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.

Nitrogen liberated via allelopathy can promote harmful algal blooms.

Allelopathy is a biological mechanism that can promote harmful algal blooms (HAB) via the inhibition of sympatric phytoplankton. While nutrient loading can also promote HABs, the ability of allelopathy to stimulate HABs via the regeneration of nutrients has yet to be explored. To examine the impacts of allelopathically liberated N on HAB species, a series of experiments were performed using multiple allelopathic HAB species including the dinoflagellates Alexandrium catenella and Margalefidinium polykrikoides, and the pelagophyte, Aureoumbra lagunensis. These HAB species were paired with the cosmopolitan dinoflagellate, Akashiwo sanguinea, that was labeled with 15NO3- or 15NH4+, allowing the release and transfer of N to be traced as a time course during allelopathic interactions. During all experiments, the allelopathic inhibition of Akashiwo was accompanied by increases in cell densities, growth rates, and the δ15N content of the HAB species, evidencing the transfer of N liberated from Akashiwo. The cellular transfer of 15N and release of dissolved N was higher when Akashiwo was grown with 15NO3- compared to 15NH4+ suggesting a differential subcellular-compartmentalization of N sources. Regardless of the type of N, HAB species obtained 60 - 100% of their cellular N from lysed Akashiwo cells and there was an enrichment of the δ15N content of the dissolved NH4+ pool post-lysis of Akashiwo. Collectively, the results demonstrate that beyond facilitating species succession, allelopathy can supply HABs with N and, therefore, is likely important for promoting and sustaining HABs. Given that allelopathy is known to be a dose-dependent process, allelopathy may induce a positive feedback loop, whereby competitors are lysed, N is liberated, HABs are intensified and, in turn, become more strongly allelopathic.

Algicidal activity of a novel bacterium, Qipengyuania sp. 3-20A1M, against harmful Margalefidinium polykrikoides: Effects of its active compound.

Margalefidinium polykrikoides causes significant economic losses in the aquaculture industry by red tide formation. Algicidal bacteria have attracted research interests as a potential bloom control approach without secondary pollution. Qipengyuania sp. 3-20A1M, isolated from surface seawater, exerted an algicidal effect on M. polykrikoides. However, it exhibited a significantly lower algicidal activity toward other microalgae. It reduced photosynthetic efficiency of M. polykrikoides and induced lipid peroxidation and cell disruption. The growth inhibition of M. polykrikoides reached 64.9 % after 24 h of co-culturing, and expression of photosynthesis-related genes was suppressed. It killed M. polykrikoides indirectly by secreting algicidal compounds. The algicide was purified and identified as pyrrole-2-carboxylic acid. After 24 h of treatment with pyrrole-2-carboxylic acid (20 µg/mL), 60.8 % of the M. polykrikoides cells were destroyed. Overall, our results demonstrated the potential utility of Qipengyuania sp. 3-20A1M and its algicidal compound in controlling M. polykrikoides blooms in the marine ecosystem.

Blooms of the harmful algae Margalefidinium polykrikoides and Alexandrium monilatum alter the York River Estuary microbiome.

Harmful algal blooms (HABs) cause damage to fisheries, aquaculture, and human health around the globe. However, the impact of HABs on water column microbiomes and biogeochemistry is poorly understood. This study examined the impacts of consecutive blooms of the ichthyotoxic dinoflagellates Margalefidinium polykrikoides and Alexandrium monilatum on the water microbiome in the York River Estuary, Chesapeake Bay, USA. The samples dominated by single dinoflagellate species and by a mix of the two dinoflagellates had different microbiome compositions than the ones with low levels of both species. The M. polykrikoides bloom was co-dominated by Winogradskyella and had increased concentrations of dissolved organic carbon. The A. monilatum bloom had little impact on the prokaryotic portion of the whole community but was associated with a specific group of prokaryotes in the particle-attached (>3 µm) fraction including Candidatus Nitrosopumilus, Candidatus Actinomarina, SAR11 Clade Ia, Candidatus Bealeia, and Rhodobacteraceae HIMB11. Thus, blooms of these two algal species impacted the estuarine microbiome in different ways, likely leading to shifts in estuarine carbon and nutrient cycling, with M. polykrikoides potentially having a greater impact on carbon cycling in the estuarine ecosystem than A. monilatum.

Detection and Quantification of the Harmful Dinoflagellate Margalefidinium polykrikoides (East Asian Ribotype) in the Coastal Waters of China.

As a marine ichthyotoxic dinoflagellate, Margalefidinium polykrikoides, previously named Cochlodinium polykrikoides, have caused mass mortalities of fish worldwide during blooms. Rapid detection of target species is a prerequisite for the timely monitoring and early warning of harmful algal blooms (HABs). However, it is difficult to achieve rapid identification with traditional methods. The technology of using quantitative real-time PCR (qPCR) to detect and quantify microalgae is relatively mature. Based on the accuracy, rapidity, and sensitivity of qPCR technology, it can be used in the monitoring and development of early warning systems for HABs. From 2017 to 2020, samples were collected from 15 locations off the Chinese coast or from local sea areas. Based on the qPCR detection and analysis, the target species, M. polykrikoides (East Asian ribotype, EAr), was found in samples from Tianjin, Yangtze River estuary, and offshore Fujian (East China Sea). This is the first time that M. polykrikoides (EAr) was detected in the coastal waters of Tianjin. The results reveal a distributive pattern of M. polykrikoides (EAr) along Chinese coastal waters. It is helpful to predict the future diffusion trend of M. polykrikoides (EAr) in the China Sea and provides a practical case for the future construction of monitoring and warning systems for M. polykrikoides and HABs.

Algal blooms historical outbreaks in the northern coastal waters of the Persian Gulf and Oman Sea (1980-2015).

Harmful algal blooms (HABs) have been serious environmental problems in the Persian Gulf and Oman Sea in recent years. There has been an increase in occurrence of HABs in coastal waters of Hormuzgan Province (to the north of the Persian Gulf and Oman Sea) in recent decade, due to eutrophication as a result of human activities. In this review, HAB phenomena in coastal waters of Hormuzgan Province are summarized, including, causative species, timing and the location of blooms during a 35-year time span from 1980 to 2015. This review illustrates that 436 algal blooms have been recorded in the north of the Persian Gulf, formed by 17 species of phytoplankton; 270 of harmful dinoflagellate (Margalefidinium polykrikoides) blooms have led to huge catastrophic impacts on the economy, environment, and society. In addition, most algal blooms (49%) have occurred in the coasts of Bandar Abbas. The data in this review suggest supporting the establishment of an algal bloom monitoring and control program in the coastal waters of the northern part of the Persian Gulf and Oman Sea (Hormuzgan Province).

Understanding controls on Margalefidinium polykrikoides blooms in the lower Chesapeake Bay.

A time-dependent model of Margalefidinium polykrikoides, a mixotrophic dinoflagellate, cell growth was implemented to assess controls on blooms in the Lafayette River, a shallow, tidal sub-tributary of the lower Chesapeake Bay. Simulated cell growth included autotrophic and heterotrophic contributions. Autotrophic cell growth with no nutrient limitation resulted in a bloom but produced chlorophyll concentrations that were 45% less than observed bloom concentrations (~80 mg Chl m-3 vs. 145 mg Chl m-3) and a bloom progression that did not match observations. Excystment (cyst germination) was important for bloom initiation, but did not influence the development of algal biomass or bloom duration. Encystment (cyst formation) resulted in small losses of biomass throughout the bloom but similarly, did not influence M. polykrikoides cell density or the duration of blooms. In contrast, the degree of heterotrophy significantly impacted cell densities achieved and bloom duration. When heterotrophy contributed a constant 30% to cell growth, and dissolved inorganic nitrogen was not limiting, simulated chlorophyll concentrations were within those observed during blooms (maximum ~140 mg Chl m-3). However, nitrogen limitation quenched the maximum chlorophyll concentration by a factor of three. Specifying heterotrophy as an increasing function of nutrient limitation, allowing it to contribute up to 50% and 70% of total growth, resulted in simulated maximum chlorophyll concentrations of 90 mg Chl m-3 and 180 mg Chl m-3, respectively. This suggested that blooms of M. polykrikoides in the Lafayette River are fortified and maintained by substantial heterotrophic nutritional inputs. The timing and progression of the simulated bloom was controlled by the temperature range, 23 °C to 28 °C, that supports M. polykrikoides growth. Temperature increases of 0.5 °C and 1.0 °C, consistent with current warming trends in the lower Chesapeake Bay due to climate change, shifted the timing of bloom initiation to be earlier and extended the duration of blooms; maximum bloom magnitude was reduced by 50% and 65%, respectively. Warming by 5 °C suppressed the summer bloom. The simulations suggested that the timing of M. polykrikoides blooms in the Lafayette River is controlled by temperature and the bloom magnitude is determined by trade-offs between the severity of nutrient limitation and the relative contribution of mixotrophy to cell growth.

High Spatial-Resolution Red Tide Detection in the Southern Coast of Korea Using U-Net from PlanetScope Imagery.

Red tides caused by Margalefidinium polykrikoides occur continuously along the southern coast of Korea, where there are many aquaculture cages, and therefore, prompt monitoring of bloom water is required to prevent considerable damage. Satellite-based ocean-color sensors are widely used for detecting red tide blooms, but their low spatial resolution restricts coastal observations. Contrarily, terrestrial sensors with a high spatial resolution are good candidate sensors, despite the lack of spectral resolution and bands for red tide detection. In this study, we developed a U-Net deep learning model for detecting M. polykrikoides blooms along the southern coast of Korea from PlanetScope imagery with a high spatial resolution of 3 m. The U-Net model was trained with four different datasets that were constructed with randomly or non-randomly chosen patches consisting of different ratios of red tide and non-red tide pixels. The qualitative and quantitative assessments of the conventional red tide index (RTI) and four U-Net models suggest that the U-Net model, which was trained with a dataset of non-randomly chosen patches including non-red tide patches, outperformed RTI in terms of sensitivity, precision, and F-measure level, accounting for an increase of 19.84%, 44.84%, and 28.52%, respectively. The M. polykrikoides map derived from U-Net provides the most reasonable red tide patterns in all water areas. Combining high spatial resolution images and deep learning approaches represents a good solution for the monitoring of red tides over coastal regions.

Developing a 3D mechanistic model for examining factors contributing to harmful blooms of Margalefidinium polykrikoides in a temperate estuary.

Blooms of Margalefidinium (previously Cochlodinium) polykrikoides occur almost annually in summer in the lower Chesapeake Bay and its tributaries (e.g., the James and York Rivers). The Lafayette River, a sub-tributary of the lower James River, has been recognized as an initiation location for blooms in this region. The timing of bloom initiation varies interannually, ranging from late June to early August. To fully understand critical environmental factors controlling bloom initiation and interactions between physical and biological processes, a numerical module simulating M. polykrikoides blooms was developed with a focus on the bloom initiation. The module also includes life cycle and behavioral strategies such as mixotrophy, vertical migration, cyst dynamics and grazing suppression. Parameterizations for these behaviors were assigned based on published laboratory culture experiments. The module was coupled with a 3D physical-biogeochemical model for the James River that examined the contribution of each environmental factor and behavioral strategy to bloom initiation and development. Model simulation results highlight the importance of mixotrophy in maintaining high growth rates for M. polykrikoides in this region. Model results suggest that while many factors contribute to the initiation process, temperature, physical transport processes, and cyst germination are the three dominant factors controlling the interannual variability in the timing of bloom initiation.

Allelopathic effects of Margalefidinium polykrikoides and Gymnodinium impudicum in the growth of Gymnodinium catenatum.

Harmful algae blooms (HABs) are characterized for the coexistence of phytoplankton species with dynamic and complex biotic interactions (e.g., competition, symbiosis, predation, parasitism, allelopathy), that occur at fine temporal and spatial scales, and are relevant to understand the role that different species of phytoplankton play in the regulation of HABs. In this work the allelopathic effects of Margalefidinium polykrikoides (=Cochlodinium polykrikoides) and Gymnodinium impudicum on Gymnodinium catenatum were evaluated. The allelopathic abilities of M. polykrikoides and G. impudicum were investigated in bi-algal culture experiments and in trials in which target species were co-cultured, separated by a 10 µm membrane to prevent a direct cell-to-cell contact; and also by the addition of different volumes of culture media without cells. For all trials, cells of each species were harvested during exponential phase and cultured together by triplicate at three relative abundances: 1:1 (200 Cells mL-1 of each species, G. catenatum and M. polykrikoides or G. impudicum), 2:1 (400 Cells mL-1 of G. catenatum and 200 Cells mL-1 of M. polykrikoides or G. impudicum), and 1:2 (200 cells mL-1 of G. catenatum and 400 Cells mL-1 of M. polykrikoides or G. impudicum). All bioassays were carried out by triplicate in 250 mL Erlenmeyer flasks with 150 mL of modified GSe medium with an initial inoculum of 200 or 400 Cells mL-1. During experiments G. catenatum abundances were enumerated daily. In bi-algal culture experiments mortalities of G. catenatum were from 50% to 100% after 48 h of cell contact with M. polykrikoides or G. impudicum. In the case of culture media without cells, only M. polykrikoides caused a decrease in the cell abundance and growth rate of G. catenatum. Morphological changes occurred in G. catenatum when in contact with M. polykrikoides and G. impudicum, such as membrane shedding, prominent nucleus, loss of flagella, cell lysis, as well as the separation of long chains into individual cells. These results suggest that in the natural environment M. polykrikoides and G. impudicum have allelopathic interactions in G. catenatum, which could negatively affect its growth and survival, indicating that these species could displace blooms of G. catenatum.

An overview of management and monitoring of harmful algal blooms in the northern part of the Persian Gulf and Oman Sea (Hormuzgan Province).

Algal bloom as a common phenomenon in the Persian Gulf and the Sea of Oman had catastrophic effects on environmental, social, economic, and human health aspects from 2008 to 2009. Since 2008, the Persian Gulf and the Sea of Oman Ecological Research Institute (PGSOERI) has monitored and managed algal blooms in the Hormuzgan coast along the northern Persian Gulf and the Sea of Oman. Management strategies have included regular monitoring of chlorophyll, water quality, and remote sensing. In this regard, relevant departments and the Regional Organization for Protection of Marine Environment (ROPME) collaborated with each other to prevent and forecast algal blooms. We reviewed historical and current monitoring, mitigation, and management systems of algal blooms in the Hormuzgan coast. In addition, complications and challenges of algal bloom monitoring and management were also discussed. Documenting algal bloom monitoring and research, improving forecasting and modeling of blooms, educating the public and fishermen, developing a cooperative monitoring framework, and controlling pollution input entering the ROPME region are the main challenges of algal bloom management in the Hormuzgan coast.

Metagenomic Sequencing Identifies Highly Diverse Assemblages of Dinoflagellate Cysts in Sediments from Ships' Ballast Tanks.

Ships' ballast tanks have long been known as vectors for the introduction of organisms. We applied next-generation sequencing to detect dinoflagellates (mainly as cysts) in 32 ballast tank sediments collected during 2001-2003 from ships entering the Great Lakes or Chesapeake Bay and subsequently archived. Seventy-three dinoflagellates were fully identified to species level by this metagenomic approach and single-cell polymerase chain reaction (PCR)-based sequencing, including 19 toxic species, 36 harmful algal bloom (HAB) forming species, 22 previously unreported as producing cysts, and 55 reported from ballast tank sediments for the first time (including 13 freshwater species), plus 545 operational taxonomic units (OTUs) not fully identified due to a lack of reference sequences, indicating tank sediments are repositories of many previously undocumented taxa. Analyses indicated great heterogeneity of species composition among samples from different sources. Light and scanning electron microscopy and single-cell PCR sequencing supported and confirmed results of the metagenomic approach. This study increases the number of fully identified dinoflagellate species from ballast tank sediments to 142 (> 50% increase). From the perspective of ballast water management, the high diversity and spatiotemporal heterogeneity of dinoflagellates in ballast tanks argues for continuing research and stringent adherence to procedures intended to prevent unintended introduction of non-indigenous toxic and HAB-forming species.

Ocean warming along temperate western boundaries of the Northern Hemisphere promotes an expansion of Cochlodinium polykrikoides blooms.

Since the early 1990s, ocean temperatures have increased and blooms of the icthyotoxic dinoflagellate Cochlodinium polykrikoides (a.k.a. Margalefidinium polykrikoides) have become more widespread across the Northern Hemisphere. This study used high-resolution (1-30 km), satellite-based sea surface temperature records since 1982 to model trends in growth and bloom season length for strains of C. polykrikoides inhabiting North American and East Asian coastlines to understand how warming has altered blooms in these regions. Methods provided approximately 180× greater spatial resolution than previous studies of the impacts of warming on harmful algae, providing novel insight into near shore, coastal environments. Along the US East Coast, significant increases in potential growth rates and bloom season length for North American ribotypes were observed with bloom-favourable conditions becoming established earlier and persisting longer from Chesapeake Bay through Cape Cod, areas where blooms have become newly established and/or intensified this century. Within the Sea of Japan, modelled mean potential growth rates and bloom season length of East Asian ribotypes displayed a significant positive correlation with rising sea surface temperatures since 1982, a period during which observed maximal cell densities of C. polykrikoides blooms have significantly increased. Results suggest that warming has contributed, in part, to altering the phenology of C. polykrikoides populations, potentially expanding its realized niche in temperate zones of the Northern Hemisphere.

The harmful algae, Cochlodinium polykrikoides and Aureococcus anophagefferens, elicit stronger transcriptomic and mortality response in larval bivalves (Argopecten irradians) than climate change stressors.

Global ocean change threatens marine life, yet a mechanistic understanding of how organisms are affected by specific stressors is poorly understood. Here, we identify and compare the unique and common transcriptomic responses of an organism experiencing widespread fisheries declines, Argopecten irradians (bay scallop) exposed to multiple stressors including high pCO2, elevated temperature, and two species of harmful algae, Cochlodinium (aka Margalefidinium) polykrikoides and Aureococcus anophagefferens using high-throughput sequencing (RNA-seq). After 48 hr of exposure, scallop transcriptomes revealed distinct expression profiles with larvae exposed to harmful algae (C. polykrikoides and A. anophagefferens) displaying broader responses in terms of significantly and differentially expressed (DE) transcripts (44,922 and 4,973; respectively) than larvae exposed to low pH or elevated temperature (559 and 467; respectively). Patterns of expression between larvae exposed to each harmful algal treatment were, however, strikingly different with larvae exposed to A. anophagefferens displaying large, significant declines in the expression of transcripts (n = 3,615; 87% of DE transcripts) whereas exposure to C. polykrikoides increased the abundance of transcripts, more than all other treatments combined (n = 43,668; 97% of DE transcripts). Larvae exposed to each stressor up-regulated a common set of 21 genes associated with protein synthesis, cellular metabolism, shell growth, and membrane transport. Larvae exposed to C. polykrikoides displayed large increases in antioxidant-associated transcripts, whereas acidification-exposed larvae increased abundance of transcripts associated with shell formation. After 10 days of exposure, each harmful algae caused declines in survival that were significantly greater than all other treatments. Collectively, this study reveals the common and unique transcriptional responses of bivalve larvae to stressors that promote population declines within coastal zones, providing insight into the means by which they promote mortality as well as traits possessed by bay scallops that enable potential resistance.

Resting Cyst Distribution and Molecular Identification of the Harmful Dinoflagellate Margalefidinium polykrikoides (Gymnodiniales, Dinophyceae) in Lampung Bay, Sumatra, Indonesia.

Margalefidinium polykrikoides, an unarmored dinoflagellate, was suspected to be the causative agent of the harmful algal blooms - associated with massive fish mortalities - that have occurred continually in Lampung Bay, Indonesia, since the first bloom event in October 2012. In this study, after examination of the morphology of putative M. polykrikoides-like cysts sampled in bottom sediments, cyst bed distribution of this harmful species was explored in the inner bay. Sediment samples showed that resting cysts, including several morphotypes previously reported as M. polykrikoides, were most abundant on the northern coast of Lampung Bay, ranging from 20.6 to 645.6 cysts g-1 dry sediment. Molecular phylogeny inferred from LSU rDNA revealed that the so-called Mediterranean ribotype was detected in the sediment while M. polykrikoides motile cells, four-cell chain forming in bloom conditions, belonged to the American-Malaysian ribotype. Moreover, hyaline cysts, exclusively in the form of four-cell chains, were also recorded. Overall, these results unequivocally show that the species M. polykrikoides is abundantly present, in the form of vegetative cells, hyaline and resting cysts in an Indonesian area.