Molecular cloning reveals co-occurring species behind red tide blooms of the harmful dinoflagellate Cochlodinium polykrikoides

Red tides caused by the marine dinoflagellate Cochlodinium polykrikoides Margalef pose significant environmental problems worldwide. Recently, the existence of severe blooms attributable to a single Cochlodinium Schütt species has been questioned by many researchers. Herein we investigated the dinoflagellate composition of harmful algal blooms (HABs) attributed to C. polykrikoides in Korean coastal waters at nine different stations (St.). The component species of Cochlodinium blooms were examined by using microscopic and gene-cloning methods. In the nine study areas, C. polykrikoides was the predominant species of HABs in St. 2, 4, 7, and St. 9. Based on the morphological identification, the bloom was initially thought to be caused only by C. polykrikoides; however, we detected additional bloom-forming dinoflagellates (Polykrikos schwartzii Bütschli and Polykrikos kofoidii Chatton), and diatoms (Pseudo-nitzschia americana (Hasle) Fryxell) along with C. polykrikoides. The parasitic dinoflagellates Amoebophrya Koeppen and Euduboscquella Coats, Bachvaroff & Delwiche were found to be co-located with Cochlodinium in our study, and for the first time, Cochlodinium fulvescens Iwataki, Kawami & Matsuoka was detected in Korea (west coast). These results suggest co-existence of multiple dinoflagellates in bloom populations of Cochlodinium and describe the composition of other dinoflagellate blooms (e.g., Polykrikos spp.) in Korean coastal regions. This co-occurrence may be considered during efforts to monitor and control HABs.     

沿岸海域富营养化与赤潮发生的关系

综述了赤潮的发生与沿岸海域富营养化的关系。近几十年来,人类活动使得天然水体的富营养化进程大大加速。营养负荷的增加与高生物量水华的增多相联系。控制营养输入后,浮游植物生物量或有害藻类水华事件也相应减少。营养的组成与浮游植物的种类组成及水华的形成有密切联系。有机营养对有害藻类水华的促进作用受到关注。营养输入时机影响浮游植物种间竞争的结果,因而对浮游植物的群落演替具有深远影响。由于浮游植物存在生理差异,因而对营养加富的反应因种而异。营养在调控某些有毒藻类的毒素产量方面也发挥着重要作用。此外,营养输入与藻类水华之间存在复杂的间接联系。当然,营养状况并非浮游植物群落演替的唯一决定因素。研究结果提示,控制营养输入、减缓水域富营养化是减少有害藻类水华发生的有效途径,而深入研究典型有害藻类的营养生理对策则为防治并最终消除有害藻类水华提供了理论基础。     

Downwelling and dominance of autochthonous dinoflagellates in the NW Iberian margin: The example of the Ría de Vigo

Dinoflagellate blooms in coastal upwelling systems are restricted to times and places with reduced exchange and mixing. The Rías Baixas of Galicia are four bays in the NW Iberian upwelling with these characteristics where harmful algal blooms (HABs) of dinoflagellates are common. These blooms are especially recurrent at the end of the upwelling season, when autumn downwelling amplifies accumulation and retention through the development of a convergence front in the interior of Rías. Because oceanic water enters the Rías during downwelling, it has been hypothesised that dinoflagellate blooms originate by the advection and subsequent accumulation of allochthonous populations. To examine this possibility, we studied the microplankton succession in relation to hydrographic variability in the Ría de Vigo (one of these four bays) along an annual cycle making use of a high sampling frequency. The results indicated that upwelling lasted from May to August, with downwelling prevailing in winter. Microplankton during upwelling, although dominated by diatoms, evidenced a progressive increase in the importance of dinoflagellates, which achieved maximum abundance at the end of the upwelling season. Thus, diatoms characterised the spring bloom, while diatoms and autochthonous dinoflagellates composed the autumn bloom. Diatoms dominated during the first moments of the autumn downwelling and dinoflagellates were more abundant later, after stronger downwelling removed diatoms from the water column. Since the dinoflagellates selected by downwelling belonged to the local community, it is concluded that advection of alien populations is not required to explain these autumn blooms.     

Spatial, seasonal and species variations of harmful algal blooms in the South Yellow Sea and East China Sea

The occurrences of harmful algal blooms (HABs), in terms of frequency and area in the Chinese coastal waters, have been increasing since 1980s and caused considerable economic losses. In the present study, we have analyzed spatial and seasonal characteristics of HAB events in the southern Yellow Sea and East China Sea along Chinese coast from 1933 to 2004. With a total 435 HAB records, the most frequent HAB occurrence area (FHA) is off the Yangtze River mouth and another two FHA areas are located south of the Yangtze River estuary along about isobaths of 30–60 m coastal water in the East China Sea. The time of HAB occurrence shifted during our study period: from autumn (August–October) before 1980s to July–August in 1980s, during May–July in 1990s, and May–June for the period of 2000–2004. Causative species were found to be different: Noctiluca scintillans and Skeletonema costatum were dominant causative species prior to 2000; and Prorocentrum donghaiense Lu was dominant from 2000 to 2004 and also caused large blooms in May. Trichodesmium sp. caused many HABs in autumn (August–October) prior to 1980s with only one HAB between 1980 and 2004. The changes of the dominant HAB species may have affected the timings of HAB occurrence, as well as the increasing HAB-affected areas in recent years.     

Molecular approaches to diagnosing nutritional physiology in harmful algae: Implications for studying the effects of eutrophication

Every year harmful algal blooms (HABs) cause serious impacts to local economies, coastal ecosystems, and human health on a global scale. It is well known that nutrient availability can influence important aspects of harmful algae biology and ecology, such as growth, toxin production, and life cycle stage, as well as bloom initiation, persistence and decline. Increases in the rate of supply of organic matter to ecosystems (eutrophication) carries many possible ramifications to coastal systems, including the potential for nutrient enrichment and the potential for stimulation of harmful algal blooms. Traditional studies on algal nutrition typically use either cultured isolates or community level assays, to examine nutrient uptake, nutrient preference, elemental composition, and other metrics of a species’ response to nutrients. In the last decade, technological advances have led to a great increase in the number of sequences available for critical harmful species. This, in turn, has led to new insights with regards to algal nutrition, and these advances highlight the promise of molecular technologies, and genomic approaches, to improving our understanding of algal nutrient acquisition and nutritional physiological ecology, in both cultures and field populations. With these developments increased monitoring of nutritional physiology in field populations of harmful algae will allow us to better discriminate how eutrophication impacts these groups.     

Distribution and dynamics of two species of Dinophyceae producing high biomass blooms over the French Atlantic Shelf

The frequency and distribution of high biomass blooms produced by two dinoflagellate species were analysed along the French continental shelf from 1998 to 2012. Two species were specifically studied: Karenia mikimotoi and Lepidodinium chlorophorum. Based on remote-sensing reflectances at six channels (410, 430, 480, 530, 550 and 670 nm), satellite indices were created to discriminate the species forming the blooms. A comparison with observations showed that the identification was good for both species in spite of a lower specificity for L. chlorophorum. The overall analysis of the satellite indices, in association with some monitoring data and cruise observations, highlights the regularity of these events and their extent on the continental shelf. L. chlorophorum blooms may occur all along the South Coast of Brittany. All the coastal areas under the influence of river plumes and the stratified northern shelf area of the Western English Channel appear to be areas of bloom events for both species. These two species are likely to be in competitive exclusion as they share the same spatial distribution and the timing of their bloom is very close. Finally, due to the scarcity of off-shore observations, these satellite indices provide useful information regarding HABs management and the development of a warning system along the French coast.     

象山港硅藻赤潮过程中真菌群落演替及互作特征

[背景] 近岸海域赤潮的发生能显著改变包括真菌在内的微生物群落组成,进而影响海洋中碳氮元素循环。真菌是海洋中重要的分解者,但赤潮过程对真菌群落的影响少有报道。[目的] 探明赤潮过程对真菌群落的影响,对进一步阐释赤潮对生态系统中物质循环的影响具有重要意义。[方法] 针对2017年春季象山港硅藻赤潮,对真菌内源转录间隔区1（Internally Transcribed Spacer 1,ITS1）序列进行高通量测序,研究赤潮过程中真菌群落响应及共现性特征。[结果] 真菌群落的α多样性指数在赤潮暴发的各阶段差异显著,而且均与磷酸盐、硝酸盐和亚硝酸盐有显著相关性（P<0.05）。子囊菌门（Ascomycota,90.2%）和担子菌门（Basidiomycota,8.27%）是本次硅藻赤潮的优势菌门,其中子囊菌门遍布赤潮暴发的4个阶段;锤舌菌纲（Leotiomycetes,16.1%）和散囊菌纲（Eurotiomycetes,9.3%）是纲水平上的主要优势类群。赤潮暴发过程中硝酸盐、亚硝酸盐、磷酸盐和温度等环境因素的改变,驱动真菌群落组成及结构发生显著变化。Helotiales、Eurotiales、Xylariales、Sacc haromycetales和Agaricostilbales构成了真菌群落共现网络的主体。[结论] 真菌群落在赤潮的各阶段具有显著的演替特征,赤潮藻生消是引起真菌群落演替的主要因素,但是环境因子也扮演着重要角色。赤潮过程中,真菌群落同类群内的互作要强于不同类群间的互作,这对维持群落的稳定性具有重要意义。     

春季东海中华哲水蚤对有害藻华物种的选择性摄食

2005年4月27日—6月5日在东海有害藻华高发区的6个典型站位采样,分析了浮游植物的群落结构,并采用现场实验研究了中华哲水蚤(Calanus sinicus)的选择性摄食.结果表明,中华哲水蚤对浮游植物的选择性摄食具有食物密度依赖性.低浮游植物细胞丰度下中华哲水蚤具有明显的选择性摄食行为,高浮游植物细胞丰度下中华哲水蚤摄食选择性由偏好转变为排斥,浮游植物细胞丰度329 cells·ml^-1是中华哲水蚤由偏好趋于排斥摄食的阈值.中华哲水蚤的选择性摄食还与食物组成有关,对于食物中的微型浮游动物,中华哲水蚤无选择性或排斥摄食.有害藻华发生区中华哲水蚤对具齿原甲藻(Prorocentrum dentatum)的偏好摄食以及对米氏凯伦藻(Karenia mikimotoi)的排斥摄食,促使藻华的物种演替最终向米氏凯伦藻水华方向发展.     

Harmful algal blooms: causes,impacts and detection

Blooms of autotrophic algae and some heterotrophic protists are increasingly frequent in coastal waters around the world and are collectively grouped as harmful algal blooms (HABs). Blooms of these organisms are attributed to two primary factors: natural processes such as circulation, upwelling relaxation, and river flow; and, anthropogenic loadings leading to eutrophication. Unfortunately, the latter is commonly assumed to be the primary cause of all blooms, which is not the case in many instances. Moreover, although it is generally acknowledged that occurrences of these phenomena are increasing throughout the world's oceans, the reasons for this apparent increase remain debated and include not only eutrophication but increased observation efforts in coastal zones of the world. There is a rapidly advancing monitoring effort resulting from the perception of increased impacts from these HABs, manifested as expanding routine coastal monitoring programs, rapid development and deployment of new detection methods for individual species, toxins, and toxicities, and expansion of coastal modeling activities towards observational forecasts of bloom landfall and eventually bloom prediction. Together, these many efforts will provide resource managers with the tools needed to develop effective strategies for the management and mitigation of HABs and their frequently devastating impacts on the coastal environment.     

Detection of bacteria associated with harmful algal blooms from coastal and microcosm environments using electronic microarrays

With the global expansion of harmful algal blooms (HABs), several measures, including molecular approaches, have been undertaken to monitor its occurrence. Many reports have indicated the significant roles of bacteria in controlling algal bloom dynamics. Attempts have been made to utilize the bacteria/harmful algae relationship in HAB monitoring. In this study, bacterial assemblages monitored during coastal HABs and bacterial communities in induced microcosm blooms were investigated. Samples were analysed using denaturing gradient gel electrophoresis (DGGE) of the 16S rRNA gene. DGGE bands with peculiar patterns before, during, and after algal blooms were isolated and identified. Probes for six ribotypes representing organisms associated with Chatonella spp., Heterocapsa circularisquama, or Heterosigma akashiwo were used for analysis on NanoChip electronic microarray. In addition, a new approach using cultured bacteria species was developed to detect longer (533 bp) polymerase chain reaction-amplified products on the electronic microarray. The use of fluorescently labelled primers allowed the detection of individual species in single or mixed DNA conditions. The developed approach enabled the detection of the presence or absence and relative abundance of the HAB-related ribotypes in coastal and microcosm blooms. This study indicates the ability of electronic microarray platform to detect or monitor bacteria in natural and induced environments.     

Recombinase-aided amplification combined with lateral flow dipstick for the rapid detection of Amphidinium carterae

Journal of Applied Phycology - In the context of global climate change, the frequency and duration of harmful algal blooms (HABs) due to eutrophic coastal waters have increased. HABs can cause...     

深圳大鹏湾海域锥状斯氏藻赤潮期间细菌群落结构变化研究

目的:近年来,赤潮在我国的发生呈增加的趋势,并造成了极大的经济损失。过去研究赤潮发生的机理主要集中在理化因素的影响,而越来越多的证据表明仅凭借营养盐等环境因素并不能解释大部分赤潮现象,藻际微生物可能发挥着重要作用。本文跟踪观测了2010年7月深圳大鹏湾海域爆发的锥状斯氏藻赤潮生消过程中细菌群落丰度种类的变化,从微生物与赤潮藻相互作用的角度探讨了赤潮的生消过程,讨论了不同时期不同关键菌群的特殊作用,为解释赤潮爆发和消亡提供了新的视角,为赤潮的监控和防治新方法的建立奠定了理论基础。方法:本文按时间顺序共采集该赤潮9次样本,利用末端限制性酶切片段长度多态性分析(T-RFLP)等分子生物学方法,通过主成分分析、克隆文库的构建,研究了微生物群落的变化过程,并探讨了特定种属的微生物在赤潮发生、发展和消亡过程中的作用。结果:从浮游细菌丰度来看,随着锥状斯氏藻细胞数量的波动,浮游细菌总数也随之呈现相应的变化。从浮游细菌的种类来看,它们主要属于变形杆菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)。从浮游细菌的动态变化过程来看,赤潮生消过程中,浮游细菌群落呈现出一定的演替现象,特别是在赤潮后期,群落主成分变化尤为显著。从关键菌群的作用来看,属于γ变形杆菌门的Alteromonas sp.一直占有较高的丰度,赤潮中后期受到菌群相互作用导致比例下降,而赤潮后期其他关键菌群的丰度的增高可能是导致赤潮消亡的重要原因。结论:本文利用T-RFLP这一DNA指纹技术分析微生物群落结构和多样性特征,通过研究赤潮生消过程中藻际浮游细菌群落的动态变化,发现随着赤潮的发展,浮游细菌群落发生着相应的变化。结果说明赤潮藻体丰度数量的改变影响着浮游细菌群落的组成。相对地,细菌群落的适应调整迅速造成赤潮藻体局部生长环境的改变,从而影响赤潮的发展过程。     

Occurrences of harmful algal blooms (HABs) associated with ocean environments in the South China Sea

Harmful algal blooms (HABs) occur frequently in the South China Sea (SCS), causing enormous economic losses in aquaculture. We analyzed historical HAB records during the period from 1980 to 2003 in SCS. We found that HABs-affected areas have expanded and the frequency of HABs varied during this period. The seasonal and annual variations, as well as causative algal species of HABs are different among the four regions. Areas with frequent HABs include the Pearl River Estuary (China), the Manila Bay (the Philippines), the Masinloc Bay (the Philippines), and the western coast of Sabah (Malaysia). HABs occurred frequently during March–May in the northern region of SCS, May–July in the eastern region, July in the western region, and year-round in the southern region. Among the species that cause HABs, Noctiluca scintillans dominated in the northern region, and Pyrodinium bahamense in the southern and eastern regions. Causative species also varied in different years for the entire SCS. Both P. bahamense and N. scintillans were the dominant species during 1980–2003. Some species not previously recorded formed blooms during 1991–2003, including Phaeocystis globosa, Scrippsiella trochoidea, Heterosigma akashiwo, and Mesodinium rubrum. Variations in HABs are related to various regional conditions, such as a reversed monsoon wind in the entire SCS, river discharges in the northern area, upwelling in Vietnam coastal waters during southwest winds and near Malaysia coastal waters during northeast winds, and eutrophication from coastal aquaculture in the Pearl River estuary, Manila Bay, and Masinloc Bay. Handling editor: D. Hamilton     

Understanding the blob bloom: Warming increases toxicity and abundance of the harmful bloom diatom Pseudo-nitzschia in California coastal waters

The toxic diatom genus Pseudo-nitzschia produces environmentally damaging harmful algal blooms (HABs) along the U.S. west coast and elsewhere, and a recent ocean warming event coincided with toxic blooms of record extent. This study examined the effects of temperature on growth, domoic acid toxin production, and competitive dominance of two Pseudo-nitzschia species from Southern California. Growth rates of cultured P. australis were maximal at 23 °C (∼0.8 d⁻¹), similar to the maximum temperature recorded during the 2014–2015 warming anomaly, and decreased to ∼0.1 d⁻¹ by 30 °C. In contrast, cellular domoic acid concentrations only became detectable at 23 °C, and increased to maximum levels at 30 °C. In two incubation experiments using natural Southern California phytoplankton communities, warming also increased the relative abundance of another potentially toxic local species, P. delicatissima. These results suggest that both the toxicity and the competitive success of particular Pseudo-nitzschia spp. can be positively correlated with temperature, and therefore there is a need to determine whether harmful blooms of this diatom genus may be increasingly prevalent in a warmer future coastal ocean.     

The Diversity of Harmful Algal Bloom-Triggering Mechanisms and the Complexity of Bloom Initiation

Mechanisms influencing initiation of harmful algal blooms (HABs) are diverse, and are not likely to be mutually exclusive. Rather, initiation of HABs is a result of interactions between processes, which result in biological, physical, and chemical conditions optimal for a bloom. Due to the complexity of some bloom initiation processes, bloom-preventative management may be possible. Results from a modeling exercise and a laboratory experiment indicated that a phytoplankton bloom could be circumvented through manipulation of the nutrient-loading mode, i.e., pulsed vs. continuous loading. These findings, should they prove consistent in more robust field experiments, may provide insights for the development of new management approaches for some HABs. Optimal bloom conditions, however, vary between HAB species. Consequently, it is unlikely that a single management solution will exist. Preventative management efforts will require early warning of HAB initiation, perhaps even before the appearance of an HAB species. An indicator based on the dynamic nature of phytoplankton succession events and phytoplankton species diversity may prove useful for this purpose. Applying this index to an existing plankton data set showed that Microcystis blooms might have been predicted months before the start of the bloom.     

Predictive Ecological Modeling of Harmful Algal Blooms

We have thus far constructed a series of coupled ecological/physical models to predict the origin and fate of harmful algal blooms of the toxic dinoflagellate, Gymnodinium breve, on the West Florida shelf. We find that (1) the maximal population growth rate of G. breve must be ～0.80 day^?1 during initiation of a red tide, but as little as 0.08 day^?1 during its decay, (2) diatoms dominate when estuarine and shelf-break supplies of nitrate are made available to a model community of small and large diatoms, coccoid cyanophytes and Trichodesmium, non-toxic and red-tide dinoflagellates, microflagellates, and coccolithophores, (3) a numerical recipe for large red tides of G. breve requires DON supplies, mediated by iron-starved, nitrogen-fixers, while small blooms may persist on sediment sources of DON, (4) selective grazing must be exerted on the non-toxic dinoflagellates, and (5) vertical migration of G. breve in relation to seasonal changes of summer downwelling and fall/winter upwelling flow fields determines the duration and intensity of red tide landfalls along the barrier islands and beaches of the west coast of Florida, once other losses are specified. Given poorly known initial and boundary conditions and the expense of shipboard monitoring programs, however, bio-optical moorings or remote sensors are the most likely sources of model validation and improved HAB forecasts. Accordingly, the bio-optical implications of our ecological models must be included in future simulation analyses of HABs on both the West Florida shelf and within other coastal regions. Of particular importance for initiation of the coupled biophysical models is an improved understanding of the relationship of remotely sensed surface signals to shade-adapted dinoflagellates, aggregating below the first optical depth of the water column.     

A large and prolonged bloom of Karenia mikimotoi in Scottish waters in 2006

A prolonged bloom of Karenia mikimotoi was observed during 2006 in Scottish waters. This bloom is thought to be unique in the region in terms of its large spatial extent. From its first detection in the west of the country, the bloom moved clockwise around the coast eventually reaching the east coast and the Shetland Isles to the north. The bloom resulted in extensive mortalities of benthic organisms including annelids and molluscs and some species of fish. Farmed fish mortalities were absent but gill damage was reported. The availability of satellite remote sensing, phytoplankton counts from multiple sites, meteorological data and some water chemistry, as well as information on the physical characteristics of the sampling sites, provided an extensive spatial and temporal data set. Analysis of remotely sensed chlorophyll-a data from Aqua-MODIS indicated that this parameter is a useful early warning indicator of K. mikimotoi in shelf waters off the Scottish west coast, and suggested that the bloom developed in this region prior to its advection to coastal waters. An earth observation (EO) based harmful bloom classifier for K. mikimotoi recognised areas of elevated K. mikimotoi cell density but generated false positives in areas of high reflectance. Data were also used to evaluate, in Scottish waters, various hypotheses that exist to explain the formation of K. mikimotoi blooms including phototaxis, nutrient availability, cell transport and elevated water temperature. Specifically, we sought to evaluate if routinely collected environmental data (water temperature, insolation, wind strength and direction, and sea-loch aspect) could be used as a predictor of bloom magnitude near aquaculture facility locations, which typically lie within fjordic sea lochs. Path analysis was used to derive intuitive models linking environmental drivers to bloom magnitude. Once the effects of latitude such as northward water cooling were taken into account, only rainfall was a significant predictor of bloom magnitude at the sampling sites. Therefore, while the offshore development and progression of a bloom may be predicted from satellite information, it is likely that local hydrodynamic influences are crucial in determining its magnitude at coastal aquaculture sites.     

High-resolution spatio-temporal distribution of a coastal phytoplankton bloom using laser in situ scattering and transmissometry (LISST)

Development of optical observation technologies provides new insights into harmful algal bloom (HAB) detection and assessment of HAB species dynamics. Based on preliminary laboratory tests, a laser in situ scattering and transmissometry instrument (LISST-100X) was used to monitor a high-biomass phytoplankton proliferation in the field. Short-term spatial and temporal changes in particle size distribution were measured during a recurrent Alexandrium taylori outbreak. Since the bloom was not monospecific, a size-fraction method to discriminate particular species from LISST-100X measurements was proposed. The results were validated to simultaneous microscopic counts of phytoplankton, and the significantly positive correlation obtained between the two methodologies confirmed the instrument's ability to discriminate phytoplankton at the group and species level. The LISST-100X obtains high-resolution in situ data, and is therefore a better alternative than the traditional microscope for assessing temporal and spatial evolution of HABs. Field observations showed high variability over a short time scale associated with diel vertical migration of A. taylori and the whole phytoplankton population (nanoplankton and microplankton). A numerical circulation model was used to investigate the influence of beach hydrodynamics in the observed horizontal variability. Simulations of the model suggested an important role of daily coastal circulation in determining the distribution of A. taylori in coastal environments.     

Harmful dinoflagellate blooms caused by Cochlodinium sp.: Global expansion and ecological strategies facilitating bloom formation

The past two decades have witnessed an expansion in the reported occurrences of harmful algal blooms (HABs) caused by the dinoflagellate Cochlodinium. Prior to 1990, blooms had been primarily reported in Southeast Asia, with South Korea alone reporting more than $100M USD in annual fisheries losses during the 1990s. Since then, time blooms have expanded across Asia, Europe, and North America, with recognition of multiple species and ribotypes that exhibit similar ecophysiological and harmful characteristics. Here, we summarize the current state of knowledge regarding taxonomy, phylogeny, detection, distribution, ecophysiology, life history, food web interactions, and mitigation of blooms formed by Cochlodinium. We review this recent expansion of Cochlodinium blooms and characterize the ecological strategies utilized by Cochlodinium populations to form HABs. Although Cochlodinium is comprised of more than 40 species, we focus primarily on the two HAB-forming species, C. polykrikoides and C. fulvescens, specifically describing their flexible nutrient acquisition strategies, inhibition of grazing by inducing rapid mortality in a diverse set of predators, and allelopathic inhibition of a broad range of competing phytoplankton. Finally, we summarize the available information on prevention, control, and mitigation strategies specific to this genus, and discuss pressing questions regarding this increasingly important HAB organism.