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Species of the marine dinoflagellate genus Karenia are known to produce various potent biotoxins and can form noxious blooms that cause mass mortalities of fish and shellfish. To date, harmful blooms of the species K. mikimotoi have been reported in Korea, but K. papilionacea was recently recorded off the southern coast of Korea. Here, we developed a quantitative real-time PCR (qRT-PCR) assay with specific primer pairs for the accurate detection and quantification of these two similar-looking unarmored species, K. mikimotoi and K. papilionacea, and investigated their distribution and dynamics in Korean coastal waters. Overall, K. papilionacea had not only a wider distribution, but also higher cell abundances (15–2553 cells L−1) than K. mikimotoi (3–122 cells L−1) in surface waters. Of 18 sampling sites, the two Karenia species were found to coexist at two sites. During monitoring at a fixed station (S5), K. papilionacea was generally predominant over K. mikimotoi; however, the two species exhibited similar dynamics and occasionally co-occurred. Both Karenia species showed similar physiological responses to temperature and salinity, requiring similar conditions for optimum growth. These results suggest that blooms of the two species may co-occur and induce a synergistic adverse effect on marine environments.

At the end of summer 2020, a moderate (~105 cells L−1) bloom of potential fish-killing Karenia spp. was detected in samples from a 24 h study focused on Dinophysis spp. in the outer reaches of the Pitipalena-Añihue Marine Protected Area. Previous Karenia events with devastating effects on caged salmon and the wild fauna of Chilean Patagonia had been restricted to offshore waters, eventually reaching the southern coasts of Chiloé Island through the channel connecting the Chiloé Inland Sea to the Pacific Ocean. This event occurred at the onset of the COVID-19 lockdown when monitoring activities were slackened. A few salmon mortalities were related to other fish-killing species (e.g., Margalefidinium polykrikoides). As in the major Karenia event in 1999, the austral summer of 2020 was characterised by negative anomalies in rainfall and river outflow and a severe drought in March. Karenia spp. appeared to have been advected in a warm (14–15 °C) surface layer of estuarine saline water (S > 21). A lack of daily vertical migration patterns and cells dispersed through the whole water column suggested a declining population. Satellite images confirmed the decline, but gave evidence of dynamic multifrontal patterns of temperature and chl a distribution. A conceptual circulation model is proposed to explain the hypothetical retention of the Karenia bloom by a coastally generated eddy coupled with the semidiurnal tides at the mouth of Pitipalena Fjord. Thermal fronts generated by (topographically induced) upwelling around the Tic Toc Seamount are proposed as hot spots for the accumulation of swimming dinoflagellates in summer in the southern Chiloé Inland Sea. The results here provide helpful information on the environmental conditions and water column structure favouring Karenia occurrence. Thermohaline properties in the surface layer in summer can be used to develop a risk index (positive if the EFW layer is thin or absent).

Quantitative PCR (qPCR) is the method of choice for specific detection and quantification of harmful algal bloom (HAB) species. Development of qPCR assay for simultaneous enumeration of species that frequently co-exist in HABs is required. A high sensitivity TaqMan qPCR assay, using probe and primers, located at ITS1–5.8S–ITS2 rDNA region, detecting, specifically, Karenia selliformis , K. bidigitata , and K. mikimotoi , was designed. ITS1–5.8S–ITS2 rDNA region copy numbers per Karenia cell genome were estimated to 217.697 ± 67.904, allowing cell quantification. An application of the designed methodology in field samples has been conducted, and it showed high sensitivity (detection of around 10 −1 cell/100 mg of bivalve mollusk tissue, equivalent to about 20 copies of the target sequence). We suggest that the optimized method could contribute to early detection of three closely related Karenia species in seafood cultivating areas to promote control quality, guarantee a fast and effective intervention, and improve public health prevention.

Little is known about the diversity and biogeography of the toxigenic dinoflagellate genera Karlodinium and Karenia because their cells are unarmoured and often small. We isolated single cells from Chinese waters to establish five Karlodinium cultures, and carried out single-cell polymerase chain reactions of eight cells of Karlodinium and Karenia isolated directly from field samples. We also obtained large-subunit (LSU) ribosomal DNA and internal transcribed spacer (ITS) sequences from the same samples used for descriptions of Karenia digitata and Karenia longicanalis. Five cultures of Karlodinium from the South China Sea and Yellow Sea of China were established and subjected to light, scanning electron, and transmission electron microscopy. These strains had a sulcal intrusion, a short apical groove, and six chloroplasts, each with an internal lenticular pyrenoid, but lacked a ventral pore. Pigment analysis showed that fucoxanthin was the main accessory pigment. On the basis of unique morphology and corroboration by molecular phylogeny, these strains were described as Karlodinium zhouanum sp. nov. LSU or ITS sequences (or both) of another eight cells revealed the presence of Karlodinium australe, Karenia longicanalis, and K. papilionacea in the East China Sea for the first time. LSU sequences showed that Karenia digitata differed from Karlodinium decipiens at only one position (99.86% similarity), and Karenia longicanalis differed from K. umbella at only four positions (99.45% similarity). Maximum-likelihood and Bayesian inference analyses supported the classification of Karenia digitata within Karlodinium, and demonstrated that presence of a ventral pore was not a reliable generic character for Karlodinium. Our results also indicated that LSU sequences might be too conservative to differentiate species of Karenia and Karlodinium.

Programmed cell death (PCD) can be induced in microalgae by many abiotic challenges via generation of reactive oxygen species (ROS). Marine phytoplankton live in a highly variable light environment, yet the potential for excess photosynthetically available radiation to trigger PCD has not been examined. On the other hand, photoprotective non-photochemical quenching (NPQ) is hypothesized to counteract intracellular ROS, potentially preventing cell death. The main objective of this study is to investigate high-light-induced death processes and their relationship with photosynthesis in bloom-forming dinoflagellate Karenia brevis . Here, we characterized the prevalence of ROS, caspase-like enzyme activity and cell death as well as photosynthetic status under acute irradiance of 500, 750 or 1000 µmol m −2  s −1 . PCD only occurred at the largest light shift. Although depressed photosynthetic capacities and oxidative stress were apparent across the stress gradient, they did not necessarily lead to cell death. NPQ exhibited dose-dependent activation with increasing light stress, which enabled cells to resist or delay PCD. These results highlight the important role of the balance between ROS generation and NPQ activation on determining cell fates in Karenia under acute irradiance stress. This research also provides insights into potential survival strategies and mechanisms of cell loss under a changeable light environment.

Most marine phytoplankton with relatively high ROS generation rates are categorized as harmful algal bloom (HAB)-forming species, among which Chattonella genera is the highest ROS-producing phytoplankton. In this review, we examined marine microalgae with ROS-producing activities, with focus on Chattonella genera. Several studies suggest that Chattonella produces superoxide via the activities of an enzyme similar to NADPH oxidase located on glycocalyx, a cell surface structure, while hydrogen peroxide is generated inside the cell by different pathways. Additionally, hydroxyl radical has been detected in Chattonella cell suspension. By the physical stimulation, such as passing through between the gill lamellas of fish, the glycocalyx is easily discharged from the flagellate cells and attached on the gill surface, where ROS are continuously produced, which might cause gill tissue damage and fish death. Comparative studies using several strains of Chattonella showed that ROS production rate and ichthyotoxicity of Chattonella is well correlated. Furthermore, significant levels of ROS have been reported in other raphidophytes and dinoflagellates, such as Cochlodinium polykrikoides and Karenia mikimotoi. Chattonella is the most extensively studied phytoplankton in terms of ROS production and its biological functions. Therefore, this review examined the potential ecophysiological roles of extracellular ROS production by marine microalgae in aquatic environment.

The environmental disaster in Kamchatka in the autumn of 2020 was caused by an extensive bloom of harmful microalgae of the genus Karenia. A spectral shape algorithm was used to detect algae on satellite imagery. The algorithm calibration of in situ species composition data made it possible to identify areas where harmful algae dominated in biomass. The algorithm allowed evaluation of the dynamics of the distribution of the algae. The state of phytoplankton was estimated based on images of the specific capacity of photosynthesis. Specific fluorescence is the ratio of the height of the fluorescence line (flh) to the concentration of chlorophyll-a (chl-a). The parameter was used to recognize the stages of the algal bloom: intensive growth, blooming, and change in the dominant algal species. In addition, an increase in the concentration of harmful substances in the coastal zone due to wind impact was analyzed. After analyzing the available data, the events that caused the ecological disaster can be summarized as follows. After the stage of intensive growth of microalgae, nutrient deficiency stimulated the production of metabolites that have a harmful effect on the environment. The change of the dominant alga species in the second half of September and the past storm contributed to a sharp increase in the concentration of metabolites and dead organic matter in the coastal zone, which caused an ecological disaster. The subsequent mass bloom of alga species of the same genus, and the regular wind impact leading to the concentration of harmful substances in the coastal zone, contributed to the development of this catastrophic phenomenon.

Continuous monitoring and early warning together represent an important mitigation strategy for harmful algal blooms (HAB). The coast of Texas experiences periodic blooms of three HAB dinoflagellates: Karenia brevis , Dinophysis ovum , and Prorocentrum texanum . A plankton image data set acquired by an Imaging FlowCytobot over a decade of operation was used to train and evaluate two new automated image classifiers. A 112 class, random forest classifier (RF_112) and a 112 class, convolutional neural network classifier (CNN_112) were developed and compared with an existing, 54 class, random forest classifier (RF_54) already in use as an early warning notification system. Both 112 class classifiers exhibited improved performance over the RF_54 classifier when tested on three different HAB species with the CNN_112 classifier producing fewer false positives and false negatives in most of the cases tested. For K. brevis and P. texanum , the current threshold of 2 cells . mL −1 was identified as the best threshold to minimize the number of false positives and false negatives. For D. ovum , a threshold of 1 cell . mL −1 was found to produce the best results with regard to the number of false positives/negatives. A lower threshold will result in earlier notification of an increase in cell concentration and will provide state health managers with increased lead time to prepare for an impending HAB.

Blooms of the dinoflagellate Karenia selliformis in Chile, often associated with massive fish kills, have been noted alongside other species from the Kareniaceae family, such as Karenia spp. and Karlodinium spp. However, the potential allelopathy impact of Chilean K. selliformis on other phytoplankton species remains unexplored. Here, we assessed the allelopathic effects of cell-free exudates from a Chilean K. selliformis strain on six phytoplankton strains representing diverse microalgal groups. The findings of these experiments offer valuable insights into the varied responses of both non-toxic and toxic microalgae to allelochemicals produced by a toxic microalga, showcasing the intricate and multifaceted nature of allelopathic interactions in microalgal communities. The study revealed species-dependent effects, with variable response in cell growth, photosynthetic efficiency (i.e., Fv/Fm), and intracellular reactive oxygen species (ROS) production. While certain strains exhibited significant growth inhibition in response to the allelochemicals, others demonstrated no apparent effect on cell proliferation, indicating varying sensitivity to specific allelochemicals or potentially distinct detoxification mechanisms. Similarly, the diverse effects on Fv/Fm highlight the complexity of allelopathic interactions, with some species showing reduced efficiency without alterations in intracellular ROS production, while others displayed increased ROS production alongside impaired photosynthesis.

Exploring mixotrophy of dinoflagellate species is critical to understanding red-tide dynamics and dinoflagellate evolution. Some species in the dinoflagellate genus Karenia have caused harmful algal blooms. Among 10 Karenia species, the mixotrophic ability of only two species, Karenia mikimotoi and Karenia brevis, has been investigated. These species have been revealed to be mixotrophic; however, the mixotrophy of the other species should be explored. Moreover, although K. mikimotoi was previously known to be mixotrophic, only a few potential prey species have been tested. We explored the mixotrophic ability of Karenia bicuneiformis, Karenia papilionacea, and Karenia selliformis and the prey spectrum of K. mikimotoi by incubating them with 16 potential prey species, including a cyanobacterium, diatom, prymnesiophyte, prasinophyte, raphidophyte, cryptophytes, and dinoflagellates. Cells of K. bicuneiformis, K. papilionacea, and K. selliformis did not feed on any tested potential prey species, indicating a lack of mixotrophy. The present study newly discovered that K. mikimotoi was able to feed on the common cryptophyte Teleaulax amphioxeia. The phylogenetic tree based on the large subunit ribosomal DNA showed that the mixotrophic species K. mikimotoi and K. brevis belonged to the same clade, but K. bicuneiformis, K. papilionacea, and K. selliformis were divided into different clades. Therefore, the presence or lack of a mixotrophic ability in this genus may be partially related to genetic characterizations. The results of this study suggest that Karenia species are not all mixotrophic, varying from the results of previous studies.