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...     

Detection of several harmful algal species by sandwich hybridization integrated with a nuclease protection assay

The frequent occurrence of harmful algal blooms (HABs) is a pressing topic in marine research. An integrated sandwich hybridization and nuclease protection assay was established to qualitatively and quantitatively detect 12 harmful algal species. This method demonstrated good reliability, specificity and accuracy for analyzing samples from individual and mixed cultures, as well as field collection, and cell volumes were positively correlated to the slopes of calibration curves. The lowest quantitative detection limits were those concentrations observed during blooms; thus, this technique provides an efficient alternative to microscopy for rapid identification and quantitation of harmful algal species and could be routinely used to monitor phytoplankton in field surveys.     

海洋赤潮水色遥感技术研究进展

海洋水色遥感已成为赤潮探测与监测的重要技术手段之一。文章回顾了赤潮卫星遥感技术的发展历程,阐述了水色遥感的原理--水体的离水辐射及其光谱特征。着重论述了高光谱卫星在赤潮水色遥感中的作用,通过大气校正减少大气和气溶胶对遥感信息的衰减,同时就赤潮发生的海洋学机理进行了探讨。综合分析后指出中分辨率成像光谱辐射计(MODIS)作为图谱合一的新一代卫星传感器将在赤潮的卫星监测研究中发挥及其重要的作用。并对今后赤潮遥感的主攻方向提出了一些建议:拓宽遥感数据来源,与地理信息系统技术结合及加强赤潮的预警研究等。     

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。     

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.     

大型海藻龙须菜与东海原甲藻间的营养竞争

生物方法作为一种新兴的赤潮防治方法,因为其作用的专一性及较少的负效应,越来越受到人们的重视。研究了龙须菜(G racilaria lem aneif orm is)与东海原甲藻(P rorocentrum d ongha iense)之间营养盐NO3-、PO43-竞争的情况。结果显示,两者共培养时,龙须菜对营养盐的快速吸收利用,使得共培养体系中营养盐迅速降低,最终导致东海原甲藻消亡,而东海原甲藻对于龙须菜的生长不构成明显的影响。在营养盐充分的条件下,1g鲜重的龙须菜对NO3-的吸收能力相当于6.0×107个东海原甲藻细胞,对PO43-的吸收相当于2.4×107个东海原甲藻细胞。相对于东海原甲藻,龙须菜对营养盐的吸收利用更具有优势。龙须菜可作为有效吸收营养盐的大型海藻,用以降低近海水域富营养化程度及有害赤潮发生的几率。     

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.     

High Resolution Spatio-temporal Detection of Potentially Harmful Dinoflagellates in Confined Waters of the NW Mediterranean

A systematic sampling programme was carried out in a large numberof confined waters (principally harbours) along the Catalancoast (NW Mediterranean) in the context of a new MonitoringProgramme. This Monitoring Programme was associated not onlywith areas subject to aquaculture activities, and thereforeunder legislation, but also with confined areas with a highrisk of harmful algal blooms (HABs) occurrence, in order toprovide an early warning of potentially widespread HABs. Thesystematic programme was performed weekly in summer and bi-monthlyin winter for five years. The main results were: (i) the detectionof many harmful species and the presence of high numbers ofharmful dinoflagellates, mainly of the genera Alexandrium andDinophysis; (ii) the detection of Alexandrium catenella, newin the study area, which had hardly ever been detected in theMediterranean Sea; (iii) the presence of some potentially harmfulspecies, including Dinophysis sacculus, present at all periodsof the year; (iv) bloom recurrence in several stations; (v)occasional coincidence of small-scale blooms, such as thoseconfined inside the harbours, with widespread blooms (mesoscaleblooms) of the same organism. The implications of this highfrequency of HAB detection is discussed in relation to the suitabilityof this sampling programme (focused on confined waters) forthe early detection of algal blooms.     

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。     

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.     

The role of dissolved organic matter (DOM) quality in the growth enhancement of Alexandrium fundyense (Dinophyceae) in laboratory culture1

Several algal species responsible for harmful algal blooms (HABs), such as Alexandrium fundyense, are mixotrophic under certain environmental conditions. The ability to switch between photosynthetic and heterotrophic modes of growth may play a role in the development of HABs in coastal regions. We examined the influence of humic dissolved organic matter (HDOM) derived from terrestrial (plant/soil) and microbial sources on the growth of A. fundyense. We found that a terrestrially derived HDOM, Suwannee River humic acid (SRHA), did enhance A. fundyense growth; however, a microbially derived HDOM, Pony Lake fulvic acid (PLFA) did not enhance growth. A. fundyense grows in association with bacteria in culture and we observed that bacterial cell densities were much lower in A. fundyense cultures than in bacteria‐only cultures, consistent with bacterial grazing by A. fundyense in culture. In bacteria‐only cultures with added algal exudates (EX), the addition of PLFA and SRHA resulted in a slight increase in bacterial cell density compared to cultures without HDOM added. Changes over time in the chemical quality of the HDOM in the A. fundyense cultures reflected contributions of microbially derived material with similar characteristics as the PLFA. Overall, these results suggest that the chemical differences between SRHA and PLFA are responsible for the greater effect of SRHA on A. fundyense growth, and that the differential effect is not a result of an effect on the growth of associated bacteria.     

Detection and quantification of cultured marine Alexandrium species by real-time PCR

The occurrence of harmful algal blooms (HABs) throughout the world has increased and poses a large threat to human health, fishery resources and tourism industries. The genus Alexandrium includes a number of toxic species associated with HABs. Therefore, it is very important to rapidly detect and monitor the harmful algae, such as Alexandrium genus. In this study, a standard curve of plasmid containing 18S rDNA-28S rDNA region from Alexandrium catenella was constructed and 5.8S rDNA sequence served as the primer of the real-time PCR. Cultured A. catenella, Alexandrium affine, Alexandrium lusitanicum and Alexandrium minutum samples were analyzed by real-time PCR using the same set of primers simultaneously. Using microscopy cells counts, 5.8S rDNA copies per cell and total DNA per cell were estimated. This assay method is promising for rapid detection of large number of Alexandrium samples.     

溶藻微生物净化富营养化水体的作用

湖泊富营养化导致的水华藻类频繁暴发已成为当今世界性的环境问题,溶藻微生物作为生物法防治有害藻类水华具有广泛的研究前景,通过查阅文献对微生物控制有害藻类水华进行概述,并探讨进一步的研究趋势和应用前景,以期对微生物溶藻方面的研究及开发应用有一定的参考价值。     

A spring poleward current and its influence on microplankton assemblages and harmful dinoflagellates on the western Iberian coast

Along-shore currents can propagate harmful algal blooms (HABs) over long distances in many coastal areas of the ocean. Harmful dinoflagellate blooms on the west coast of Iberia frequently occur when the Iberian poleward current (IPC) establishes on the continental slope. This has led to the suggestion that HABs could be transported northward by the IPC. To examine this possibility, the microplankton composition along the west coast of Iberia was studied in May 1993 coinciding with the presence of the IPC. The microplankton of the IPC was almost exclusively composed of small flagellates, with the notable absence of the harmful species usually associated with coastal waters. The primary influence of the IPC was to confine coastal microplankton populations to the shelf, where a downwelling convergence prevented their export from the coastal environment. Microplankton assemblages on the shelf revealed a north–south gradient related to different stages of succession. Earlier stages of succession in which diatoms were prominent were found on the northern shelf, whereas dinoflagellates were more abundant in the south. The toxic species Gymnodinium catenatum, which was only present in the southern shelf, did not show a northward transport associated with the IPC. It is suggested that the northward spreading of HABs along the west coast of Iberia must be related to the interaction between the IPC, which accumulates coastal populations on the shelf, and the latitudinal progress of microplankton succession that determines species composition. Thus, during the course of the season, HABs are likely to be observed in the south prior to their development in the north.     

Fiber-Optic Microarray for Simultaneous Detection of Multiple Harmful Algal Bloom Species

Harmful algal blooms (HABs) are a serious threat to coastal resources, causing a variety of impacts on public health, regional economies, and ecosystems. Plankton analysis is a valuable component of many HAB monitoring and research programs, but the diversity of plankton poses a problem in discriminating toxic from nontoxic species using conventional detection methods. Here we describe a sensitive and specific sandwich hybridization assay that combines fiber-optic microarrays with oligonucleotide probes to detect and enumerate the HAB species Alexandrium fundyense, Alexandrium ostenfeldii, and Pseudo-nitzschia australis. Microarrays were prepared by loading oligonucleotide probe-coupled microspheres (diameter, 3 μm) onto the distal ends of chemically etched imaging fiber bundles. Hybridization of target rRNA from HAB cells to immobilized probes on the microspheres was visualized using Cy3-labeled secondary probes in a sandwich-type assay format. We applied these microarrays to the detection and enumeration of HAB cells in both cultured and field samples. Our study demonstrated a detection limit of approximately 5 cells for all three target organisms within 45 min, without a separate amplification step, in both sample types. We also developed a multiplexed microarray to detect the three HAB species simultaneously, which successfully detected the target organisms, alone and in combination, without cross-reactivity. Our study suggests that fiber-optic microarrays can be used for rapid and sensitive detection and potential enumeration of HAB species in the environment.     

The effect of iron on Chilean Alexandrium catenella growth and paralytic shellfish toxin production as related to algal blooms

BioMetals - The dinoflagellate Alexandrium catenella is a well-known paralytic shellfish toxin producer that forms harmful algal blooms (HABs) worldwide. Blooms of this species have repeatedly...     

Identification of unique microbiomes associated with harmful algal blooms caused by Alexandrium fundyense and Dinophysis acuminata

Biotic interactions dominate plankton communities, yet the microbial consortia associated with harmful algal blooms (HABs) have not been well-described. Here, high-throughput amplicon sequencing of ribosomal genes was used to quantify the dynamics of bacterial (16S) and phytoplankton assemblages (18S) associated with blooms and cultures of two harmful algae, Alexandrium fundyense and Dinophysis acuminata. Experiments were performed to assess changes in natural bacterial and phytoplankton communities in response to the filtrate from cultures of these two harmful algae. Analysis of prokaryotic sequences from ecosystems, experiments, and cultures revealed statistically unique bacterial associations with each HAB. The dinoflagellate, Alexandrium, was strongly associated with multiple genera of Flavobacteria including Owenweeksia spp., Maribacter spp., and individuals within the NS5 marine group. While Flavobacteria also dominated Dinophysis-associated communities, the relative abundance of Alteromonadales bacteria strongly co-varied with Dinophysis abundances during blooms and Ulvibacter spp. (Flavobacteriales) and Arenicella spp. (Gammaproteobacteria) were associated with cells in culture. Eukaryotic sequencing facilitated the discovery of the endosymbiotic, parasitic dinoflagellate, Amoebophrya spp., that had not been regionally described but represented up to 17% of sequences during Alexandrium blooms. The presence of Alexandrium in field samples and in experiments significantly altered the relative abundances of bacterial and phytoplankton by both suppressing and promoting different taxa, while this effect was weaker in Dinophysis. Experiments specifically revealed a negative feedback loop during blooms whereby Alexandrium filtrate promoted the abundance of the parasite, Amoebophrya spp. Collectively, this study demonstrates that HABs formed by Alexandrium and Dinophysis harbor unique prokaryotic and eukaryotic microbiomes that are likely to, in turn, influence the dynamics of these HABs.     

Inhibition of harmful algal blooms caused by Aureococcus anophagefferens (Pelagophyceae) using native (Gracilaria tikvahiae) and invasive (Dasysiphonia japonica) red seaweeds from North America

Journal of Applied Phycology - Harmful algal blooms (HABs) caused by the pelagophyte Aureococcus anophagefferens have spread globally and are a threat to coastal ecosystems. Although some HAB...     

近海海洋生态系统与有害赤潮的发生综述

海洋生态系统是地球生物圈的重要组成部分。目前,人类活动已经严重破坏了海洋生态系统,导致了海洋生态系统的失衡。赤潮的发生正是这种不平衡的具体体现。文章讨论了人类活动对海洋生态系统的负面影响以及对近海海域赤潮发生的推波助澜作用;主要综述了影响赤潮发生和消亡的生态条件及环境影响因子。最后,强调了学科交叉研究在管理和防范赤潮的发生,维护近海海洋生态系统健康服务功能的必要性,并提出了一些相应的对策和措施。     

OPTICAL MONITORING AND FORECASTING SYSTEMS FOR HARMFUL ALGAL BLOOMS: POSSIBILITY OR PIPE DREAM?

Monitoring programs for harmful algal blooms (HABs) are currently reactive and provide little or no means for advance warning. Given this, the development of algal forecasting systems would be of great use because they could guide traditional monitoring programs and provide a proactive means for responding to HABs. Forecasting systems will require near real-time observational capabilities and hydrodynamic/biological models designed to run in the forecast mode. These observational networks must detect and forecast over ecologically relevant spatial/ temporal scales. One solution is to incorporate a multiplatform optical approach utilizing remote sensing and in situ moored technologies. Recent advances in instrumentation and data-assimilative modeling may provide the components necessary for building an algal forecasting system. This review will outline the utility and hurdles of optical approaches in HAB detection and monitoring. In all the approaches, the desired HAB information must be isolated and extracted from the measured bulk optical signals. Examples of strengths and weaknesses of the current approaches to deconvolve the bulk optical properties are illustrated. After the phytoplankton signal has been isolated, species-recognition algorithms will be required, and we demonstrate one approach developed for Gymnodinium breve Davis. Pattern-recognition algorithms will be species-specific, reflecting the acclimation state of the HAB species of interest.Field data will provide inputs to optically based ecosystem models, which are fused to the observational networks through data-assimilation methods. Potential model structure and data-assimilation methods are reviewed.