Harmful Algal Species Research Articles

Enhanced eDNA monitoring for detection of viable harmful algal bloom species using propidium monoazide

This study investigated the use of propidium monoazide (PMA) to improve the accuracy of environmental DNA (eDNA) monitoring by selectively detecting intracellular DNA (iDNA) from living cells, while excluding extracellular DNA (exDNA) from dead organisms. eDNA samples were collected from various depths off the coast of Tongyeong, South Korea, and analyzed alongside environmental factors, such as temperature, dissolved oxygen, turbidity, and nutrient levels. The results showed that PMA-treated iDNA provided a more accurate estimate of viable harmful algal bloom species (HABs) than total eDNA and DNase-treated iDNA. Strong correlations were found between iDNA (PMA) and environmental factors, particularly nutrient levels and turbidity, suggesting its effectiveness in biological environments. The iDNA (PMA) concentrations were higher in the surface and bottom layers, indicating that these layers were more indicative of living organisms in marine environments. The application of PMA in eDNA monitoring reduces false positives and enhances the detection accuracy of viable HAB species, representing a promising tool for real-time monitoring and management of marine ecosystems.

Differential allelopathic effects of mangrove plants Kandelia obovata and Aegiceras corniculatum on harmful algal species: Potential applications in algal bloom control

This study examined effects of mangrove plants Kandelia obovata and Aegiceras corniculatum on harmful algal species. While A. corniculatum leaf extract had no inhibitory effect, K. obovata leaf extract significantly inhibited the growth of two harmful algal species Alexandrium tamarense and Karenia mikimotoi. The inhibitory effect was concentration-dependent, with over 90 % inhibition at the highest concentration. Morphological changes and cell size reduction were observed in both microalgae. Excessive production of reactive oxygen species and damage to algal photosynthetic system were found. The allelopathic effect of K. obovata on K. mikimotoi with low-concentration repeated exposure was more effective than high-concentration single exposure. The EC50 of K. obovata (0.33 g L−1) was lower than reported values on other coastal plants. Higher inhibitory effects of K. obovata were found on naked algal species than the armoured ones. These findings suggest potential applications of K. obovata leaf extract in controlling harmful algal blooms.

Mapping the main harmful algal species in the East China Sea (Yangtze River estuary) and their possible response to the main ecological status and global climate change via a global vision

The Yangtze River Estuary (YRE) is one of the areas in China most severely affected by harmful algal blooms (HABs). This study explored the distributive patterns of HABs in the YRE and how they are influenced by the El Niño-Southern Oscillation (ENSO) and other environmental factors. Quantitative real-time PCR (qPCR) was employed to detect and quantify the four predominant HAB species in the YRE, Karenia mikimotoi, Margalefidinium polykrikoides, Prorocentrum donghaiense, and Heterosigma akashiwo. Additionally, the study analyzed how turbidity, pH, salinity, and temperature influence these algae. Distribution of the four HAB species in the YRE area shows clear geographical variations: K. mikimotoi is predominantly found in the northwest and central sea areas, M. polykrikoides (East Asian Ribotype, EAR) is mainly distributed in the southeastern part, P. donghaiense is abundant in the northern regions, and H. akashiwo is especially prevalent at stations S26 and S27 in the northeastern part of the study area. HABs dominated by H. akashiwo and P. donghaiense were observed in the northeastern sea area of the YRE on July 22, 2020. Our study reveals that K. mikimotoi, M. polykrikoides (EAR), and P. donghaiense are mainly affected by turbidity, pH, and salinity, while temperature predominantly influences the blooms of H. akashiwo. Moreover, runoff in the YRE has a certain correlation with ENSO events, which may also impact the nutrient content of the region. The findings of this study illustrate the distributive patterns of the four HAB species under various ecological conditions in the YRE and emphasize the importance of establishing practical cases for future warning systems. To better understand how climate change affects HABs, exploring the link between ENSO and HABs is essential.

Toxic effects of the emerging Alexandrium pseudogonyaulax (Dinophyceae) on multiple trophic levels of the pelagic food web

The dinoflagellate Alexandrium pseudogonyaulax, a harmful algal bloom species, is currently appearing in increasing frequency and abundance across Northern European waters, displacing other Alexandrium species. This mixotrophic alga produces goniodomins (GDs) and bioactive extracellular substances (BECs) that may pose a threat to coastal ecosystems and other marine resources. This study demonstrated the adverse effects of A. pseudogonyaulax on four marine trophic levels, including microalgae (Rhodomonas salina), microzooplankton (Polykrikos kofoidii) and mesozooplankton (Acartia tonsa), as well as fish gill cells (RTgill-W1, Oncorhynchus mykiss), ultimately leading to enhanced mortality and cell lysis. Furthermore, cell-free supernatants collected from A. pseudogonyaulax cultures caused complete loss of metabolic activity in the RTgill-W1 cell line, indicating ichthyotoxic properties, while all tested GDs were much less toxic. In addition, cell-free supernatants of A. pseudogonyaulax led to cell lysis of R. salina, while all tested GDs were non-lytic. Finally, reduced egg hatching rates of A. tonsa eggs exposed to cell-free supernatants of A. pseudogonyaulax and impaired mobility of P. kofoidii and A. tonsa exposed to A. pseudogonyaulax were also observed. Altogether, bioassay results suggest that the toxicity of A. pseudogonyaulax is mainly driven by BECs and not by GDs, although further research into factors modulating the lytic activity of Alexandrium spp. are needed.

Identification of microalgal particles using pixel feature analysis of Mueller matrix images

Fine sorting of aquatic particles is of great significance for water environment monitoring. Natural water contains aquatic particles that exhibit a high degree of diversity and complexity, and the identification of aquatic particles remains a persistent challenge in the field. In this article, we propose a new technique for identifying the target species of microalgal particles by using the pixel feature analysis of Mueller matrix images. This technique is independent of any prior knowledge or data about the existing particles in the environment, which is advantageous when applied to real-world situations. The pixel-level polarimetric features are fully leveraged to construct polarization feature templates, which can be used to characterize and filter specific microalgal particles in complex environments. This method could enable the accurate detection of harmful algal blooms species in natural water, which can facilitate early warning of algal blooms. The preliminary results show that the recall rate reached 97.2%, and the average accuracy is 98.9%, which demonstrate the effectiveness of this approach for identifying the target species of aquatic particles in natural water.

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.

Diurnal-Rhythmic Relationships between Physiological Parameters and Photosynthesis- and Antioxidant-Enzyme Genes Expression in the Raphidophyte Chattonella marina Complex.

Diurnal rhythms in physiological functions contribute to homeostasis in many organisms. Although relationships between molecular biology and diurnal rhythms have been well studied in model organisms like higher plants, those in harmful algal bloom species are poorly understood. Here we measured several physiological parameters and the expression patterns of photosynthesis-related and antioxidant-enzyme genes in the Chattonella marina complex to understand the biological meaning of diurnal rhythm. Under a light-dark cycle, Fv/Fm and expression of psbA, psbD, and 2-Cys prx showed significant increases in the light and decreases during the dark. These rhythms remained even under continuous dark conditions. DCMU suppressed the induction of psbA, psbD, and 2-Cys prx expression under both light regimes. Oxidative stress levels and H2O2 scavenging activities were relatively stable, and there was no significant correlation between H2O2 scavenging activities and antioxidant-enzyme gene expression. These results indicate that the Chattonella marina complex has developed mechanisms for efficient photosynthetic energy production in the light. Our results showed that this species has a diurnal rhythm and a biological clock. These phenomena are thought to contribute to the efficiency of physiological activities centered on photosynthesis and cell growth related to the diurnal vertical movement of this species.

Detecting Margalefidinium polykrikoides through high-frequency imagery: Example of a bloom formation, environmental conditions, and phytoplankton community composition changes

In August 2018, the harmful algae species Margalefidinium polykrikoides bloomed to levels previously unobserved in the open waters of Narragansett Bay, Rhode Island, in a transient but intense bloom. Detected by an Imaging FlowCytobot providing hourly data, it is characterized by a time span of less than a week and patchiness with sub-daily oscillations in concentration. The highest concentrations are recorded at lower salinity and higher temperature, suggesting the bloom may have developed in the upper bay and was transported south. The proportion of chains increased during the height of the bloom, and many of the images contained 4-cells per chain. The development of the bloom was favored by optimal temperature and salinity conditions as well as increased nitrogen coincident with greater precipitation and river flow. The period preceding bloom formation also saw a sharp decrease in the dominating large chain-forming diatom Eucampia sp. and highly abundant Skeletonema spp., thus reducing competition over resources for the slow-growing M. polykrikoides. The height of the bloom was reached during the lowest tidal range of the month when the turbulence and water displacement were lower. This time series highlights an out-of-the-ordinary bloom's environmental and biological conditions and the importance of frequent sampling during known favorable conditions.

Development of a Rapid Detection Method to Prorocentrum lima by Loop-Mediated Isothermal Amplification with Hydroxy Naphthol Blue.

Prorocentrum lima, a widely distributed dinoflagellate known for its production of harmful biotoxins, poses a significant threat to humans, aquaculture, and marine ecosystems. As a result, the detection of this toxic alga in coastal waters has become an urgent research focus. In this study, a rapid, sensitive, and cost-effective detection method based on loop-mediated isothermal amplification (LAMP) was developed to identify P. lima. In this method, cell extracts of P. lima were diluted and used directly as templates for amplification, eliminating the need for nucleic acid purification and simplifying the detection process. Hydroxy naphthol blue (HNB) was incorporated into the reaction mix to facilitate result interpretation, enabling visual determination of the amplification outcome with the naked eye. The entire detection process, from DNA extraction to template amplification and product detection, could be completed within 80min using a simple constant temperature-control device. This LAMP-based detection method demonstrated excellent reliability, specificity, and a low detection limit of 5.87 cells/mL for DNA crude extract. The assay offered an efficient alternative to PCR for rapid detection of P. lima. By streamlining the detection process and offering a visual readout, this technique holds promise for efficient and routine monitoring of harmful algal species, benefitting both research efforts and environmental management strategies.

Metabarcoding analysis of potential HABs in the Maowei Sea, China: A baseline information for the “Pinglu Canal Waterway” mega-waterway project

Maowei Sea is one of the cleanest sea areas in China, with relatively little anthropogenic activity. The Maowei Sea is rich in biodiversity and is the largest natural ground for the Hong Kong oyster Crassostrea hongkongensis seed collection. In recent years, the Chinese government has launched a mega project called “Pinglu Canal Waterway”, which will connect the main river system to the Maowei Sea. This project is expected to alter the physical and chemical properties of the Maowei Sea and affect the structure of the phytoplankton community. This study was conducted to screen potential harmful algal bloom species (HABs) in Maowei Sea using metabarcoding technique. In general, nine potential HABs were identified, including six toxin producers (Chattonella spp., Heterosigma akashiwo, Heterocapsa rotundata, Heterocapsa pygmaea, Karenia mikimotoi and Karlodinium veneficum) and three non-toxin producers (Gymnodinium impudicum, Skeletonema costatum and Ulva prolifera). The occurrence and relative abundance of these potential HABs increases with increasing water salinity. The findings of this study not only provide information on the occurrence of potential HABs in Maowei Sea, but also provide valuable information that contributes to the development of aquaculture and fishery management plans to be implemented in Maowei Sea.

Tracking Vibrio: population dynamics and ecology of Vibrio parahaemolyticus and V. vulnificus in an Alabama estuary.

Vibrio spp. are bacteria found in estuaries worldwide; some species can cause illness and infections in humans. Relationships between Vibrio spp. abundance, salinity, and temperature are well documented, but correlations to other environmental parameters are less understood. This study identifies unique correlates (e.g., E-W wind scalar and harmful algal species) that could potentially inform the next iteration of predictive Vibrio models for the EMSS region. Additionally, these correlates may allow existing environmental monitoring efforts to be leveraged in providing data inputs for future Vibrio risk models. An observed correlation between salinity and V. vulnificus/particle-size associations suggests that predicted environmental changes may affect the abundance of Vibrio spp. in certain reservoirs, which may alter which vectors present the greatest vibrio risk.

Abundance of Vibrio aestuarianus, water temperature, and stocking density are associated with summer mortality of Pacific oysters in suspended culture

High mortality rates of cultured Pacific oysters (Crassostrea gigas) during the summer months have regularly occurred on oyster farms in British Columbia, Canada over the last 10 years, but little is known about the microbial and environmental conditions that contribute to such mortality events. The objective of the study was to determine correlative factors associated with the onset of a summer mortality event in oysters (mean ± SD shell height: 14.2 ± 0.5 mm) grown in suspended culture at four stocking densities (150, 300, 450, 600 oysters tray−1) from May 11 to September 17, 2018. Variables examined included both biotic (oyster growth, mortality, reproductive development, and microbiome (approximately every week); Vibrio and harmful algal species abundance) and abiotic (temperature, salinity, turbidity, dissolved oxygen, pCO2, pH, and aragonite saturation) ones. Both the absolute abundance of V. aestuarianus and the relative abundance of Vibrio spp. increased with observed oyster mortality and declining health. Mortality was highest on August 12 and associated with a prior period of elevated temperatures (i.e., increasing temperatures from early July to early August) and increased oyster growth/reproductive development. At that time, systemic mixed microbial infections and necrotic gill tissue in histological cross sections were observed in 19% of oysters that appeared healthy macroscopically. Cumulative percent mortalities per tray ranged from 34 to 75%, the highest-density trays having significantly less mortality and smaller shell width, shell length, and gonad length than lower-density trays. This study demonstrates the significant impact of summer mortality on Pacific oysters and highlights the biotic (host growth, reproductive development, and microbiome composition as well as Vibrio spp. abundance) and abiotic (water temperature) factors associated with the observed mortality in this region.

Critical factors driving spatiotemporal variability in the phytoplankton community structure of the coral habitat in Dongshan Bay, China.

This study investigated the spatiotemporal distribution of the phytoplankton in the coral habitat of Dongshan Bay (China), along with critical factors affecting the distribution, during June, August, and December 2022. Phytoplankton abundance in Dongshan Bay exhibited considerably temporal variation, peaking in June 2022, gradually decreasing thereafter, and reaching its lowest point in December 2022. The abundance of bottom-layer phytoplankton consistently exceeded that of the surface layer throughout all seasons. The average phytoplankton abundance in the coral habitat of Dongshan Bay was lower than that in non-coral habitat areas. Fluctuations in the Zhangjiang River and coastal upwelling influenced the diversity and community structure of the phytoplankton. Critical factors causing spatiotemporal variability in phytoplankton community structure included nutrient concentrations and seawater temperature. Nutrients played key roles in influencing various phytoplankton groups. Dominant diatom species, such as Thalassionema nitzschioides and Thalassiosira diporocyclus, were positively correlated with ammonia nitrogen, seawater salinity, coral cover, and the number of coral species present. In winter, Calanus sinicus exhibited a negative correlation with harmful algal bloom species. Additionally, it was found that both in the coral habitat and surrounding open sea, currents, nutrients, and zooplankton may play crucial roles in determining the spatiotemporal variability in the phytoplankton community structure.

Responses of microeukaryotic community structure to a Phaeocystis globosa bloom in a semi-enclosed subtropical bay

The occurrence of Phaeocystis globosa, a harmful algal bloom species in Chinese coastal waters, has significant impacts on marine organisms and poses a threat to the safety of coastal nuclear power plants. Although previous studies have established a close association between P. globosa blooms and the bacterial community, the relationship between the microeukaryotic community and P. globosa blooms remains poorly understood. In this study, the variations in the microeukaryotic community resulting from a P. globosa bloom were analyzed using 18S rRNA gene amplicon sequencing. The results indicated that the diversity of the microeukaryotic community during the bloom phase was significantly higher than that during the dissipation phase. The microeukaryotic community compositions varied significantly between the two phases of the P. globosa bloom. During the bloom phase, the dominant microeukaryotic was Viridiplantae, which was then replaced by Dinoflagellata during the dissipation phase. Co-occurrence network analysis showed that the relationship among the microeukaryotic community during the bloom phase was more complex than that during the dissipation phase, and the keystone taxa varied as the bloom progressed. Additionally, microeukaryotic community assembly was primarily driven by stochastic processes during the bloom phase based on the β-nearest taxon distance, whereas it was driven by both deterministic processes and stochastic processes during the dissipation phase. Overall, our findings provide novel insight into the mechanisms and interactions involved in microeukaryotic community dynamics in environments disturbed by P. globosa blooms.

Rapid three-dimensional detection of harmful algae using holographic microimaging

Holographic microimaging technology is widely used in the plankton detection due to its large depth of field and ability to record three-dimensional (3D) information. However, traditional methods involve tedious and time-consuming steps, e.g., region of interest (ROI) extraction, dense reconstruction, focused evaluation, and focused ROI recognition. As a result, traditional methods are difficult to detect in real time. In this paper, we propose a neural network-based 3D detection method, which for the first time uses the holograms to obtain 3D sampling data for algae detection, and innovatively designs a convolutional neural network (DHCNN) to improve the speed and accuracy of detection. A dataset containing 3D samples of eight harmful algae species is established, and the trained DHCNN model achieves a mean average precision of 99.2 % on the test set. We find that accuracy is higher with 3D sampled data, which contains more valid information than with autofocused ROIs. Moreover, detection rate is 99.5 %, which means it could detect attached algae and algae abundance statistics are more accurate. And detection time of the proposed method is only 0.066 s, which is 20 times faster than the traditional method. We believe that the proposed method represents an effective approach for real-time algae monitoring.

Effects of polystyrene nanoplastics on growth and hemolysin production of microalgae Karlodinium veneficum

There are few studies on the effects of nanoplastics on growth and hemolysin production of harmful algal bloom species at present. In this study, Karlodinium veneficum was exposed to different concentrations (0, 5, 25, 50, 75 mg/L) of polystyrene nanoplastics (PS-NPs, 100 nm) for 96 h. The effects of PS-NPs on growth of K. veneficum were investigated by measuring algal cell abundance, growth inhibition rate (IR), total protein (TP), malondialdehyde (MDA), glutathione reductase (GSH), superoxide dismutase (SOD), ATPase activity (Na+/K+ ATPase and Ca2+/Mg2+ ATPase). Scanning electron microscope and transmission electron microscope (SEM and TEM) images of microalgae with or without nanoplastics were also observed. The effects of PS-NPs on hemolysin production of K. veneficum were studied by measuring the changes of hemolytic toxin production of K. veneficum exposed to PS-NPs on 1, 3, 5 and 7 days. High concentrations (50 and 75 mg/L) of PS-NPs seriously affected the growth of K. veneficum and different degrees of damage to cell morphology and ultrastructure were found. Excessive free radicals and other oxidants were produced in the cells, which disrupted the intracellular redox balance state and caused oxidative damage to the cells, and the basic activities such as photosynthesis and energy metabolism were weakened. The athletic ability of K. veneficum was decreased, but the ability to produce hemolysin was enhanced. It was suggested that the presence of nanoplastics in seawater may strengthen the threat of harmful algal bloom species to aquatic ecosystems and human health.

Multi-omics analysis reveals the molecular response to heat stress in a “red tide” dinoflagellate

Background“Red tides” are harmful algal blooms caused by dinoflagellate microalgae that accumulate toxins lethal to other organisms, including humans via consumption of contaminated seafood. These algal blooms are driven by a combination of environmental factors including nutrient enrichment, particularly in warm waters, and are increasingly frequent. The molecular, regulatory, and evolutionary mechanisms that underlie the heat stress response in these harmful bloom-forming algal species remain little understood, due in part to the limited genomic resources from dinoflagellates, complicated by the large sizes of genomes, exhibiting features atypical of eukaryotes.ResultsWe present the de novo assembled genome (~ 4.75 Gbp with 85,849 protein-coding genes), transcriptome, proteome, and metabolome from Prorocentrum cordatum, a globally abundant, bloom-forming dinoflagellate. Using axenic algal cultures, we study the molecular mechanisms that underpin the algal response to heat stress, which is relevant to current ocean warming trends. We present the first evidence of a complementary interplay between RNA editing and exon usage that regulates the expression and functional diversity of biomolecules, reflected by reduction in photosynthesis, central metabolism, and protein synthesis. These results reveal genomic signatures and post-transcriptional regulation for the first time in a pelagic dinoflagellate.ConclusionsOur multi-omics analyses uncover the molecular response to heat stress in an important bloom-forming algal species, which is driven by complex gene structures in a large, high-G+C genome, combined with multi-level transcriptional regulation. The dynamics and interplay of molecular regulatory mechanisms may explain in part how dinoflagellates diversified to become some of the most ecologically successful organisms on Earth.

Phytoplankton communities of the west coast of Florida – multiyear and seasonal responses to nutrient enrichment

Blooms of the harmful algae species Karenia brevis are frequent off the southwest coast of Florida despite having relatively slow growth rates. The regional frequency of these harmful algal blooms led to the examination of the dominant estuarine outflows for effects on both K. brevis and the phytoplankton community in general. There is comparatively little information on the growth rates of non-Karenia taxonomic groups other than diatoms. A seasonally based series (Fall, Winter, and Spring) of bioassay experiments were conducted to determine the nutrient response of the coastal phytoplankton community. Treatments included estuarine waters (Tampa Bay, Charlotte Harbor, and the Caloosahatchee River) applied in a 1:25 dilution added to coastal water to mimic the influence of estuarine water in a coastal environment. Other treatments were 5–15 μM additions of nitrogen (N), phosphorus (P), and silica (Si) species, amino acids, and N (urea) + P added to coastal water. Incubations were conducted under ambient conditions with shading for 48 h. Analyses of dissolved and particulate nutrients were coupled with HPLC analysis of characteristic photopigments and taxonomic assignments of biomass via CHEMTAX. The coastal phytoplankton community, dominated by diatoms, cyanophytes and prasinophytes, was significantly different both by bioassay and by season, indicating little seasonal fidelity in composition. Specific growth rates of chlorophyll a indicated no significant difference between any controls, any estuarine treatment, P, or Si treatments. Conditions were uniformly N-limited with the highest growth rates in diatom biomass. Despite differing initial communities, however, there were seasonally reproducible changes in community due to the persistent growth or decline of the various taxa, including haptophytes, cyanophytes, and cryptophytes. For the one bioassay in which K. brevis was present, the slow growth of K. brevis relative to diatoms in a mixed community was evident, indicating that identifying the seasonally based behavior of other taxa in response to nutrients is critical for the simulation of phytoplankton competition and the successful prediction of the region's 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.

Diversity and distribution of benthic dinoflagellates in Tonga include the potentially harmful genera Gambierdiscus and Fukuyoa

Benthic dinoflagellates that can cause illness, such as ciguatera poisoning (CP), are prevalent around the Pacific but are poorly described in many locations. This study represents the first ecological assessment of benthic harmful algae species in the Kingdom of Tonga, a country where CP occurs regularly. Surveys were conducted in June 2016 in the Tongatapu island group, and in June 2017 across three island groups: Ha'apai, Vava'u, and Tongatapu. Shallow subtidal coastal habitats were investigated by measuring water quality parameters and conducting quadrat surveys. Microalgae samples were collected using either macrophyte collection or the artificial substrate method. Benthic dinoflagellates (Gambierdiscus and/or Fukuyoa, Ostreopsis, and Prorocentrum) were counted using light microscopy, followed by molecular analyses (real-time PCR in 2016 and high throughput sequencing (metabarcoding) in 2017) to identify Gambierdiscus and Fukuyoa to species level. Six species were detected from the Tongatapu island group in 2016 (G. australes, G. carpenteri, G. honu, G. pacificus, F. paulensis, and F. ruetzleri) using real-time PCR. Using the metabarcoding approach in 2017, a total of eight species (G. australes, G. carpenteri, G. honu, G. pacificus, G. cheloniae, G. lewisii, G. polynesiensis, and F. yasumotoi) were detected. Species were detected in mixed assemblages of up to six species, with G. pacificus and G. carpenteri being the most frequently observed. Ha'apai had the highest diversity with eight species detected, which identifies this area as a Gambierdiscus diversity ‘hotspot’. Vava'u and Tongatapu had three and six species found respectively. Gambierdiscus polynesiensis, a described ciguatoxin producer and proposed causative agent of CP was found only in Ha'apai and Vava'u in 2017, but not in Tongatapu in either year. Ostreopsis spp. and Prorocentrum spp. were also frequently observed, with Prorocentrum most abundant at the majority of sites. In 2016, the highest number of Gambierdiscus and/or Fukuyoa cells were observed on seagrass (Halodule uninervis) from Sopu, Tongatapu. In 2017, the highest numbers of Gambierdiscus and/or Fukuyoa from artificial substrate samples were recorded in the Halimeda dominant habitat at Neiafu Tahi, Vava'u, a low energy site. This raised the question of the effect of wave motion or currents on abundance measurements from artificial substrates. Differences in detection were noticed between macrophytes and artificial substrates, with higher numbers of species found on artificial substrates. This study provides a baseline of benthic dinoflagellate distributions and diversity for Tonga that may be used for future studies and the development of monitoring programmes.