Harmful blooms of cyanobacteria (Oscillatoriaceae) and dinoflagellates (Gymnodiniaceae) in the Golfo de Nicoya, Costa Rica

Recently, the Pacific coast of Costa Rica has experienced an increase in both magnitude and frequency of harmful algae blooms (HAB). The lack of data regarding the dynamics of these events in the area, and the species of microalgae that produce them, are themes of great interest. The blooms have produced negative impacts on fishery resources and on human health in Costa Rica. In May 2002 a HAB left a large number of dead fish along the central Pacific coast. Water samples were collected using a phytoplankton net and fixed for subsequent processing by electron microscopy. In addition, a one liter sample of surface water was taken for later cell count. In the observed HAB, the dominating organisms found were the cyanobacteria Trichodesmiun erythraeum surrounded by high concentrations of Gram-bacteria and the dinoflagellate Cochlodinium cf. polykrikoides. T. erythraeum, is one of the most important N2 fixing cyanobacteria in marine waters that has been associated with HAB events in diverse parts of the world as well as with symptoms that produce contact dermatitis and other discomforts. C. cf. polykrikoides is a dinoflagellete associated with fish kills; although the type of associated toxins are unknown. In a national newspaper 17 cases of intoxication in humans were reported during this same period, which presented respiratory disorders and burning of the eyes. This is the first report in Costa Rica where a cyanobacteria and a dinoflagellate were observed together producing HAB.     

Development of an oligonucleotide microarray for the detection and monitoring of marine dinoflagellates

Harmful Algal Blooms (HABs), mainly caused by dinoflagellates and diatoms, have great economic and sanitary implications. An important contribution for the comprehension of HAB phenomena and for the identification of risks related to toxic algal species is given by the monitoring programs. In the microscopy-based monitoring methods, harmful species are distinguished through their morphological characteristics. This can be time consuming and requires great taxonomic expertise due to the existence of morphologically close-related species. The high throughput, automation possibility and specificity of microarray-based detection assay, makes this technology very promising for qualitative detection of HAB species. In this study, an oligonucleotide microarray targeted to the ITS1-5.8S-ITS2 rDNA region of nine toxic dinoflagellate species/clades was designed and evaluated. Two probes (45-47 nucleotides in length) were designed for each species/clade to reduce the potential for false positives. The specificity and sensitivity of the probes were evaluated with ITS1-5.8S-ITS2 PCR amplicons obtained from 20 dinoflagellates cultured strains. Cross hybridization experiments confirmed the probe specificity; moreover, the assay showed a good sensitivity, allowing the detection of up to 2 ng of labeled PCR product. The applicability of the assay with field samples was demonstrated using net concentrated seawater samples, un-spiked or spiked with known amounts of cultured cells. Despite the general application of microarray technology for harmful algae detection is not new, a peculiar group of target species/clades has been included in this new-format assay. Moreover, novelties regarding mainly the probes and the target rDNA region have allowed sensitivity improvements in comparison to previously published studies.     

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 mum) 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.     

Developing a two-step method for retrieving cyanobacteria abundance from inland eutrophic lakes using MERIS data

Cyanobacteria are the main dominant species in inland eutrophic lakes during algae blooms, and measures of cyanobacteria abundance can be used for monitoring and early detection of algal blooms by remote sensing. During May 2013 and August 2016, a total 137 water samples were collected from Lake Taihu and Lake Chaohu. Remote-sensing reflectance was measured, surface water was collected in the field, and chlorophyll-a concentration, phycocyanin concentration, suspended-matter concentration and phytoplankton pigment absorption parameters were measured in the laboratory. The composition and density of planktonic algae were also detected by microscope examination. The remote-sensing reflectance at 15 MERIS bands was simulated based on our measured spectral data, and a two-step method for detecting cyanobacteria abundance using the partial least squares model based on 5 MERIS bands was developed. The results showed that the estimation algorithm can predict cyanobacteria abundance in inland eutrophic lakes with satisfactory accuracy, with RMSE of 7.56 and MAPE of 13.44 %. This algorithm was successfully applied to the MERIS image acquired on August 12, 2010, and showed a reasonable spatial distribution of cyanobacteria abundance in Lake Taihu. It demonstrated that the developed estimation method was an effective way to monitor cyanobacteria abundance in water with a potential to be successfully applied to Sentinel-3 images.     

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.     

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。     

Harmful algal bloom (HAB) in the East Sea identified by the Geostationary Ocean Color Imager (GOCI)

Harmful Cochlodinium polykrikoides blooms have frequently appeared and caused fatal harm to aquaculture in Korean coastal waters since 1995. We investigated the applicability of GOCI, the world's first Geostationary Ocean Color Imager, in monitoring the distribution and temporal movement of a harmful algal bloom (HAB) that was discovered in the East Sea near the Korean peninsula in August 2013. We identified the existence of C. polykrikoides at a maximum cell abundance of over 6000 cells/mL and a chlorophyll a concentration of over 400 mg/m³. In areas of C. polykrikoides blooms, GOCI remote sensing reflectance (R_rs) spectra demonstrated the typical radiometric features of a HAB, and from the diurnal variations using GOCI-derived chlorophyll concentration images, we were able to identify the vertical migration of the red tide species. We also found that the formation and propagation of the HAB had relations with cold water mass in the coastal region. GOCI can be effectively applied to the monitoring of short-term and long-term movements of red tides.     

A low-density oligonucleotide array study for parallel detection of harmful algal species using hybridization of consensus PCR products of LSU rDNA D2 domain

A low-density oligonucleotide array approach based on the hybridization of consensus PCR products of LSU rDNA was developed in order to simultaneously detect various harmful algae. A set of oligonucleotide probes for the hybridization of specific LSU rDNA D2 regions was developed for the identification of 10 representative harmful microalgae. Each probe was spotted onto a streptoavidin-coated glass slide by pipetting. Universal primers were designed within the conserved regions adjacent to the D2 regions of all harmful algae and used to PCR amplify the complete D2 regions. The PCR products were hybridized to the oligonucleotides arrayed on the slide. The array produced unique hybridization patterns for each species of harmful algae and allowed us to differentiate the closely related species. Furthermore, we were able to simultaneously detect several predominant HAB species from a mixture of culture strains and from a natural sample. These results show that DNA microarray can be a new technical platform for parallel discrimination of harmful algae and has great potential to alter the manner in which researchers monitor these microorganisms.     

山东海域赤潮灾害特征及预警报管理

掌握赤潮灾害时空分布规律是科学设计赤潮监控方案、提高防灾管理效率的基础和依据。本文统计分析了1990—2010年间山东海域的76次赤潮灾害事件数据。结果表明:赤潮灾害频率显著地分为4个波段,且峰值呈递增趋势;灾害面积呈现周期性倒U型曲线变化;每年的5—10月是赤潮灾害的多发期,其中,8月的灾害次数和灾害面积均占全年总数的30%以上;灾害空间分布集中在莱州湾海域、青岛近海、庙岛群岛的北隍城海域,烟台四十里湾的赤潮灾害最频繁;夜光藻(Noctiluca scintillans)和球型棕囊藻(Phaeocystis globosa)引发的赤潮灾害面积最高,夜光藻和红色裸甲藻(Gymnodinium sanguineum)致灾次数最多。本文从赤潮监测与信息采集、赤潮预报方法以及赤潮灾害预警报管理机制3个方面分析总结了山东海域赤潮灾害预警报业务化管理现状,结合山东海域赤潮灾害特征和预警报管理实践,提出了以控制污染和修复生态环境为根本的赤潮防控建议。     

Comparative composition and dynamics of harmful dinoflagellates in Mediterranean salt marshes and nearby external marine waters

The taxonomic structure of phytoplankton populations in two Mediterranean coastal lagoons were compared with those of nearby marine waters (external waters). Mediterranean confined lagoons remain isolated for most the year and concentrate phytoplankton to a very high biomass. Coastal lagoons on the Mediterranean may, therefore, act as accumulators of neritic phytoplankton (including species related to harmful algal blooms). We examined whether coastal lagoons act as concentrators of marine toxic dinoflagellates during confinement periods, and the common environmental factors that favour growth of specific harmful species in the two ecosystems considered: coastal lagoons and external waters. An alternation between the dominance of diatoms and dinoflagellates was observed, coinciding with that described in Margalef's mandala, occurring in external waters as well as in coastal lagoons. Moreover, the temporal patter was different in the two ecosystems. Dinoflagellate species composition and their bloom period were highly variable in time and space, thus, species had to be analysed individually. Most of the dinoflagellate species found in this study were potentially harmful and high biomass producers. Harmful dinoflagellate species performed well in both, external waters and lagoons, but the specific species-dependent affinity to each of these environments determined which organisms bloom there. Thus, expansion of harmful algal blooms (HAB) to inland waters is not likely and some environmental factors such as the oxidised state of available nitrogen, became determinant to the success and bloom of a species in the coastal lagoon ecosystem.     

Development of a quantitative PCR method to explore the historical occurrence of a nuisance microalga under expansion

A number of marine and freshwater harmful algal bloom (HAB) species have colonized new areas and expanded their habitat range in recent years. Nevertheless it is notoriously difficult to establish when colonization first occurred, what the dispersal routes are, and to separate recent invasion from increases in existent but small populations. The freshwater raphidophyte Gonyostomum semen is a nuisance species that has expanded its habitat range and increased in abundance in northern Europe during the past decades. To evaluate to what extent sediments can be used for determining historic occurrence of G. semen, a quantitative real-time PCR method for detecting cysts of this algae was developed. This paper presents a qPCR protocol with a set of primers that are specific to Gonyostomum and with PCR conditions optimized for sediment samples from humic lakes, which are the common habitat of G. semen. With this sensitive method as few as 1.6 cysts per PCR reaction could be reliably quantified, corresponding to 320 cysts per g wet weight sediment. Cysts were present in sediments with ages ranging from years to decades and their persistence allows detection of historic populations up to at least 50 years old. With this qPCR assay it will be possible to trace the presence of G. semen in environments prior to the onset of algae-specific monitoring programs as well as for quantification in water column samples.     

A model for the prediction of harmful algae blooms in the Vietnamese upwelling area

Satellite pictures and in situ observations indicate strong phytoplankton blooms including harmful algae blooms (HABs) during southwest (SW) summer monsoon in the Vietnamese upwelling area. In this period, nutrients are provided by coastal upwelling and by the very high river runoff from the Mekong River. During SW monsoon, in general two circulation patterns exist which allow the prediction of advection and diffusion of HAB patches. A Lagrangian HAB model that is driven by a circulation model and applied to HABs in Vietnamese waters is presented. Advection which is the most complicated part in modelling transport of passive substances is validated with a Lagrangian sediment trap experiment. The model produces realistic results compared to in situ observations and satellite images and might be used for real time forecast in the future.     

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.     

陆源物质输送对赤潮高发区的影响-以铝为例

近年来长江口附近海域富营养化问题日趋严重,赤潮爆发次数增多,而富含营养物质的长江水的输入可能是海内富营养化的主要物质来源．Al在海洋中停留时间较短且不易受到人为活动的影响,常用Al作为陆源输入及不同水团运动的有效示踪元素．结果表明。研究海域中溶解态Al的浓度秋季高于夏季,且与水体的盐度和悬浮颗粒物浓度表现出一定的关系．赤潮高发区内溶解态Al的浓度一般大于0．04pmol.L-1,明显表现出受到陆源输入的影响．赤潮发生时由于其自身的絮凝及赤潮生物颗粒物表面的吸附作用使水体中的溶解态Al含量平均下降约40％,而在赤潮发生过程中溶解态Al形态变化及机理尚需进一步深入研究．     

In vivo fluorometric method for early detection of cyanobacterial waterblooms

A specific method was developed for monitoring the concentration of cyanobacteria (blue-green algae) before waterblooms, based on their characteristics ofin vivo fluorescence. The excitation and emission spectra of cyanobacteria are very different from those of eukaryotic algae, due to the importance of phycocyanin, rather than chlorophylla, in determining the fluorescence characteristics. Our results, based on four cyanobacteria:Microcystis aeruginosa, Anabaena cylindrica, Phormidium tenue andSpirulina platensis, indicate that excitation at 620 nm and its emission at 645 nm is a sensitive and specific method for their detection. Furthermore, the addition of 10 M photosynthesis inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) induced only 3% increase in phycocyanin fluorescence, suggesting that this measurement is almost independent of the ongoing rate of photosynthesis.Author for correspondence     

Red to red - the marine bacterium Hahella chejuensis and its product prodigiosin for mitigation of harmful algal blooms

Harmful algal blooms (HABs), commonly called red tides, are caused by some toxic phytoplanktons, and have made massive economic losses as well as marine environmental disturbances. As an effective and environment-friendly strategy to control HAB outbreaks, biological methods using marine bacteria capable of killing the harmful algae or algicidal extracellular compounds from them have been given attention. A new member of the gamma-Proteobacteria, Hahella chejuensis KCTC 2396, was originally isolated from the Korean seashore for its ability to secrete industrially useful polysaccharides, and was characterized to produce a red pigment. This pigment later was identified as an alkaloid compound, prodigiosin. During the past several decades, prodigiosin has been extensively studied for its medical potential as immunosuppressants and antitumor agents, owing to its antibiotic and cytotoxic activities. The lytic activity of this marvelous molecule against Cochlodinium polykrikoides cells at very low concentrations (1 ppb) was serendipitously detected, making H. chejuensis a strong candidate among the biological agents for HAB control. This review provides a brief overview of algicidal marine bacteria and their products, and describes in detail the algicidal characteristics, biosynthetic process, and genetic regulation of prodigiosin as a model among the compounds active against red-tide organisms from the biochemical and genetic viewpoints.     

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.     

EVALUATION OF PIGMENT EXTRACTION METHODS AND A RECOMMENDED PROTOCOL FOR PERIPHYTON CHLOROPHYLL a DETERMINATION AND CHEMOTAXONOMIC ASSESSMENT1

Many methods have been proposed to extract and quantify algal pigments. Comparative studies have found that pigment extraction efficiency varies among solvent and mechanical disruption protocols due to differential cellular resistance, thereby, leading to potential misinterpretation of pigment data. When the type or resistance of algae are unknown, a method is required that efficiently extract pigments from all taxonomic groups. The objective of this study was to develop a simple and efficient one stage periphyton pigment extraction protocol by comparing the extractability of four solvents (acetone, methanol, methanol/acetone, and methanol/acetone/N,N‐dimethylformamide), the effects of grinding, and the effects of freeze‐drying. The best overall extraction was obtained using freeze‐dried samples extracted with methanol/acetone/DMF/water (MAD). Eighty‐six percent more chlorophyll was extracted when the sample was freeze‐dried relative to fresh/frozen samples extracted with 90% acetone. Freeze‐drying greatly improved the extraction of both polar and non‐polar (lipophilic/hydrophobic) pigments while MAD increased the extractability of polar pigments and improved peak resolution of all pigments. Chemotaxonomic assessment differed between samples that were fresh/frozen or freeze‐dried before extraction. The relative abundance of cyanobacteria was greater for freeze‐dried material compared with fresh/frozen due to the improved extractability of cyanobacterial pigments. Based on the results of this study, the traditional approach of 90% acetone as a solvent is not recommended for periphyton samples containing cyanobacteria or when the composition of the mat is unknown. The combination of freeze‐drying and MAD was sufficient for the extraction of pigments from a periphyton mat containing filamentous cyanobacteria, green algae, and diatoms.     

Promotion of harmful algal blooms by zooplankton predatory activity

The relationship between algae and their zooplanktonic predators typically involves consumption of nutrients by algae, grazing of the algae by zooplankton which in turn enhances predator biomass, controls algal growth and regenerates nutrients. Eutrophication raises nutrient levels, but does not simply increase normal predator-prey activity; rather, harmful algal bloom (HAB) events develop often with serious ecological and aesthetic implications. Generally, HAB species are outwardly poor competitors for nutrients, while their development of grazing deterrents during nutrient stress ostensibly occurs too late, after the nutrients have largely been consumed already by fast-growing non-HAB species. A new mechanism is presented to explain HAB dynamics under these circumstances. Using a multi-nutrient predator-prey model, it is demonstrated that these blooms can develop through the self-propagating failure of normal predator-prey activity, resulting in the transfer of nutrients into HAB growth at the expense of competing algal species. Rate limitation of this transfer provides a continual level of nutrient stress that results in HAB species exhibiting grazing deterrents protecting them from top-down control. This process is self-stabilizing as long as nutrient demand exceeds supply, maintaining the unpalatable status of HABs; such events are most likely under eutrophic conditions with skewed nutrient ratios.     

基于感光色素吸收信号的太湖藻类识别

铜绿微囊藻和斜生栅藻是太湖"水华"的主要藻种,基于室内纯铜绿微囊藻、斜生栅藻组成色素的吸收系数,通过四阶微分、标准化系数等方法对太湖水体浮游植物中铜绿微囊藻和斜生栅藻进行识别,并确定其组成比例。结果表明,纯藻组成色素的吸收系数应用于其在太湖水体浮游植物中比例的确定和识别中,能够全面考虑辅助色素的识别作用,较好地避免非色素物质吸收信号的干扰,具有较好的识别效果。太湖水体浮游色素中铜绿微囊藻比例最高,斜生栅藻次之,由夏季向冬季过度中,铜绿微囊藻比例不断减小,斜生栅藻的比例逐渐升高,但铜绿微囊藻比例仍略高于斜生栅藻;铜绿微囊藻的区域分布差异性较小,但时间差异性相对较大,而斜生栅藻恰恰相反,空间分布差异性较大而时间分布差异性较小。