Dynamics and scale variations of blooms of Phaeocystis globosa "giant colony" ecotype and key environmental factors in the Beibu Gulf, China.

The Beibu Gulf has experienced blooms of Phaeocystis globosa "giant colony" ecotype (PGGCE), with noticeable variations in bloom scale across years. However, driving environmental factors and their roles remain poorly understood. In this study, we quantified dynamics of PGGCE cells in 2016-2017 and 2018-2019, and analyzed their correlations with environment factors. The results revealed that PGGCE blooms primarily occurred in Guangxi coast and western waters of Leizhou Peninsula during winter months, exhibiting distinct developmental processes. Bloom intensity, duration, and distribution differed significantly between two bloom events. In 2016-2017, peak PGGCE density exceeded 2.0 × 105 cells L-1 nearly double that of 2018-2019. Furthermore, bloom sustained five months during 2016-2017, compared to three months during 2018-2019. Prolonged period of low temperatures and elevated nitrate concentrations favored PGGCE growth and colony formation, resulting in a larger scale bloom during winter 2016 as opposed to winter 2018.

Potential roles of hydroxybenzoate paralytic shellfish toxins in modulating toxin biokinetics in scallops.

Paralytic shellfish toxins (PSTs) produced by some marine dinoflagellates can cause severe human intoxication via vectors like bivalves. Toxic dinoflagellate Gymnodinium catenatum produce a novel group of hydroxybenzoate PSTs named GC toxins, but their biokinetics in bivalves haven't been well examined. In this experiment, we analyzed PSTs in bay scallops Argopecten irradians exposed to G. catenatum (strain MEL11) to determine their accumulation, elimination, anatomical distribution, and biotransformation. To our surprise, up to 30% of the PSTs were accumulated in the adductor muscle of scallops at the end of the experiment, and the toxicity of adductor muscle exceeded the regulatory limit of 800 µg STXeq/kg in only 6 days. High concentration of toxins in the adductor muscle are likely linked to the rapid transfer of GC toxins from viscera to other tissues. Moreover, most GC toxins in scallops were found rapidly transformed to decarbamoyl toxins through enzyme-mediated hydrolysis, which was further supported by the in vitro incubation experiments. Our study demonstrates that GC toxins actively participate in toxin distribution and transformation in scallops, which may increase the risks of food poisoning associated with the consumption of scallop adductor muscle. ENVIRONMENTAL IMPLICATION: The negative impacts of harmful algal blooms (HABs) have become a global environmental concern under the joint effects of cultural eutrophication and climate change. Our study, targeted on the biokinetics of paralytic shellfish toxins in scallops exposed to Gymnodinium catenatum producing unique GC toxins, aims to elucidate potential risks of seafood poisoning associated with GC toxins. The findings of this study will help us to understand the roles of GC toxins in seafood poisoning, and to develop effective management strategies against toxic algal blooms and phycotoxins.

Ultrasensitive fluorescence detection of gonyautoxins in seawater using a novel molecularly imprinted nanoprobe.

Gonyautoxins (GTXs), a group of potent neurotoxins belonging to paralytic shellfish toxins (PSTs), are often associated with harmful algal blooms of toxic dinoflagellates in the sea and represent serious health and ecological concerns worldwide. In the study, a highly selective and sensitive fluorescence nanoprobe was constructed based on photoinduced electron transfer recognition mechanism to rapidly detect GTXs in seawater, using specific entrapment of molecularly imprinted polymers (MIPs) combined with fluorescence analyses. The green emissive fluorescein isothiocyanate was grafted in a silicate matrix as a signal transducer and fluorescence intensity of the nanoprobe with a core-shell structure exhibited a strong enhancement due to efficient analyte blockage in a short response time. Under optimal conditions, the developed MIPs nanoprobe presented an excellent analytical performance for spiked seawater samples including a recovery from 94.44 % to 98.23 %, a linear range between 0.018 nmol L-1 and 0.36 nmol L-1, as well as good accuracy. Furthermore, the method had extremely high sensitivity, with limit of detection obtained as 0.005 nmol L-1 for GTXs and GTX2/3. Finally, the nanoprobe was applied for the determination of GTXs in seven natural seawater samples with GTXs mixture (0.035-0.058 nmol L-1) or single GTX2/3 (0.033-0.050 nmol L-1), and the results agreed well with those of a UPLC-MS/MS method. The findings of our study suggest that the constructed MIPs-based fluorescence enhancement nanoprobe was suitable for rapid, selective and ultrasensitive detection of GTXs, particular GTX2/3, in natural seawater samples.

Seasonal occurrence of toxic phytoplankton and phycotoxins at a mussel aquaculture site in Nova Scotia, Canada.

A three-year field study at a mussel (Mytilus edulis) aquaculture site in Ship Harbour, Nova Scotia, Canada was carried out between 2004 and 2006 to detect toxic phytoplankton species and dissolved lipophilic phycotoxins and domoic acid. A combination of plankton monitoring and solid phase adsorption toxin tracking (SPATT) techniques were used. Net tow and pipe phytoplankton samples were taken weekly to determine the abundance of potentially toxic species and SPATT samplers were deployed weekly for phycotoxin analysis. Mussels were also collected for toxin analysis in 2005. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to analyse the samples for spirolides (SPXs), pectenotoxins (PTXs), okadaic acid group toxins (OA, DTXs) and domoic acid (DA). Phycotoxins were detected with SPATT samplers beginning from the time of deployment until after the producing organisms were no longer observed in pipe samples. Seasonal changes in toxin composition occurred over the sampling period and were related to changes in cell concentrations of Alexandrium Halim, Dinophysis Ehrenberg and Pseudo-nitzschia (Hasle) Hasle. Spirolides peaked in late spring and early summer, followed by DA in mid-July. Okadaic acid, DTX1 and PTXs occurred throughout the field season but peaked in late summer. Concentrations of some phycotoxins detected in SPATT samplers deployed within the area where mussels were suspended on lines were lower than in those deployed outside the mussel farm. The SPATT samplers provided a useful tool to detect the presence of phycotoxins and to establish trends in their appearance in the Ship Harbour estuary.

A dinoflagellate bloom caused by multiple species of Kareniaceae in the coastal waters of Fujian in June 2022 and its adverse impacts on Brachionus plicatilis and Artemia salina.

Recently, dinoflagellate blooms have frequently occurred in the coastal waters of Fujian, East China Sea. In June 2022, a fish-killing bloom of Kareniaceae species occurred in this region. In this study, four species of Kareniaceae, namely, Karenia longicanalis, K. papilionacea, Karlodinium veneficum, and Karl. digitatum were identified from this bloom event based on the results of single-cell PCR and clone libraries, and intraspecies genetic diversity was found in the Karl. veneficum population. The results of acute toxicity assays of the bloom water to two zooplankton species (Brachionus plicatilis and Artemia salina) demonstrated this bloom event strongly inhibited their swimming capacities and survival. The results of this study suggested that the bloom events caused by multiple species of Kareniaceae in the Fujian coastal waters had adverse impacts on the local fishery resources and zooplankton community.

Fourier Transform Infrared Spectrometry Detection of Phaeodactylum tricornutum Biomacromolecules in Response to Environmental Changes.

Environmental factors play an important role in the lipid, protein, and carbohydrate compositions of microalgae, wherein temperature and light are key influencing factors. Fourier transform infrared (FTIR) spectrometry was used in this study to detect biomacromolecules in Phaeodactylum tricornutum cells under different temperatures (10, 15, 20, and 25 °C) and different illumination conditions (1000, 2000, 3000, and 4000 lx) to study the corresponding changes in lipid, protein, and carbohydrate contents. Results indicate that the biomacromolecule content at different temperatures has different patterns. Specifically, the patterns at 15 and 25 °C are similar to each other and the contents accumulate with extended culture time. However, the pattern at 20 °C is different. The carbohydrate and protein contents peaked during the early stage of the exponential phase, whereas lipid accumulation lagged behind the former two, peaking during the middle of the culture stage and then decreasing. Lipid content was analyzed by transmission electron microscopy (TEM), which revealed that the highest lipid content was observed at 15 °C. Results also show that all of the lipid, protein, and carbohydrate contents in cells were the highest when the illumination was at 2000 lx and that the contents decreased with increasing illumination. By using FTIR, less samples were needed as compared to the traditional chemical quantitative detection methods. Moreover, the relative content changes of various biomacromolecules during the growth of P. tricornutum could be accurately determined by a single detection, thereby providing a new technique for the further study of metabolic mechanisms.

Marked shifts of harmful algal blooms in the Bohai Sea linked with combined impacts of environmental changes.

The Bohai Sea, a semi-enclosed inland sea in China and an important mariculture region, has experienced extensive harmful algal blooms (HABs) and their negative impacts for several decades. To investigate the changes of HABs and their potential drivers over time and space, a dataset of 230 HAB events (1952-2017), along with corresponding environmental data (1990-2017) was compiled. The frequency of HAB events in the Bohai Sea has increased over time but plateaued in the last decade, and our analysis showed that history of HABs in the Bohai Sea could be categorized into three periods based on their frequency, scale, and HAB-forming species. The seasonal window of HAB events has started earlier and lasted longer, and the main hotspot has moved from Bohai Bay to coastal waters of Qinhuangdao over time. There were marked shifts in the representative HAB-forming microalgae, from dinoflagellates in the first period (before 2000) to haptophytes in the second period (2000-2009), and pelagophytes in the third period (2009 onwards). These community changes are accompanied by a trend toward diversification of HAB-forming microalgae, decrease in cell-size, and increase in negative impacts. Statistical analyses indicate that long-term changes in HABs in the Bohai Sea are linked with the combined effects of climate change, eutrophication and mariculture development. The results of the present study require to refine future monitoring programs, develop adaptive management strategies and predictive models for HABs in the Bohai Sea.

Brown tides linked to the unique nutrient profile in coastal waters of Qinhuangdao, China.

Brown tides caused by the pelagophyte Aureococcus anophagefferens have frequently occurred in the Bohai Sea since 2009 and have led to a dramatic collapse of the local scallop culture. To determine why brown tides occurred in the Bohai Sea rather than in other eutrophic coastal waters of China, phytoplankton communities and nutrients were evaluated and nutrient addition experiments were conducted in the Qinhuangdao coastal area. The concentration of dissolved organic nitrogen (DON) was nearly five times higher than that of dissolved inorganic nitrogen (DIN) during brown tides. High levels of phytoplankton biomass and nutrients were observed in the inshore waters, and the patterns of different nutrients were heterogeneous, which could be due to the uneven distribution of pelagophytes and non-brown tide phytoplankton populations (NBTP). The nutrient enrichment results indicated that the growth of the phytoplankton community was nitrogen-limited. Enrichment of DON, especially urea, could promote the growth of pelagophytes during the development stages of the brown tide. In brief, the results of this study imply that the unique nutrient profile (rich in DON but deficient in DIN) could support the outbreak of brown tides in the inshore waters of Qinhuangdao.

An investigation on bloom dynamics of Alexandrium catenella and A. pacificum and toxin accumulation in shellfish along the coast of Qinhuangdao, China.

The dinoflagellate genus Alexandrium comprises most of the toxic bloom-forming species producing paralytic shellfish toxins (PSTs) in the sea. Recently, repeated paralytic shellfish poisoning episodes have been recorded in Qinhuangdao located at the west coast of the Bohai Sea. To elucidate the relationship between toxic Alexandrium blooms and the poisoning episodes, a year-round investigation was carried out in this region from July 2020 to July 2021. Two qPCR assays were used to detect A. catenella and A. pacificum, and LC-MS/MS was applied to analyze PSTs in phytoplankton and shellfish samples. The blooms of A. catenella and A. pacificum were found in April and July, respectively, and PST content in three bivalves exhibited notable increase following the bloom of A. catenella. The results revealed bloom dynamics of the two toxic Alexandrium species in the Bohai Sea for the first time, and further confirmed A. catenella as the causative agent of poisoning episodes.

A Massive Green Tide in the Yellow Sea in 2021: Field Investigation and Analysis.

A massive green tide occurred in the Southern Yellow Sea (SYS) in 2021. As in previous years, its high biomass caused trouble to the coastal environment and landscape in 2021. Unusually, the 2021 green tide was unexpectedly massive. Thus, field surveys and remote sensing were conducted in the SYS from December 2020 to July 2021. Compared to pure satellite observations, it revealed the initial development of the green tide more clearly. Given the effects of temperature and nutrient changes on green tide biomass, we compared the environmental parameters in recent years and found no significant increase in nutrient concentrations or changes in temperature of the SYS in 2021. Relative to 2020, the 2021 green tide exhibited a wide distribution, high biomass, and prolonged duration. It was mainly affected by (1) reduced implementation of source-control measures and (2) limited Sargassum biomass, which reduced competition for Ulva prolifera. Strengthening the implementation of source control measures in Subei Shoal is recommended, which is currently the most effective way to control green tides.

Evolution of harmful algal blooms in the East China Sea under eutrophication and warming scenarios.

Harmful algal blooms (HABs) worldwide are experiencing obvious changes under the combined impacts of global warming, eutrophication, and other driving forces. In the East China Sea (ECS), large-scale blooms caused by dinoflagellates occurred since 2000 and there has been an apparent shift of bloom-causative microalgae from diatoms to dinoflagellates. To predict the future evolution of HABs in this region, a model was developed based on the competition between diatoms and dinoflagellates, which would serve to reproduce the seasonal succession of microalgal blooms driven by multiple environmental factors. The evolution features of HABs were then projected under different scenarios of eutrophication and global warming. Under the 'business as usual' scenario, dinoflagellate blooms are expected to become more frequent with higher peak biomass concentrations over the next 30 years. Changes in nutrient composition of the Changjiang riverine discharge may largely give rise to this phenomenon, and accelerated warming associated with climate change may result in earlier occurrence of dinoflagellate blooms. To prevent further intensification of dinoflagellate blooms, efforts could be made to reduce nitrogen inputs and maintain or even increase silicate inputs from the Changjiang river.

Intense blooms of Phaeocystis globosa in the South China Sea are caused by a unique "giant-colony" ecotype.

The haptophyte Phaeocystis globosa, an important causative agent of harmful algal blooms globally, exhibits varying morphological and physiological features and high genetic diversity, yet the relationship among these has never been elucidated. In this study, colony sizes and pigment profiles of 19 P. globosa isolates from the Pacific and Atlantic Oceans were determined. Genetic divergence of these strains was analyzed using the chloroplast rbcS-rpl27 intergenic spacer, a novel high-resolution molecular marker. Strains could be divided into four genetic clades based on these sequences, or two groups based on colony size and the identity of diagnostic pigments (19'-hexanoyloxyfucoxanthin, hex-fuco, and 19'-butanoyloxyfucoxanthin, but-fuco). Three strains from the South China Sea (SCS), all belonging to the same genetic clade, have unique biological features in forming giant colonies and possessing but-fuco as their diagnostic pigment. Based on these findings, we propose that these SCS strains should be a unique "giant-colony" ecotype of P. globosa. During the period 2016-2021, more than 1000 rbcS-rpl27 sequences were obtained from 16 P. globosa colony samples and 18 phytoplankton samples containing solitary P. globosa cells in the SCS. Phylogenetic analysis indicated that >95% of the sequences from P. globosa colonies in the SCS were comprised of the "giant-colony" ecotype, whereas the genetic diversity of solitary cells was much higher. Results demonstrated that intense blooms of P. globosa featuring giant colonies in the SCS were mainly caused by this giant-colony P. globosa ecotype.

Co-occurrence of Alexandrium minutum (Dinophyceae) ribotypes from the Chinese and Malaysian coastal waters and their toxin production.

The bloom-forming dinophyte Alexandrium minutum comprises biogeographic inferred, global and Pacific clades with both toxic and nontoxic strains reported. A. minutum has a wide distribution in the Western Pacific, but to date only a few strains have available DNA sequences. To fully understand its genetic diversity, sampling was undertaken from the Yellow Sea, the East and South China Sea, and five strains of A. minutum and two strains of its sister species, A. tamutum, were established. Their morphology was examined by light and scanning electron microscopy. In addition, sequences were obtained from both large subunit (LSU) ribosomal DNA and/or internal transcribed spacer (ITS) region. Strains of A. minutum are morphologically indistinguishable, characterized by a smaller cell size and a narrow sixth precingular plate. In contrast, A. tamutum has a wider sixth precingular plate. High nucleotide divergences of LSU (D1-D3) rDNA and ITS were revealed amongst strains of A. minutum (10% and 25%, respectively), and A. tamutum (3% and 13%, respectively). Molecular phylogenies based on LSU rDNA and ITS revealed three ribotypes (B-D) of A. minutum, and two ribotypes of A. tamutum in the Western Pacific. Seasonal sampling in the East China Sea to detect A. minutum using the DNA metabarcoding targeting ITS1 region was also performed. Our results showed that the ribotypes B and C of A. minutum co-occurred in the water. Paralytic shellfish toxin (PSTs) of all seven strains was analysed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). PSTs were detected only in A. minutum ribotypes B and C with predominance of gonyautoxins 1/4. Our results suggest high diversity and risk potential of this toxic species in this region.

Rising CO2 will increase toxicity of marine dinoflagellate Alexandrium minutum.

Ocean acidification caused by increasing emission of carbon dioxide (CO2) is expected to have profound impacts on marine ecological processes, including the formation and evolution of harmful algal blooms (HABs). We designed a set of experiments in the laboratory to examine the effects of increasing CO2 on the growth and toxicity of a toxic dinoflagellate Alexandrium minutum producing paralytic shellfish toxins (PSTs). It was found that high levels of CO2 (800 and 1200 ppm) significantly promoted the growth of A. minutum compared to the group (400 ppm) representing the current CO2 level. The total yields of PSTs by A. minutum, including both intracellular and extracellular toxins, were significantly enhanced, probably due to the induction of core enzyme activity and key amino acids synthesis for PST production. More interestingly, high level of CO2 promoted the transformation from gonyautoxin2&3 to gonyautoxin1&4 and depressed the release of PSTs from inside to outside of the cells. All these processes collectively led to an apparent increase of A. minutum toxicity. Our study demonstrated that rising CO2 would increase the risk of toxic A. minutum based on the comprehensive analyses of different processes including algal growth and toxin synthesis, transformation and release.

First record of a Takayama bloom in Haizhou Bay in response to dissolved organic nitrogen and phosphorus.

Since 1990s, harmful algal blooms (HABs) of Kareniaceae, primarily caused by species of Karenia and Karlodinium and rarely by Takayama species, have been substantially increasing in frequency and duration in the coastal waters of China. In this study, we recorded a bloom of high abundance of T. acrotrocha in the Haizhou Bay, the Yellow Sea in September 2020, which is the first record of a Takayama bloom in the temperate coastal waters of China. We found that high concentrations of DON and DOP accelerated the proliferation of T. acrotrocha in the Haizhou Bay. Intensive mariculture, and terrestrial nitrogen and phosphorus input may be responsible for the eutrophication in the Haizhou Bay featuring high concentrations of DON and DOP, and high DIN/DIP ratios. The results suggested that, under ocean warming, the HABs of Kareniaceae are becoming increasingly dominant in eutrophic temperate coasts with intensive mariculture activities.

Toxin production of dinoflagellate Gymnodinium catenatum isolated from the East China Sea.

Dinoflagellate Gymnodinium catenatum is an important producer of paralytic shellfish toxins (PSTs), including a novel group of hydroxybenzoate derivatives named GC toxins. In the East China Sea, G. catenatum has been considered as the causative agent for several paralytic shellfish poisoning (PSP) episodes, yet the knowledge on their toxin production was still quite limited. In this study, toxins produced by a strain of G. catenatum (MEL11) isolated from the East China Sea were determined, using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). Changes of toxin profile in the stain MEL11 in response to nutrient and temperature variations were also examined. A total of 11 PST components dominated by hydroxybenzoate analogs and N-sulfocarbamoyl toxins were detected, which was different from other G. catenatum strains previously established in the East China Sea in the presence of GC5 and the lack of dcGTX2&3. Cellular toxin composition and content of the strain had no apparent change within a range of temperature from 20°C to 26°C. In contrast, nutrient limitation and nitrogen source had notable impacts on toxin production. The molar percentage of GC toxins decreased remarkably at the stationary growth phase under nutrient-deprived conditions of both nitrogen (N) and phosphorus (P). The replacement of nitrate with ammonium as the source of N significantly promoted PST production by G. catenatum. The study revealed the potential diversity of toxin profiles of G. catenatum in the East China Sea, and highlighted the effects of nutrients on production of GC toxins by G. catenatum.

Green Tides in the Yellow Sea Promoted the Proliferation of Pelagophyte Aureococcus anophagefferens.

Harmful algal blooms formed by fast-growing, ephemeral macroalgae have expanded worldwide, yet there is limited knowledge of their potential ecological consequences. Here, we select intense green tides formed by Ulva prolifera in the Yellow Sea, China, to examine the ecological consequences of these blooms. Using 28-isofucosterol in the surface sediment as a biomarker of green algae, we identified the settlement region of massive floating green algae in the area southeast of the Shandong Peninsula in the southern Yellow Sea. The responses of the phytoplankton assemblage from the deep chlorophyll-a maximum layer were then resolved using high-throughput sequencing. We found striking changes in the phytoplankton community in the settlement region after an intensive green tide in 2016, characterized by a remarkable increase in the abundance of the pelagophyte Aureococcus anophagefferens, the causative species of ecosystem disruptive brown tides. Our study strongly suggests that the occurrence of massive macroalgal blooms may promote blooms of specific groups of microalgae through alteration of the marine environment.

Toxic effects, mechanisms, and ecological impacts of harmful algal blooms in China.

Over the last 30 years, harmful algal blooms (HABs) have occurred frequently in the coastal waters of China, resulting in financial losses of over 5.9 billion yuan (about 0.87 billion US dollars) due to massive fish and shellfish mortalities and negative impacts on tourism. To better understand HABs in China, herein we summarized bloom events with massive fish/shellfish mortalities and/or economic losses. Our results suggest that the diversity of HAB species has increased over the last 30 years, with the main causative species shifting from the raphidophyte Chattonella marina and dinoflagellates Gymnodinium spp. to various other species, including the dinoflagellates Karenia mikimotoi and Prorocentrum donghaiense, the haptophyte Phaeocystis globosa, and the pelagophyte Aureococcus anophagefferens. In addition, new types of HABs, such as macroalgal blooms, emerged with severe ecological impacts. We also reviewed the toxic effects, mechanisms, and ecological impacts of common HAB causative species in China. Analysis of the toxic effects of three types of harmful algae (toxin-producing, fish killing, and ecosystem disruptive algae) on marine organisms commonly found in China at different trophic levels revealed that HABs often had toxic effects on multiple organisms in addition to fish or shellfish, with species-specific impacts. Common mechanisms of intoxication include shifting environmental parameters, shellfish poisoning, reactive oxygen species, and haemolytic/cytotoxic toxins. The main mechanism appears to vary with the type of HAB species, and for some notorious algae such as K. mikimotoi and C. marina, further investigations are needed to identify their intoxication mechanism.

Progress on the investigation and monitoring of marine phycotoxins in China.

Marine phycotoxins associated with paralytic shellfish poisoning (PSP), diarrhetic shellfish poisoning (DSP), amnesic shellfish poisoning (ASP), neurotoxic shellfish poisoning (NSP), ciguatera fish poisoning (CFP), tetrodotoxin (TTX), palytoxin (PLTX) and neurotoxin ß-N-methylamino-L-alanine (BMAA) have been investigated and routinely monitored along the coast of China. The mouse bioassay for monitoring of marine toxins has been progressively replaced by the enzyme-linked immunosorbent assay (ELISA) and liquid chromatography tandem mass spectrometry (LC-MS/MS), which led to the discovery of many new hydrophilic and lipophilic marine toxins. PSP toxins have been detected in the whole of coastal waters of China, where they are the most serious marine toxins. PSP events in the Northern Yellow Sea, the Bohai Sea and the East China Sea are a cause of severe public health concern. Okadaic acid (OA) and dinophysistoxin-1 (DTX1), which are major toxin components associated with DSP, were mainly found in coastal waters of Zhejiang and Fujian provinces, and other lipophilic toxins, such as pectenotoxins, yessotoxins, azaspiracids, cyclic imines, and dinophysistoxin-2(DTX2) were detected in bivalves, seawater, sediment, as well as phytoplankton. CFP events mainly occurred in the South China Sea, while TTX events mainly occurred in Jiangsu, Zhejiang and Fujian provinces. Microalgae that produce PLTX and BMAA were found in the phytoplankton community along the coastal waters of China.

Rapid and sensitive detection of Karenia mikimotoi by loop-mediated isothermal amplification combined with a lateral flow dipstick.

Harmful algal blooms frequently occur in various coastal regions worldwide, deteriorating marine ecology and causing huge economic losses. Therefore, developing a potential method for rapid detection of harmful algae species is highly necessitated. In this study, a loop-mediated isothermal amplification (LAMP) method coupled with a lateral flow dipstick (LFD) was developed for detecting the harmful algae Karenia mikimotoi. In this method, the internal transcribed spacer (ITS) sequence of K. mikimotoi was used as the template, and the corresponding specific primers were designed by the online software PrimerExplorer V5. Biotin was labeled on the 5' end of forward inner primer (FIP), and the LAMP reaction was performed under the determined optimal conditions of 63℃ and 60 min. The lowest concentration of K. mikimotoi DNA tested using LAMP was 3.3 × 10-1 pg/µL. Additionally, a 6-FAM-labeled probe was designed and displayed on the LFD after hybridization of the amplified product with the probe. The results demonstrated that LAMP-LFD could be a promising approach for detecting and monitoring harmful algae due to its high sensitivity and specificity.