固定化微藻对虾池弧菌数量动态的影响

引入固定化波吉卵囊藻(Oocystis borgei)和微绿球藻(Nannochloris oculata)于凡纳对虾(Litop Penaeus vannamei)养殖环境中,检测水体、对虾胃和后肠中弧菌的数量变化,研究固定化微藻对虾池弧数量动态影响。结果表明:波吉卵囊藻培养液中9d后不能检测出弧菌,微绿球藻培养液中15d后不能检测出弧菌。引入固定化波吉卵囊藻和微绿球藻的褐藻胶藻珠能抑制弧菌的生长,实验组养殖水体、对虾胃和后肠中弧菌的数量都比对照组低;抑制效果是固定化波吉卵囊藻和微绿球藻混合固定化波吉卵囊藻固定化微绿球藻;试验后期实验组弧菌的数量明显低于试验前期。试验期间固定化波吉卵囊藻和微绿球藻的生物量分别增加了约10倍和17倍,证明它们的生理活性不会因固定化而受干扰。因此,固定化微藻可应用于虾池微生态调控防病。       

虾池常见微藻的光照强度、温度和盐度适应性

通过Smith生态位宽度指数和Pianka生态位重叠指数分析了虾池常见微藻种群(啮蚀隐藻、新月菱形藻、微绿球藻和蛋白核小球藻)在光照强度、温度和盐度资源上的生态位宽度和生态位重叠特征.结果表明:新月菱形藻和蛋白核小球藻具有较大的生态位宽度值,啮蚀隐藻和微绿球藻的生态位宽度值则相对较小.蛋白核小球藻和微绿球藻在光照强度、温度和盐度资源上的生态位重叠值均为最大,啮蚀隐藻在各资源与其他微藻的重叠值最小.新月菱形藻与蛋白核小球藻适应光照强度的范围较广.当水温达16.9℃,可定向培育新月菱形藻;当水温达25℃,可定向培育新月菱形藻和啮蚀隐藻;当水温达30℃时,可定向培育新月菱形藻、蛋白核小球藻和微绿球藻.养殖水体盐度处于9～26,可引入蛋白核小球藻与微绿球藻;处于9～17.5,应引入啮蚀隐藻;高盐水体,应引入新月菱形藻.蛋白核小球藻和微绿球藻在光照强度、温度和盐度资源上的生态位重叠值均为最大,因此微藻定向培育,不可同时引入蛋白核小球藻与微绿球藻.     

虾池常见微藻种群温度、盐度和氮、磷含量生态位

就温度、盐度、氮磷含量设计试验,对4种虾池常见微藻种群新月菱形藻、啮蚀隐藻、微绿球藻、蛋白核小球藻的生态位进行研究.结果显示：啮蚀隐藻在温度和盐度资源上的生态位宽度值最大,分别达0.980和0.988,新月菱形藻在氮、磷含量资源上的生态位宽度值最大,达0.990,蛋白核小球藻在各资源的生态位宽度值均最小,平均只有0.926;啮蚀隐藻与蛋白核小球藻在温度和盐度资源上的生态位重叠值最小,分别为0.809和0.702,啮蚀隐藻与新月菱形藻在氮、磷含量资源上的生态位重叠值最小,只有0.829,蛋白核小球藻与微绿球藻在温度、盐度和氮、磷含量资源上的生态位重叠值均最大,分别为0.986、0.974和0.989.表明在养殖水环境调控过程中,啮蚀隐藻适宜与新月菱形藻、微绿球藻或蛋白核小球藻共同培育,而蛋白核小球藻和微绿球藻对资源的竞争性明显,不适合同时引入同一池塘进行培育.     

虾-鱼-贝-藻养殖结构优化试验研究

本研究建立了一种虾、鱼、贝、藻优化养殖结构及水质调控系统,该系统包括对虾养殖、鱼类养殖、贝类养殖、大型海藻栽培等4个功能不同的养殖区,1个益生菌及微藻培养区和1个水处理区以及1条应急排水渠。通过在封闭循环水养殖系统内不同池塘中放养生态位互补的对虾、口孵非鲫、牡蛎、江蓠等经济动植物,对虾池水质环境进行生物调控与自我修复。结果表明,循环水养殖系统内虾池水层悬浮物数量、COD值、氨态氮和总氮含量比对照组对虾单养池明显降低。养殖后排放水不处于富营养化状态(E<1)。投入每千克对虾饲料生产0.667kg对虾,同时生产0.037kg口孵非鲫、0.738kg牡蛎、0.437kg江蓠,相当于净增产值2.01元。投入与产出比,对虾单养为0.671,优化生态养殖为0.235,明显降低。该养殖模式不仅实现了虾池水质环境的自我修复与调控及养殖期间用水的零排放,而且饲料利用率大幅度提高,经济效益显著增加,还具有防病性、环保性、高效性等优点。     

亚硝酸盐氮对凡纳滨对虾毒性和抗病相关因子影响

用常规生物毒性实验方法,在不同盐度下进行亚硝酸盐氮对凡纳滨对虾的急性毒性实验;并加亚硝酸盐氮于凡纳滨对虾的养殖环境中,检测与抗病力相关因子的变化。研究亚硝酸盐氮对凡纳滨对虾的毒性和抗病力相关因子的影响。结果表明:盐度对亚硝酸盐氮的毒性有较大影响。盐度为31时,24hLC50、48hLC50、72hLC50和96hLC50分别为314.9mg/L1、75.3mg/L1、00.2mg/L和89.0mg/L;盐度为17时,分别为132.3mg/L、65.6mg/L、51.3mg/L和39.5mg/L。盐度31实验组的亚硝酸盐氮半致死浓度均显著(P<0.05)高于盐度17实验组。在低盐度条件下亚硝酸盐氮的毒性较强。亚硝酸盐氮对凡纳滨对虾抗病力相关因子有显著的影响。亚硝酸盐氮浓度为4.0mg/L和8.0mg/L时,其血细胞数、超氧化物歧化酶(SOD)活力、酚氧化酶(PO)活力、抗菌活力(Ua)、溶菌活力(UL)和血清蛋白含量均显著(P<0.05)低于对照组;不吸污组抗病力相关因子活性均显著(P<0.05)低于吸污组。低浓度的亚硝酸盐氮可降低凡纳滨对虾抗病能力,亚硝酸盐氮浓度越高,其抗病能力越弱。     

基于模糊综合评价的产生物柴油微藻藻种筛选

产生物柴油微藻大规模培养对微藻藻种的性能要求较高。从丰富的藻种资源中筛选到高品质的藻种一直是个亟待解决的问题。通过研究3株产油微藻,从系统工程的角度综合整个微藻生物柴油的技术工艺,建立了以生长速率、含油率、油脂组成等18种指标的二级评价体系,采用二级模糊综合评价的模糊数学方法对产生物柴油微藻的性能进行综和分析、筛选。最终确定供评价的三株微藻二级模糊综合评价集：小球藻LICME001［0.360 0.315 0.192 0.069 0.064］,微绿球藻LICME002［0.277 0.331 0.236 0.104 0.052］和葡萄藻LICME003［0.325 0.371 0.232 0.071 0.060］。根据最大隶属度法则分析得：小球藻LICM001株产生物柴油微藻品质为优等级别,适合产生物柴油的技术工艺要求;微绿球藻LICME002和葡萄藻LICME003为良等级别的产生物柴油藻种。     

单生卵囊藻(Oocystis solitaria)和月牙藻(Selenastrumsp.)的培养条件及其细胞组成

从海滨的淡水池沼中分离得到单生卵囊藻(Oocystis solitaria)和月牙藻(Selenastrum sp.),分别研究了温度、光照强度、盐度对2种微藻生长繁殖的影响,分析了2种微藻的细胞组成、脂肪酸组成及盐度对2种微藻脂肪积累的影响.结果表明:单生卵囊藻和月牙藻的适宜生长温度分别为35.9℃～40.5℃和29.7℃～32.8℃;单生卵囊藻和月牙藻的适宜光照强度分别为46～70μmol · m-2·s-1和17 ～54 μmol·m-2·s-1;单生卵囊藻在淡水培养液中生长最好,月牙藻在盐度为2的半成水培养液中生长最好;在适宜培养条件下,单生卵囊藻细胞蛋白、总糖和总脂肪分别为27.61％、22.00％和3.84％;月牙藻细胞的蛋白、总糖和总脂肪分别为28.06％、21.99％和12.53％;单生卵囊藻的脂肪酸组成中含有丰富的18∶3n3;月牙藻的脂肪酸组成中含有DHA;盐度影响2种微藻的总脂肪含量;单生卵囊藻和月牙藻分别在盐度4和10的半咸水培养液中细胞总脂含量最高.     

富油能源微藻的筛选及产油性能评价

为了筛选具有产油潜力的能源微藻,以实验室保藏的20株淡水和海洋微藻(绿藻门18株,真眼点藻纲1株,硅藻纲1株)为研究对象,利用光径为3 cm柱状光生物反应器通气分批培养,通过测定微藻培养物的生物量和总脂含量等指标,从中筛选生长速度快、生物量和总脂含量高的微藻。结果表明:20株微藻的生物量和总脂含量分别在1.81~7.88g/L和16.0%~55.9% dw(% Dry weight)之间,筛选得到具有产油潜力的微藻9株,分别是栅藻(Scenedesmus sp.)(6.34g/L,55.9% dw)、麻织绿球藻(Chlorococcum tatrense)(5.93g/L,46.9% dw)、眼点拟微绿球藻(Nannochloropsis oculata)(7.88g/L,35.0% dw)、油面绿球藻(Chlorococcum oleofaciens)(5.58g/L,45.9% dw)、多形拟绿球藻(Pseudochlorococcum polymorphum)(6.10g/L,40.0% dw)、八月衣藻(Chlamydomonas augustae)(5.78g/L,40.5% dw)、椭圆小球藻(Chlorella ellipsoidea)(5.56g/L,40.7%dw)、椭圆绿球藻(Chlorococcum ellipsoideum)(5.41g/L,38.0% dw)、雪绿球藻(Chlorococcum nivale)(5.55g/L,36.3% dw),其中最具产业化潜力的微藻为栅藻(Scenedesmus sp.),其总脂收获量和单位体积总脂产率分别为3.5 g/L和218.7mg/L·d。     

夏季混养鳙和银鲫对凡纳滨对虾低盐养殖池塘浮游藻类群落结构的影响

基于2011年6-8月在浙江省慈溪市庵东镇凡纳滨对虾低盐(2-3.2)养殖塘调查取得的数据, 分析了混养鳙和银鲫对凡纳滨对虾低盐养殖塘浮游藻类群落结构的影响。结果显示: (1) 调查期间共检出浮游藻类5门32属48种, 其中蓝藻门16种, 绿藻门20种, 硅藻门5种, 裸藻门5种, 隐藻门2种, 主要优势种为螺旋藻Spirulinasp.)、湖泊鞘丝藻(Planktolyngbya limnetica)和湖生卵囊藻(Oocystis lacustris)等。(2) 混养鳙和银鲫的虾塘浮游藻类平均生物量(22.7 mg·L−1)要低于仅混养银鲫的(32.6 mg·L−1)和单养对虾的(34.0 mg·L−1), 其中混养鳙和银鲫与仅混养银鲫的蓝藻生物量所占比例分别为46.4%与45.1%, 要低于对虾单养池塘的68.1%。(3) 混养鳙和银鲫的虾塘浮游藻类群落相似度在中、后期较高,而仅混养银鲫的虾塘在后期相似度变化较大。因此, 混养适当密度的鳙和鲫能够改善虾塘藻相, 防止藻过度增长, 促进养殖期间生态系统的稳定性。     

高位虾池养殖过程浮游植物群落的演替

通过对湛江东海岛北寮村和庵里村各2个高位虾池养殖过程的浮游植物演替和主要理化因子进行调查,研究浮游植物群落的演替过程对高位虾池的健康养殖的影响.结果表明:4池共检出浮游植物7门76属140种;对虾生长正常的北寮高位虾池演替优势种主要有螺旋环沟藻、椭圆扁胞藻、海链藻、湖泊束球藻密胞变种、裸甲藻、卵囊藻、微囊藻、锥形斯克里普藻、色球藻和尖尾蓝隐藻等,而对虾发病的庵里高位虾池演替优势种主要有铜绿微囊藻、柔弱布纹藻、咖啡形双眉藻、小席藻、卷曲鱼腥藻、水生集胞藻、衣藻、尖尾蓝隐藻和螺旋环沟藻等,北寮正常虾池浮游植物种类(包括优势种)较对虾发病的庵里虾池多而复杂;对虾生长正常池优势种的演替快而门类交替,而对虾发病池呈现硅藻到蓝藻优势的演替;对虾生长正常池浮游植物密度随养殖过程延伸呈上升态势,而对虾发病池浮游植物密度初期增加病害后下降;养殖过程浮游植物密度与主要理化因子的相关关系不显著;对虾生长正常的北寮高位池多样性指数与均匀度高于对虾发病的庵里高位池,而优势度却较低.水体理化因子变化平缓,浮游植物有较高的多样性指数和均匀度、较低的优势度对于稳定虾池养殖水环境有重要作用.     

Serial optimization of biomass production using microalga Nannochloris oculata and corresponding lipid biosynthesis

As energy and environment have become urgent issues, there has been increasing needs to develop alternative energy source, such as microalgal bio-fuel. In this study, we investigated the growth and lipid contents of microalgae Nannochloris oculata under various environmental conditions for biodiesel production. Our results indicated that biomass productivities of N. oculata were correlated with increasing nitrogen concentrations up to 37.5 ppm. High irradiance using 230-250 μmol/m(2) led to higher biomass yields than low irradiance of 160-180 μmol/m(2). Biomass productivities increased further by manipulating surface to volume ratio (S/V), which in turn enhanced light penetration. Finally, optimal biomass productivities (1.04 g/l day) could be achieved by the supplementation of yeast extract. Lipid contents and fatty acid profiles of N. oculata were affected by the different growth conditions. Lipid contents of N. oculata decreased as nitrogen concentration increased. Lower temperature (15 °C) resulted in higher lipid content than higher temperature (25 °C). Fatty acid profiles of N. oculata indicated that palmitic acid (C16:0) and linoleic acid (C18:2) were the two most abundant fatty acids, but the supplementation of yeast extract increased linolenic acid (C18:3) content. Our results suggested the feasibility of N. oculata for the biodiesel production.     

Critical micelle concentrations of allelopathic substances produced by Nannochloris oculata which affect a red tide organism, Gymnodinium breve

Laboratory cultures of the green algae Nannochloris oculata and Nannochloris eucaryotum are known to cause lysis of Gymnodinium breve, which is Florida's red tide organism. Two cytolytic agents were previously identified as methyl palmitate and methyl stearate. In this study, the critical micelle concentrations of these substances were determined by ultraviolet light and turbidimetric methods to be 3.5 +/- 0.3 ppm (methyl stearate) and 4.3 +/- 0.6 (methyl palmitate). There were no significant differences in results obtained using the two methods.     

Bioassays of APONIN-3 and -4 with rabbit erythrocytes

Rabbit erythrocytes in methanolic phosphate medium were used to bioassay the activity of authentic samples of methyl stearate and methyl palmitate (in 10% methanol:90% water, v/v), which had been identified as apparent oceanic naturally occurring cytolins (APONIN-3 and -4) produced by Nannochloris oculata. The two natural products are notable for cytolytic activity toward the unarmoured dinoflagellate, Gymnodinium breve Davis, an organism responsible for red tides consisting of harmful algal blooms in the Gulf of Mexico and along the eastern coast of the United States. Bioassays were done with heparinized rabbit blood. The absorbance at 540 nm was observed for 15 min in comparison with a sample treated with haemolysing agent. The results indicated that at reasonable concentrations of 1-10 ppm, neither was a haemolysin, although such concentrations caused cytolysis of G. breve cultures.     

维生素C对凡纳滨对虾生长及抗病力的影响

以不同水平维生素C 2 磷酸酯 (添加量分别为 0、75、15 0、30 0和 6 0 0mg/kg)的饲料喂养凡纳滨对虾 10周 ,研究维生素C 2 磷酸酯对凡纳滨对虾生长及抗病力的影响。结果显示 :在养殖前 4周 ,饲料中添加维生素C 2 磷酸酯显著促进凡纳滨对虾的生长 ,然而对对虾的成活以及饲料利用不产生影响 (P >0 0 5 ) ;而到实验后期添加维生素C 2 磷酸酯不能促进凡纳滨对虾的生长 ,却显著提高凡纳滨对虾的成活率 (P <0 0 5 )。维生素C 2 磷酸酯对对虾体水分、脂肪、蛋白质和维生素C在肝胰脏中的积累量的影响显著 (P <0 0 5 ) ,对对虾体灰分影响不显著 (P >0 0 5 )。维生素C 2 磷酸酯对对虾血清中超氧化物歧化酶活力无显著影响 ,饲料中未添加维生素C或过量添加 (超过 30 0mg/kg饲料 )均导致血清中酚氧化酶活力、血细胞总数和溶菌酶活力的显著下降。以生长、成活和酚氧化酶活力为指标 ,饲料中维生素C 2 磷酸酯的适宜添加量为 15 0mg/kg。     

洋河水库浮游植物组成及优势种演替规律研究

在洋河水库设置6个采样点, 对浮游植物进行了周年研究, 并在夏季进行了每周一次的加密采样, 以揭示水华期间藻类优势种演替规律。结果表明洋河水库全年共检测到浮游植物8门41属49种, 群落季节变化与温度密切相关。春季隐藻门的啮齿隐藻(Cryptomonas erosa)为优势, 夏季初期表层水温在25℃以下时, 绿藻门的波吉卵囊藻(Oocystis borgei)占主要优势; 当表层水温升至25℃以上, 微囊藻(Microcystis spp.)迅速取代其成为绝对优势。秋季硅藻门的克洛脆杆藻(Fragiaria crotomensis)和隐藻门的啮齿隐藻(C. erosa)为优势。空间分布上水库北部浅水区域隐藻和硅藻生物量普遍高于南部; 受东南风影响, 蓝藻生物量在西洋河口S2点位最大。CCA分析表明夏季水华主体微囊藻的生物量与氮浓度正相关, 螺旋鱼腥藻在夏季仅作为第二优势种短暂出现于西洋河口处, 其出现与否受到磷营养盐的限制。       

Population growth dynamics of the rotifer Brachionus plicatilis cultured in non-limiting food condition

Population growth parameters in batch culture of Brachionus plicatilis under a continuous supply of freeze-dried microalgae powder have been determined. Two B. plicatilis strains (L- and S-types) and four microalgae species (Nannochloropsis oculata, Nannochloropsis gaditana, Nannochloris oculata and Tetraselmis suecica) have been tested, establishing the dynamics of growth at different daily food rations. Cultures showed a short lag phase, an exponential growth phase, a long post-exponential growth phase and long decline with episodic increases. In both rotifer strains, the best growth was obtained with Nannochloropsis oculata and the poorest with Nannochloris oculata.     

Enhancement of BODIPY505/515 lipid fluorescence method for applications in biofuel-directed microalgae production

This paper describes a microalgal cell lipid fluorescence enhancement method using BODIPY(505/515), which can be used to screen for lipids in wild-type microalgae and to monitor lipid content within microalgae production processes to determine optimal harvesting time. The study was based on four microalgae species (Dunaliella teteriolecta, Tetraselmis suecica, Nannochloropsis oculata, and Nannochloris atomus) selected because of their inherent high lipid content. An extended analysis was carried out with N. oculata due to the depressed fluorescence observed when compared with the other experimental strains. BODIPY(505/515) lipid fluorescence was determined for two solvent pre-treatment methods (DMSO and glycerol) and four staining condition parameters (analysis time, staining temperature, dye concentration, and algal cell concentration). It was found that lipid fluorescence of thick cell-walled microalgae, such as N. oculata, is significantly enhanced by both the pre-treatment methods and staining condition parameters, thereby significantly enhancing lipid fluorescence by ca. 800 times the base autofluorescence. The lipid fluorescence enhancement method provides a quick and simple index for in vivo Flow Cytometry quantification of total lipid contents for purposes of species screening or whole culture monitoring in biofuel-directed microalgae production.     

Biosorption of chromium (Cr(III)/Cr(VI)) on the residual microalga Nannochloris oculata after lipid extraction for biodiesel production

In order to increase the economic feasibility of biodiesel production from microalgae, the residual biomass after biodiesel production can be utilized as biosorbent for heavy metal removal. In this study, biosorption of chromium by residual Nannochloris oculata after lipid extraction was investigated. Increased surface area of N. oculata was observed after lipid extraction. Cr(III) removal increased as the pH increased from 2 to 6, while Cr(VI) removal was highest at pH 2 and it decreased with the increase in pH. Cr(VI) was reduced to Cr(III) in the presence of biomass under acidic conditions; X-ray photoelectron spectroscopy revealed that the converted Cr(III) was bound to the biomass. Chromium removal was significantly enhanced at high chromium concentrations, which indicates that surface reactions may occur at high chromium/biomass ratios. FTIR study indicated that phosphate and carboxyl functional groups of the biomass were mainly responsible for chromium binding.