Alexandrium Sp Research Articles

Dinoflagellate cyst assemblages in surface sediment from Jakarta Bay

Dinoflagellate cysts play an essential role in the initiation, recurrence and geographical expansion of harmful algal blooms (HABs). The horizontal distribution and abundance of dinoflagellate cysts in marine sediments provide essential information in giving early warnings of the presence of toxic species and possible continuing recurrence of HABs in a given area. This research carried out in the waters of Jakarta Bay. Sediment samples collected from thirteen sampling stations using TFO gravity corer 2.1 cm diameter from sediment thickness 0-4 cm. The results showed that resting cysts belong to Gonyaulax, Alexandrium, Scrippsiella, Protoperidinium, Gymnodinium, Gyrodinium. Two of them, namely Gymnodinium sp and Alexandrium sp, known as toxic species. Cysts in the sediment thickness 0-4 cm mainly dominated by the group of Gonyaulax (Gonyaulacoid). Cyst abundance in the sediment depth-layer 0-2 cm is higher than in depth-layer 2-4 cm. The number of dino-cysts in sediment layer 0-2 cm ranged from 2.984 – 22.298 cysts cm−3 while in the layer 2-4 cm the amount varies from 1.063 – 8.132 cysts cm−3 of wet sediment. The result revealed that dino-cyst abundance is higher in the deposit collected from locations near to the coast. The assemblages of dinoflagellate resting cysts seemed to have a relation with some hydrological parameters.

Identifying the Source Organisms Producing Paralytic Shellfish Toxins in a Subtropical Bay in the South China Sea.

Identifying the exact phytoplanktonic sources of paralytic shellfish toxins (PSTs) is crucial for monitoring and preventing the buildup of toxin pollution, especially for causative species occurring at low levels. Phytoplankton and shellfish samples were simultaneously collected from representative mariculture zones in Daya Bay, China. Low concentration/low toxicity PSTs predominated with N-sulfocarbamoyl toxins 1, 2 (C1/2) being detected in phytoplankton (≤6.25 pmol L-1) and shellfish (≤0.21 μg STXeq g-1), which pose a potential risk of seafood poisoning. High-throughput sequencing of the phytoplankton samples based on 18S rDNA V4 regions identified 93 genera in 445 operational taxonomic units (OTUs). Five OTUs were assigned to Alexandrium hiranoi, Ambicodamus leei, Alexandrium pacificum, Alexandrium minutum, and an uncertain Alexandrium sp. A. pacificum and A. minutum are candidate PST producers and observed under the light microscope with densities of 66-972 cells L-1. Three strains of toxigenic species were successfully isolated and identified as A. pacificum and A. minutum based on their 18S rDNA V4 regions. The predominant toxins in A. pacificum were C1/2 (43.9-53.6 fmol cell-1) and resembled the toxins found in field samples predominated with C1/2. A. minutum produced only gonyautoxins 2/3 (8.03 fmol cell-1). Therefore, A. pacificum was identified as the predominant PST contributor in this area. This research makes a valuable contribution to the understanding of the traceability of phycotoxins in marine waters.

Harmful micro-algal blooms and local environmental conditions of the coastal fringe: Moroccan Mediterranean coasts as a case study

The dynamic study of harmful phytoplankton, conducted from June 2013 to May 2014, has tracked the evolution of the microalgae community at four sites located along the Moroccan western Mediterranean coast. 91 species and genera of phytoplankton were identified, 8 of which are recognized as potentially toxic. The genus Pseudo-nitzschia, a diatom known to produce domoic acid, is quite prevalent in all four sites, particularly in spring. The toxic dinoflagellate species detected in our study are Gymnodinium catenatum, Alexandrium sp PSP producers, Prorocentruml Lima, Dinophysis acuminata, D. caudata, D. fortii DSP producer, and Ostreopsis sp known palytoxin producer. The species Gymnodinium catenatum proliferates intensively from S1 to S3 in winter and S4 in summer, while Alexandrium sp proliferates mainly during spring and late summer with an exceedance of the threshold. Significant blooms of Dinophysis sp were recorded during the summer at all sites. On the other hand, the abundance of Ostreopsis sp was noted during the spring summer period with low concentrations. Discriminant analysis (DFA) of nutrients, toxic species identifies perfect seasonal discrimination. Winter and fall are characterized by high nutrient inputs, but algal biomass is low. On the contrary, spring and summer are characterized by a depletion of nutrients in the environment following the assimilation of these elements by phytoplankton.

Spatial Distribution of Dissolved Inorganic Nutrients and Phytoplankton around Kota Kinabalu Wetland, Sabah, Malaysia

Kota Kinabalu Wetland is importantly habited of mangrove, diverse aquatic flora and fauna as well as feeding stop for migratory birds. This wetland is inundated with the tidal flow, as connected with a small river and nearby coastal areas, thus. A study was carried out to determine the spatial distribution of dissolved inorganic nutrients and phytoplankton diversity at Kota Kinabalu (KK) Wetlands. Five stations, in which river mouth of Likas Bay, river channel (two stations) and inundated area (two stations) in KK Wetland were selected for this study. In-situ parameters of water, water for nutrients and phytoplankton samples were collected from May 2019 until October 2019. The highest concentration of nitrate (0.115 mg/L) was recorded at inundated area of wetland (S5) while the lowest nitrate concentration (0.0047 mg/L) was found at river (S3) flowing towards wetland. The concentrations of ammonia (0.2004 to 2.311 mg/L) were recorded relatively higher at every station compared to other dissolved inorganic nutrients (DIN). The concentration of phosphate was determined in the ranges of 0.0089 - 0.0513 mg/L. Nitrate, ammonia and phosphate showed no significant difference (P = 0.737) in terms of DIN concentration at all five sampling stations during the study period. Twenty-four genera of phytoplankton were identified, dominated by diatoms (55.29%), followed by dinoflagellates (24.95%), Chrysophyta (11.15%), Spirotrichea (5.28%) and Cyanophyta (3.33%). Dominating species throughout the study period include Chaetoceros sp., Pseduo-nitzschia sp., and Cylindrotheca closterium, Peridinium quinquecorne and Alexandrium sp. Phytoplankton species compositions were observed the highest in river mouth area in July with the highest density of 12.115 × 104 cells/mL. The study showed that nutrient concentration was insignificant (P = 0.614) in altering the phytoplankton density, as influences with the tidal water.

The Genus Alexandrium (Dinophyceae, Dinophyta) in Brazilian Coastal Waters

A review of the dinoflagellate genus Alexandrium occurring in Brazilian coastal waters is presented based on both published information and new data. Seven Alexandrium species have been recorded from Brazil so far: A. catenella, A. fraterculus, A. gaardnerae, A. kutnerae, A. tamiyavanichi, A. tamutum, and Alexandrium sp. While A. gaardnerae and A. kutnerae were identified based only on morphological characteristics, phylogenetic analysis (ITS and LSU rDNA) were performed for the remaining species based on cultures and/or field populations. Monoclonal cultures of the analyzed species were isolated from field samples obtained from Bahia (A. tamiyavanichi, two strains), Rio de Janeiro (A. tamutum, three strains; Alexandrium sp., two strains), Santa Catarina (A. fraterculus, one strain) and Rio Grande do Sul (A. tamarense, three strains). This is the first record of A. tamutum for the South Atlantic. In addition, molecular data for Brazilian strains of A. fraterculus are presented for the first time, as well as sequences from the ITS region for A. catenella (previously reported as A. tamarense) from Brazilian coastal waters. The morphological characters of the three species corresponded to those typically recorded in the literature and their identification was confirmed by molecular analysis. Based on the LSU rDNA and ITS regions, the three strains of A. catenella showed a high degree of similarity with strains from Southern Chile and North America. The implications and limitations of these findings for the monitoring protocols within the global and regional context are discussed.

Ship traffic and the introduction of diatoms and dinoflagellates via ballast water in the port of Annaba, Algeria

We present here the first study on the role of ship traffic in the introduction of potentially harmful and/or non-indigenous species in the port of Annaba (Algeria). A total of 25 ships of two different types (general cargo and bulk carriers) were sampled and separated into two categories: oceanic and Mediterranean ships. We estimated propagule pressure of high-risk coastal phytoplankton delivered in ballast water to the port of Annaba. We identified 40 diatom and 38 dinoflagellate taxa, among which, 11 harmful/toxic taxa: Pseudo-nitzschia spp., Alexandrium tamarense, Alexandrium sp., Dinophysis acuminata, Dinophysis rotundata, Dinophysis sp., Gonyaulax spinifera, Gymnodinium catenatum, Lingulodinium polyedrum, Protoceratium reticulatum and cyst of Alexandrium sp. In addition, 8 taxa (5 diatoms, 1 dinoflagellate and 2 dinoflagellate cysts) never observed in the Annaba region were considered as potentially non-indigenous: Actinoptychus splendens, Coscinodiscus asteromphalus, Coscinodiscus lineatus, Odentella granulata, Thalassiosira cf. decipiens, Prorocentrum scutellum, cyst of Polykrikos kofoidii and Islandinium minutum. Several factors were examined, including ship routes, ballast water age and the volume of ballast water discharged. Our analyses revealed that diatom and dinoflagellate abundances decreased with ballast water age, possibly as a result of mortality of species due to voyage length and lack of light in ballast tanks. Estimates of actual propagule pressure, diatoms and dinoflagellates abundances varied from 1 to 4×108 cells/ship. The results of this study could serve as the baseline for the development and implementation of monitoring and ballast water management programs in ports of Algeria.

Effects of environmental parameters and nutrients on phytoplankton communities around the shrimp farm complexes in Bushehr Province, in the Persian Gulf

The aim of this work was to be informed and aware of the red tide conditions before the entering of algal blooms in seawater supply canals of the shrimp farm and hatchery complexes in Bushehr Province. Field investigations and monthly samplings have been carried out to determine environmental parameters, nutrients, chlorophyll–a and phytoplankton in the southern part of the input water channels of Mond, Delvar and Heleh farmed shrimp complexes from April to December 2011. The identified phytoplankton belonged to three classes of Bacillariophyceae, Dinophyceae and Cyanophyceae. 12 genera belonging to Dinophyceae, 25 genera to Bacillariophyceae and two genera to Cyanophyceae were observed during the study. The highest average density of total phytoplankton was recorded at Heleh station at 18374 cells/lit. The maximum density of phytoplankton was at Delvar station in December. The highest density of Dinophyceae was observed in August. Alexandrium sp ., Ornithocercus and Prorocentrum sp. were the predominant species of Dinophyceae class. Bacillariophyceaes are thermo-tolerant and halo-tolerant while Dinophyceaes and Cyanophyceaes are thermo-intolerant and halo-intolerant. Bacillariophyceaes are silica limited while Dinophyceaes are phosphorus limited phytoplankton

HARMFUL MICROALGAE ASSEMBLAGE IN THE AQUACULTURE AREA OF AMAN ISLAND, NORTHEN STRAIT OF MALACCA

A field survey was carried out in Aman Island, the northern Strait of Malacca, to investigate the occurrence of harmful microalgae. The area is known for marine finfish cage-culture and blood cockle farming. Plankton samples were collected fortnightly between July 2011 and September 2013. A total of 58 phytoplankton genera were recorded; 37 belong to the diatoms, and 21 genera were dinoflagellates. Harmful microalgae species were identified microscopically, of which nine are bloom-forming species that responsible for fish kills. These included Akashiwo sanguinea, Chaetoceros affinis, Cha. curvisetus, Cha. laevis, Ceratium furca, C. fusus, Cochlodinium sp., Karlodinium australe and Noctiluca scintillans. Three Alexandrium species were identified from the samples, i.e. A. leei, A. affine and Alexandrium sp. Pulau Aman; some species in the genus have been known to associate with Paralytic Shellfish Poisoning (PSP). Diarrhetic Shellfish Poisoning (DSP)-toxin producers were also encountered in the water samples, viz. Dinophysis acuminata, D. caudata, Dinophysis sp. and Prorocentrum micans. The harmful diatom, Pseudo-nitzschia spp. that associated with Amnesic Shellfish Poisoning (ASP), was commonly found in the water samples. Cell abundance of each harmful microalga was enumerated. The results showed that cell densities of harmful microalgae were relatively low throughout the sampling period. The microalgae community was dominated by Ceratium furca in May 2012, with cell densities exceeding 70,000 cells L-1; no fish kill event was reported during the time. Even though no fish kill or shellfish poisoning was reported in the area, he presence of harmful microalgae albeit at low cell densities, may pose a potential threat to human health and seafood biosecurity. Therefore, regular phytoplankton monitoring is necessary to safeguard human health and seafood industries in the country. Keywords: Aman Island; blood cockle; fish-cage; Harmful Algal Bloom; Malaysia; Strait of Malacca

Occurrence of Amoebophrya spp. infection in planktonic dinoflagellates in Changjiang (Yangtze River) Estuary, China

The parasitic dinoflagellates in the genus of Amoebophrya can infect broad ranges of planktonic dinoflagellates, and transform algal biomass into organic matter that can be recycled within the planktonic community. The ecological significance of Amoebophrya spp. during harmful algal bloom (HAB) events was gradually recognized along with revelation of its host specificity and diversity in picoplankton communities. The eutrophicated coastal waters of China are frequently affected by HABs, particularly in Changjiang (Yangtze River) estuary and the adjacent East China Sea; while, no research has been conducted to explore the ecological roles of parasitism during HAB events and the related dinoflagellate bloom dynamics. For the first time, we confirmed the presence of Amoebophrya infections in the planktonic community of this region; six species of dinoflagellates were infected, including Ceratium tripos, Scrippsiella trochoidea, Gonyaulax spinifera, Gymnodinium sp., Gonyaulax sp. and an Alexandrium sp. Molecular sequences retrieved from environmental water samples revealed high genetic diversity of Amoebophryidae-like organisms in the water column. Amoebophrya-infected dinoflagellates were only observed in high salinity (>20) stations suggesting that salinity may be a factor limiting the distribution of Amoebophyra infections in natural environment. Whereas, no evidence of Amoebophrya infection was observed in the bloom-forming species Karenia mikimotoi, suggesting that K. mikimotoi in this region was likely free of Amoebophridae infection.

Impact of economic development on the dynamics of phytoplankton and physic-chemical quality of Dakhla Bay (South of Morocco)

A detailed monitoring of physic-chemical parameters and a complete inven-torization of phytoplankton species was conducted from May 2011 to May 2012 in Dakhla bay in south of Morocco. This study comes to bridge the gap of environmental data which has started since 2000 by Moroccan national institute of fisheries research (INRH) in this area. The results showed a differ-ence in the distribution of physic-chemical parameters (especially nutrient and dissolved oxygen). In addition, inventory of phytoplankton species has allowed identifying three different groups: diatoms (Rhizosolenia spp, Lepto-cylindrus spp, Thalassionema spp, Pseudo- nizschia spp.,...) represented 92% of the total phytoplankton flora, dinoflagellates (Scrippsiella sp, Prorocen-trum sp, Gymnodinium sp.,...) with 7%, while Silicoflagellates (Dictyocha sp) contributed with 1%. Seasonal fluctuation of these groups was governed, in first order, by the importance of upwelling in the region during the summer (intense phytoplankton activity). Yet this activity was much stronger in the PK25 site for tourism activity that suggested a remarkable anthropogenic input. An occurrence of Scrippsiella species (45% Hoja Iliamera growing area) and Prorocentrum sp (41% Duna Blanca aquaculture area), recognized as species of colored waters, was permanent at the study sites. Alexandrium sp represented 1% of the total dinoflagellates in Duna Blanca (<1% in the other sites), whereas Dinophysis sp (<1%) was observed in Lassarga (the entrance bay) and PK25 only. The latter two species are potentially the cause of PSP toxins's production (Paralytic Shellfish Poisoning) and DSP (Diarrheic Shellfish Poisoning) source of contamination of bivalves in the Bay during 2004 and 2006.

SURFACTANT PROMOTION OF THE INHIBITORY EFFECTS OF CUPRIC GLUTAMATE ON THE DINOFLAGELLATE ALEXANDRIUM(1).

We studied cupric glutamate as a novel algicide for marine harmful algae and hexadecyltrimethyleamine bromide (HDTMAB) as an accelerant. Cupric glutamate had an excellent ability to inhibit the growth of Alexandrium sp. LC3, but the inhibition efficiency did not increase with higher cupric glutamate concentration. The studies on the inhibition ofAlexandrium sp. LC3 by cupric sulfate or cupric glutamate showed that cupric glutamate had a higher inhibition rate than cupric sulfate (P < 0.05). HDTMAB could significantly enhance the inhibition by cupric glutamate (P < 0.05). Ultrastructural changes of Alexandrium sp. LC3 under cupric sulfate, cupric glutamate, and cupric glutamate-HDTMAB combined treatment were studied with TEM. Under these stresses, the integrity of the cell plasma membranes (cell plasma membrane, chloroplast and mitochondria membranes) was destroyed. The degree of damage under cupric glutamate-HDTMAB combined treatment was more severe than under the other stresses. These results indicated that mechanistically cupric glutamate inhibits algal growth by destroying the cell membranes, and that HDTMAB promotes this process, which induced mass extravasation of intracellular components and more copper ion entry into the plasma.

Characterization of Cupric Glutamate Extinguishing Mechanism of Alexandrium sp. LC3 with Two-dimensional Electrophoresis and MALDI-TOF MS

Mechanisms by which cupric glutamate, a novel algicide, extinguishes Alexandrium sp. LC3 are shown in this study. We show that cupric glutamate not only stimulated the production of malonaldehyde (MDA) and dramatically promoted cell plasma membrane permeability (p < 0.01) but also remarkably reduced sulfhydryl (SH) group content (p < 0.01). Analysis of protein expression profiles by two-dimensional electrophoresis (2-DE) indicated that only 47 protein spots were detected in both control and cupric glutamate treated cells. Three reliable spots were identified by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) as ribulose-bisphosphate carboxylase large subunit precursor, RNA polymerase beta chain, and hypothetical protein, which can be well correlated with cupric glutamate stress. Based on above results, we hypothesize that the extinguishing mechanisms include (1) the cell membrane being damaged by cupric glutamate; (2) cupric glutamate probably induced denaturation and disintegration of intracellular protein, which led to inhibition of cell growth.

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

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.

Fourteen FITC-conjugated lectins as a tool for the recognition and differentiation of some harmful algae in Chinese coastal waters

In order to test the use of lectins as a tool for the differentiation of harmful algal species, 13 species and 23 strains of algae were tested with 14 fluorescein isothiocyanate (FITC)-conjugated lectins, and the results examined using flow cytometry (FCM), epifluorescence microscopy (EFM) and spectrofluorometry (SFM). The lectin probes SBA, WGA, GSL I, DBA and PHA-E could distinguish between morphologically similar Gymnodinium-like species, such as Karenia mikimotoi (GMDH01), Takayama pulchellum (TPXM01) and Gymnodinium sp. (GspXM01), by their different binding activities. With the precise quantitative measurements of binding obtained using SFM and FCM, lectins appeared to be useful in distinguishing different strains of the same species. The results also showed that PHA-E could differentiate Alexandrium tamarense (ATDH04) from other strains of this species, and SJA could distinguish A. tamarense (ATMJ02) from other strains of this species (including ATMJ01). Similarly, PNA could identify A. tamarense (ATDH01, 02, 03); UEA I could recognize A. tamarense (ATCI01-JN, ATCI01); and RCA120 could differentiate Alexandrium sp (AspGX01) from strain AspGX02, which was shown to produce different levels of paralytic shellfish poisoning toxin. Lectin probes could also bind these target cells in mixed algal samples. Positive cells identified by FCM were clearer than negative cells thus, in EFM, both GspXM01 and TPXM01 labeled with a WGA lectin probe could be distinguished from target cells of K. mikimotoi, Prorocentrum donghaiense and P. minimum (PMDH01, PMXM01) in mixed algal samples. FCM, EFM and SFM analysis could clearly distinguish lectin-probe-bound cells from negative cells in culture.

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

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

Promotion of hexadecyltrimethyleamine bromide to the damage of Alexandrium sp. LC3 by cupric glutamate

The effect of hexadecyltrimethyleamine bromide (HDTMAB) on the removal of Alexandrium sp. LC3 under cupric glutamate stress was investigated. Toxic effect of cupric glutamate on Alexandrium sp. LC3 was significantly promoted in the presence of HDTMAB, especially at 3.0 cmc of HDTMAB. It was found that the sulfhydryl group content of the cell decreased, while the malonaldehyde content and membrane permeability increased when Alexandrium sp. LC3 was treated with HDTMAB and cupric glutamate complex, compared with cupric glutamate alone. The data suggest that HDTMAB might stimulate the damage of Alexaundrium sp. LC3 by enhancing the membrane permeability.