紫菜壳孢子萌发过程及其世代差异研究

紫菜是一类海洋藻类研究的模式系统，具有重要的经济价值和理论研究意义。本研究基于紫菜的世代交替生活史，对其壳孢子萌发过程进行了研究，并对丝状孢子体与叶状配子体的世代差异进行了分析，主要包括以下三部分： 1） 对紫菜丝状孢子体与叶状配子体的连接枢纽——壳孢子的发育过程及其光影响进行了研究与讨论，发现壳孢子只有在附着后才能形成细胞壁并发育，说明壳孢子的附着是触发了壳孢子细胞壁的形成以及后期发育的开关，亦即附着是触发紫菜世代交替过程中配子体转录组表达的开关；纤维素酶和果胶酶均能抑制壳孢子附着，但影响机制各不相同，推测果胶质主要介导壳孢子的初始附着，而纤维素则与永久附着相关；波长≥580 nm的高强度（200 μmol•m-2•s-1）可见光有利于壳孢子早期发育。 2） 结合现有藻类数据，对坛紫菜丝状孢子体阶段11000 EST数据进行了大规模的生物信息学分析，结果首次发现坛紫菜丝状孢子体中可能存在PCK型C4光合固碳途径，并筛选出44条在紫菜孢子体中表达上调的代表基因。 3） 结合坛紫菜、条斑紫菜、海带和红毛菜，对红藻和褐藻等大型海藻孢子体与配子体阶段代表基因Rubisco的表达与羧化酶活性差异进行了研究分析，结果表明Rubisco的表达量和初始羧化酶活在其配子体中均显著高于其孢子体世代，即与藻体不同世代的相对复杂度无关，而与染色体倍性相关，说明Rubisco的世代差异极有可能与染色体倍性相连锁，因而可能是海藻世代交替过程中的重要功能基因。

藻体对水环境中N、P及重金属Cu2+、Pb2+、Cd2+、Cr6+的吸附特征研究

随着全球生态环境的恶化，各国日益重视对水体中各种污染物的治理。利用藻类的吸收、富集和降解作用，可以去除污水中的营养物质、重金属离子和有机污染物，与其他物理、化学及工程的方法相比，该技术具有以下优点：成本低、能耗小、治理效果较好，对环境污染小，有利于资源化，有利于整体生态环境的改善，是治理水质污染的新途径。 本文利用几种大型海藻对富营养化海水进行处理，结果发现孔石莼、刚毛藻均有很强的吸收N、P的能力，吸收能力依次为褶曲刚毛藻>束生刚毛藻>孔石莼。水体中褶曲刚毛藻3 g/L含量，在3～5小时之内，可把中等以上富营养化海水中的N、P降低至一类海水水平。利用刚毛藻处理富营养化地下海水和养殖废水，进行海参和大菱鲆养殖试验，探索藻类净化水质和废水循环利用的新模式，使水体保持较低的营养盐状态，减轻养殖废水对环境的影响，实现了海水养殖业与环境的可持续发展。 刚毛藻在我国近海滩涂分布广泛，利用它来处理富营养化水体，并和水产养殖业相结合，既净化水体，使养殖废水能循环利用，满足水产养殖的需求，又改善水产业生态环境。同时，将回收藻体生产优质饲料、食品和药物等，实现藻类资源的高值利用。刚毛藻营养丰富，用其替代鼠尾藻作海参饲料，资源丰富，成本低，效果好，是一种值得加以开发利用的宝贵资源，具有广泛的应用前景。 生物吸附法是一种经济有效的移除废水中有害重金属离子的方法。由于藻类细胞壁中的多聚糖可提供吸附重金属的位点，廉价而蕴藏丰富的海藻对多种重金属表现出很强的吸附能力。所以本文通过分批实验，研究了非活体刚毛藻对水体中重金属Cu2+、Pb2+和Cd2+的吸附影响因子、吸附热力学、吸附动力学及吸附机理，得到了平衡等温线及动力学数据。吸附过程的最佳pH值为5.0，吸附量随温度的升高而增加，水体中常见的Na+、K+、Ca2+、Mg2+阳离子及Cl-、NO3-、SO42-、C2O42-等阴离子的存在对吸附的影响并不显著。EDTA存在时，吸附百分率大大降低。吸附等温线符合Langmuir和Freundlich方程。刚毛藻对重金属Cu2+、Pb2+和Cd2+的吸附容量很高，25℃时，对Cu2+、Pb2+和Cd2+的最大吸附容量分别为1.61 mmol/g、0.96 mmol/g和0.98 mmol/g，且吸附过程为吸热反应。刚毛藻对重金属Cu2+、Pb2+和Cd2+的吸附过程为化学吸附，在吸附过程中藻体表面的官能团可能与金属离子发生了螯合作用。吸附动力学过程符合pseudo-二级动力学模型，在初始的30min内，吸附速率很快，随后速率逐渐降低。解吸试验表明，用EDTA可以对重金属进行回收，刚毛藻可以循环利用。实验结果表明刚毛藻是一种高效、经济实用的生物吸附材料，可用来吸附回收水体中的重金属Cu2+、Pb2+和Cd2+等。 通过非活体刚毛藻对重金属Cr6+的吸附影响因子、吸附动力学、吸附机理的研究发现，刚毛藻对Cr6+具有很强的还原能力，对电镀废水中的Cr6+的还原去除提供了非常好的方法。吸附过程的最佳pH值为2～3，实际电镀废水通常在此pH范围，因此处理实际废水时，首先在原酸性条件下，对Cr6+进行还原去除，然后调废水pH至5.0，继续进行吸附，去除其他二价离子及被还原的三价Cr离子，实现了利用同一材料还原Cr6+为Cr3+，并将Cr3+和其他重金属离子同时去除。通过对机理的讨论，认为刚毛藻对Cr6+的生物吸附过程不是一个简单的“离子交换过程”，而是一个“吸附还原过程”。在海藻量足够的前提下，只要时间足够长，Cr6+可被彻底还原去除。 利用工业废弃物褐藻渣，对水体中重金属离子Cu2+、Pb2+、Cd2+及Cr6+的生物吸附特性分别进行了讨论，结果表明褐藻渣对重金属离子的吸附特性与刚毛藻一致，吸附等温线符合Langmuir和Freundlich方程，在25℃时，pH为5.0时，由Langmuir方程求出褐藻渣对Cu2+、Pb2+和Cd2+的最大吸附容量分别为4.20 mmol/g、3.13 mmol/g和2.97 mmol/g。褐藻渣对低、高浓度的重金属Cr6+都具有很强的吸附能力，且移除效果比较彻底。实际应用结果表明，褐藻渣是一种高效、经济实用的生物吸附材料，可用来吸附回收水体中的重金属离子，具有广泛的应用前景。

海洋微藻产生溶解有机物及东海溶解有机物三维荧光特征研究

本文通过胶州湾围隔实验、微藻培养实验研究了海洋微藻产生溶解有机物的三维荧光特性，并初步利用经验正交函数对三维荧光光谱图进行了主成分分析，通过东海取样研究了东海荧光溶解有机物的时空分布特征，并对荧光溶解有机物的来源进行了分析。得到了一些初步结论： 胶州湾围隔实验中不同营养盐培养结果表明浮游植物可产生类蛋白和类腐殖质荧光，类蛋白荧光峰由类酪氨酸(tyrosine-like)荧光峰和类色氨酸(tryptophan-like)荧光峰组成，主要位置为Exmax/Emmax=270nm/290~310nm，Exmax/Emmax= 270~290/320~350的荧光峰强度比较弱；在Exmax/Emmax=250~260/380~480nm(A峰)、Exmax/Emmax=310~320/380~420nm(C峰)和Exmax/Emmax=330~350/420~480nm(M峰)位置均出现零散的类腐殖质荧光峰，其中以A峰为主。类酪氨酸荧光强度明显高于类腐殖质荧光强度。浮游植物量降低时，类酪氨酸荧光强度与叶绿素a浓度呈明显的负相关。硅藻和甲藻产生的类酪氨酸和类色氨酸荧光强度之间具有较好的相关性，两者来源相似, 并且甲藻与硅藻相比能够产生更多的类蛋白荧光物质。不同环境下类腐殖质混合物的组分比例不同，甲藻生长环境下相对于硅藻具有较低的A:C比值。 在实验室培养中肋骨条藻（Skeletonema costatum）、塔玛亚历山大藻（Alexandrium tamarense）、微小亚历山大藻（Alexandrium mimutum）、锥状斯氏藻（Scrippsiella trochoidea）、东海原甲藻（Prorocentrum donghaiense）及海洋原甲藻（Prorocentrum micans）的实验结果表明，微藻在生长过程中产生出荧光溶解有机物，中肋骨条藻为代表的硅藻主要产生类腐殖质荧光物质，而甲藻在指数增长期主要产生类蛋白荧光物质。进入消亡期后类蛋白荧光和类腐殖质荧光强度迅速增大，原因可能是衰老、死亡藻细胞的破碎释放出大量的荧光有机物质所致，此外还有细菌对非荧光有机物进一步降解。塔玛亚历山大藻、微小亚历山大藻、东海原甲藻及海洋原甲藻的类蛋白荧光强度在消亡后期由于细菌降解或光降解等因素而降低。同属微藻产生的荧光物质相似，例如塔玛亚历山大藻与微小亚历山藻、东海原甲藻与海洋原甲藻，但具体荧光峰位置有所不同。利用经验正交函数能够对三维荧光光谱谱图进行主成份分析。 在利用三维荧光光谱法研究长江口海域台风前后不同站位荧光溶解有机物荧光特性及分布特征的结果表明，长江口海区主要的荧光溶解有机物荧光峰为T峰、S峰和A峰。风前和风后的类色氨酸分别来源于相似物质。风前，在表层浮游植物能够产生类蛋白荧光物质，而底层类蛋白荧光物质不受浮游植物的影响，长江冲淡水能够带来部分类蛋白荧光物质；表层的类腐殖质不受浮游植物的影响，而底层的浮游植物在降解过程中能够产生一部分类腐殖质，并且长江冲淡水对表层和底层的类腐殖质来源均有很大贡献。风后，表层的类色氨酸与叶绿素a浓度不呈相关性，而底层却呈正相关，另外表层和底层的类色氨酸均受到长江冲淡水的影响；表层和底层的类腐殖质与叶绿素a均没有相关性，但受陆源影响显著，长江冲淡水能带来类腐殖质。

两株海藻内生真菌次生代谢产物及其生物活性研究

海洋微生物拥有丰富多样的次生代谢途径，其中海洋生物内生真菌次生代谢产物研究日益受到天然产物化学界的重视。本论文以菌丝体生物量、发酵产物重量、抗菌与细胞毒活性、薄层色谱分析结果以及高效液相色谱分析结果等为评价依据对采自青岛沿海的13株海藻内生真菌在四种液体培养基上的静置发酵产物进行了综合评价，并从中选择了黑曲霉Aspergillus niger EN-13（分离自褐藻囊藻Colpomenia sinuosa）和杂色曲霉A. versicolor EN-7（分离自褐藻鼠尾藻Sargassum thunbergii）两株真菌进行了30升规模发酵（分别采用GPYM培养基和PDB培养）和化学成分的研究，对分离得到的大部分化合物进行了初步的生物活性筛选。 发酵提取物采用常规的硅胶柱层析、反相硅胶柱层析，凝胶Sephadex LH-20柱层析、制备薄层层析、半制备高效液相色谱以及重结晶等分离手段，得到单体化合物。利用各种现代波谱技术(IR、UV、EI-MS、FAB-MS、HR-ESI-MS、1H-NMR、13C-NMR、DEPT、1H-1H COSY、HSQC、HMBC等)并结合化学方法从两种菌株发酵提取物中鉴定了55个化合物的结构。其中从菌株A. niger EN-13分离鉴定了31个化合物，发现9个新化合物，包括2个鞘酯类化合物（AN-1~2）、3个萘并-γ-吡喃酮类化合物（AN-3~5）、3个苯乙基取代的α-吡喃酮类化合物（AN-17, AN-19~20）和1个甾体Diels-Alder加成产物（AN-21），另有1个新的天然环二肽（AN-27）被分离鉴定；从菌株A. versicolor EN-7分离鉴定了24个化合物，发现2个新化合物，为蒽醌AV-12与AV-17，另外，从前一菌株（A. niger EN-13）中鉴定的2个新鞘酯类化合物（AN-1~2）在A. versicolor EN-7中也被再次分离到。 对大部分单体化合物进行了抗菌活性、DPPH自由基清除活性和细胞毒活性测试。结果显示新化合物AN-1、AN-5和AN-20具有弱或中等强度的抑制白色念珠菌生长的活性，AN-4、AN-5、AN-21显示了弱或中等强度的抑制黑曲霉生长的活性，AV-12、AV-17显示了弱的抑制大肠杆菌生长的活性。在DPPH自由基清除活性筛选中，AN-5显示了中等强度的活性，其EC50为109.3 mM，与阳性对照BHT相近（EC50为81.8 mM）。其它部分已知化合物在抗菌和DPPH自由基清除活性的筛选中也显示了弱或中等强度的活性。在针对人肝癌细胞株SMMC-7721和人肺腺癌细胞株A549的体外细胞毒活性筛选中，所测样品均未显示显著活性。

三种海洋红藻和两株放线菌次级代谢产物研究

海洋是一个巨大的天然产物宝库，约占地球表面积70%的海洋蕴藏着80%的生物资源。由于海洋生态环境的特殊性，导致海洋生物能够产生大量结构独特多变和活性特殊多样的代谢产物。我国海域辽阔，海洋资源丰富，为寻找结构新颖、生理活性独特的先导化合物，加强对海洋资源的开发利用，本论文对中国沿海的三种海洋红藻和两株放线菌次生代谢产物以及生物活性进行研究，为新药研究与开发提供模式结构和药物前体。 对红藻似瘤凹顶藻Laurencia similis乙酸乙酯萃取物进行分离纯化，从中得到单体化合物35个，通过波谱学方法（IR、MS、NMR等）鉴定了他们的结构。分别为：2, 2, 5, 5, 6, 6-sixibromo-3, 3-bi-1H-indole (1)，3,5-dibromo- 1-methyl-indole (2)，3',5',6,6'-tetrabromo-2,4-dimmethyldiphenyl ether (3)，1,2,5- tribromo-3-bromoamino-7-bromomethylnaphthalene (4)，2,5,8-tribromo-3-bromo- amino-7-bromomethylnaphthalene (5)，2,5,6-tribromo-3-bromoamino-7-bromo- methylnaphthalene (6), 2,5,6,5',6'-pentabromo-3,4,3',4'-tetramethoxybenzophenone (7), (4E)-1-bromo-5-[(1'S*,3'R*)-3'-bromo-2',2'-dimethyl-6'-methylenecyclohexyl] -3-methylpent-4-ene-2,3-diol (8)，4-hydroxy-Palisadin C (9)，Isopalisol (10)，Luzonensol (11)，Palisadin B (12)，Aplysistatin (13)，Palisadin A (14)，5-Acetoxypalisadin B (15)，Aristolan-1(10)- en-9-ol (16)，Aristol-8-en-1-one (17)，Aristolan-9-en-1-one (18)，Aristolan-1(10)-en- 9-one (19)，Aristofone (20)，Aristolan-1(10)-8-diene (21)，Aristolan-1,9-diene (22)，10-Hydroxyaristolan-9-one (23)，7,11,15-trimethyl-3-methylene-hexadecan-1,2-diol (24)，3β-Hydroxyergosta- 5,24(28)-dien-7-one (25)，Isofucosterol (26)，β-sitosterol (27)，豆甾-4-烯-3α,6β-二醇 (28)，Cholesta-5-en-3β-ol (29)，Stigmasterol (30)，2,3,5,6-四溴-吲哚 (31)，2,3,6-tribromo-1H-indole (32)，3,5,6-tribromo-1-methylindole (33)，3,5,6-tribromo -1H-indole (34)，2,3,5-tribromo-1-methylindole (35)，其中化合物1-9为新化合物，化合物10-15、20和化合物24-30均为首次从该种海藻中得到。对新化合物1-9进行PTP1B酶抑制剂活性筛选，新化合物1、3、7显示强的PTP1B酶抑制活性。 对红藻齐藤凹顶藻Laurencia saitoi乙酸乙酯萃取物进行分离纯化，从中得到单体化合物11个，通过波谱学方法（IR、MS、NMR等）鉴定了他们的结构，分别为：2-hydroxyl-Luzofuranone (1)，2-hydroxyl-Luzofuranone B (2)，4-hydroxyl-Palisudin C (3)，2-bromo-γ-ionone (4)，Aplysistatin (5)，5-Acetoxypalisadin B (6)，Palisadin B (7)，Palisadin A (8)，Pacifigorgiol (9)，豆甾-4-烯-3α,6β-二醇 (10)，2, 3, 5, 6-四溴-吲哚 (11)，其中化合物1-4为新化合物，所有化合物均为首次从该种海藻中得到。通过MTT法对分离得到的新化合物1-4进行肿瘤细胞毒活性筛选，结果显示4个新化合物对所测肿瘤细胞株均无明显的活性。 对红藻瘤状软骨凹顶藻Chondrophycus papillous乙酸乙酯萃取物进行分离纯化，从中得到单体化合物5个，通过波谱学方法（MS、NMR等）鉴定了他们的结构，分别为邻苯二甲酸二丁酯 (1)，邻苯二甲酸二异辛酯 (2)，胆甾醇 (3)，3,7,11,15-tetramethyl-hexadec-2-en-1-ol (4)，4-羟基苯甲醛 (5)，所有化合物均为首次从该种海藻中得到。 对海洋放线菌M159乙酸乙酯萃取物进行分离纯化，从中得到单体化合物13个，通过波谱学方法（MS、NMR等）鉴定了他们的结构，分别为：5-(4',6'-dihydroxy-6-methyloctyl)furan-2(5H)-one (A)，phenethyl alcohol (1)，4-羟基苯甲醛(2)，anthranilic acid (3)，4-Hydroxy-3-methoxy- phenyl-propionic acid (4)，5-(6,7-dihydroxy-6-methyloctyl)furan-2(5H)-one (5)，p-Hydroxyphenylethyl alcohol (6)，3-Indoleacrylic acid (7)，Indol-3-carboxylic acid (8)，Adenine cordyceposide (9)，腺嘌呤核苷(10)，尿嘧啶核苷(11)，Thymidine (12)，其中化合物A为新化合物。所有化合物均为首次从该株放线菌中得到。 对海洋放线菌L211乙酸乙酯萃取物进行分离纯化，从中得到单体化合物15个，通过波谱学方法（MS、NMR等）鉴定了7个结构，分别为：spatozoate (1)，anthranilic acid (2)，3-Indolylethanol (3)，1-Acetyl-β-carbolin (4)，p-Hydroxyphen- ylethyl alcohol (5)，Indole-3-acetic acid (6)，Indol-3-carboxylic acid (7)，所有化合物均为首次从该株放线菌中得到。

Effects of single and mixed diatom diets on the reproduction of copepod Calanus sinicus

Under laboratory conditions, the potential influence of diatom diets on reproduction of zoo-plankton Calanus sinicus was studied. Four diatom diet ingredients: Skeletonema costatum (SC), Chaetoceros muelleri (CM), Phaeodactylum tricornutum (PT), diatom mixture (MIX) and a control diet: the flagellate Platymonas subordiformis (PS), were used at the same carbon concentrations of 2.0 mu g mL(-1) C. In a period of 17-day laboratory experiment, the effects of these algae diets on egg production and hatching success of the copepod Calanus sinicus were examined. The diets were analyzed for fatty acid content as an indicator of food quality. The results showed that the female survival of all treatments reached more than 80% except PT. Comparing to the initial value, egg production of Calanus sinicus was reduced in diatom diets (PT, CM), but remained in normal level in SC and MIX, indicating that some single diatom diets had a negative effect on the egg production of Calanus sinicus. Feeding with mixed food however can eliminate this negative effect. Among all the treatments, hatching success in filtered seawater was significantly higher than in algal exudates, indicating that not only diatoms but also other phytoplankton in certain concentration can release extracelluar substance that may inhibit eggs from hatching. Fatty acid analysis showed that both egg production rate and hatching success were negatively correlated to the ratio of 20:5 omega 3 and 14:0 in fatty acid composition.

Typhoon effects on DOC dynamics in a phosphate-limited reservoir

To explore typhoon effects on dissolved organic carbon (DOC) dynamics, field investigations (tributary and dam site) and laboratory experiments (bioassay and DOC consumption) were conducted in a subtropical reservoir. A tributary survey indicated that after typhoon disruption, upstream areas were the sources of phosphate (P) but not DOC for the dam site located downstream. Bioassay experiments verified P-limitation on bacteria and phytoplankton during summer stratification, and bacteria showed a faster response than algae to added P. Experiments indicated that DOC consumption was determined by the availability of P. The 4 yr typhoon period (June-September) data of the dam site denoted that DOC concentration (27 to 270 mu M C) and its rate of change (-13 to 24 mu M C d(-1)) varied more dramatically in the weak (2006 and 2007) than in the strong (2004 and 2005) typhoon years. The negative correlation of DOC with the ratio of bacterial production (BP) to primary production (PP) in the euphotic zone (0 to 10 m) signified the interactive effects of auto- and heterotrophic processes on DOC variation. In the aphotic zone, the variation of DOC could be ascribed to the change of BP, which showed a positive correlation with P concentrations. This study documents that DOC concentration in the studied system varied at multiple time scales. Such variation can be explained by the decoupling between BP and PP, which is believed to be a function of the limiting nutrient's availability. More importantly, this study suggests that the P supply introduced by strong typhoons might have substantiated a tighter coupling between BP and PP, so that the amplitude of DOC oscillation during the summer period was effectively reduced.

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 16S rDNA sequences of the isolates shows four obvious clusters, Halomonas venusta, Vibrio tasmaniensis, Vibrio lentus, and Vibrio splendidus. Isolates from the surface of P. urceolata are more abundant and diverse, of which strains P9 and P28 have a 16S rDNA sequence very similar (97.5%-99.8%) to that of V. splendidus. On the contrary, the isolates from the surfaces of G textorii, U. pertusa and L. japonica are quite simple and distribute on different branches of the phylogenetic tree. In overall, the results of this study indicate that the genetic relationships among the isolates are quite close and display a certain level of host species specificity, and alga-associated bacteria species are algal species specific.

Morphological and genetic study of Prorocentrum donghaiense Lu from the East China Sea, and comparison with some related Prorocentrum species

In this paper, the detailed morphology of Prorocentrum donghaiense Lu from both field samples and cultures was examined, and a taxonomic comparison was made between P donghaiense and some related Prorocentrum spp. using morphological and molecular data and other published information. There were distinct differences among these species in morphological characteristics that historically have been presented as conservative features. The discrepancies extended beyond that of individual variations within the same species due to environmental factors. Therefore, these morphological features may not be conservative but, rather, polymorphic depending on environmental conditions. Based on this analysis, we suggest that the high-biomass bloom-forming species in the East China Sea, previously reported as Prorocentrum dentatum Stein, is P donghaiense Lu. The species reported from the East China Sea and Japanese and Korean waters appear to be the same species. Molecular data also suggest that P. dentatum (CCMP1517) and P. donghaiense are genetically identical. Therefore, the geographic distribution of P. donghaiense may be much wider than expected. (C) 2004 Elsevier B.V. All rights reserved.

Experimental study on the impact of dinoflagellate Alexandrium species on populations of the rotifer Brachionus plicatilis

To investigate harmful effects of the dinoflagellate Alexandrium species on microzooplankton, the rotifer Brachionus plicatilis was chosen as an assay species, and tested with 10 strains of Alexandrium including one known non-PSP-producer (Alexandrium tamarense, AT-6). HPLC analysis confirmed the PSP-content of the various strains: Alexandrium lusitanicum, Alexandrium minutum and Alexandrium tamarense (ATHK, AT5-1, AT5-3, ATC102, ATC103) used in the experiment were PSP-producers. No PSP toxins were detected in the strains Alexandrium sp1, Alexandrium sp2. Exposing rotifer populations to the densities of 2000 cells ml(-1) of each of these 10 Alexandrium strains revealed that the (non-PSP) A. tarnarense (AT-6) and two other PSP-producing algae: A. lusitanicum, A. minutum, did not appear to adversely impact rotifer populations. Rotifers exposed to these three strains were able to maintain their population numbers, and in some cases, increase them. Although some increases in rotifer population growth following exposures to these three algal species were noted, the rate was less than for the non-exposed control rotifer groups. In contrast, the remaining seven algal strains (A. tamarense ATHK, AT5-1, AT5-3, ATC102, ATC103; also Alexandrium sp1 and Alexandrium sp2) all have adverse effects on the rotifers. Dosing rotifers with respective algal cell densities of 2000 cells ml-1 each, for Alexandrium spl, Alexandrium sp2, and A. tamarense strains ATHK and ATC103 showed mean lethal time (LT50) on rotifer populations of 21, 28, 29, and 36h, respectively. The remaining three species (A. tamarense strains AT5-1, AT5-3, ATC102) caused respective mean rotifer LT50S of 56, 56, and 71 h, compared to 160 h for the unexposed "starved control" rotifers. Experiments to determine ingestion rates for the rotifers, based on changes in their Chlorophyll a content, showed that the rotifers could feed on A. lusitanicum, A. minutum and A. tamarense strain AT-6, but could graze to little or no extent upon algal cells of the other seven strains. The effects on rotifers exposed to different cell densities, fractions, and growth phases of A. tamarense algal culture were respectively compared. It was found that only the whole algal cells had lethal effects, with strongest impact being shown by the early exponential growth phase of A. tamarense. The results indicate that some toxic mechanism(s), other than PSP and present in whole algal cells, might be responsible for the adverse effects on the exposed rotifers. (C) 2004 Elsevier B.V. All rights reserved.

A preliminary study on the mechanism of harmful algal bloom mitigation by use of sophorolipid treatment

in order to investigate a new method of mitigating the deleterious effect of harmful algal blooms (HABs), the inhibition of the glycolipid biosurfactant sophorolipid on three common harmful algae Alexandrium tamarense, Heterosigma akashiwo and Cochlodinium polykrikoides was studied. The optimum preparation condition for sophorolipid, the inhibition capability of sophorolipid and the interaction mechanism of sophorolipid on the three algal species were investigated. Results showed that sophorolipid prepared by extraction with ethyl acetate exhibited the most prominent inhibition effect and that storage time of one year had little influence on the inhibition effect of sophorolipid. The optimum concentration of 10-20 mg/l sophorolipid inhibited the motility of about 90% of the tested harmful algal cells without recovery. Investigation of the algicidal process revealed that sophorolipid induced ecdysis of A. tamarense, quick lysis of H. akashiwo and swelling of C. polykrikoides in a relatively short time. Investigation of the nucleotides showed that more than 15% of the nucleotides were released from the cytoplasm under the effect of 10-20 mg/l sophorolipid, indicating the irreversible damage on the cellular membrane, which resulted in the disintegration of the harmful algal cells. (C) 2004 Elsevier B.V. All rights reserved.

Application of Neomysis awatschensis as a standard marine toxicity test organism in China

The small mysid crustacean Neomysis awatschensis was collected in the west coast of Jiaozhou Bay, Qingdao, China in 1992 and acclimated and cultured in laboratory conditions since then. Standard acute toxicity tests using 4-6 d juvenile mysids of this species were conducted and the results were compared with Mysidopsis bahia, a standard toxicity test organism used in the US in terms of their sensitivities to reference toxins, as well as their taxonomy, morphology and geographic distributions. Because of its wide distribution along the Chinese coast, similar sensitivity to pollutants as M. bahia, short life history, small size and the case of handling, this study intended to use N. awatschensis as one of the standard marine organisms for toxicity testing in China. The species were applied to acute toxicity evaluations of drilling fluid and its additives I organotin TPT and toxic algae, and to chronic ( life cycle) toxicity assays of organotin TPT and a toxic dinofalgellate Alexandrium tamarense, respectively. Using N, awatschensis as a standard toxicity testing organism in marine pollution assessment in China is suggested.

A comparison of future increased CO2 and temperature effects on sympatric Heterosigma akashiwo and Prorocentrum minimum

Very little is known about how global anthropogenic changes will affect major harmful algal bloom groups. Shifts in the growth and physiology of HAB species like the raphidophyte Heterosigma akashiwo and the dinoflagellate Prorocentrum minimum due to rising CO2 and temperature could alter their relative abundance and environmental impacts in estuaries where both form blooms, such as the Delaware Inland Bays (DIB). We grew semi-continuous cultures of sympatric DIB isolates of these two species under four conditions: (1) 20 degrees C and 375 ppm CO2 (ambient control), (2)20 degrees C and 750 ppm CO2 (high CO2),(3) 24 degrees C and 375 ppm CO2 (high temperature), and (4) 24 degrees C and 750 ppm CO2 (combined). Elevated CO2 alone or in concert with temperature stimulated Heterosigma growth, but had no significant effect on Prorocentrum growth. P-Bmax (the maximum biomass-normalized light-saturated carbon fixation rate) in Heterosigma was increased only by simultaneous CO2 and temperature increases, whereas P-Bmax in Prorocentrum responded significantly to CO2 enrichment, with or without increased temperature. CO2 and temperature affected photosynthetic parameters alpha, Phi(max), E-k, and Delta F/F'(m) in both species. Increased temperature decreased and increased the Chl a content of Heterosigma and M Prorocentrum, respectively. CO2 availability and temperature had pronounced effects on cellular quotas of C and N in Heterosigma, but not in Prorocentrum. Ratios of C:P and N:P increased with elevated carbon dioxide in Heterosigma but not in Prorocentrum. These changes in cellular nutrient quotas and ratios imply that Heterosigma could be more vulnerable to N limitation but less vulnerable to P-limitation than Prorocentrum under future environmental conditions. In general, Heterosigma growth and physiology showed a much greater positive response to elevated CO2 and temperature compared to Prorocentrum, consistent with what is known about their respective carbon acquisition mechanisms. Hence, rising temperature and CO2 either alone or in combination with other limiting factors could significantly alter the relative dominance of these two co-existing HAB species over the next century. (c) 2007 Elsevier B.V. All rights reserved.

Ocean urea fertilization for carbon credits poses high ecological risks

The proposed plan for enrichment of the Sulu Sea, Philippines, a region of rich marine biodiversity, with thousands of tonnes of urea in order to stimulate algal blooms and sequester carbon is flawed for multiple reasons. Urea is preferentially used as a nitrogen source by some cyanobacteria and dinoflagellates, many of which are neutrally or positively buoyant. Biological pumps to the deep sea are classically leaky, and the inefficient burial of new biomass makes the estimation of a net loss of carbon from the atmosphere questionable at best. The potential for growth of toxic dinoflagellates is also high, as many grow well on urea and some even increase their toxicity when grown on urea. Many toxic dinoflagellates form cysts which can settle to the sediment and germinate in subsequent years, forming new blooms even without further fertilization. If large-scale blooms do occur, it is likely that they will contribute to hypoxia in the bottom waters upon decomposition. Lastly, urea production requires fossil fuel usage, further limiting the potential for net carbon sequestration. The environmental and economic impacts are potentially great and need to be rigorously assessed. (C) 2008 Elsevier Ltd. All rights reserved.

Is inorganic nutrient enrichment a driving force for the formation of red tides? A case study of the dinoflagellate Scrippsiella trochoidea in an embayment

Red tides (high biomass phytoplankton blooms) have frequently occurred in Hong Kong waters, but most red tides occurred in waters which are not very eutrophic. For example, Port Shelter, a semi-enclosed bay in the northeast of Hong Kong, is one of hot spots for red tides. Concentrations of ambient inorganic nutrients (e.g. N, P), are not high enough to form the high biomass of chlorophyll a (chl a) in a red tide when chl a is converted to its particulate organic nutrient (N) (which should equal the inorganic nutrient, N). When a red tide of the dinoflagellate Scrippsiella trochoidea occurred in the bay, we found that the red tide patch along the shore had a high cell density of 15,000 cells ml(-1), and high chl a (56 mu g l(-1)), and pH reached 8.6 at the surface (8.2 at the bottom), indicating active photosynthesis in situ. Ambient inorganic nutrients (NO3, PO4, SiO4, and NH4) were all low in the waters and deep waters surrounding the red tide patch, suggesting that the nutrients were not high enough to support the high chl a >50 mu g l(-1) in the red tide. Nutrient addition experiments showed that the addition of all of the inorganic nutrients to a non-red-tide water sample containing low concentrations of Scrippsiella trochoidea did not produce cell density of Scrippsiella trochoidea as high as in the red tide patch, suggesting that nutrients were not an initializing factor for this red tide. During the incubation of the red tide water sample without any nutrient addition, the phytoplankton biomass decreased gradually over 9 days. However, with a N addition, the phytoplankton biomass increased steadily until day 7, which suggested that nitrogen addition was able to sustain the high biomass of the red tide for a week with and without nutrients. In contrast, the red tide in the bay disappeared on the sampling day when the wind direction changed. These results indicated that initiation, maintenance and disappearance of the dinoflagellate Scrippsiella trochoidea red tide in the bay were not directly driven by changes in nutrients. Therefore, how nutrients are linked to the formation of red tides in coastal waters need to be further examined, particularly in relation to dissolved organic nutrients. (C) 2008 Elsevier B.V. All rights reserved.