Search Results (10)

The dinoflagellate genus Karlodinium J. Larsen is well known to form harmful algal blooms (HABs), some of which can produce karlotoxins or other ichthyotoxins and thus cause fish-killing events. Among the 16 currently accepted species of Karlodinium (about half of which are reported to be toxic), six species (K. australe, K. decipiens, K. digitatum, K. elegans, K. veneficum, and K. zhouanum) have been reported or described in the coastal waters of China. However, a fine morphological and molecular characterization of the seldom-observed species K. decipiens has not been conducted; moreover, the negative effects of this species on aquatic animals have not been investigated. This work reports the morphological and phylogenetic characterization of a strain of K. decipiens isolated from Jiaozhou Bay, China, in 2019. The characterization of the strain was conducted using light and scanning electron microscopy, LSU, SSU rDNA, and ITS sequences-based systematic analyses, pigment analysis, and a detailed investigation of its potential toxic/harmful activity on aquatic animals. We observed the typical diagnostic features of K. decipiens, including its relatively large size, ellipsoidal or ovoid cell shape, ventral pore, ventral ridge connecting the two displaced ends of the cingulum, cingulum with a displacement of about one-third of the cell length, numerous polyhedral or slightly elongated chloroplasts distributed peripherally, and large nucleus located centrally. However, we also observed a large amphiesmal vesicle at the dorsal end of the ASC at the dorsal epicone, which is a novel feature that has never been reported from any species of the genus. Based on the results of this study, it is not clear whether this feature is a specific structure of the species or a common characteristic of the genus; therefore, this novel feature is worthy of further examination. Fucoxanthin was the most abundant pigment among all the carotenoids detected. The phylogenies inferred using Bayesian inference (BI) and maximum likelihood (ML) techniques confirmed the conspecificity of our isolate with the holotype K. decipiens (accession no. EF469236). In molecular trees, K. decipiens and K. antarcticum form a separate clade from other species of Karlodinium, and it should be examined whether a large amphiesma vesicle may be a characteristic of this clade. The exposure bioassays using brine shrimp (Artemia salina) indicated that K. decipiens exhibited toxicity to zooplankton, with 100% and 68% mortality observed in brine shrimp using live cell cultures and cell culture lysates over 120 h, respectively. Our work provides a detailed morphological and molecular characterization of K. decipiens from China. The results of this study broaden the known geographical distribution of this species and demonstrate it to be a harmful dinoflagellate. Full article

Ammonium and polyamines are essential nitrogen metabolites in all living organisms. Crosstalk between ammonium and polyamines through their metabolic pathways has been demonstrated in plants and animals, while no research has been directed to explore this relationship in algae or to investigate the underlying molecular mechanisms. Previous research demonstrated that high concentrations of ammonium and putrescine were among the active substances in bacteria-derived algicide targeting dinoflagellates, suggesting that the biochemical inter-connection and/or interaction of these nitrogen compounds play an essential role in controlling these ecologically important algal species. In this research, putrescine, ammonium, or a combination of putrescine and ammonium was added to cultures of three dinoflagellate species to explore their effects. The results demonstrated the dose-dependent and species-specific synergistic effects of putrescine and ammonium on these species. To further explore the molecular mechanisms behind the synergistic effects, transcriptome analysis was conducted on dinoflagellate Karlodinium veneficum treated with putrescine or ammonium vs. a combination of putrescine and ammonium. The results suggested that the synergistic effects of putrescine and ammonium disrupted polyamine homeostasis and reduced ammonium tolerance, which may have contributed to the cell death of K. veneficum. There was also transcriptomic evidence of damage to chloroplasts and impaired photosynthesis of K. veneficum. This research illustrates the molecular mechanisms underlying the synergistic effects of the major nitrogen metabolites, ammonium and putrescine, in dinoflagellates and provides direction for future studies on polyamine biology in algal species. Full article

In recent years, the role of lectins in the feed selection of bivalve has become hot research topic. Manila clam Ruditapes philippinarum is a species of marine bivalve with important economic value. A new C-type lectin (Rpcl) from the clam was obtained and its potential role in feeding and feed selection was studied. Rpcl cDNA was 929 bp in length and had 720 bp of open reading frame. Rpcl encoded 235 amino acids, comprising a carbohydrate recognition domain (CRD) as well as an N-terminal signal peptide. Rpcl contained a conserved CRD disulfide bond including five cysteine residues (Cys¹²⁵, Cys¹⁴², Cys²¹³, Cys²¹⁹, and Cys²³⁷) and the QPN motif (GLN²⁰⁴-PRO²⁰⁵-ASN²⁰⁶). Phylogenetic analysis revealed that the amino acid sequence of Rpcl was closely related to that of Vpclec-1 from R. philippinarum and C-type lectin from Mercenaria. The qPCR analysis indicated that Rpcl expression was observed in all examined tissues and was the highest in gills followed by in the hepatopancreas, and to a lesser extent in the mantle and lip. The in vitro agglutination experiments showed that, the purified Rpcl protein could selectively agglutinate with different microalgae. The strongest agglutinating effect with Chlorella sp. was observed, followed by Karlodinium veneficum and Chaetoceros debilis Cleve. No agglutination with Prorocentrum minimum was observed. In the feeding experiment, compared with that of the starvation group, Rpcl expression in the lip and gill of the clam fed with C. debilis and K. veneficum showed a significant upward trend with the change of time. In addition, it was found that the changes in the expression of the Rpcl gene in the gill and lip, the main feeding tissues, were consistent with the slope of the decrease in the number of algal cells in the water body. In summary, the structure of a new C-type lectin (Rpcl) was reported in this study and its correlation with the feeding and feed selection in R. philippinarum was confirmed. Full article

In numerous studies, researchers have explored the interactions between fungi and their hosting biota in terrestrial systems, while much less attention has been paid to the counterpart interactions in aquatic, and particularly marine, ecosystems. Despite the growing recognition of the potential functions of fungi in structuring phytoplankton communities, the current insights were mostly derived from phytoplankton hosts, such as diatoms, green microalgae, and cyanobacteria. Dinoflagellates are the second most abundant group of phytoplankton in coastal marine ecosystems, and they are notorious for causing harmful algal blooms (HABs). In this study, we used high-throughput amplicon sequencing to capture global snapshots of specific fungal assemblages associated with laboratory-cultured marine dinoflagellate. We investigated a total of 13 clonal cultures of the dinoflagellate Karlodinium veneficum that were previously isolated from 5 geographic origins and have been maintained in our laboratory from several months to more than 14 years. The total recovered fungal microbiome, which consisted of 349 ASVs (amplicon sequencing variants, sequences clustered at a 100% sequence identity), could be assigned to 4 phyla, 18 classes, 37 orders, 65 families, 97 genera, and 131 species. The fungal consortium displayed high diversity and was dominated by filamentous fungi and ascomycetous and basidiomycetous yeasts. A core set of three genera among all the detected fungi was constitutively present in the K. veneficum strains isolated from geographically distant regions, with the top two most abundant genera, Thyridium and Pseudeurotium, capable of using hydrocarbons as the sole or major source of carbon and energy. In addition, fungal taxa previously documented as endophytes in other hosts were also found in all tested strains of K. veneficum. Because host–endophyte interactions are highly variable and strongly case-dependent, these fungal taxa were not necessarily genuine endosymbionts of K. veneficum; instead, it raised the possibility that dinoflagellates could potentially serve as an alternative ecological niche for the colonization of fungal endophytes. Our findings lay the foundation for further investigations into the potential roles or functions of fungi in the regulation of the growth dynamics and HABs of marine dinoflagellates in the field. Full article

Numerous potentially toxic plankton species commonly occur in the Black Sea, and phycotoxins have been reported. However, the taxonomy, phycotoxin profiles, and distribution of harmful microalgae in the basin are still understudied. An integrated microscopic (light microscopy) and molecular (18S rRNA gene metabarcoding and qPCR) approach complemented with toxin analysis was applied at 41 stations in the northwestern part of the Black Sea for better taxonomic coverage and toxin profiling in natural populations. The combined dataset included 20 potentially toxic species, some of which (Dinophysis acuminata, Dinophysis acuta, Gonyaulax spinifera, and Karlodinium veneficum) were detected in over 95% of the stations. In parallel, pectenotoxins (PTX-2 as a major toxin) were registered in all samples, and yessotoxins were present at most of the sampling points. PTX-1 and PTX-13, as well as some YTX variants, were recorded for the first time in the basin. A positive correlation was found between the cell abundance of Dinophysis acuta and pectenotoxins, and between Lingulodinium polyedra and Protoceratium reticulatum and yessotoxins. Toxic microalgae and toxin variant abundance and spatial distribution was associated with environmental parameters. Despite the low levels of the identified phycotoxins and their low oral toxicity, chronic toxic exposure could represent an ecosystem and human health hazard. Full article

A monthly survey of the feeding selectivity of Ruditapes philippinarum in the Yalu River Estuary in 2020–2021 was conducted using high-throughput sequencing identification and visual grading technology. The results showed that the most-dominant species in the water of the shellfish culture area and in the stomachs of R. philippinarum was Karlodinium veneficum in those years. The selectivity index (E) indicated that R. philippinarum avoided consuming Bacillariophyta, Chrysophyta and Cryptophyta throughout the year and preferentially consumed Dinophyta and Chlorophyta. In 2020, the annual average biomass of Dinophyta, Bacillariophyta, Chlorophyta, Dictyochophyta, Cryptophyta and Chrysophyta in the stomach contents of R. philippinarum was 54:14:16:1:10:4; it was 41:12:28:0:1:17 in 2021. The annual average biomass ratio of picophytoplankton, nanophytoplankton and microphytoplankton in the stomachs of R. philippinarum was 13:48:39 in 2020; it was 14:66:20 in 2021. R. philippinarum actively fed on nanophytoplankton and avoided picophytoplankton. Among the phytoplankton of different sizes and groups that R. philippinarum prefer to feed, chemical oxygen demand (COD) and organic phosphorus (DOP) have a significant negative effect on the nanophytoplankton community, pH has a positive effect on the Dictyochophyta community and COD and the inorganic nitrogen to phosphorus ratio (DI-N/P) have a significant positive effect on the Chlorophyta community. Full article

Shewanella sp. IRI-160 is an algicidal bacterium that secretes an algicide, IRI-160AA. This algicide specifically targets dinoflagellates, while having no adverse effects on other algal species tested. Dinoflagellates exposed to IRI-160AA exhibited increased production of reactive oxygen species (ROS), DNA damage, and cell cycle arrest, implying a programmed pathway leading to cell death (PCD). Here, a metabolomic analysis was conducted on dinoflagellate Karlodinium veneficum and a control cryptophyte species Rhodomonas exposed to IRI-160AA to investigate the cellular mechanisms behind the physiological effects and the specificity of this algicide. Results of this research supported previous observations about physiological responses to the algicide. A suite of metabolites was identified that increased in the cell pellets of K. veneficum but not in Rhodomonas, including oxidative stress biomarkers, antioxidants, and compounds involved in DNA damage and PCD. Overall, the results of this study illustrated the metabolomic mechanisms underlying the algicidal effects of IRI-160AA on dinoflagellates. This research also provided insights and future directions for studies on the cellular response of dinoflagellates exposed to antagonistic bacteria in the environment. Full article

Biosurfactant has potential application value in the removal of microalgal blooms, but the ecological risks require more research. In this paper, the effects of surfactin on the toxic dinoflagellate Karlodinium veneficum were studied. The coaction of surfactin and K. veneficum was also evaluated through toxicological experiments on Artemia and juvenile clams. The results showed that: (1) in the concentration range of 0–10 mg/L, surfactin significantly killed algal cells in a dose-dependent manner within 48 h; the 24 h EC₅₀ was 3.065 mg/L; (2) K. veneficum had the ability to restore population growth after stress reduction and the restored proliferation was positively correlated with the initial surfactin concentration; (3) the ability to restore population growth was associated with protection afforded by the promotion of antioxidant enzymes, including catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD), whose increase was positively correlated with the surfactin concentration; (4) the toxicity of the coculture of surfactin and K. veneficum was significantly greater than that of the K. veneficum culture or surfactin alone and was dose and time dependent. The potential ecological risks should be considered when applying biosurfactants, such as surfactin, in the removal of harmful algal blooms. Full article

As the number of mixotrophic protists has been increasingly documented, “mixoplankton”, a third category separated from the traditional categorization of plankton into “phytoplankton” and “zooplankton”, has become a new paradigm and research hotspot in aquatic plankton ecology. While species of dinoflagellates are a dominant group among all recorded members of mixoplankton, the trophic modes of Karlodinium, a genus constituted of cosmopolitan toxic species, were reviewed due to their representative features as mixoplankton and harmful algal blooms (HABs)-causing dinoflagellates. Among at least 15 reported species in the genus, three have been intensively studied for their trophic modes, and all found to be phagotrophic. Their phagotrophy exhibits multiple characteristics: (1) omnivority, i.e., they can ingest a variety of preys in many forms; (2) flexibility in phagotrophic mechanisms, i.e., they can ingest small preys by direct engulfment and much bigger preys by myzocytosis using a peduncle; (3) cannibalism, i.e., species including at least K. veneficum can ingest the dead cells of their own species. However, for some recently described and barely studied species, their tropical modes still need to be investigated further regarding all of the above-mentioned aspects. Mixotrophy of Karlodinium plays a significant role in the population dynamics and the formation of HABs in many ways, which thus deserves further investigation in the aspects of physiological ecology, environmental triggers (e.g., levels of inorganic nutrients and/or presence of preys), energetics, molecular (genes and gene expression regulations) and biochemical (e.g., relevant enzymes and signal molecules) bases, origins, and evaluation of the advantages of being a phagotroph. Full article

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the detection and quantitation of karlotoxins in the selected reaction monitoring (SRM) mode. This novel method was based upon the analysis of purified karlotoxins (KcTx-1, KmTx-2, 44-oxo-KmTx-2, KmTx-5), one amphidinol (AM-18), and unpurified extracts of bulk cultures of the marine dinoflagellate Karlodinium veneficum strain CCMP2936 from Delaware (Eastern USA), which produces KmTx-1 and KmTx-3. The limit of detection of the SRM method for KmTx-2 was determined as 2.5 ng on-column. Collision induced dissociation (CID) spectra of all putative karlotoxins were recorded to present fragmentation patterns of each compound for their unambiguous identification. Bulk cultures of K. veneficum strain K10 isolated from an embayment of the Ebro Delta, NW Mediterranean, yielded five previously unreported putative karlotoxins with molecular masses 1280, 1298, 1332, 1356, and 1400 Da, and similar fragments to KmTx-5. Analysis of several isolates of K. veneficum from the Ebro Delta revealed small-scale diversity in the karlotoxin spectrum in that one isolate from Fangar Bay produced KmTx-5, whereas the five putative novel karlotoxins were found among several isolates from nearby, but hydrographically distinct Alfacs Bay. Application of this LC-MS/MS method represents an incremental advance in the determination of putative karlotoxins, particularly in the absence of a complete spectrum of purified analytical standards of known specific potency. Full article