三种常见原生动物对褐潮藻种抑食金球藻(Aureococcus anophagefferens)的摄食

微型浮游动物在抑食金球藻(Aureococcus anophagefferens)引发褐潮时表现的摄食压力可潜在控制褐潮的爆发和消亡。本研究就三种海洋常见原生动物——海洋尖尾藻(Oxyrrhis marina)、海洋尾丝虫(Uronema marinum)和扇形游仆虫(Euplotes vannus)——对单种饵料及混合饵料中抑食金球藻中国株的摄食进行了研究。单种抑食金球藻指数期细胞喂食的三种原生动物的生长率和摄食率呈现米氏方程变化趋势。比较三种原生动物摄食抑食金球藻的最大摄食率, 发现其随动物粒径的增大而增大, 但仅为摄食球等鞭金藻(Isochrysis galbana)的30%~59%。海洋尖尾藻和海洋尾丝虫的最大生长率(μ_max)与饵料种类无关, 扇形游仆虫摄食抑食金球藻时的μ_max值小于摄食球等鞭金藻的个体。海洋尖尾藻、海洋尾丝虫和扇形游仆虫摄食抑食金球藻时的毛生长率(gross growth efficiency, GGE)分别为65.8%、35.2%和49.1%。三种原生动物摄食抑食金球藻指数期细胞和球等鞭金藻以不同比例混合的饵料时表现出对抑食金球藻的选择倾向; 在含有抑食金球藻稳定期细胞的混合饵料喂食的情况下, 三种原生动物避食抑食金球藻或不表现明显摄食倾向性。抑食金球藻释放胞外聚合物(extracellular polymeric substance, EPS)的测定结果显示, 细胞从指数期生长至稳定期释放出的EPS的水平显著上升(P<0.05), 可能与原生动物对不同生长期藻细胞具有不同选择偏好有关。     

核酸指标对桡足类安氏伪镖水蚤摄食率、生长率及产卵率的指示作用

核酸指标可表征桡足类营养及生长状态, 然而多物种和多指标比较研究的相对缺乏却限制了现场海区调查中对核酸标志物的广泛应用。本研究测定了系列饵料浓度(0.2~2.0μg C·mL^-1)喂食的安氏伪镖水蚤Pseudodiaptomus annandalei的桡足幼体Ⅱ期个体的摄食率、生长率及成体产卵率, 并就生化组成, 包括RNA含量、DNA含量、蛋白含量(以下简写为PRO)、RNA:DNA、RNA:PRO、DNA:PRO及元素组成(N含量、P含量、C:N 和C:P比值)进行定量分析和计算。摄食率、生长率、产卵率及桡足幼体的RNA相关的核酸指标随饵料浓度的变化趋势均符合双曲线模型, 而雌性成体中仅RNA:DNA比值符合此模型。对RNA相关指标与各生理参数的相关性分析显示, 桡足幼体的RNA相关指标均与摄食率及生长率呈显著线性相关性, 显著程度的顺序为RNA:DNA>RNA:PRO>RNA含量; 雌性成体中RNA:DNA及RNA:PRO比值与产卵率显著相关, 前者的相关性远较后者显著。此结果表明, 在常用的现场浮游桡足类调查的标志比值(RNA:DNA比值或RNA含量)之外, RNA:PRO比值亦可作为指示桡足类生长状态的标志比值, 但是当指示体生长时, 桡足类必须处于同一生长阶段。RNA相关指标与饵料浓度之间稳固的双曲线相关性提示我们, RNA相关指标可帮助评估桡足类的饵料环境。     

日本虎斑猛水蚤在抑食金球藻中存活、发育和繁殖的实验研究

抑食金球藻(Aureococus anophagefferens)可以形成褐潮, 并对贝类、浮游动物等多种生物均能造成不利影响。为进一步探究抑食金球藻对浮游动物的影响, 本文以日本虎斑猛水蚤(Tigriopus japonicus)为实验生物, 研究抑食金球藻对日本虎斑猛水蚤摄食、存活、生长发育以及繁殖的影响。日本虎斑猛水蚤具有易于在实验室培养、生长周期短、雌雄异体等优点, 是海洋毒性污染物检测的模式生物。实验利用高效液相色谱分析方法, 在日本虎斑猛水蚤体内色素中检测到了抑食金球藻的特征色素19’-丁酰氧基岩藻黄素(But-fuco), 表明日本虎斑猛水蚤能够摄食抑食金球藻。当微藻生物量(相对碳含量)分别同为0.7 μg/mL和7.2 μg/mL时, 在抑食金球藻中无节幼体发育至桡足幼体及成体的存活率均高于以青岛大扁藻为饵料的对照组, 但无显著差异, 并且在前者中无节幼体的发育时间显著低于后者, 12 d内雌体的产卵次数与产卵量显著高于后者。结果表明, 日本虎斑猛水蚤在抑食金球藻中能进行正常的生命活动, 并且是首次报导的一种能够在抑食金球藻中正常摄食、生存、生长发育和繁殖的浮游动物。因此, 当褐潮发生时, 由于贝类幼体等生物会受到显著的不利影响, 日本虎斑猛水蚤等抗性较强的生物可能会成为优势种, 从而会影响浮游动物群落结构的组成, 进而可能会使整个海洋生态系统发生变化。本研究有助于全面了解褐潮对海洋生态系统的影响。     

三角褐指藻与抑食金球藻的竞争及化感作用研究

为探明三角褐指藻（Phaeodactylum tricornutum）对抑食金球藻（Aureococcus anophagefferens）是否具有化感作用,本研究以三角褐指藻和抑食金球藻为实验材料,研究了抑食金球藻在单培养和共培养条件下的生长情况及三角褐指藻培养滤液对其生长和叶绿素荧光参数的影响。结果表明:三角褐指藻对抑食金球藻有明显的化感抑制作用。在单培养体系中,抑食金球藻的生长曲线可用逻辑斯谛增长模型拟合,随着起始密度的增加,环境容量（K）逐渐减小,而抑食金球藻的种群瞬间增长率（r）、进入拐点时间及稳定期细胞密度均较为接近;当三角褐指藻与抑食金球藻以不同起始密度比共同培养时,抑食金球藻的生长均受到了显著地抑制（P<0.05）,但其抑制作用并未与三角褐指藻的密度呈明显的线性关系;滤液培养实验发现,10 mL和15 mL三角褐指藻培养滤液的加入可对抑食金球藻的生长产生显著影响（P<0.05）,对其叶绿素荧光参数没有影响（P>0.05）,25 mL和35 mL三角褐指藻培养滤液的加入可对抑食金球藻的生长和叶绿素荧光参数均产生显著的抑制作用（P<0.05）,叶绿素荧光参数F_v/F_m、ΦPSⅡ、alpha的值降低,Ik的值增加,PSⅡ受到损害。     

小球藻与褐潮原因种-抑食金球藻间的相互作用研究

近年来渤海海域频繁暴发以抑食金球藻(Aureococcus anophagefferens)为主的褐潮对海水贝类养殖造成巨大的危害。筛选出既能强烈抑制抑食金球藻生长又可为养殖微环境中贝类提供饵料的微藻,并将其浓缩液缓释于贝类养殖的微环境中,将对降低褐潮的危害,提高养殖经济效益具有非常重要的实际应用价值。本实验以小球藻(Chlorella vulgaris)和抑食金球藻为实验材料,采用荧光定量PCR技术监测抑食金球藻的细胞密度,研究了抑食金球藻在单培养体系中的生长情况、与小球藻共培养时两种微藻生长情况的变化以及小球藻培养滤液对其生长和叶绿素荧光参数的影响。研究结果表明:(1)在单培养体系中,抑食金球藻的生长曲线可用逻辑斯谛增长模型拟合,随着起始密度的增加,环境容量(K)逐渐减小;而抑食金球藻的种群瞬间增长率(r)、进入拐点时间及稳定期细胞密度均较为接近;(2)当小球藻与抑食金球藻以不同起始密度比共同培养时,抑食金球藻的生长均受到了显著地抑制(P0.05),且随着小球藻比例的升高抑制作用加强;(3)滤液培养实验发现,≥10m L小球藻培养滤液的加入可对抑食金球藻的生长和叶绿素荧光参数产生显著的影响(P0.05),生长速度减缓或滞长,叶绿素荧光参数Fv/Fm、ΦPSⅡ、alpha的值降低,Ik的值增加,PSⅡ受到损害,随着滤液加入量的增加,损害程度加深,并且不可恢复。     

赤潮藻对桡足类摄食、产卵及孵化影响的研究进展

本文对近年来有关赤潮藻对桡足类摄食、产卵及孵化的影响研究动态进行了综述。一般认为，海洋桡足类对赤潮藻存在着潜在的调控作用，但近年来的研究发现，某些赤潮藻反过来对桡足类的生长、发育、产卵等生理状态也存在着负面影响。文中提出在今后赤潮研究中，应加强对有毒赤潮藻和桡足类相互作用和特异性的研究。     

不同温度下抑食金球藻、中肋骨条藻和海洋卡盾藻间的相互作用

本文选取抑食金球藻(Aureococcus anophagefferens)、中肋骨条藻(Skeletonema costatum)和海洋卡盾藻(Chattonella marina)三种藻在20℃和25℃下进行单种、两种或三种藻间的培养实验, 探究在不同温度下藻种间的相互作用和竞争优势。单种培养结果显示抑食金球藻和中肋骨条藻对温度25℃敏感, 其环境容量(K)被显著降低; 而海洋卡盾藻的內禀增长率(r)在25℃下显著升高, 但K保持不变。在共培养中, 海洋卡盾藻在20℃下因抑食金球藻添加致死, 但在25℃处理下其K值上升120%。虽然中肋骨条藻的r在两个温度下均受海洋卡盾藻添加的促进, 但其K值从20℃下的上升43%转变为25℃下的降低48%。温度变化对于抑食金球藻和中肋骨条藻共培养的结果无明显影响, 中肋骨条藻的K均上升40%, 而抑食金球藻的K值均下降60%~70%。结果说明, 种间竞争除了共培养藻固有的相互作用关系因素外, 也受到微藻间温度适应性差异的影响。在20℃条件下三种藻混合, 抑食金球藻K的抑制率达到最高为79%, 而中肋骨条藻K的促进率达到最高为108%, 这可能表明三者之间的相互作用存在协同效应。     

应用实时定量PCR技术检测秦皇岛近岸海域褐潮原因种——抑食金球藻(Aureococcus anophagefferens)

Aureococcus anophagefferens, a small pelagophyte algae, has caused brown tide blooms in coastal waters of Qinhuangdao in recent years, presenting significant negative impacts on the shellfish mariculture industry. Under standard light microscopy, it is visually indistinguishable from other small algae in field samples due to its extremely small size. In this study, quantitative polymerase chain reaction(q PCR) based on 18 S r DNA sequences was developed and used to detect and enumerate A. anophagefferens. A linear regression(R2 = 0.91) was generated based on cycle thresholds value(Ct) versus known concentrations of A. anophagefferens. Twenty-two field samples collected in coastal waters of Qinhuangdao were subjected to DNA extraction and then analyzed using q PCR. Results showed that A. anophagefferens had a wide distribution in coastal waters along Qinhuangdao. Elevated A. anophagefferens abundance, category 3 brown tide blooms(200 000 cells/m L) occurred at Dongshan Beach and Tiger-stone Beach in August in 2013. In shellfish mariculture areas along coastal waters of Qinhuangdao, 4 stations had category 3 blooms, and 6 stations had category 2 blooms(35 000–200 000 cells/m L) in August and all stations had category 1 blooms(0 to ≤35 000 cells/m L) in October. Quantitative PCR allows for detection of A. anophagefferens cells at low levels in filed samples, which is essential to effective management and prediction of brown tide blooms.     

微塑料对海洋桡足类摄食、排泄及生殖的影响

微塑料污染目前成为海洋污染普遍关注的一个研究热点。本文在实验室内将青岛近海常见的海洋桡足类猛水蚤暴露于不同浓度的微塑料尼龙6中,研究了猛水蚤的摄食、排泄以及生殖的变化。研究结果表明,微塑料尼龙6对猛水蚤的摄食、排泄、生殖均产生不利的影响,并且存在剂量-效应关系。微塑料尼龙6对猛水蚤摄食率、滤水率、排粪率的24 h·EC 50分别为67.7、62.2、84.1 mg·L^-1,对猛水蚤抱卵率的144 h·EC 50为30.3 mg·L^-1。“饱食感”造成猛水蚤摄食率降低,从而能量和营养摄入不足可能是导致猛水蚤抱卵率降低的原因。猛水蚤对微塑料的摄食,导致猛水蚤排泄的粪便颗粒小型化,由长椭球体变为短小椭球体,可能与其粘度或物理结构的改变有关。暴露于尼龙6的猛水蚤的粪便体积和沉降速率显著低于未暴露微塑料的对照组。本实验结果对于研究微塑料对海洋桡足类以及滤食性浮游动物的生态毒理影响具有一定的帮助。     

Grazing effects by Nereis diversicolor on development and growth of green algal mats

Nereis diversicolor is generally considered to be a predator and deposit feeder, but have also been found to graze on benthic algae in shallow coastal areas. In this study we investigated the grazing effects on the development and growth of green algae, Ulva spp. Algal growth was studied in an experiment including two levels of sediment thickness; 100 mm sediment including macrofauna and 5 mm sediment without macrofauna, and three treatments of varying algal biomass; sediment with propagules, sediment with low algal biomass (120 g dry weight (dwt) m^− 2) and sediment with high algal biomass (240 g dwt m^− 2). In the 100 mm sediment, with a natural population of macrofauna, N. diversicolor was the dominating (60% of total biomass) species. After three weeks of experimentation the result showed that N. diversicolor was able to prevent initial algal growth, affect growth capacity and also partly reduce full-grown algal mats. The weight of N. diversicolor was significantly higher for polychaetes in treatments with algae added compared to non-algal treatments. There were also indications that a rich nutrient supply per algae biomass counteracted the grazing capacity of N. diversicolor.     

Effects of residual chlorine on the mortality, grazing and respiration of Labidocera euchaeta (Copepoda)

In this study,the authors investigated the effects of residual chlorine on mortality,grazing and respiration of Labidocera euchaeta in laboratory.The grazing rate was evaluated by subtraction method of food concentration and respiration rate was measured using oxygen electrode.It was found that the lethal effect of residual chlorine on L.euchaeta increased with enhanced concentration and prolonged duration.The medium lethal concentration (LC 50 ) of chlorine for L.euchaeta in 24 h was about 0.58 mg/L and the safe concentration was about 0.21 mg/L.However,the grazing and respiration of L.euchaeta decreased by 32.6% and 18.9% when exposed to 0.2 mg/L residual chlorine for 4 h.It indicated that the physiological activities of zooplankton could be suppressed by the residual chlorine less than the safety concentration.Therefore,both survival and physiological activities of the organisms living in the thermal (nuclear) power plant discharging waters should be considered when carrying out the ecological risks assessment.     

中国近海浮游桡足类中华哲水蚤（Calanussinicus Brodsky）的分子系统地理学和种群遗传结构研究

Planktonic copepod Calanus sinicus is the dominant meso-zooplankton in the Northwest Pacific Ocean. To better understand its population dynamics and phylogeographic patterns, 243 C. sinicus individuals were collected from seven locations across the shelf waters of China and its population genetics was studied by mitochondrial DNA cytochrome oxidase I(mtCOI) sequences analyses. Thirty-nine different sequences, or haplotypes, were detected with moderate haplotype diversity(h=0.749) and low nucleotide diversity(π=0.003) for all populations. The evolutionary divergence between geographic populations varied from 0.24% to 0.37%, indicative of very limited genetic differentiation. Visualized minimum spanning network(MSN) and phylogenetic analysis of all the detected haplotypes did not reveal any clear phylogeographic pattern. Furthermore, AMOVA data showed no significant spatial population differentiation existed among the individuals collected across China shelf waters. Pairwise FST values showed that population collected from northwest of the East China Sea(ECS) displayed a low difference to other populations. Mismatch distribution analyses and neutrality tests indicated that C. sinicus might undergo a demographic/population expansion. No significant population genetic structuring was detected, indicating an extensive gene flow among the C. sinicus populations. Our results provide molecular evidence for the hypothesis that C. sinicus in the northwestern South China Sea in winter is transported from the East China Sea and the Yellow Sea by the China Coastal Current during the northeast monsoon period.     

Impacts of solar UV radiation on grazing, lipids oxidation and survival of Acartia pacifica Steuer (Copepod)

UV radiation is known to affect aquatic primary producers and their grazers. However, little has been documented on its effects on zooplankton grazing. In this study, the authors investigated the effects of photosynthetically active radiation (PAR, 400-700 nm), ultraviolet-A (UV-A, 320-400 nm) and ultraviolet-B (UV-B, 280-320 nm) radiation on grazing, mortality and lipids oxidation of the copepod Acartia pacifica collected from the Xiamen Bay. After 30 min of the exposures, the copepod was fed in darkness with the diatom Phaeodactylum tricornutum at two cell concentrations (2.5 × 10 4 and 2.5 × 10 5 cells/ml). At the low cell concentration, the individuals pre-exposed to PAR (218.0 W/m 2 )+UV-A (48.2 W/m 2 ) or PAR+UV-A+UV-B (2.1 W/m 2 ) showed suppressed clearance and grazing activities compared with those receiving PAR alone, by 22.7% and 17.1% for clearance and by 22.6% and 5.5% for grazing rates, respectively. However, the suppression on clearance and grazing became indistinctive at the high food concentration. Exposures to UV-A and UV-B led to increased lipid oxidation and higher mortality, furthermore, the mortality linearly increased with enhanced oxidation of lipid.     

Grazing experiments and model simulations of the role of zooplankton in Phaeocystis food webs

A combined empirical and modelling study was conducted to further examine the potential importance of grazing by zooplankton in pelagic food webs in which Phaeocystis is a significant or dominant component. Laboratory experiments were designed to measure ingestion of Phaeocystis and other potential prey items which co-occur with Phaeocystis. Grazers included copepods and ciliates, and prey included Phaeocystis colonies and solitary cells, diatoms, ciliates, bacteria, and detritus. These data were expressed in the model currency of nitrogen units, and fit to hyperbolic tangent equations which included minimum prey thresholds. These equations and literature data were used to constrain a food web model whose purpose was to investigate trophic interactions rather than to mimic actual events. Nevertheless, the model output was similar to the general pattern and magnitude of development of Phaeocystis–diatom communities in some environments where they occur, e.g. north Norwegian waters. The model included three forms of nitrogen, three phytoplankton groups, bacteria, two zooplankton groups, and detritus, with detailed flows between compartments. An important component of the model was inclusion of variable prey preferences for zooplankton. The experiments and model simulations suggest several salient conclusions. Phaeocystis globosa colonies were eaten by a medium-sized copepod species, but ingestion appeared to be strongly dependent upon a proper size match between grazer and prey. If not, colonies were eaten little if at all. Phaeocystis solitary cells were ingested rapidly by ciliate microzooplankton, in agreement with prior literature observations. In contrast, detritus was eaten comparatively slowly by both ciliates and copepods. Both types of zooplankton exhibited apparent minimum prey thresholds below which grazing did not occur or was inconsequential. Model simulations implied that transitions between life cycle stages of Phaeocystis may potentially be important to phytoplankton–zooplankton interactions, and that relative rates of ingestion of Phaeocystis by various zooplankton may have significant impacts upon material fluxes through and out of Phaeocystis–diatom ecosystems. Indirect effects of trophic interactions appear to be equally significant as direct effects.     

龙须菜(Gracilariopsis)/江蓠(Gracilaria)的生长和种质特性分析

龙须菜(Gracilariopsis lemaneiformis)是具有重要经济价值和生态效益的大型红藻, 主要用作琼胶提取原料和鲍的饵料。本研究利用生理生化、液相色谱-质谱联用和氨基酸分析等方法, 比较了龙须菜(Gp. lemaneiformis) (wt、981、Gl-1、Gl-s、Gl-g)、异枝龙须菜(Gp. heterocla, Gh)和细基江蓠繁枝变种(Gracilaria tenuistipitata var. liu, Gt)的生长及藻胆蛋白、琼胶、红藻糖苷和氨基酸等的差异, 以期为龙须菜/江蓠栽培中的种质区分及选择提供参考。结果表明Gt在23 ℃和30 ℃条件下生长均最快, 其相对生长速率分别为野生型龙须菜(wt)的2.19倍和2.49倍。龙须菜Gl-s中藻胆蛋白浓度最高, 为wt的1.91倍。除了Gt之外, 其余6种藻中琼胶产率较高(16.22%~18.91%)。Gt中红藻糖苷和海藻糖积累最多, 分别为wt的3.50倍和1.81倍。Gl-g、Gl-s、Gt和Gh多糖丰富, 在36.89%~40.23%; 龙须菜981、Gl-1、wt和Gl-s总氨基酸浓度较高, 在152.35~161.32 mg·g^–1干质量之间, 并且981、Gl-1、Gl-s氨基酸评分较优。综合以上结果, 龙须菜981、Gl-1和Gl-s的藻胆蛋白、琼胶和氨基酸等均显著优于其他藻, 并且生长较快, 可用于琼胶、藻胆蛋白及多糖的提取或鲍的饵料; 而细基江蓠繁枝变种生长快、红藻糖苷和海藻糖丰富, 可大规模栽培用作鲍的饵料。该研究为丰富及开发利用中国大型海藻种质资源提供了重要的资料。     

糖原含量与糖原磷酸化酶基因(GPH)和己糖激酶基因(HK)的表达在福建牡蛎生殖周期中的相关性分析

Glycogen, a polymer of glucose, is an important means of storing energy. It is degraded by glycogen phosphorylase (GPH) and hexokinase (HK), glycogen phosphorylase, and hexokinase cDNAs (Ca-GPH andCa-H...