菹草石油醚组分中抑藻活性物质的分离纯化和抑藻活性

菹草石油醚组分能显著抑制赤潮微藻米氏凯伦藻的生长,为了获悉该组分抑制米氏凯伦藻的物质基础,采用硅胶柱层析和制备薄层层析等方法,对菹草石油醚组分中的抑藻活性物质进行分离纯化。进一步应用质谱、核磁共振碳谱和氢谱等技术,鉴定抑藻活性物质。实验从菹草石油醚组分中分离纯化到4种抑藻活性物质。当浓度为16μg/m L时,它们对米氏凯伦藻表现出一定的抑藻活性。结构鉴定表明,此4种抑藻活性物质分别为二十五烷醇、邻苯二甲酸二(2-甲基己基)酯、棕榈酸和邻苯二甲酸二(5,5-甲基己基)酯。此4种化合物为首次从菹草中分离得到。抑藻活性分析表明,当浓度为50μg/m L时,4个样品均能显著抑制赤潮异弯藻的生长。其中,二十五烷醇和邻苯二甲酸二(5,5-甲基己基)酯还能较明显地抑制东海原甲藻和球形棕囊藻的生长。     

条斑紫菜提取物对4种赤潮微藻生长的抑制作用

研究条斑紫菜水溶性抽提液对前沟藻、中肋骨条藻、米氏凯伦藻和塔玛亚历山大藻等4种赤潮微藻生长的影响,在此基础上,利用甲醇、丙酮、乙酸乙酯、氯仿和石油醚等有机溶剂浸泡条斑紫菜干粉,经抑藻圈方法检测条斑紫菜水溶性抽提液的抑藻活性。通过测定藻细胞密度和细胞体积,观察藻细胞形态,分析藻细胞内叶绿素、蛋白质和多糖等生理生化指标的变化,对抑藻活性最大的提取物对前沟藻、米氏凯伦藻、中肋骨条藻和塔玛亚历山大藻赤潮微藻生长的抑制作用进行分析,并依次以石油醚、乙酸乙酯和正丁醇为提取溶剂,采用液液分离法对此提取物做了进一步分离。结果表明,当条斑紫菜水溶性抽提液浓度超过16g/L时能显著抑制4种赤潮微藻的生长,尤其是对前沟藻和米氏凯伦藻具有很强的抑制作用。在5种有机溶剂提取物中,甲醇提取物的抑制作用最为明显。进一步研究发现此提取物对4种赤潮微藻的生长抑制显著且具有浓度效应,在16g/L时,此提取物对前沟藻、米氏凯伦藻、中肋骨条藻和塔玛亚历山大藻的生长抑制率分别为70.5%、79.9%、67.1%和65.1%。同时,致使米氏凯伦藻、中肋骨条藻和塔玛亚历山大藻等3种赤潮微藻体积变小,运动能力下降,藻细胞出现空洞、细胞破碎和色素减褪等现象;...     

不同氮磷比对福建沿海米氏凯伦藻生长的影响

本文在不同氮磷比(N∶P=4∶1、8∶1、16∶1、32∶1、80∶1)培养条件下,对福建沿海赤潮海域分离的米氏凯伦藻(Karenia mikimotoi)进行培养,研究其生长特性。实验结果表明:不同的氮磷比对米氏凯伦藻的生长有明显的影响。过高或过低的氮磷比均不适合米氏凯伦藻的生长,该藻在N∶P=32∶1条件下比生长率最快,为0.33 d-1。米氏凯伦藻对氮的需求高于磷,在适当的磷限制环境中能够维持更长的生长周期。引发赤潮的主要原因并不是由于米氏凯伦藻赤潮暴发海域的低氮磷比,而是赤潮暴发过程中,米氏凯伦藻对营养盐的大量消耗,尤其是对氮的消耗。     

福建沿海米氏凯伦藻赤潮的初步研究

本文初步研究了2003年5～6月发生于福建连江海域的米氏凯伦藻(Karenia mikimotoi Hansen)赤潮.研究发现强降雨、充足的光照、稳定的气温和良好的海况是米氏凯伦藻赤潮发生和发展的必要条件.米氏凯伦藻适宜的温度范围为20～24℃,适宜的盐度范围为27.9～30.5.米氏凯伦藻细胞密度与营养指数以及磷酸盐浓度呈正相关性.米氏凯伦藻赤潮的发生发展过程中常伴随出现其它赤潮生物,在条件适合的情况下会向其它种类的赤潮转化.米氏凯伦藻赤潮对鲍鱼等海洋生物的危害较大,会造成严重的渔业经济损失.     

两种常见外来入侵赤潮藻的PCR鉴定

米氏凯伦藻与环状异帽藻为日本海域流入我国的两种外来入侵赤潮藻。利用核糖体ITS区分别设计出针对该两种藻的特异性PCR引物。通过prmier5.0软件设计多对引物,经PCR扩增,琼脂糖凝胶电泳检测、以筛选目标藻的特异性引物,并以链状亚历山大藻、立玛原甲藻、牟氏角毛藻、赤潮异弯藻作为阴性对照,做进一步PCR验证。筛选到米氏凯伦藻最佳引物Ki1F3/Ki1B3和环状异帽藻最佳引物YiF3a/YiB3a。两对特异性引物成功鉴定了两种外来入侵藻,而对其它藻种则是阴性反应,可为赤潮的预测预报提供分子鉴定基础。     

米氏凯伦藻对海洋生物致毒作用的研究进展

近年来,米氏凯伦藻(Karenia mikimotoi)已成为我国海域主要的有毒赤潮藻。仅2012年在我国浙江和福建海域就连续爆发了12起米氏凯伦藻赤潮,造成大量养殖鲍死亡,经济损失超过20亿元人民币。本研究通过收集国内、外最新报道,探讨了米氏凯伦藻赤潮对各类海洋生物,主要包括海洋浮游动物、贝类、鱼类以及哺乳类毒性作用的研究进展。研究发现,溶血毒素是米氏凯伦藻赤潮引发海洋生物大量死亡的主要原因;直接接触活的米氏凯伦藻细胞是导致浮游动物死亡的主要途径;米氏凯伦藻产生的溶血毒素和鱼毒素,可溶解鱼类的鳃组织,从而造成鱼类死亡。然而,关于米氏凯伦藻产生的溶血毒素是否会在贝类体内累积、在贝类体内的迁移和转化规律、米氏凯伦藻赤潮对鲍鱼死亡的致死途径与致死机制、溶血毒素在生物链传递过程中是否会发生改变、这些毒素对人类健康的影响机制与综合作用等问题还不了解,这些问题亟待深入的研究。米氏凯伦藻溶血毒素的研究报道还处于起步阶段,毒素以何种方式导致红细胞破裂、不同红细胞膜上组分的差异和毒素的溶血活性是否存在对应关系等问题也都需要开展进一步的研究。     

碳、氮、磷营养对7种海洋微藻种群增长的影响研究

文章采用批次培养方式研究双胞旋沟藻(Cochlodinium geminatum)、米氏凯伦藻(Karenia mikimotoi)、中肋骨条藻(Skeletonema costatum)、牟氏角毛藻(Chaetoceros muelleri)、球形棕囊藻(Phaeocystis globosa)、亚心形扁藻(Platymonas subcordiformis)和眼点拟微绿球藻(Nannochloropsis oculata)在不同碳(C)、氮(N)、磷(P)条件下的生长特性,以期为养殖尾水的原位处理筛选优良藻种(株)。结果表明,这7种微藻的比生长速率与C、N、P浓度总体呈正相关。相较对照组,C对旋沟藻、骨条藻和拟微绿球藻比生长速率的促进作用分别为35%、19%和19%,P对比生长速率的最大促进作用介于25.01%~446.60%,硝酸盐为16.54%~77.52%,铵盐为15.79%~88.82%,尿素为25.16%~71.43%。7种微藻的最大比生长速率与细胞体积总体呈负相关,棕囊藻的比生长速率最高,拟微绿球藻和扁藻最大比生长速率与其他藻华种相比无显著差别。微藻的氮磷吸收速率随细胞体积增加呈升高趋势,单位氮磷利用效率随细胞体积增加呈降低趋势。培养液中P吸收速率显著高于N,氮磷比均呈升高趋势。眼点拟微绿球藻和亚心形扁藻可用于养殖尾水氮磷的快速去除。     

6种微藻生长及脂肪累积对氮含量变化的响应

研究了绿色巴夫藻、米氏凯伦藻、球等鞭金藻、三角褐指藻、海水小球藻、衣藻的生长及脂肪累积对氮含量变化的响应。试验结果表明,6种微藻生长及总脂对不同氮含量均有一定程度的响应。微藻生长,绿色巴夫藻、米氏凯伦藻、球等鞭金藻3种微藻响应明显,绿色巴夫藻以氮含量100%组生长最快,第10d时,比氮含量40%组快512%,比氮含量20%组及60%组分别快62%、61%;米氏凯伦藻氮含量80%组生长最快,第10d达到最高值,超出其他试验组约60%;球等鞭金藻氮含量80%组最快,第10d生长量超过100%试验组67%;三角褐指藻、海水小球藻、衣藻响应程度小于其他微藻。总脂积累,三角褐指藻氮含量80%组总脂含量最高,达细胞干质量的80.77%,是氮含量100%组2.5倍;绿色巴夫藻氮含量60%组最高,达细胞干质量的73.68%,是氮含量100%组3.4倍;其他4种微藻在降低氮含量后,总脂均超过细胞干质量的30%。三角褐指藻在生物柴油制备方向极具开发前景。     

浙江海域米氏凯伦藻赤潮成因的初步研究

通过对2005年5月30日发生在浙江海域米氏凯伦藻赤潮的连续监测分析,初步研究了本次赤潮发生的成因。结果表明:24℃左右的气温、非降雨的天气和良好的海况是赤潮发生及维持的重要因素。米氏凯伦藻生长所需最佳水温为23.4~23.8℃,富营养化是赤潮发生的基本条件,而赤潮生物的数量与水体中的无机氮呈负相关关系,与磷酸盐呈正相关关系。     

营养因子对赤潮异弯藻生长的影响

以赤潮异弯藻为材料,采用正交试验方法,研究了氮、磷、微量元素混合液和维生素混合液对赤潮异弯藻生长的影响。试验结果表明,在对数期和稳定期,4种营养因子中对赤潮异弯藻生长影响的顺序为磷>维生素>氮≥微量元素;在衰亡期,营养因子中对赤潮异弯藻生长影响的顺序为氮>磷>维生素>微量元素。在对数期,氮、磷、微量元素和维生素等营养因子的最适浓度分别为880、18、10.55、0.32μmol/L;在稳定期和衰亡期,最适浓度分别为2640、108、21.1、0.96μmol/L。各营养因子在不同时期均对赤潮异弯藻的增殖具有显著影响(P<0.01)。     

Neurotoxins in a toxic red tide of Heterosigma akashiwo (Raphidophyceae) in Kagoshima Bay, Japan

In April 1995, an unusual large-scale red tide of Heterosigma akashiwo (Hada) Hada occurred, causing massive fish kills in Kagoshima Bay, Japan. Four neurotoxic components, HaTx-i , HaTx-ii a, HaTx-ii b and HaTx-iii , which corresponded to brevetoxin components. PbTx-2. PbTx-9, PbTx-3 and oxidized PbTx-2, were inferred from analysis of Heterosigma red tide toxins on TLC and HPLC.     

浒苔对赤潮异湾藻的克生作用

研究了浒苔对赤潮微藻—— 赤潮异湾藻生长的克生效应。结果表明, 浒苔新鲜组织对赤潮异湾藻的生长具有强烈的克生效应; 一次性添加浒苔培养过滤液对赤潮异湾藻生长没有显著的抑制作用, 而半连续添加浒苔培养过滤液对赤潮异湾藻生长具有显著的克生作用; 浒苔干粉末和甲醇抽提液均对赤潮异湾藻的生长产生显著的抑制作用。试验结果表明, 抑制赤潮异湾藻生长的物质很可能更多地存在于浒苔组织内, 向环境中分泌的克生物质较少。因而, 克生物质的连续分泌是有效克制赤潮异湾藻生长的关键。同时, 克生作用存在着明显的浓度效应, 当抑制物浓度达到一定阈值才表现出明显的克生作用, 在阈值之上浓度越高克生作用越强。     

Characteristics of the marine environment and algal blooms in Gamak Bay

This study was conducted to gain insight into the characteristics of algal blooms in relation to the marine environment of Gamak Bay. From the first known occurrence of algal blooms in 1984 until 2006, 23 causal species have been identified, the most common ones being Prorocentrum sp., Cochlodinium polykrikoides, Chaetoceros sp., Skeletonema costatum, and Heterosigma akashiwo. A principal component analysis indicated that blooms of these species develop under different conditions in terms of water temperature, salinity, precipitation, and insolation. Field data showed Cochlodinium polykrikoides to be favored by high temperatures and to be euryhaline, whereas Skeletonema costatum appeared to be eurythermal and euryhaline. Prorocentrum sp. and Chaetoceros sp. appeared to be stenothermal and stenohaline. Finally, Heterosigma akashiwo appeared at the lowest temperature and highest salinity of the five species and was classified here as stenothermal and stenohaline.     

Effect of probiotic (Bacillus spp.) addition during larvae and postlarvae culture of the white shrimp Litopenaeus vannamei

The effect of the addition of Bacillus probiotic during larvae and postlarvae culture of the white shrimp Litopenaeus vannamei was examined in three separate experiments: (I) Nauplius_4‐5 to Zoea₃, which were exposed to probiotic in the water (P_w), only in the microalgae (P_m), in the water and microalgae (P_wm) and a control with no probiotic (C); (II) Mysis₁ to Mysis₃, which were exposed to probiotic in the water (P_w), only in Artemia (P_a), in the water and Artemia (P_wa) and a control (C); (III) PL₁ to PL₁₀, which were exposed to similar treatments for mysis experiment. The use of probiotic Bacillus spp. resulted in an increase in the survival and growth of zoea and mysis phases, especially when the probiotic was added only in the water. For postlarvae, the use of the probiotic had no influence on the zootechnical parameters, however, there was a reduction in the count of presumptive Vibrio both for water and shrimp.     

Isolation and characterization of Noctiluca-killing bacteria from a shrimp aquaculture pond in Thailand

To control harmful algal blooms (HABs), in particular dinoflagellate Noctiluca scintillans, which causes damage to shrimp production, algicidal bacteria were isolated from shrimp pond water and screened for their killing effect against N. scintillans under laboratory condition. Among 260 bacterial isolates, 10 showed killing effects on N. scintillans. Out of 5 strains showing relatively stronger killing activity, 4 strains belonged to the Gammaproteobacteria group. Among these, strain BS2 (identified as Marinobacteria salsuginis) showed the strongest Noctiluca-killing activity. In the present study, we also investigated bacterial effects against growth of other phytoplankton species. Strain BS2 killed only N. scintillans but not any other phytoplankton (Heterosigma akashiwo NIES-6, Chatonella antique NIES-1, Chaetoceros ceratosporum, Prorocentrum lima, and Dunaliella sp.). Results of the present study lead us to expect prevention of damage by HABs, in particular dinoflagellate N. scintillans, in shrimp farming and mitigation for shrimp production in ponds using Noctiluca-killing bacteria.     

星康吉鳗形态性状与体重的通径分析及生长曲线拟合

为探明星康吉鳗（Conger myriaster）形态性状和体重的关系，测定了体重（Y）及全长（X₁）、肛长（X₂）、尾长（X₃）、头长（X₄）、吻长（X₅）、眼径（X₆）、眼后头长（X₇）、背鳍前长（X₈）、躯干长（X₉）、头宽（X₁₀）、眼间距（X₁₁）等11个形态性状，通过相关分析、通径分析、回归分析等方法研究了形态性状对体重的影响；通过曲线拟合获得4个形态性状与体重的最佳拟合模型。结果表明，星康吉鳗各形态性状之间以及形态性状与体重之间均呈极显著正相关关系（P< 0.01）。相关分析发现肛长（X₂）与体重的相关系数最大（0.970）；通径分析发现肛长（X₂）对体重的直接作用最大（0.450），头宽（X₁₀）通过肛长（X₂）对体重的间接作用最大（0.431）；决定系数分析发现，肛长（X₂）对体重的直接决定系数最大（0.203），肛长（X₂）和头宽（X₁₀）的共同决定系数最大（0.172）。形态性状（X）与体重（Y）的多元回归方程为Y=-113.859+6.481X₂+49.213X₆+11.514X₇+20.145X₁₀（R²=0.954）。星康吉鳗肛长（X₂）、眼后头长（X₇）、头宽（X₁₀）与体重的最佳拟合模型为指数函数，模型方程分别为Y=1.466e^0.287X、Y=1.435e^1.328X、Y=1.970e^1.974X，眼径（X₆）与体重的最佳拟合模型为线性，方程为Y=-77.460+222.093X。结论认为，在星康吉鳗选育时，应以肛长（X₂）和头宽（X₁₀）为主要选择性状，以眼径（X₆）和眼后头长（X₇）为辅助选择性状。     

Bioprospection of Microalgae and Cyanobacteria as Biocontrol Agents Against Vibrio campbellii and Their Use in White Shrimp Litopenaeus vannamei Culture

Vibrio spp. are the most common and harmful shrimp pathogenic bacteria; however, microalgae and cyanobacteria have the ability to produce antimicrobial substances against these species. In this study, the organic and aqueous extracts of 28 species of marine microalgae and cyanobacteria were screened against Vibrio campbellii M1. Two of these phytoplankton species with antibacterial activity in aqueous extracts (Dunaliella tertiolecta and Skeletonema costatum) and nontoxic to brine shrimp Artemia franciscana nauplii were used to evaluate their anti‐Vibrio effect when used as green‐water cultures in Vibrio‐challenged white shrimp Litopenaeus vannamei cultures. No differences in mortality of juvenile L. vannamei were observed between treatments tested, suggesting that the pathogenicity of V. campbellii could be related to the growth stage of shrimp. The proximal composition of D. tertiolecta and S. costatum was in the recommended range for penaeid shrimp nutrition, allowing shrimp supplemented with these microalgae to have significantly greater total length and weight than control shrimp. Shrimp supplemented with S. costatum presented the highest values of organic mass (11.48 mg/organism) and growth rate (0.31 mg/d) in comparison to D. tertiolecta. These results indicate that microalgae are not only capable of producing antibacterial compounds against Vibrio but can also help shrimp nutrition.     

Analysis of Phytosterol,Fatty Acid,and Carotenoid Composition of 19 Microalgae and 6 Bivalve Species

ABSTRACT

In this study, the functional components of 19 microalgae and 6 bivalve species were investigated in the context of the application in bivalve feeding and human health food. Principal component analysis was performed to detect any association between the functional components and individual microalgal species or taxonomic group. The proportions of the functional components differed depending on the taxonomic group or species of microalga. The genera Cheatoceros, Thalassiosira, and Isochrysis contained high concentrations of fucosterol and fucoxanthin, which are present in large amounts in brown algae. Diatoms, haptophytes, and eustigmatophytes, which are used as feed for bivalves, were rich in fucosterol and eicosapentenoic acid (EPA); further, haptophytes were rich in docosahexaenoic acid (DHA). In addition, the microalgae associated with red tide, i.e., the raphidophytes, were found to be rich in fucoxanthin, β-sitosterol, and EPA, whereas dinoflagellates were rich in DHA. Seven bivalve species also contained high concentrations of fucosterol, EPA, and DHA, as did microalgae, which were used to feed by bivalves. These results are useful in selecting microalgae effectively as feed of the bivalves.     

Effect of microalgae and temperature on absorption efficiency of razor clam (Sinonovacula constricta Lamark, 1818)

The razor clam (Sinonovacula constricta) is a bivalve species living in the lower to mid intertidal zones along the coasts of China, Japan and Korea. In this study, the effects of temperature, microalgae species and concentration on the absorption efficiency (AE) of cultured adult S Constricta fed with six species of microalgae, including Chlorella sp., Pavlova viridis, Nitzschia closterium f. minutissima, Platymonas subcordiformis, Nannochloropsis oculata and Isochrysis zhanjiangensis was examined. The clams were exposed to three different temperatures (15, 20 and 25°C) and five microalgae concentrations (0.16, 0.27, 0.37, 0.45 and 0.53 mg L^?1). The results indicated that under different temperatures, there is significant difference (P < 0.05) in AE and the efficiency peaks at the water temperature of 20°C. Under different microalgae concentrations, the absorption efficiencies were also significantly different (P < 0.05) and there was a negative correlation between AE and microalgae concentration. At different combinations of temperature and microalgae concentration, the absorption efficiencies of Chlorella sp. and N. oculata were lower than those of other microalgae. The interaction of temperature and microalgae concentration affected the AE significantly (P < 0.05).     

Selectivity of the rotifer Brachionus plicatilis fed mixtures of algal species with various cell volumes and cell densities

The ability for food selectivity of rotifer Brachionus plicatiliswas studied in the laboratory by feeding mixtures of microalgae with various cell volumes. Chlorella sp. (≈22 µm³) was the reference algal species, and Asteromonas gracilis, (Chlorophyta) (≈2150 µm³), Tetraselmis suesica (≈268 µm³), Dunaliella salina (≈52 µm³) and Chaetoceros sp. (≈150 µm³) the experimental species. Each was mixed with Chlorella and fed in three experiments. In the first experiment, filtration and ingestion rates of rotifers each fed with algae revealed that the highest values were measured with the mixture of Chlorella + Asteromonas, and the lowest for Chlorella + Chaetoceros. In the second and the third experiments, by using several combinations of algal densities with the mixture of Chlorella + Asteromonas, a selectivity ability of the rotifers for Asteromonas was found. A hypothesis is presented that accounts for the preference of rotifers for Asteromonas, which is suggested as a new candidate species for use in live food production of fish hatcheries.