两种微藻改善虾池环境增强凡纳对虾抗病力的研究

人工引入波吉卵囊藻和微绿球藻于凡纳对虾养殖环境中,检测与凡纳对虾抗病力有关因子变化和测定主要水质因子,研究微藻生态调控对凡纳对虾抗病力的影响。结果表明,引入波吉卵囊藻和微绿球藻能改善养殖水体的水质,凡纳对虾的血细胞数目,血清蛋白的含量以及酚氧化酶、超氧化物歧化酶,溶菌酶,抗菌酶的活性都较对照组显提高。因此,微藻生态调控是防止对虾疾病的重要技术措施。     

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.     

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.     

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.     

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.     

虾池拟柱胞藻爆发的生态因子调查

2002～2003年,在广东珠三角地区凡纳滨对虾(Litopenaeus vannamei)低盐度养殖虾池水体生态的调查过程中发现部分虾池爆发拟柱胞藻(Cylindrospermopsis raciborskii).在调查的低盐度虾池中,共统计20个拟柱胞藻爆发的虾池,其种群密度平均高达7.74×10⁷ filaments·L^-1,平均优势度达47.0%.虾池水体平均水温为30.3℃,pH为8.9,盐度1.4,透明度19cm,化学耗氧量为24.3mg·L^-1.拟柱胞藻爆发的虾池水体环境表现为高水温、高pH、高有机质、低盐度且水体不断被搅动;通常爆发在对虾养殖的中后期富营养化水体.调查发现大量的拟柱胞藻对虾池其它藻类和微生物菌群表现一定的抑制作用,蓝纤维藻和小环藻等少数藻种可与拟柱胞藻形成共优势种.拟柱胞藻为广东珠三角凡纳滨对虾低盐度虾池重要优势藻种,且以螺旋型为主.     

凡纳滨对虾低盐度高产虾池环境微生物生态研究

对广东珠三角地区,凡纳滨对虾低盐度高产虾池环境微生物调查结果,养殖水体异养菌平均数量为5.15×10⁴cfu·mL^-1,致病性弧菌为5.00×10³cfu·mL^-1,池底泥浆中异养细菌平均数量为2.41×10⁶cfu·mL^-1,致病性弧菌为1.45×10⁵cfu·mL^-1。不同虾池异养细菌的数量差别小而稳定,致病性弧菌的数量差别及波动大。淤泥较深的老化虾池,水体和泥浆中异养细菌平均分别为6.10×10⁴cfu·mL^-1和2.89×10⁶cfu·mL^-1,致病性弧菌为6.00×10³cfu·mL^-1和2.14×10⁵cfu·mL^-1;无淤泥虾池水体和泥浆中异养细菌平均分别为4.53×10⁴cfu·mL^-1和2.08×10⁶cfu·mL^-1,致病性弧菌为4.34×10³cfu·mL^-1和9.86×10⁴cfu·mL^-1。老化虾池底泥致病性弧菌明显偏高,成为老化虾池易爆发虾病的重要原因。低盐度虾池水体异养菌和致病性弧菌的数量变化,表现为养殖前期高,中后期低而稳定;池底泥浆中异养菌和致病性弧菌的数量变化,呈不规则波动。高的养殖水温对养殖环境异养菌和致病性弧菌表现抑制作用,养殖环境微生物与水体浮游藻类也有一定的关系。     

微生物制剂对斑节对虾亲虾池异养细菌的影响

研究了施用10 mg·kg-1光合细菌以及1、2和3 mg·kg-1芽孢杆菌制剂对斑节对虾亲虾培育池水体异养细菌和弧菌数量的影响.结果表明:在试验初期,试验组和对照组水体总异养细菌和弧菌数量都明显上升;试验后期,试验池水体异养细菌和弧菌数量均明显下降,明显低于对照池;施用10 mg·kg-1光合细菌能显著抑制亲虾池水体异养细菌的生长(P＜0.01);施用3 mg·kg-1芽孢杆菌对亲虾池水体弧菌数量的抑制效果最好(P＜0.01).     

基于16S rRNA基因扩增子测序分析日本囊对虾肠道菌群结构与功能的特征

【背景】肠道菌群在对虾的生理活动中起关键作用。日本囊对虾是我国海水养殖虾类中的主要品种之一,迄今为止有关其肠道菌群结构与功能的研究还鲜有报道。【目的】利用高通量测序技术探究日本囊对虾肠道菌群的组成结构与功能作用,揭示虾体肠道菌群与外源菌群结构间的相关性。【方法】60 d的养殖周期结束后,分别采集日本囊对虾肠道样品(归为虾肠组,n=3)、养殖水体样品(归为水体组,n=3)和对虾饲料样品(归为饲料组,n=3),提取各样品总DNA进行16SrRNA基因扩增子测序,基于生物信息学方法分析与比较样品间的菌群结构特征,并使用PICRUSt软件预测日本囊对虾肠道菌群功能。【结果】3组样品测序共获得822 713条有效序列,抽平处理后可聚类为3 416个OTU。虾肠组样品中有28.49%、59.30%的OTU可以依次在水体组、饲料组样品中检测到。门水平上,虾肠组样品中的优势菌门为变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和梭杆菌门(Fusobacteria)。水体组、饲料组与虾肠组样品中的优势菌门结构不尽相同,但均由变形菌门和拟杆菌门组成。属水平上,虾肠组样品中的优势菌属包括弧菌属(Vibrio)、另类弧菌属(Aliivibrio)、假交替单胞菌属(Pseudoalteromonas)、假黄棕杆菌属(Pseudofulvibacter)、科尔韦尔氏菌属(Colwellia)、小纺锤状菌属(Fusibacter)、发光杆菌属(Photobacterium)、脱硫弧菌属(Desulfovibrio)、嗜冷杆菌属(Psychrobacter)以及弓形杆菌属(Arcobacter)。水体组和饲料组中检出的核心菌属结构与虾肠组相比有明显差异,其中海命菌属(Marivita)和假单胞菌属(Pseudomonas)分别为养殖水体及对虾饲料样品中的最优势菌属。PICRUSt预测结果显示,日本囊对虾肠道菌群的基因功能主要与新陈代谢类功能有关,包含氨基酸代谢、碳水化合物代谢与能量代谢等。【结论】日本囊对虾肠道菌群与其他种类对虾肠道菌群的结构间存在共性,其形成在一定程度上受到了外源菌群的干预,并在虾体的日常代谢活动中发挥了一定的作用。     

广东湛江地区几种海洋微藻中的脂肪酸检测

本文检测了广东湛江地区6种典型的海洋经济微藻中绿藻纲的微绿球藻(Nannochloropsis oculata)和小球藻(Chlorella sp.)、金藻纲的湛江等鞭金藻     

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.     

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.     

THE BIOCHEMICAL COMPOSITION OF MARINE MICROALGAE FROM THE CLASS EUSTIGMATOPHYCEAE1

The biochemical composition of four strains of microalgae from the class Eustigmatophyceae was determined to assess their usefulness as live feeds for mariculture and to establish characteristic features for use in chemotaxonomic studies. We studied Nannochloropsis salina (strain CS-190) from Scotland, two strains of Nannochloropsis oculata (CS-179 and CS-216) from Japan, and an unnamed eustigmatophyte (CS-246) isolated from Queensland waters that appears to be closely related to N. oculata. Gross compositional features were similar: total carbohydrate ranged from 5.2% (N. oculata CS-179) to 8.9% (N. salina) of cell dry weight. Polysaccharide comprised 74% (N. oculata CS-179) to 88% (CS-246) of this total. Glucose was the principal polysaccharide sugar (45.2–66.2% of total sugars). Other sugars included fucose, galactose, mannose, rhamnose, ribose, and xylose (2.0–14.0%). Arabinose was a minor constituent in all species (0.6–1.7%). Protein varied from 17.8% (N. salina) to 22.1% (N. oculata CS-216) of the cell dry weight. The major amino acids were arginine, glutamate, and asparatate (7.2–10.4% of total amino acids), with methionine, cystine, histidine, tryptophan, hydroxy-proline, ornithine, and γ-aminobutyric acid much less abundant (0.03–2.6%). Lipid content ranged from 8.2% (N. oculata CS-216) to 16.9% (N. salina) of cell dry weight, the latter value reflecting enhanced concentrations of triacylglycerols in N. salina. The major fatty acids were palmitic acid (16:0), palmitoleic acid [16:1(n-7)], and eicosapentaenoic acid [20:5(n-3)] with lesser amounts of lauric acid (14:0), linoleic acid [18:2(n-6)], and others. The sterols consisted almost entirely of cholesterol, which is an essential constituent of crustacean diets. Chlorophyll a ranged from 0.6% (N. oculata CS-216) to 1.7% (N. oculata CS-179 and N. salina) of cell dry weight. Chlorophylls b and c were not detected. All strains contained a characteristic pattern of carotenoid pigments, which included violaxanthin, β-carotene, zeaxanthin, and a pigment tentatively identified as vaucheriaxanthin-ester. The distinctive pigment and lipid compositional data can be used as chemotaxonomic markers for Nannochloropsis and for assigning microalgae to the class Eustigmatophyceae. Nannochloropsis oculata is widely used as an algal feed in mariculture, and based on the similarity of the biochemical data, both N. salina and the unnamed tropical species should also prove to be nutritionally valuable live algal feedstocks. Feeding trials will be needed to confirm this.     

Increased pigment and lipid content,lipid variety,and cell and population size of the microalgae Chlorella spp. when co-immobilized in alginate beads with the microalgae-growth-promoting bacterium Azospirillum brasilense

Three strains of the freshwater microalgae used for wastewater treatment, Chlorella vulgaris and Chlorella sorokiniana co-immobilized separately in alginate beads with the microalgae-growth-promoting bacterium Azospirillum brasilense Cd, resulted in significant changes in microalgal-population size, cell size, cell cytology, pigment, lipid content, and the variety of fatty acids produced in comparison with microalgae immobilized in alginate without the bacterium. Cells of C. vulgaris UTEX 2714 did not change in size, but the population size within the beads significantly increased. On the other hand, C. vulgaris UTEX 395 cells grew 62% larger, but their numbers did not increase. The population of C. sorokiniana UTEX 1602 increased, but not their cell size. The content of pigments chlorophyll a and b, lutein, and violoaxanthin increased in all microalgal species. The lipid content also significantly increased in all three strains, and the number of different fatty acids in the microalgae increased from four to eight. This study indicates that the microalgae-growth-promoting bacterium induced significant changes in the metabolism of the microalgae.     

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.     

Increased growth of the microalga Chlorella vulgaris when coimmobilized and cocultured in alginate beads with the plant-growth-promoting bacterium Azospirillum brasilense

Coimmobilization of the freshwater microalga Chlorella vulgaris and the plant-growth-promoting bacterium Azospirillum brasilense in small alginate beads resulted in a significantly increased growth of the microalga. Dry and fresh weight, total number of cells, size of the microalgal clusters (colonies) within the bead, number of microalgal cells per cluster, and the levels of microalgal pigments significantly increased. Light microscopy revealed that both microorganisms colonized the same cavities inside the beads, though the microalgae tended to concentrate in the more aerated periphery while the bacteria colonized the entire bead. The effect of indole-3-acetic acid addition to microalgal culture prior to immobilization of microorganisms in alginate beads partially imitated the effect of A. brasilense. We propose that coimmobilization of microalgae and plant-growth-promoting bacteria is an effective means of increasing microalgal populations within confined environments.     

固定化微绿球藻去除NH4+-N、PO43——P效果的研究

为了探究固定化微绿球藻（Nannochloropsis oculata）去除污水中NH4+-N、PO43--P的效果,采用海藻酸钠固定化包埋技术进行实验。开展了固定化藻球大小、藻细胞包埋密度、藻球投放质量及充气培养条件对NH4+-N、PO43--P去除效果的单因子试验研究。结果表明,固定化藻球大小、藻细胞包埋密度、藻球投放质量和充气培养条件对NH4+-N、PO43--P的去除效果影响显著（P0.05）。藻球直径3.5 mm时生长速率（K）值最大（0.3320.002）,同时NH4+-N、PO43--P去除率效果最佳,分别为（75.083.83）%和（80.803.81）%;藻细胞包埋密度100104 cells/ball时K值最大（0.3300.033）,而NH4+-N、PO43--P去除率则以藻细胞包埋密度300104 cells/ball组为佳,分别达（87.200.43）%和（82.581.72）%,但考虑单位藻细胞去除率,包埋密度以100104 cells/ball为宜;随着藻球用量的增加K值下降,10 g/L组K值最大（0.3010.02）、50 g/L组K值最小（0.1930.01）,投放量30和50 g/L时NH4+-N去除率较高分别为（84.120.78）%和（84.630.45）%,30 g/L组PO43--P去除率最高达（77.131.43）%。综合考虑,藻球投放量选用30 g/L为宜;充气条件培养K值、NH4+-N和PO43--P去除率显著（P0.05）高于不充气,K值分别为（0.3060.006）和（0.1770.010）;NH4+-N去除率分别为（85.930.45）%和（49.320.45）%;PO43--P去除率分别为（66.665.00）%和（46.292.12）%。研究优化了微绿球藻固定化条件:固定化微绿球藻应进行充气培养,藻球规格3.5 mm、藻细胞包埋密度100104 cells/ball、藻球投放量30 g/L。     

二氧化碳减排产柴油微藻培养体系研究进展

污水资源化、二氧化碳减排及微藻生物柴油是当前能源与环境领域的前沿课题。以下围绕污水及烟道气资源化培养产油微藻的培养体系,就藻种、营养条件、培养方式、培养环境及微藻生物反应器等影响产油微藻培养的因素研究进展进行了综述。在综述的基础上提出：由于微藻具有特殊营养方式,通过藻种筛选、微藻营养条件和培养环境的优化以及高效光生物反应器和生产工艺等的创新,可利用污水进行产油微藻生产,以获得生物柴油等高附加值产品,实现微藻生物能源、污水资源化处理和CO2减排三者高度耦合的产油微藻生产体系,从而减少微藻培养费用及污水处理费用,因此,该体系具有重要的环境、社会、经济价值和商业化应用前景。