HPLC pigment profiles of 31 harmful algal bloom species isolated from the coastal sea areas of China

Chemotaxonomy based on diagnostic pigments is now a routine tool for macroscopic determination of the composition and abundance of phytoplankton in various aquatic environments. Since the taxonomic capability of this method depends on the relationships between diagnostic pigments and chlorophyll a of classified groups, it is critical to calibrate it by using pigment relationships obtained from representative and/or dominant species local to targeted investigation area. In this study, pigment profiles of 31 harmful algal bloom(HAB) species isolated from the coastal sea areas of China were analyzed with high performance liquid chromatography(HPLC). Pigment compositions, cellular pigment densities and ratios of pigments to chlorophyll a were determined and calculated. Among all these species, 25 kinds of pigments were detected, of which fucoxanthin, peridinin, 19'-butanoyloxyfucoxanthin, 19'-hexanoyloxyfucoxanthin, violaxanthin, and antheraxanthin were diagnostic pigments. Cellular pigment density was basically independent of species and environmental conditions, and therefore was recommended as a bridge to compare the results of HPLC-CHEMTAX technique with the traditional microscopy method. Pigment ratios of algal species isolated from the coast of China, especially the diagnostic pigment ratios, were higher than those from other locations. According to these results, pigment ratio ranges of four classes of phytoplankton common off the coast of China were summarized for using in the current chemotaxonomic method. Moreover, the differences of pigments ratios among different species under the same culturing conditions were consistent with their biological differences. Such differences have the potential to be used to classify the phytoplankton below class, which is meaningful for monitoring HABs by HPLC-CHEMTAX.     

Effects of modified clay on cysts of Scrippsiella trochoidea for harmful algal bloom control

We present results on the effect of modified clay on cyst formation of Scrippsiella trochoidea in harmful algal bloom(HAB).Modified clay(in concentration of 0,0.1,0.5,and 1.0 g/L) were added to cultures,and observations were made on cysts of 5.trochoidea under controlled laboratory conditions.Results indicate that the removal rate of algal cells reached 97.7%at the clay concentration of 1.0 g/L.The cyst formation rate increased from 4.6%to 24.6%when the concentration of clay was increased from 0to 1.0 g/L.Two cyst metamorphs were observed:spinal calcareous cysts and smooth noncalcareous ones.The proportion of the spinal cysts decreased from 76.9%to 24.1%when clay concentration increased from0 to 1.0 g/L.In addition,modified clay affected cyst germination.The germination rate decreased with the increases in the clay concentrations.Non-calcareous cysts had a lower germination rate with a longer germination time.We conclude that modified clay could depress algal cell multiplication and promote formation of temporal cysts of 5.trochoidea,which may help in controlling HAB outbreaks.     

Laboratory simulation of dissolved oxygen reduction and ammonia nitrogen generation in the decay stage of harmful algae bloom

To evaluate how the decay of bloom-forming algae affect the coastal dissolved oxygen,a laboratory simulation was conducted in terms of three typical harmful algae,Alexandrium catenella,Prorocentrum donghaiense,and Skeletonema costatum.Algae of same biomass(55 μg/mL) were conducted in lightproof columns,and the cell density,dissolved oxygen(DO),and ammonia nitrogen of different layers were monitored at certain time series.Results show that the decomposition of algae significantly decreased the DO,and increased the ammonia nitrogen in all layers;and significant deference between different species was observed.The A.catenella treatment showed the lowest DO(average concentration of 3.4 mg/L) and the highest ammonia nitrogen(average concentration of 0.98 mg/L) at the end of test,followed by P.donghaiense; and the S.costatum showed relatively high DO and low ammonia nitrogen due to slow decay rate.Results indicate that decomposition of harmful bloom algae,especially dinoflagellate,would cause significantly DO depletion and toxic ammonia nitrogen increase,which will detrimentally affect both pelagic and benthic ecosystem.     

Effects of temperature,salinity, and irradiance on the growth of harmful algal bloom species Phaeocystis globosa Scherffel(Prymnesiophyceae) isolated from the South China Sea

Blooms of Phaeocystis globosa have been frequently reported in Chinese coastal waters, causing serious damage to marine ecosystems. To better understand the ecological characteristics of P. globosa in Chinese coastal waters that facilitate its rapid expansion, the effects of temperature, salinity and irradiance on the growth of P. globosa from the South China Sea were examined in the laboratory. The saturating irradiance for the growth of P. globosa( I s) was 60 μmol/(m 2 ?s), which was lower than those of other harmful algal species(70–114 μmol/(m~2 ?s)). A moderate growth rate of 0.22/d was observed at 2 μmol/(m~2 ?s)(the minimum irradiance in the experiment), and photo-inhibition did not occur at 230 μmol/(m~2 ?s)(the maximum irradiance in the experiment). Exposed to 42 different combinations of temperatures(10–31°C) and salinities(10–40) under saturating irradiance, P. globosa exhibited its maximum specific growth rate of 0.80/d at the combinations of 24°C and 35, and 27°C and 40. The optimum growth rates(0.80/d) were observed at temperatures ranging from 24 to 27°C and salinities from 35 to 40. While P. globosa was able to grow well at temperatures from 20°C to 31°C and salinities from 20 to 40, it could not grow at temperatures lower than 15°C or salinities lower than 15. Factorial analysis revealed that temperature and salinity has similar influences on the growth of this species. This strain of P. globosa not only prefers higher temperatures and higher salinity, but also possesses a flexible nutrient competing strategy, adapted to lower irradiance. Therefore, the P. globosa population from South China Sea should belong to a new ecotype. There is also a potentially high risk of blooms developing in this area throughout the year.     

THE AVAILABILITY OF DISSOLVED ORGANIC PHOSPHORUS COMPOUNDS TO MARINE PHYTOPLANKTON

The availability of three dissolved organic phosphorus (DOP) compounds as nutrient sources for experimentalcultare of three algae was studied. Results indicated that these compounds could be utilized byalgae, and that dissolved inorganic phosphorus (DIP) was first to be uptaken when various forms ofphosphorus (DIP and DOP) co-existed. Dicrateria zhanjiangensis' uptake of sodium glycerophosphatewas faster than that of D-ribose-5-phosphate. The increase of sodium glycerphosphate had little effecton the maximum uptake rate(V_(max)) of Chlorella sp., but increased the semisaturation constant(K_s) remarkably;the photosynthesis rates(PR) of Dicrateria zhanjiangensis and Chlorella sp. were rarely affected byusing various forms of phosphorus in the culture experiments. The possible DOP pathways utilizedby algae are discussed.     

Identification of the toxic compounds produced by Sargassum thunbergii to red tide microalgae

The inhibitory effects of methanol extracts from the tissues of three macroalgal species on the growths of three marine red tide microalgae were assessed under laboratory conditions.Extracts of Sargassum thunbergii(Mertens ex Roth) Kuntz tissue had stronger inhibitory effects than those of either Sargassum pallidum(Turner) C.Agardh or Sargassum kjellmanianum Yendo on the growths of Heterosigma akashiwo(Hada) Hada,Skeletonema costatum(Grev.) Grev,and Prorocentrum micans Ehrenberg.Methanol extracts of S.thunbergii were further divided into petroleum ether,ethyl acetate,butanol,and distilled water phases by liquid-liquid fractionation.The petroleum ether and ethyl acetate fractions had strong algicidal effects on the microalgae.Gas chromatography-mass spectrometry analyses of these two phases identified nine fatty acids,most of which were unsaturated fatty acids.In addition,pure compounds of four of the nine unsaturated fatty acids had effective concentrations below 5 mg/L.Therefore,unsaturated fatty acids are a component of the allelochemicals in S.thunbergii tissue.     

Effects of modified clay on cysts of <Emphasis Type="Italic">Scrippsiella trochoidea</Emphasis> for harmful algal bloom control

We present results on the effect of modified clay on cyst formation of Scrippsiella trochoidea in harmful algal bloom （HAB）. Modified clay （in concentration of 0, 0.1, 0.5, and 1.0 g/L） were added to cultures, and observations were made on cysts of S. trochoidea under controlled laboratory conditions. Results indicate that the removal rate of algal cells reached 97.7% at the clay concentration of 1.0 g/L. The cyst formation rate increased from 4.6% to 24.6% when the concentration of clay was increased from 0 to 1.0 g/L. Two cyst metamorphs were observed： spinal calcareous cysts and smooth noncalcareous ones. The proportion of the spinal cysts decreased from 76.9% to 24.1% when clay concentration increased from 0 to 1.0 g/L. In addition, modified clay affected cyst germination. The germination rate decreased with the increases in the clay concentrations. Non-calcareous cysts had a lower germination rate with a longer germination time. We conclude that modified clay could depress algal cell multiplication and promote formation of temporal cysts of S. trochoidea, which may help in controlling HAB outbreaks.     

Temporal and spatial characteristics of harmful algal blooms in Qingdao Waters,China

Qingdao waters,including both the semi-enclosed Jiaozhou Bay(JB) and the adjacent water out of JB(OJB),have been the areas that are most frequently affected by harmful algal blooms(HABs) in the western Yellow Sea(west of 124°E).In this research,HAB occurrences in Qingdao waters from 1990 to 2009 were investigated using spatial tools in geographic information system(GIS) and are discussed in terms of their connection to temporal variation.Additionally,the effects of each HAB occurrence were further evaluated using a simple model.The calculated results were then visualized using a GIS software to indicate the effects of HABs in Qingdao waters during the entire period.As a result,the OJB was proven to be responsible for the frequent HAB occurrences in Qingdao waters after 2000,although JB was traditionally believed to be the principle source of HAB occurrences in Qingdao waters.In addition,increasing nitrogen and N/P structure imbalance were essential for increasing HAB occurrences in Qingdao waters throughout the entire period,especially for the recent HAB occurrences in the OJB.The results of this research would improve the current understanding on HAB occurrences in Qingdao waters,which would benefit HAB monitoring and the implementation of a control strategy in China as well.     

Development and evaluation of a DNA microarray assay for the simultaneous detection of nine harmful algal species in ship ballast and seaport waters

Rapid, high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae, which are responsible for algal blooms, such as red and green tides. In this study, we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously, namely Alexandrium tamarense, Gyrodinium instriatum, Heterosigma akashiwo, Karenia mikimotoi, Prorocentrum donghaiense, Prorocentrum minimum, Ulva compressa, Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection (LOD) of 0.5 ng of genomic DNA (orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether, 230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%, respectively, relative to conventional morphological methods. This indicated that this high-throughput, automatic, and specific method is well suited for the detection of algae in water samples.     

Increased diversity and environmental threat of harmful algal blooms in the Southern Yellow Sea,China

Journal of Oceanology and Limnology - Harmful algal blooms (HABs) in the Southern Yellow Sea (SYS) have shown a trend of increasing diversity and detrimental effects. While the Bohai Sea, East...     

Influence of dimethylsulfide produced by marine algae on the global climate

DMS (dimethylsulfide), a breakdown product of cellular solutes of many species of macroalgae andphytoplankton plays an important role in regulating global climate and counteracting partly the "greenhouse" effect.In this paper, the advance and prospects of DMS study are reviewed and discussed with respectto DMS sample storage, measurement and importance in regulating global climate and the acidity ofrain and aerosol.     

Seasonal changes of dissolved and particulate organic carbon in Donghu Lake, China

Concentrations of dissolved and particulate organic carbon (DOC and POC) were documented in 1996–1997 at 4 different trophic state stations in Donghu Lake, a typical shallow eutrophic lake along the Changjiang River's middle reaches. The mean concentrations of DOC were 15.11±3.26, 15.19±4.24, 14.27±3.43, and 13.31±3.30 mg/L in Station I, II, III, and IV, respectively. The DOC concentrations of the studied area were very similar to that in other lakes along the Changjiang River's middle reaches. The POC mean of the whole lake was 5.01 mg/L due to the large amount of organic detritus of both allochthonous and autochthonous origin. Significant linear relationship was found between POC and chlorophyll a at all 4 stations, which presumably reflect that phytoplankton, its exudates and its metabolic products were the main contributors to the POC pool in the water column. The slope of such linear relationship at Station IV was significantly steeper than that at Station I, II and III. In addition, the DOC/POC ratios (mean value: 4.40) indicated that the organic detritus was the most important component of the particulate organic matter; in other works, next to organic detritus, phytoplankton dominated the particulate organic matter in Donghu Lake. Project 39770146, 39430101 supported by NSFC and the State Key Laboratory of Freshwater Ecology and Biotechnology, affiliated to the Institute of Hydrobiology, Chinese Academy of Sciences.     

Decomposition of algal lipids in clay-enriched marine sediment under oxic and anoxic conditions

A series of laboratory incubation experiments were conducted to examine the decomposition of algal organic matter in clay-enriched marine sediment under oxic and anoxic conditions. During the 245-day incubation period, changes in the concentrations of TOC, major algal fatty acid components (14:0, 16:0, 16:1, 18:1 and 20:5), and n-alkanes (C₁₆–C₂₃) were quantified in the samples. Our results indicate that the organic matters were degraded more rapidly in oxic than anoxic conditions. Adsorption of fatty acids onto clay minerals was a rapid and reversible process. Using a simple G model, we calculated the decomposition rate constants for TOC, n-alkanes and fatty acids which ranged from 0.017–0.024 d⁻¹, 0.049–0.103 d⁻¹ and 0.011 to 0.069 d⁻¹, respectively. Algal organic matter degraded in two stages characterized by a fast and a slow degradation processes. The addition of clay minerals montmorillonite and kaolinite to the sediments showed significant influence affecting the decomposition processes of algal TOC and fatty acids by adsorption and incorporation of the compounds with clay particles. Adsorption/association of fatty acids by clay minerals was rapid but appeared to be a slow reversible process. In addition to the sediment redox and clay influence, the structure of the compounds also played important roles in affecting their degradation dynamic in sediments.     

Measurement and feature analysis of absorption spectra of four algal species

Two methods for particulate pigments(i.e.,quantitative filter technique,QFT,and in vivo measurement,InVivo,respectively)and two methods for dissolved pigments(i.e.,Acetone Extracts,AceEx,and high-performance liquid chromatography,HPLC,respectively) were used to obtain the optical absorption coefficient spectra for cultures of four typical algal species.Through normalization and analysis of the spectra,it is shown that(1) the four methods are able to measure optical absorption spectra of particulate and/or dissolved pigments;(2)that the optical absorption spectra of particulate and dissolved pigments were consistent in terms of the peak position in the blue wavelength,and the difference of the peak position in the near infrared wavelength was ~10nm between each other;and(3)that the leveling effect of the absorption spectra of particulate pigments was significant.These four methods can all effectively measure the absorption coefficients of phytoplankton pigments,while each one has its unique advantages in different applications.Therefore,appropriate method should be carefully selected for various application due to their intrinsic difference.     

Decomposition of algal lipids in clay-enriched marine sediment under oxic and anoxic conditions

A series of laboratory incubation experiments were conducted to examine the decomposition of algal organic matter in clay-enriched marine sediment under oxic and anoxic conditions.During the 245-day incubation period,changes in the concentrations of TOC,major algal fatty acid components (14:0,16:0,16:1,18:1 and 20:5),and n-alkanes (C16-C23) were quantified in the samples.Our results indicate that the organic matters were degraded more rapidly in oxic than anoxic conditions.Adsorption of fatty acids onto cla...     

中国西南喀斯特地区水库溶解态与颗粒态碳研究进展

为了阐明喀斯特水库对河流水体碳循环造成的影响,总结了近年来喀斯特流域筑坝作用对不同形态碳迁移转化和环境影响的研究进展。通过光谱学、稳定及放射性同位素等手段等对喀斯特地区河流-水库系统中的溶解无机碳(DIC)、溶解有机碳(DOC)、颗粒无机碳(PIC)和颗粒有机碳(POC)迁移转化及其控制机制的研究,发现喀斯特水库碳循环展现出明显的季节性分布特征以及梯级水库群可能会进一步放大单个水库的生态效应,这一结果不仅极大地促进了人们对水库碳循环的认识,还有利于探索河流碳循环中“遗失”的碳汇以及更加准确的评估岩溶水库在全球河流碳循环中扮演的角色。喀斯特水库相比非喀斯特水库对人为活动加剧的影响可能具有更强的响应强度,这也意味着喀斯特水库在全球变暖的趋势中发挥的作用需要得到更准确的评估,而在未来的研究中,通过不同分析手段从微观到宏观系统性的总结不同形态碳迁移转化特点将更准确的回答该问题。     

Isolation and identification of bacteria associated with the surfaces of several algal species

We conducted this study to assess the diversity of bacteria associated with the surfaces of algae based on 16S rDNA sequence analyses.Twelve strains of bacteria were obtained from the surfaces of the following four species of algae:Gracilaria textorii,Ulva pertusa,Laminaria japonica,and Polysiphonia urceolata.The isolated strains of bacteria can be divided into two groups:Halomonas and Vibrio,in physiology,biochemical characteristics and 16S rDNA sequence analyses.The phylogenetic tree constructed based on ...     

Comparative study of hydrographic conditions for algal bloom formation in the coastal waters of east and west of Hong Kong during 1998

Phytoplankton abundance was found to be positively correlated with seasonal changes of seawater temperature in Port Shelter and Lamma Channel, Hong Kong in 1998. Rising water temperature from around 20°C to 25°C coincided with an increase in phytoplankton abundance at both locations. Heavy rains from June to September reduced salinity from 30 to 20, but the decrease in salinity was not correlated with a decline in phytoplankton abundance. In spring 1998, over 0.6×106 cells dm-3 and 0.1×106 cells dm-3 of the...     

Seston and dissolved organic carbon dynamics duringMicrocystis bloom with special reference to the effects of three Chinese domestic fishes in experimental enclosures

TheMicrocystis bloom that once disappeared from Donghu Lake, a shallow, eutrophic Chinese lake, was successfully resumed with the use of the enclosure method. Seston and dissolved organic carbon (DOC) dynamics during theMicrocystis bloom were investigated with particular emphasis on the effect of three Chinese domestic fishes (silver carp, bighead carp and grass carp). The results showed that the change of chlorophyll a obviously related to the fish species and its stocking density. Due to the release of extracellular ogranic carbon and increased bacterial activity, DOC increased apparently after silver and bighead carps were introduced. POC/DOC dropped considerably in the enclosures containing silver and bighead carps and in the surrounding lake water, which implied that the bacterial activity was more pronounced in such ecosystems. Statistical analysis indicated that the linear relationship between POC and chlorophyll a could reflect the state of algal growth and nutrient metabolism. It was expected that this study would provide some information on biomanipulation in the shallow, eutrophic lakes. This work was supported in part by a Grant-In-Aid from the Chinese Academy of Sciences, State Key Laboratory of Freshwater Ecology and Biotechnology (FEBL) affiliated to the Institute of Hydrobiology for the project “Control Ecosystem: An Approach to Lake Restoration”.