solvent system of acetone-water (89:11, v/v). HPLC analysis showed that the purity of chlorophyll a from the second step was over 83%, and after the third it was over 99%. Thirty milligrams of chlorophyll a was isolated from a crude sample of 250 mg of chlorophylls, and its structure was identified by analyzing its MS, 1H NMR and 13C NMR spectra.

A New Unsaturated Glycoglycerolipid from a Cultured Marine Dinoflagellate Amphidinium carterae.

AbstractFor Abstract see ChemInform Abstract in Full Text.

A New Unsaturated Glycoglycerolipid from a Cultured Marine Dinoflagellate Amphidinium carterae

From the cultured marine dinoflagellate Amphidinium carterae, a new unsaturated glycoglycerolipid (2S)-1,2-O-6,9,12,15-dioctadecatetraenoyl-3-O-[alpha-D-galactopyranosyl-(1''''-->6''')-O-beta-D-galactopyranosyl]-glycerol (1), has been isolated together with two known saturated ones, (2S)-1,2-distearoyl-3-O-(6-sulpho-alpha-D-quinovopyranosyl)-glycerol (2) and (2S)-1-stearoyl-3-O-(6-sulpho-alpha-D-quinovopyranosyl)-glycerol (3). Their structures were elucidated on the basis of chemical and spectral data.

Flow cytometric analysis of chronic and acute toxicity of copper(II) on the marine dinoflagellate Amphidinium carterae.

Copper(II) is a heavy metal whose levels have increased in some marine ecosystems to polluting levels. Dinoflagellates, an important phytoplankton group, are at the base of aquatic food chains and bioaccumulation of copper by these microorganisms can result in complex ecosystem alterations, so we investigated how copper disturbs those cells. Cytotoxic effects of sublethal and lethal copper concentrations ranging from 4.2 nM (control condition) to 3.13 microM estimated labile copper were studied in batch cultures of Amphidinium carterae. Cell morphology, motility, autofluorescence, and fluorescein diacetate (FDA)-dependent fluorescence generation were evaluated by flow cytometry (FCM) and microscopy. Exposure of A. carterae to toxic levels of copper impaired cell mobility, delayed cell proliferation, led to increased green autofluorescence, and at 3.13 microM labile copper also induced encystment and death. Chlorophyll fluorescence, however, was not affected. Kinetic FCM assay of FDA-dependent fluorescence generation showed a dose-dependent enhancement of fluorescein fluorescence immediately after copper addition and in cultures with sustained exposure to this toxicant. Our data suggest that copper toxicity occurs quickly at the membrane level in relation to oxidative stress generation. Based on fluorescence kinetic studies, the Na(+)/H(+) antiporter seemed to be affected by copper, thereby affecting intracellular pH.

Production of extracellular Cu complexing ligands by eucaryotic phytoplankton in response to Cu stress

Organic chelators that are thought to be of recent biological origin control copper speciation in seawater. Previous studies have often resolved these copper binding ligands into two classes, class 1 ligands (conditional stability constants 1012 –1014 M−1) and class 2 ligands (conditional stability constants 109–1012 M−1). At present the sources and structures of these chelators are unknown. Previous work has shown that in response to copper toxicity, cultures of the marine cyanobacteria Synechococcus spp. produce a strong ligand, similar in binding strength to class 1 ligands. We extended this work by surveying a suite of copper‐stressed marine eucaryotic phytoplankton for the production of copper binding ligands. Most eucaryotic species we studied did not produce ligands with the binding strength of class 1 ligands, but many produced class 2 ligands. A marine dinoflagellate Amphidinium carterae was found to produce very strong ligands; however, the low abundance of dinoflagellates in the open ocean may mean that this source of class 1 ligands is more important in coastal waters. This survey shows that there are many biological sources of copper binding ligands, but that procaryotes such as cyanobacteria are more plausible sources of class 1 ligands in the open ocean than eucaryotes.

Influence of bacteria on growth and hemolysin production by the marine dinoflagellate Amphidinium carterae

The effect of various bacteria on the growth of and hemolysin production by Amphidinium carterae was studied. The algal culture had an indigenous bacterial flora of Moraxella and Pseudomonas and these could not be eliminated by treatment with bactericidal antibiotics like gentamicin. Various bacteria like Micrococcus, Aeromonas, Vibrio, and Moraxella-like bacteria were added to A. carterae cultures to study their effect on growth and hemolysin production. Micrococcus and Aeromonas were found to improve the growth marginally. Hemolysin titres were considerably higher in A. carterae cultures supplemented with bacteria.

Photoinhibition at low quantum flux densities in a marine dinoflagellate (Amphidinium carterae)

Growth and photosynthetic properties of the marine dinoflagellate Amphidinium carterae Hulbert were examined under continuous illumination in batch cultures at four different irradiances between 2 and 150 μE m-2 s-1. The slope of both cell- and Chl a-based photosynthesis versus the irradiance curves was greatest for cells grown at 15 μE m-2 s-1. The relative Chl a values cell-1 were 1, 1.5 and 2 for cultures grown at 150, 80 and 15 μE m-2 s-1, respectively. A low-temperature (-196°C) fluorescence technique was used to examine cells for photoinhibiton. Photoinhibition was greatest for cells grown at 150 μE m-2 s-1. However, significant photoinhibition of this species was noted even at 80 μE m-2 s-1. No significant difference in the fluorescence pattern was found between cells grown at 2 and 15 μE m-2 s-1. Time course studies indicate that photoinhibition may occur within 2 h following exposure to 350 μE m-2 s-1 in cells grown at 15 μE m-2 s-1 and is reversible when light levels are lowered within 4 h. The ecological significance of phytoplankton unable to cope with excess photosynthetic excitation energy is discussed.

Peridinin-Chlorophyll a Proteins of the Dinoflagellate Amphidinium carterae (Plymouth 450)

The marine dinoflagellate Amphidinium carterae (Plymouth 450) releases several water-soluble peridinin-chlorophyll a proteins after freezethawing. These chromoproteins have a molecular weight of 39.2 x 10(3) and are comprised of noncovalently bound peridinin and chlorophyll a and a nonoligomeric protein. They have distinct isoelectric points and may be resolved into six components by either isoelectric focusing on polyacrylamide gel or ion exchange chromatography. The predominant chromoprotein, which has a pI of 7.5, constitutes about 90% of the extractable peridinin-chlorophyll a protein. It consists of an alanine-rich apoprotein of molecular weight 31.8 x 10(3) stoichiometrically associated with 9 peridinin and 2 chlorophyll a molecules. Additionally, the peridinin-chlorophyll a proteins with pI values of 7.6 and 6.4 were purified and found to have amino acid and chromophore composition essentially identical with the pI 7.5 protein. Peridinin-chlorophyll a protein, pI 7.5, after treatment at alkaline pH was transformed into several more acid pI forms of the protein, strongly suggesting that the naturally occurring proteins are deamidation products of a single protein. Fluorescence excitation and emission spectra demonstrate that light energy absorbed by peridinin induces chlorophyll a fluorescence presumably by intramolecular energy transfer. The peridinin-chlorophyll a proteins presumably function in vivo as photosynthetic light-harvesting pigments.