Influence of multi-stressor combinations of pCO2, temperature, and salinity on the toxicity of Heterosigma akashiwo (Raphidophyceae), a fish-killing flagellate.

Climate change and global warming have led to more frequent harmful algal blooms in the last decade. Among these blooms, Heterosigma akashiwo, a golden-brown phytoflagellate, is one of the 40 species with a high potential to form harmful blooms, leading to significant fish mortality. Climate change leads to rising atmospheric and ocean temperatures. These changes, along with altered rainfall patterns and meltwater input, can cause fluctuations in ocean salinity. Elevated atmospheric carbon dioxide (CO2) levels increase water acidity as oceans absorb CO2. This study investigated the effects of temperature, salinity, and CO2 levels on lipid production, hemolytic activity, and toxicity of H. akashiwo using the design of experiment approach, which can be used to investigate the effect of two or more factors on the same response simultaneously in a precise manner with fewer experiments and materials but in a larger region of the factor space. The lipid content was measured using a high-throughput Nile Red method, and the highest level of lipid content was detected at 25°C, a salinity of 30, and a CO2 concentration of 400 ppm. Hemolytic activity was assessed using rabbit blood erythrocytes in a 96-well plate, and the optimal conditions for achieving the highest hemolytic activity were determined at 15°C, a salinity of 10, and a CO2 concentration of 400 ppm. As the chemical structure of the toxin is not known, we used the toxicity against the cell line RTgill-W1 as the cell toxicity proxy. The maximum toxicity was identified at 15°C, a salinity of 10, and a CO2 level of 700 ppm.

Investigating the molecular mechanisms of Pseudalteromonas sp. LD-B1's algicidal effects on the harmful alga Heterosigma akashiwo.

Heterosigma akashiwo is a harmful algal bloom species that causes significant detrimental effects on marine ecosystems worldwide. The algicidal bacterium Pseudalteromonas sp. LD-B1 has demonstrated potential effectiveness in mitigating these blooms. However, the molecular mechanisms underlying LD-B1's inhibitory effects on H. akashiwo remain poorly understood. In this study, we employed the comprehensive methodology, including morphological observation, assessment of photosynthetic efficiency (Fv/Fm), and transcriptomic analysis, to investigate the response of H. akashiwo to LD-B1. Exposure to LD-B1 resulted in a rapid decline of H. akashiwo's Fv/Fm ratio, with cells transitioning to a rounded shape within 2 hours, subsequently undergoing structural collapse and cytoplasmic leakage. Transcriptomic data revealed sustained downregulation of photosynthetic genes, indicating impaired functionality of the photosynthetic system. Additionally, genes related to the respiratory electron transfer chain and antioxidant defenses were consistently downregulated, suggesting prolonged oxidative stress beyond the cellular antioxidative capacity. Notably, upregulation of autophagy-related genes was observed, indicating autophagic responses in the algal cells. This study elucidates the molecular basis of LD-B1's algicidal effects on H. akashiwo, advancing our understanding of algicidal mechanisms and contributing to the development of effective strategies for controlling harmful algal blooms.

Microplastic size-dependent biochemical and molecular effects in alga Heterosigma akashiwo.

Micro- and nano-plastics (MNPs) are increasingly prevalent contaminants in marine ecosystems and have a variety of negative impacts on marine organisms. While their toxic impact on freshwater microalgae has been well-documented, limited research has been conducted on the influence of MNPs on marine red tide algae, despite their significant implications for human health and coastal ecological stability. This study investigated the physiological, biochemical and molecular reactions of the common harmful algal species, Heterosigma akashiwo, when exposed to polystyrene (PS) MNPs of 80 nm and 1 µm in size with the concentrations of 0, 1, 10, and 20 mg L-1 in 12 days. The results showed that 80 nm-sized MNPs (at concentrations of 10 mg L-1 and 20 mg L-1) inhibited algal growth. Despite the increased superoxide dismutase (SOD) activity and up-regulation of glutathione metabolism, exposure-induced oxidative stress remained the main cause of the inhibition. Up-regulation of aminoacyl-tRNA biosynthesis and amino acid biosynthesis pathways provide the necessary amino acid feedstock for the synthesis of antioxidant enzymes such as SOD. 1 µm sized PS MNPs increased chlorophyll a (Chl-a) content without significant effects on other parameters. In addition, H. akashiwo have an effective self-regulation ability to defend against two sized MNPs stress at concentrations of 1 mg L-1 by upregulating gene expression related to endocytosis, biotin metabolism, and oxidative phosphorylation. These results provided evidence that H. akashiwo was able to resist exposure to 1 µm MPs, whereas 80 nm NPs exerted a toxic effect on H. akashiwo. This study deepens our understanding of the interaction between MNPs and marine harmful algal at the transcriptional level, providing valuable insights for further evaluating the potential impact of PS MNPs on harmful algal blooms in marine ecosystems.

Insights into Cellular Localization and Environmental Influences on the Toxicity of Marine Fish-Killing Flagellate, Heterosigma akashiwo.

Heterosigma akashiwo is a unicellular microalga which can cause massive mortality in both wild and cultivated fish worldwide, resulting in substantial economic losses. Environmental parameters such as salinity, light, and temperature showed a significant effect on bloom initiation and the toxicity of H. akashiwo. While in previous studies a one-factor-at-a-time (OFAT) approach was utilized, which only changes one variable at a time while keeping others constant, in the current study a more precise and effective design of experiment (DOE) approach, was used to investigate the simultaneous effect of three factors and their interactions. The study employed a central composite design (CCD) to investigate the effect of salinity, light intensity, and temperature on the toxicity, lipid, and protein production of H. akashiwo. A yeast cell assay was developed to assess toxicity, which offers rapid and convenient cytotoxicity measurements using a lower volume of samples compared to conventional methods using the whole organism. The obtained results showed that the optimum condition for toxicity of H. akashiwo was 25 °C, a salinity of 17.5, and a light intensity of 250 µmol photons m-2 s-1. The highest amount of lipid and protein was found at 25 °C, a salinity of 30, and a light intensity of 250 µmol photons m-2 s-1. Consequently, the combination of warm water mixing with lower salinity river input has the potential to enhance H. akashiwo toxicity, which aligns with environmental reports that establish a correlation between warm summers and extensive runoff conditions that indicate the greatest concern for aquaculture facilities.

Effect of water accommodated fractions of fuel oil on fixed carbon and nitrogen by microalgae: Implication by stable isotope analysis.

Effect of water accommodated fractions (WAF) of #180 fuel oil on fixed carbon and nitrogen in microalgae was studied by stable isotopes. Platymonas helgolandica, Heterosigma akashiwo and Nitzschia closterium were exposed to five WAF concentrations for 96 h. The δ13C value of microalgae was significantly lower than that of the control group, indicated that carbon was limited in the WAF concentrations. The δ13C value of microalgae appeared peak valley at 48 h in control group, corresponding to the enhanced capacity in carbon fixation during microalgae photosynthesis. The physiological acclimation capacity of microalgae was revealed by the occurrence time when the δ13C value was in peak valley, and thus the physiological acclimation capacity of microalgae decreased in the order of Nitzschia closterium > Heterosigma akashiwo > Platymonas helgolandica. Principal component analysis (PCA) were applied to the δ13C value in order to verify the "hormesis" phenomenon in microalgae. The δ13C value could discriminate between stimulatory effects at low doses and inhibitory effects at high doses. In addition, the present study also investigated the effect of the nitrogen on microalgae growth. Because microalgae could still absorb the NO3-N and release of NO2-N and NH4-N in present study, the nitrogen cycle in microalgae was in the equilibrium status. The δ15N value in microalgae exhibited no obvious change with the increasing of WAF concentrations at the same time. However, due to the enrichment of nitrogen, the δ15N value first increased gradually with the time and finally was stable. Overall, the fractionation of carbon and nitrogen stable isotopes illustrated that the effect of carbon on the growth of microalgae was more prominent than nitrogen. Stable isotopes was used to investigate the influence of WAF on fixed carbon and nitrogen in microalgae growth, providing a fundamental theoretical guidance for risk assessment of marine ecological environment.

Phylogeographic characteristics of hypervariable regions in the mitochondrial genome of a cosmopolitan, bloom-forming raphidophyte, Heterosigma akashiwo.

The global distribution of phytoplankton is defined by many events, including long-term evolutionary processes and shorter time span processes (e.g., global climate change). Furthermore, human-assisted, unintentional dispersion, including the transport of live fish and spat for aquaculture, and transfer of aquatic microorganisms contained in ship ballast water, may aid the spread of phytoplankton. To understand the phylogeographic history of a species, the development of useful molecular markers is crucial. We previously reported a hypervariable mitochondrial gene in the cosmopolitan bloom-forming alga, Heterosigma akashiwo. In this study, we identified two additional hypervariable segments in the H. akashiwo mitochondrial genome, one a protein coding sequence, and the other an intergenic region, by comparing the whole mitogenomes of strains obtained from various geographic origins. Interestingly, the newly identified hypervariable protein coding sequence was a paralogue of the previously identified gene, and both sequences showed tendencies to correlate with latitude of geographic origin. However, the hypervariable intergenic sequence did not show a clear correlation with origin. Our results demonstrated that the protein coding sequences may serve as useful tools for understanding the phylogeographic history of H. akashiwo, and they may crucially function in adaptation of the species to the environment.

Effects of carbon and silicon nanotubes and carbon nanofibers on marine microalgae Heterosigma akashiwo.

The effect of carbon and silicon nanotubes (CNTs and SiNTs) and carbon nanofibers (CNFs) to microscopic marine algae Heterosigma akashiwo was studied, using algal growth inhibition for 3 days (acute effect) and 7 days (chronic effect) as toxicity endpoints. The criterion of the toxic effect was the statistically significant reduction of the number of algal cells in the exposed samples compared to the control. Samples did not demonstrate toxic effects at doses 1 mg/l and 10 mg/l. CNTs and SiNTs samples at 100 mg/l exhibited both acute and chronic toxic effects. We assume that the main cause of cell death in these samples was related to the mechanical damage of cell integrity. CNFs at concentrations of 100 mg/l did not inhibit algal growth, but cells with irregular shapes were observed, which were not observed after exposure to CNTs and SiNTs. Nickel impurities present in CNFs samples are presumably the main cause of observed cell deformations.

A hypervariable mitochondrial protein coding sequence associated with geographical origin in a cosmopolitan bloom-forming alga, Heterosigma akashiwo.

Geographical distributions of phytoplankton species can be defined by events on both evolutionary time and shorter scales, e.g. recent climate changes. Additionally, modern industrial activity, including the transport of live fish and spat for aquaculture and aquatic microorganisms in ship ballast water, may aid the spread of phytoplankton. Obtaining a reliable marker is key to gaining insight into the phylogeographic history of a species. Here, we report a hypervariable mitochondrial gene in the cosmopolitan bloom-forming alga, Heterosigma akashiwo We compared the entire mitochondrial genome sequences of seven H. akashiwo strains from Japanese and North American coastal waters and identified a hypervariable segment. The region codes for a hypothetical protein with no defined function, and its variations between Japanese and North American isolates were prominent, while the sequences were more conserved among Japanese strains and North American isolates. Comparison of the sequence in isolates obtained from different geographical points in the Northern Hemisphere revealed that the sequence variations largely correlated with latitude and longitude (i.e. Pacific/Atlantic oceans). Our results demonstrate the usefulness of the sequence in determining the phylogeographic history of H. akashiwo.

The effects of salinity on the cellular permeability and cytotoxicity of Heterosigma akashiwo.

A laboratory study using the fish-killing raphidophyte Heterosigma akashiwo was conducted to examine its capability to grow at salinities below oceanic, and to test the perceived relationship between reduced salinities and increased cytotoxicity. A nonaxenic strain of H. akashiwo isolated from the U.S. Pacific Northwest was exposed to a combination of three salinity (32, 20, and 10) and five temperature (14.7°C, 18.4°C, 21.4°C, 24.4°C and 27.8°C) conditions. Our results demonstrate that cell permeability and cytotoxicity are strongly correlated in unialgal cultures of H. akashiwo, which both increased as salinity decreased from 32 to 10. Furthermore, over a broad median range of salinities (10 and 20), neither temperature nor specific growth rate was correlated with cytotoxicity. However, in cultures grown at the salinity of 32, both temperature and specific growth rate were inversely proportional to toxicity; this relationship was likely due to the effect of contamination by an unidentified species of Skeletonema in those cultures. The presence of Skeletonema sp. resulted in a cytotoxic response from H. akashiwo that was greater than the response caused by salinity alone. These laboratory results reveal the capability of H. akashiwo to become more toxic not only at reduced salinities but also in competition with another algal species. Changes in cell permeability in response to salinity may be an acclimation mechanism by which H. akashiwo is able to respond rapidly to different salinities. Furthermore, due to its strong positive correlation with cytotoxicity, cellular permeability is potentially associated with the ichthyotoxic pathway of this raphytophyte.

Evaluation of rapid cell division in non-uniform cell cycles.

To better understand the mechanisms of development of harmful algal blooms (HABs), accurate estimates of species-specific in situ growth rates are needed. HABs are caused by rapid cell division by the causative microorganisms. To accurately estimate the in situ growth rates of harmful algae having non-uniform and/or irregular cell cycles, we modified a standard equation based on the cell cycle, and calculated the in situ growth rate to describe the process of bloom development in nature. Sampling of a developing bloom of Heterosigma akashiwo in Pohang Bay, Korea, was conducted every 3 h from 15:00 on August 2 to 07:00 on August 4, 2006. The amount of H. akashiwo DNA was measured using flow cytometry following tyramide signal amplification-fluorescence in situ hybridization. On August 2, the percentage of G1 phase cells decreased from 15:00 to 19:00 then increased until 22:00; it then decreased until 07:00 on August 3, followed by an increase to 10:00. This indicates the ability of the cells in nature to undergo more than one round of division per day. During the following night two rounds of division did not occur. The in situ growth rates estimated using the modified equation ranged from 0.31 to 0.53 d(-1) . We conclude that the use of this equation enables more accurate estimates of bloom formation by rapidly dividing cells.

The allelopathic effects of Heterosigma akashiwo on Skeletonema costatum: Insights from gene expression and metabolomics analysis.

The globally distributed harmful algal blooms (HAB) species, Heterosigma akashiwo, has been found to exhibit ichthyotoxicity. Previous studies have shown that H. akashiwo achieves a competitive edge during bloom occurrences by inhibiting the growth of a coexisting diatom, Skeletonema costatum, through allelopathy. However, the specific allelopathic mechanisms underlying the allelopathic effects of H. akashiwo on S. costatum remain unknown. To bridge this gap, our study utilized a combination of quantitative real-time PCR and metabolomics to examine the allelopathic processes of H. akashiwo on S. costatum. Our results demonstrate that the growth of S. costatum is hindered when co-cultured with H. akashiwo (initial cell concentration, 2 × 104 cell/mL). Gene expression investigation showed a substantial reduction in the mRNA levels of cytochrome b6, ribulose bisphosphate carboxylase large chain, and silicon transporter in S. costatum when grown in co-culture conditions. Furthermore, metabolic pathway analysis suggested that the allelopathic effects of H. akashiwo disrupted several vital metabolic pathways in S. costatum, including a reduction in purine and pyrimidine metabolism and an increase in fatty acid biosynthesis. Our investigation has revealed the intricate and substantial involvement of allelopathy in the formation of H. akashiwo blooms, demonstrating the complexity of the allelopathic interaction between H. akashiwo and S. costatum. These insights also contribute significantly to our understanding of the dynamics within HAB species.

The novel application of violacein produced by a marine Duganella strain as a promising agent for controlling Heterosigma akashiwo bloom: Algicidal mechanism, fermentation optimization and agent formulation.

Controlling harmful algal blooms with algicidal bacteria is thought to be an efficient and eco-friendly way but lack of comprehensive studies from theory to practice limited the field application. Here we presented a purple bacterial strain Duganella sp. A3 capable of killing several harmful algae, including Heterosigma akashiwo, a world-wide fish-killing microalga. A bioactivity-guided purification and identification approach revealed the major algicidal compound of A3 as the pigment violacein, which was never reported for its algicidal potential before. Violacein rapidly disrupted cell permeability, caused long-term oxidative stress, but mildly affected algal photosystem, which might explain its highly species-specific activity against unarmored H. akashiwo. To explore the application potential of violacein, a fermentation optimization approach combing single-factor and multi-factor experiments was conducted to increase the violacein yield, which finally reached 0.4199 g/L just using a simple medium formula beneficial for compound purification. Finally, taking advantages of the physical and chemical stabilities, we successfully developed the novel application of violacein as a sustained-releasing and easy-to-preserve algicidal agent using alginate-acacia-gum-chitosan encapsulation, which paved the path for its future application in controlling H. akashiwo bloom.

Transcriptome analysis of the harmful alga Heterosigma akashiwo under a 24-hour light-dark cycle.

The photoperiod, which is defined as the period of time within a 24-hour time frame that light is available, is an important environmental regulator of several physiological processes in phytoplankton, including harmful bloom-forming phytoplankton. The ichthyotoxic raphidophyte Heterosigma akashiwo is a globally distributed bloom-forming phytoplankton. Despite extensive studies on the ecological impact of H. akashiwo, the influence of the photoperiod on crucial biological processes of this species remains unclear. In this study, gene expression in H. akashiwo was analyzed over a 24-hour light-dark (14:10) treatment period. Approximately 36 % of unigenes in H. akashiwo were differentially expressed during this 24-hour treatment period, which is indicative of their involvement in the response to light-dark variation. Notably, the number of differentially expressed genes exhibited an initial increase followed by a subsequent decrease as the sampling time progressed (T0 vs. other time points). Unigenes associated with photosynthesis and photoprotection reached their peak expression levels after 2-4 h of illumination (T12-T14). In contrast, the expression of unigenes associated with DNA replication peaked at the starting point of the dark period (T0). Furthermore, although several unigenes annotated to photoreceptors displayed potential diel periodicity, genes from various photoreceptor families (such as phytochrome and cryptochrome) showed unique expression patterns. Collectively, our findings offer novel perspectives on the response of H. akashiwo to the light-dark cycle, serving as a valuable resource for investigating the physiology and ecology of this species.

Allelochemicals of Alexandrium tamarense and its algicidal mechanism for Prorocentrum donghaiense and Heterosigma akashiwo.

The effects of culture filtrate of Alexandrium tamarense on Prorocentrum donghaiense and Heterosigma akashiwo were investigated, including determination of algal density, photosynthesis, intracellular enzyme content and activity. The filtrate of A. tamarense had a stronger inhibitory effect on P. donghaiense than H. akashiwo, and the inhibitory effect decreased with higher temperature treatment of the filtrate. Instantaneous fluorescence (Ft) and maximum quantum yield of photosystem II (Fv/Fm) values of both kinds of target algae were reduced as exposed to the filtrate of A. tamarense, which proved that allelopathy could inhibit the normal operation of photosynthetic system. The increase of Malondialdehyde (MDA) content of the two kinds of target algae indicated that the cell membrane was seriously damaged by allelochemicals released by A. tamarense. The different responses of Superoxide Dismutase (SOD) and Catalase (CAT) activity in two kinds of target algae demonstrated the complexity and diversity of allelopathic mechanism. The filtrate of A. tamarense also influenced the metabolic function (ATPases) of P. donghaiense and H. akashiwo, and the influence on P. donghaiense was greater. Liquid-liquid extraction was used to extract and isolate allelochemicals from the filtrate of A. tamarense. It was found that only component I with molecular weight of 424.2573 and 434.2857 could inhibit the growth of P. donghaiense by HPLC-MS.

Taxonomic difference in marine bloom-forming phytoplanktonic species affects the dynamics of both bloom-responding prokaryotes and prokaryotic viruses.

The production of dissolved organic matter during phytoplankton blooms and consumption by heterotrophic prokaryotes promote marine carbon biogeochemical cycling. Although prokaryotic viruses presumably affect this process, their dynamics during blooms are not fully understood. Here, we investigated the effects of taxonomic difference in bloom-forming phytoplankton on prokaryotes and their viruses. We analyzed the dynamics of coastal prokaryotic communities and viruses under the addition of dissolved intracellular fractions from taxonomically distinct phytoplankton, the diatom Chaetoceros sp. (CIF) and the raphidophycean alga Heterosigma akashiwo (HIF), using microcosm experiments. Ribosomal RNA gene amplicon and viral metagenomic analyses revealed that particular prokaryotes and prokaryotic viruses specifically increased in either CIF or HIF, indicating that taxonomic difference in bloom-forming phytoplankton promotes distinct dynamics of not only the prokaryotic community but also prokaryotic viruses. Furthermore, combining our microcosm experiments with publicly available environmental data mining, we identified both known and novel possible host-virus pairs. In particular, the growth of prokaryotes associating with phytoplanktonic organic matter, such as Bacteroidetes (Polaribacter and NS9 marine group), Vibrio spp., and Rhodobacteriales (Nereida and Planktomarina), was accompanied by an increase in viruses predicted to infect Bacteroidetes, Vibrio, and Rhodobacteriales, respectively. Collectively, our findings suggest that changes in bloom-forming species can be followed by an increase in a specific group of prokaryotes and their viruses and that elucidating these tripartite relationships among specific phytoplankton, prokaryotes, and prokaryotic viruses improves our understanding of coastal biogeochemical cycling in blooms.IMPORTANCEThe primary production during marine phytoplankton bloom and the consumption of the produced organic matter by heterotrophic prokaryotes significantly contribute to coastal biogeochemical cycles. While the activities of those heterotrophic prokaryotes are presumably affected by viral infection, the dynamics of their viruses during blooms are not fully understood. In this study, we experimentally demonstrated that intracellular fractions of taxonomically distinct bloom-forming phytoplankton species, the diatom Chaetoceros sp. and the raphidophycean alga Heterosigma akashiwo, promoted the growth of taxonomically different prokaryotes and prokaryotic viruses. Based on their dynamics and predicted hosts of those viruses, we succeeded in detecting already-known and novel possible host-virus pairs associating with either phytoplankton species. Altogether, we propose that the succession of bloom-forming phytoplankton would change the composition of the abundant prokaryotes, resulting in an increase in their viruses. These changes in viral composition, depending on bloom-forming species, would alter the dynamics and metabolism of prokaryotes, affecting biogeochemical cycling in blooms.

Salinity as an Abiotic Stressor for Eliciting Bioactive Compounds in Marine Microalgae.

This study investigated the impact of culture medium salinity (5-50 PSU) on the growth and maximum photochemical yield of photosystem II (Fv/Fm) and the composition of carotenoids, fatty acids, and bioactive substances in three marine microalgae (Chrysochromulina rotalis, Amphidinium carterae, and Heterosigma akashiwo). The microalgae were photoautotrophically cultured in discontinuous mode in a single stage (S1) and a two-stage culture with salt shock (S2). A growth model was developed to link biomass productivity with salinity for each species. C. rotalis achieved a maximum biomass productivity (Pmax) of 15.85 ± 0.32 mg·L-1·day-1 in S1 and 16.12 ± 0.13 mg·L-1·day-1 in S2. The salt shock in S2 notably enhanced carotenoid production, particularly in C. rotalis and H. akashiwo, where fucoxanthin was the main carotenoid, while peridinin dominated in A. carterae. H. akashiwo also exhibited increased fatty acid productivity in S2. Salinity changes affected the proportions of saturated, monounsaturated, and polyunsaturated fatty acids in all three species. Additionally, hyposaline conditions boosted the production of haemolytic substances in A. carterae and C. rotalis.

The effects of extracellular algicidal compounds of Bacillus sp. B1 on Heterosigma akashiwo: a metabolomics approach.

Heterosigma akashiwo (H. akashiwo), a harmful algal species, has been a global environmental problem. Extracellular algicidal compounds (EACs) extracted from Bacillus sp. B1 exhibited algicidal effects against H. akashiwo. However, little is known about the algicidal mechanism and metabolic process. In this study, metabolomics and physiological analyses were combined to investigate the cellular responses of H. akashiwo when treated with EACs. The results indicated that EACs at 10% (vEACs/vsample) showed more than 90% inhibition of H. akashiwo. EAC treatment resulted in excessive reactive oxygen species (ROS) production in algal cells, causing stress responses such as inhibition of photosynthetic pigment synthesis, reduction of sugar synthesis, imbalance of osmotic pressure in the cell membrane, disruption of cell size and morphology, and eventual cell death. The results reveal the underlying mechanism of the algicidal process and provide new insights into algae-bacteria interactions and the application of metabolomics to algal research.

Full-Length Transcriptome Analysis of the Ichthyotoxic Harmful Alga Heterosigma akashiwo (Raphidophyceae) Using Single-Molecule Real-Time Sequencing.

The raphidophyte Heterosigma akashiwo is a harmful algal species. The bloom of this organism has been associated with the massive mortality of fish in many coastal waters. To investigate the molecular mechanism of H. akashiwo blooms, having a reliable reference transcriptome of this species is essential. Therefore, in this study, a full-length transcriptome of H. akashiwo was obtained by single-molecule real-time sequencing. In total, 45.44 Gb subread bases were generated, and 16,668 unigenes were obtained after the sequencing data processing. A total of 8666 (52.00%) unigenes were successfully annotated using seven public databases. Among them, mostly phosphorus and nitrogen metabolism genes were detected. Moreover, there were 300 putative transcription factors, 4392 putative long non-coding RNAs, and 7851 simple sequence repeats predicted. This study provides a valuable reference transcriptome for understanding how H. akashiwo blooms at a molecular level.

Inactivation of Heterosigma akashiwo under UV/peroxydisulfate advanced disinfection system in marine waters.

Heterosigma akashiwo (H. akashiwo) is recognized as a harmful algal bloom (HABs) species with a global distribution, capable of posing significant threats to marine ecosystems, particularly when spread through ship ballast water. This investigation focused on elucidating the inactivation kinetics and underlying mechanism of H. akashiwo through a combined ultraviolet irradiation and peroxydisulfate (UV/PDS) process. The results demonstrated a strong synergistic effect within the UV/PDS system, resulting in an inactivation of 0.78-ln and 2.67-ln within 40 min of UV and UV/PDS processes. The principal agents accountable for inactivation were identified as sulfate radicals (•SO4-) and hydroxyl radical (•OH), which exhibited a synergistic effect in the UV/PDS process. Furthermore, the study observed a negatively impact of seawater pH and salinity on the efficiency of inactivation. UV/PDS caused oxidative stress on algal cells, initially involving the participation of antioxidant enzymes in counteracting cellular damage, but this protective mechanism diminished as the reaction duration extended. The UV/PDS treatment not only inflicted damage upon H. akashiwo's photosynthetic system but also caused the extracellular release of DNA and algal organic matter (AOM) due to damaged cell membranes. Transcriptome analysis provided a molecular biology perspective on the cellular inactivation process. Upregulation of genes linked to photosynthesis and oxidative phosphorylation suggested a potential elevation in energy metabolism. In contrast, genes associated with cellular and metabolic processes, including glycolysis and the tricarboxylic acid cycle (TCA cycle), exhibited downregulation. Moreover, this treatment exerted an inhibitory influence on RNA polymerase and protein synthesis, resulting in the reduced expression of genetic information.

Body condition of larval roundherring, Gilchristella aestuaria (family Clupeidae), in relation to harmful algal blooms in a warm-temperate estuary.

Eutrophication-driven harmful algal blooms (HABs) can have secondary effects on larval fishes that rely on estuaries as nurseries. However, few studies worldwide have quantified these effects despite the global rise in eutrophication. This study presents a novel approach using biochemical body condition analyses to evaluate the impact of HABs on the growth and body condition of the larvae of an estuarine resident fish. Recurrent phytoplankton blooms of Heterosigma akashiwo occur in the warm-temperate Sundays Estuary on the southeast coast of South Africa. The response in body condition and assemblage structure on larval estuarine roundherring (Gilchristella aestuaria) was measured in conjunction with bloom conditions, water quality and zooplanktonic prey and predators. Larvae and early juveniles were sampled during varying intensity levels, duration and frequency of hypereutrophic blooms. This study demonstrated that extensive HABs could significantly impact larval roundherring, G. aestuaria, by decreasing larval nutritional condition and limiting their growth, resulting in poor grow-out into the juvenile phase. Poor condition and growth may likely affect recruitment success to adult populations, and since G. aestuaria is an important forage fish and zooplanktivore, poor recruitment will hold consequences for estuarine food webs.