Kinetic studies on the effects of rare earth elements (REES) on the growth of Microcystis and their accumulation by Microcystis

In the paper the kinetic effects of La3+ and Ce4+ on the growth of Microcystis and the accumulation kinetics of Microcystis in the single and combined systems of La3+ and Ce4+ were studied. The mechanism of the effects of La3+ and Ce4+ on the growth of Microcystis and their accumulation kinetics were also discussed. In the single system, La3+ stimulated the growth of Microcystis at initial concentrations below 2 mg / 1, but inhibited it above 2 mg / 1. Ce4+ accelerated the growth of Microcystis at initial concentrations below 0.2 mg / 1 and inhibited at above 0.2 mg /l. Furthermore, the stimulation weakened with the increase of initial concentrations of La3+ and Ce4+. In the combined system, the growth of Microcystis was accelerated in the over all cases. In the single system, the amount of La3+ and Ce4+ uptake was more at higher initial concentrations than at lower ones. At the same initial concentrations, La3+ and Ce4+ uptake in the combined system was less than that in the single system. The kinetic process of La3+ and Ce4+ adsorpted by Microcystis can be explained with the second order kinetics adsorption model.

The effects of fresh and decomposed Microcystis aeruginosa on cladocerans from a subtropic Chinese Lake

We studied in the laboratory the population growth rates of four cladocerans fed both with decomposed Microcystis aeruginosa and with a mixture of fresh colonial M. aeruginosa and Scenedesmus obliquus. The neonates of Diqphanosoma brachyurum and Daphnia carinata were able to develop into adults when they were fed with <64mum decomposed M. aeruginosa, while those of Moina micrura could not use decomposed M. aeruginosa. The population growth rate of the largest species, D. carinata, was less affected by the presence of fresh colonial M. aeruginosa than the other three species. D. carinata obtained the highest growth rate at a biomass level of 10 mg L-1 fresh colonial M. aeruginosa, indicating that, to some extent, it can use colonial M. aeruginosa at a size range of 64-112mum. The population growth rate of M. micrura was negatively correlated with fresh colonial M. aeruginosa within a range of 10-100 mg L-1. The population growth rates of D. brachyurum and Ceriodaphnia cornuta were remarkably decreased by fresh colonial M. aeruginosa, although no significant difference was found within the M. aeruginosa biomass range of 10-100 mg L-1 for either cladoceran. At a biomass level of 50 mg L-1 M. aeruginosa, the population growth rates of the four cladocerans positively correlated with S. obliquus biomass within a range of 0.1-5.0 mg L-1. Our results indicate that the zooplankton community under bloom condition is shaped by the quantity of both M. aeruginosa and other edible algae.

Detection of hepatotoxic Microcystis strains by PCR with intact cells from both culture and environmental samples

Microcystins are small hepatotoxic peptides produced by a number of cyanobacteria. They are synthesized non-ribosomally by multifunctional enzyme complex synthetases encoded by the mcy genes. Primers deduced from mcy genes were designed to discriminate between toxic microcystin-producing strains and non-toxic strains. Thus, PCR-mediated detection of mcy genes could be a simple and efficient means to identify potentially harmful genotypes among cyanobacterial populations in bodies of water. We surveyed the distribution of the mcyB gene in different Microcystis strains isolated from Chinese bodies of water and confirmed that PCR can be reliably used to identify toxic strains. By omitting any DNA purification steps, the modified PCR protocol can greatly simplify the process. Cyanobacterial cells enriched from cultures, field samples, or even sediment samples could be used in the PCR assay. This method proved sensitive enough to detect mcyB genes in samples with less than 2,000 Microcystis cells per ml. Its accuracy, specificity and applicability were confirmed by sequencing selected DNA amplicons, as well as by HPLC, ELISA and mouse bioassay as controls for toxin production of every strain used.

Effects of CO2 enrichment on the bloom-forming cyanobacterium Microcystis aeruginosa (Cyanophyceae): Physiological responses and relationships with the availability of dissolved inorganic carbon

Microcystis aeruginosa Kutz. 7820 was cultured at 350 and 700 muL.L-1 CO2 to assess the impacts of doubled atmospheric CO2 concentration on this bloom-forming cyanobacterium. Doubling Of CO2 concentration in the airflow enhanced its growth by 52%-77%, with pH values decreased and dissolved inorganic carbon (DIC) increased in the medium. Photosynthetic efficiencies and dark respiratory rates expressed per unit chl a tended to increase with the doubling of CO2. However, saturating irradiances for photosynthesis and light-saturated photosynthetic rates normalized to cell number tended to decrease with the increase of DIC in the medium. Doubling of CO2 concentration in the airflow had less effect on DIC-saturated photosynthetic rates and apparent photosynthetic affinities for DIC. In the exponential phase, CO2 and HCO3- levels in the medium were higher than those required to saturate photosynthesis. Cultures with surface aeration were DIC limited in the stationary phase. The rate of CO2 dissolution into the liquid increased proportionally when CO2 in air was raised from 350 to 700 muL.L-1, thus increasing the availability of DIC in the medium and enhancing the rate of photosynthesis. Doubled CO2 could enhance CO2 dissolution, lower pH values, and influence the ionization fractions of various DIC species even when the photosynthesis was not DIC limited. Consequently, HCO3- concentrations in cultures were significantly higher than in controls, and the photosynthetic energy cost for the operation of CO2 concentrating mechanism might decrease.

Long-term (1956-1999) dynamics of phosphorus in a shallow, subtropical Chinese lake with the possible effects of cyanobacterial blooms

This paper describes the long-term dynamics of phosphorus concentrations in both the lake water and the sediment in a subtropical Chinese lake, Lake Donghu. The total phosphorus (TP) concentration in the lake water experienced an upward trend from the 1950s, and peaked in 1983/1984, but declined obviously afterwards. From the 1950s to the 1990s, TP content in the upper 10 cm sediment of the lake increased steadily from 0.307 to 1.68 mg Pg DW-1 at Station I and from 0.151 to 0.89 mg Pg DW-1 at Station II, respectively. The TP increase in the lake water before mid-1980s was mainly attributed to the massive input of sewage P. The outbreak of cyanobacterial blooms coincided with the peaks of TP and Orthophosphate (PO4-P) in the water in mid-1980s, and the maximum TP of the water reached as high as 1.349 mg/1 at Station I and 0.757 mg/l at Station II (in 1984), respectively. The declines of TP and PO4-P in the water after mid-1980s was coincident with the disappearance of cyanobacterial bloom. (C) 2001 Elsevier Science Ltd. All rights reserved.

溶藻菌群对铜绿微囊藻的溶藻效应及其作用机理研究

水华暴发是一个世界性的问题，近年来在发展中国家显得尤其严重。水华暴发给环境和公众健康带来巨大灾难，一些蓝藻产生的毒素可以造成鱼类、鸟禽和家畜的死亡，而臭名昭著的微囊藻产生的微囊藻毒素更是有强烈致癌效应。因此，寻找控制水华藻类的有效方法非常迫切。在利用物理和化学方法处理不甚理想的情况下，利用溶藻细菌控藻成为一个新的研究方向。溶藻细菌一般直接从富营养化水体中分离，杀藻活力对有害蓝藻具有较强的选择性而不危害其它生物，尤其适合在水华发生初期使用，可以在短时间内达到阻止藻类增殖的效果。本研究富集分离到一个高效溶解铜绿微囊藻的溶藻菌群，对其溶藻效应和溶藻机制进行了探索研究。 1溶藻菌群的富集筛选及其溶微囊藻效果 富集筛选得到一个有明显抑藻效果的菌群，它对铜绿微囊藻有显著溶藻效果。与对照组相比，加入富集的溶藻菌后，第4 d开始出现溶藻现象，6~8 d出现明显的溶藻效果，8 d后测得叶绿素去除率在85%以上。 2 溶藻菌群的作用范围及溶藻特性 富集分离到的溶藻菌群对铜绿微囊藻和念珠藻有显著溶藻作用，对水华微囊藻和其它几株受试微囊藻没有明显溶藻效应。该溶藻菌群不仅可以在液体中溶解铜绿微囊藻，生长在固体平板上的藻苔也有一定的溶藻效应，生成溶藻空斑。保证快速溶藻的最大稀释度可以达到1/100, 000。 3 环境因子对菌群溶藻效力的影响 试验发现，不同的pH、温度、和光照条件下，溶藻菌群溶藻效力明显不同，且不同种类的氮源对其溶藻作用也有一定影响。这些条件对该菌群溶藻作用的影响，在相当的程度上可能取决于它们对藻和细菌两者的生长状况的影响综合。 4 溶藻菌群的溶藻作用机理 溶藻菌液过滤除菌和煮沸灭菌处理后溶藻液，未见明显的溶藻效果，只有原液具有很好的溶藻效果。因此可初步确定，蓝藻细胞的溶解可能是由溶藻菌直接接触藻细胞产生的作用效果。显微镜观察发现，细菌在溶藻的过程中频繁地接触藻细胞并侵入藻细胞，破坏进而裂解杀死藻细胞。这也进一步说明了此溶藻菌是通过直接方式杀藻。 5 溶藻菌群的菌群结构解析 分离有溶藻效果的纯菌的多次尝试都没有成功。结合DGGE和16S rDNA文库综合分析发现：Rubritepida菌，假单胞菌和鞘氨醇单胞菌是存在于铜绿微囊藻中的三种伴生细菌。加入富集的溶藻菌群后，菌群结构发生明显的变化，Rubritepida菌、假单胞菌消失，混合菌群则包含未培养黄杆菌，鞘氨醇单胞菌和噬氢菌，其中黄杆菌是优势菌群，并且细菌种群结构的变化与藻细胞消亡之间有显著的相关性。通过菌种的分离鉴定与DGGE和16S rDNA文库的测序结果比较，一些未培养菌可能在溶藻过程中起重要调控作用。 6 溶藻细菌控藻应用基础 (1) 扩大规模的模拟水华实验进一步确定了细菌对微囊藻的强烈溶解作用。 (2) 铜绿微囊藻(Microcystis aeruginosa 905, zc)、微囊藻(Microcystis spp., zd)和溶藻菌群共培养试验表明，zc可以抑制zd生长，而溶藻菌群可以溶zc。 本研究是第一次报道混合菌群的溶藻效应。该溶藻菌群对带有藻际细菌的铜绿微囊藻具有高效的溶藻效力，表明它对自然界中存在的带菌铜绿微囊藻和其它一些蓝藻的生消具有一定的控制作用。对进一步研究菌藻关系与生态学作用，以及对富营养化湖泊和水库水体中蓝藻暴发的防控，该菌群具有一定的应用潜力。 Cyanobacterial blooms break out frequently all over the world, especially in developing countries. Blooms create enormous disasters to public health and to the environment. Some cyanobacterial blooms produce extremely toxic substances that have killed fish, domestic animals and birds. It has been well known that microcystins, a hepatoxin produced by Microcystis, can promote tumors in humans. So it is very important to find an effective method for controlling the growth of the bloom-forming algae. Measures for controlling such kind of algae include physical, chemic and biologic means, but the former two may damage the aquatic environment and require high-energy inputs. The alternative approach for the elimination of nuisance algae involves the application of algicidal bacteria. The algicidal bacteria, which are nontoxic to other organisms and most of which are isolated from the eutrophic lake in situ, may be potential microbial algaecides. In the initial stages of the water blooms, they are able to restrain the biomass or multiplication of the bloom-forming algae in a short time. In order to use algicidal bacteria to suppress blooms of M. aeruginosa, we isolated a bacterial culture capable of lysing the noxious cyanobacteria M. aeruginosa. In this paper we described some properties of the bacterial culture and its growth-inhibiting or algicidal effects on the growth of M. aeruginosa, and investigated its algicidal mechanisms. 1 Enrichment of a microbial culture that lyses Microcystis aeruginosa A mixed bacterial culture was isolated from a hypereutrophic pond and showed significant algicidal activity against the noxious Microcystis aeruginosa. Algae lysis would be seen obviously 4 days later when the algae culture was killed and became yellow contrast to no-addition controls, and chlorophyll a (chl-a) reduction went beyond 85% 8 days later. 2 The host range and some other algicidal feature of the mixed algicidal culture. Microcystis aeruginosa, Nostoc sp., were susceptible to the mixed algicidal culture, while the lytic effects of this mixed culture on Microcystis flos-aquae and some other tested Microcystis were feeble.The algicidal culture can not only lyse M. aeruginosa in liquid media, but aslo lyse M. aeruginosa lawns on soft agar plates and form plaques. The maximun dilution of the mixed culture required for rapid Microcystis lysis is 1/100, 000. 3 Influences of environmental factors such as pH, temperature, illumination, and the nitrogen source on the lytic activity of the mixed bacterial culture on Microcystis aeruginosa. In our investigations, it was shown that the lytic activity of the mixed bacterial culture on Microcystis aeruginosa was straightly correlated with pH, temperature, illumination, as well as the nitrogen source in the medium. The impacts of these environmental factors on the algicidal activity of the mixed bacterial culture, to a certain extent, may depend on both the algal and the bacterial growth rates under the tested environmental conditions. 4 The mechanisms of algal cell lysis by the algicidal bacteria Death was detected when the mixed bacterial culture was added to the algal culture, but not when only the culture filtrate or autoclaved bacterial culture was added. This indicates that the mixed bacterial culture did not release extracellular products inhibitory to Microcystis aeruginosa. In addition, under the microscope, we observed frequent contacts btween bacteria and algae cells, and some bacteria can even penetrate into target algal cells and destroyed them. These results may suggest that the bacterium kill the alga by direct contact. 5 Molecular Characterization of the algicidal bacterial culture Attempts for isolation of pure bacterium or bacteria from the enrichment culture responsible for Microcystis lysis have so far been failed. Based on PCR-DGGE (denaturing gradient gel electrophoresis) and 16S rDNA clone library analysis, Rubritepida sp., Pseudomonas sp. and Sphingomonas sp., as accompanying bacteria, were existed in M. aeruginosa. The bacterial community in M. aeruginosa showed significant change after adding the enrichment culture, where uncultured Flavorbacterium sp., Sphingomonas sp. and Hydrogenophaga sp. were observed, and the uncultured Flavorbacterium sp. became a dominant species. The obvious correlation can be seen between change of bacterial population and extinction of M. aeruginosa. Compared identification of pure bacterium with sequencing of DGGE bands and the clone distribution of the clone libraries, it was inferred that some uncultured bacteria were probably play an important role in controlling the growth and abundance of M. aeruginosa. This report is the first example of a mixed bacterial culture with the ability to lyse M. aeruginosa. 6 Further study for algae control by applications of algicidal bacteria (1) Algae lysis would be seen obviously 6 days later when the algae culture was killed and became yellow contrast to no-addition controls, and chlorophyll a (chl-a) was reducted to a low level 20 days later in the simulated water bloom experiments. (2) The growth of Microcystis sp. (zd) was restrained by Microcystis aeruginosa 905 (zc) when they were co-cultured together, and zc was lysed by the algicidal bacterial culture. This report is the first example of a mixed bacterial culture with the ability to lyse M. aeruginosa, and its algicidal activity remained high against non-axenic tested M. aeruginosa, suggesting that bacteria in the natural environment could play a role in controlling the growth and abundance of M. aeruginosa and other cyanobacteria. Such bacteria could also potentially be used as agents to prevent the mass development of cyanobacteria in eutrophic lakes and reservoirs.

Ulva diversity in the Yellow Sea during the large-scale green algal blooms in 2008-2009

P>In the Yellow Sea of China, large-scale green tides have broken out for three consecutive years from 2007 to 2009. As part of the efforts to localize the algal source, two cruises were conducted in the early stage and the outbreak stage of the bloom in 2009. We analyzed the morphological and genetic diversity of drifting Ulva specimens and culture-derived isolates from seawater sampled in different localities. For phylogenetic analyses, the nuclear encoded ribosomal DNA internal transcribed spacer region (ITS nrDNA) and the plastid encoded large subunit of ribulose-1, 5-bisphosphate carboxylase/oxgenase gene (rbcL) were used. Our molecular and morphological data indicate that the dominant free-floating Ulva species in 2008 and 2009 possibly belonged to a single strain of the U. linza-procera-prolifera (LPP) clade. The ITS sequences from bloom-forming algal samples with dense branches were identical to those from U. linza-like specimens without branches derived from the Yellow Sea. Microscopic individuals of the dominant Ulva strain were detected in eight stations, revealing that spore dispersal in the water helped to enlarge biomass in the water during the outbreak stage of green tide in the Yellow Sea.

Relationships between nitrogen and phosphorus forms and ratios and the development of dinoflagellate blooms in the East China Sea

During late spring and early summer of 2005, large-scale (> 15 000 km(2)), mixed dinoflagellate blooms developed along the the coast of the East China Sea. Karenia mikimotoi was the dominant harmful algal bloom species in the first stage of the bloom (late May) and was succeeded by Prorocentrum donghaiense approximately 2 wk later. Samples were collected from different stations along both north-south and west-east transects, from the Changjiang River estuary to the south Zhejiang coast, during 3 cruises of the Chinese Ecology and Oceanography of Harmful Algal Blooms Program, before and during the bloom progression. Nitrogen isotope tracer techniques were used to measure rates of NO3-, NH4+, urea, and glycine uptake during the blooms. High inorganic nitrogen (N), but low phosphorus (P) loading from the Changjiang River led to high dissolved inorganic N:dissolved inorganic P ratios in the sampling area and indicate the development of P limitation. The rates of N-15-uptake experiments enriched with PO43- were enhanced compared to unamended samples, suggesting P limitation of the N-uptake rates. The bloom progression was related to the change in availability of both organic and inorganic N and P. Reduced N forms, especially NH4+, were preferentially taken up during the blooms, but different bloom species had different rates of uptake of organic N substrates. K mikimotoi had higher rates of urea uptake, while P. donghaiense had higher rates of glycine uptake. Changes in the availability of reduced N and the ratios of N:P in inorganic and organic forms were suggested to be important in the bloom succession. Nutrient ratios and specific uptake rates of urea were similar when compared to analogous blooms on the West Florida Shelf.

Screening of surfactants for harmful algal blooms mitigation

Screening experiments were conducted in order to find promising synthetic surfactants for harmful algal blooms (HABs) mitigation. The chemically synthesized surfactant cocamidopropyl betaine (CAPB) showed characteristics of relatively high inhibition efficiency, high biodegradability and low cost. The motility inhibition ratios of 10 mg/L CAPB on Cochlodinium polykrikoides and Alexandrium tamarense were about 60% after 5 min. The biodegradation test indicated that the half-life of CAPB in seawater was shorter than one day and 90% was biodegraded after five days under the initial concentration of 100 mg/L at 25degreesC. Further cell lysis experiments revealed the selective lysis effect of CAPB on different HAB organisms. More than 90% of C. polykrikoides lysed at the concentration of 10 mg/L CAPB after 24 h and at 15 mg/L CAPB after 4 h, whereas the lysis effect of CAPB on A. tamarense was slight, no more than 10% after 2 h interaction with 50 mg/L CAPB. This research provided preliminary data for CAPB as a candidate in harmful algal blooms mitigation and pointed out unresolved problems for its practical application in the meantime. (C) 2003 Elsevier Ltd. All rights reserved.

Synergistic effect of sophorolipid and loess combination in harmful algal blooms mitigation

The inhibition effect of sophorolipid and removal efficiency of loess on Cochlodinium polykrikoides and Alexandrium tamarense was investigated separately in the laboratory. Based on this, the combination of sophorolipid and loess for harmful algal bloom mitigation was proposed. Algal sedimentation tests in the laboratory and in the field revealed that the combination of sophorolipid and loess showed synergistic effects both on the removal efficiencies and on the mitigation cost. The concentration of 1 g/l loess and 5 mg/l sophorolipid was determined as the optimum ratio for C polykrikoides mitigation. In the field test, the effective concentration of loess and sophorolipid in the combination group was reduced to 10% and 25%, respectively, compared to the non-combination group, and the cost decreased more than 60%. The combination of loess and sophorolipid was considered as a promising novel method in harmful algal bloom mitigation. (C) 2003 Elsevier Ltd. All rights reserved.

Temporal and spatial variability in nitrogen uptake kinetics during harmful dinoflagellate blooms in the East China Sea

Temporal and spatial variability in the kinetic parameters of uptake of nitrate (NO3-), ammonium (NH4+), urea, and glycine was measured during dinoflagellate blooms in Changjiang River estuary and East China Sea coast, 2005. Karenia mikimotoi was the dominant species in the early stage of the blooms and was succeeded by Prorocentrum donghaiense. The uptake of nitrogen (N) was determined using N-15 tracer techniques. The results of comparison kinetic parameters with ambient nutrients confirmed that different N forms were preferentially taken up during different stages of the bloom. NO3- (V-max 0.044 h(-1); K-s 60.8 mu M-N) was an important N source before it was depleted. NH4+ (V-max 0.049 h(-1); K-s 2.15 mu M-N) was generally the preferred N. Between the 2 organic N sources, urea was more preferred when K. mikimotoi dominated the bloom (V-max 0.020 h(-1); K-s 1.35 mu M-N) and glycine, considered as a dominant amino acid, was more preferred when P. donghaiense dominated the bloom (V-max 0.025 h(-1); K-s 1.76 mu M-N). The change of N uptake preference by the bloom-forming algae was also related to the variation in ambient N concentrations. Published by Elsevier B.V.

改性当地岸沙治理有害赤潮的研究

本论文从生态环境可持续发展观出发，选用了两种天然高分子聚合物壳聚糖与黄原胶作为改性剂，考察了它们在海水体系中的絮凝性能以及改性当地岸沙絮凝除藻的性能，同时探讨了赤泥复合剂絮凝除藻的性能。研究结果表明： 海水体系中的离子强度阻碍了壳聚糖高分子链的舒展，有效除藻时投加量大于50 mg/L；聚合氯化铝的加入，降低了壳聚糖的投加量，并且10 mg/L壳聚糖与10 mg/L聚合氯化铝协同对100 mg/L当地岸沙进行改性，改性后沙子具有较强的除藻效果，3 min后，强壮前沟藻与海洋小球藻的去除率为80%，沉淀4 h后，两种藻的去除率高达92-96%。 黄原胶单独使用时强壮前沟藻的去除率为32%-55%；氢氧化钙的加入，提高了黄原胶的絮凝活性；当黄原胶:氢氧化钙:土壤/沙子=1:5:15时，黄原胶投加量为20 mg/L时，30 min后，强壮前沟藻的去除率为83%-89%。 赤

滇池铜绿微囊藻(M．aeruginosa zosa Kutz)的分离培养与总DNA提取的改进

利用稀释法结台平板分离培养法将采自滇池水华中的铜绿微囊藻进行了成功的 分离纯化对比纯化培养前后，发现纯化后该藻的生长速度明显加快，密度大为提高，约为纯化前 的5倍用乙醇乙醚混合液剐该藻作预处理后，破坏了其胶质被．再经常规方法提取总DNA，其得 率提高3 4倍