萱藻丝状体的种质保存技术

采用液体保存法和胶囊化低温保存法对萱藻丝状体进行保存,探讨了液体培养法中温度、光照强度以及胶囊化低温保存法中胶球含水量对萱藻丝状体种质保存的影响。结果显示:(1)在液体保存法中,温度对萱藻丝状体的保存影响较小,当光强为5.4μmol/(m2·s)时,6~18℃均可实现萱藻丝状体的长期保存;(2)光照强度对萱藻丝状体的液体保存影响显著,16.2μmol/(m2·s)及以上的光强不利于种质的保存,5.4μmol/(m2·s)对萱藻丝状体的液体保存最有利;(3)在6~18℃,5.4μmol/(m2·s)条件下保存萱藻丝状体60 d后,萱藻丝状体细胞状态良好且存活率仍在97%以上;(4)胶球含水量过高或过低均会降低萱藻丝状体胶囊化低温保存后的存活率,15%是萱藻丝状体胶球的最适含水量,在该条件下,萱藻丝状体保存1个月后的存活率仍在50%以上。研究证明,在适宜条件下,液体保存法和胶囊化低温保存法均可实现萱藻丝状体的种质保存。     

光照对中国南北方萱藻丝状体生长发育影响的比较

通过分离、培养我国北方海域(烟台、青岛)和南方海域(舟山、温州)的萱藻丝状体,比较研究了不同光强[7.2、21.6、36.0、50.4、64.8、86.4、108.0和129.6μmol/(m2·s)]和不同光周期(8L:16D、10L:14D、12L:12D、14L:10D和16L:8D)对我国南北方萱藻丝状体生长及发育的影响。结果表明:(1)过高[≥108.0μmol/(m2·s)]或过低[≤21.6μmol/(m2·s)]光强条件均不利于我国南北方萱藻丝状体的快速生长;(2)我国南北方萱藻丝状体生长的最适光强条件相同,在64.8μmol/(m2·s)条件下培养20 d后,其丝状体的增重倍比和日均增长率均达到最大值;(3)我国南北方萱藻丝状体发育的最适光强条件均为36.0μmol/(m2·s),培养20 d后,烟台、青岛、舟山和温州的萱藻丝状体的孢子囊枝比例均达到最大值,分别为34.72%±0.91%、35.06%±1.17%、35.37%±0.59%和34.33%±0.41%;(4)我国南北方萱藻丝状体生长的最适光周期条件不同,培养20 d后,北方萱藻丝状体的增重倍比和日均增长率均在14L:10D条件下达到最大值,分别为82.75%±2.39%、3.00%±0.36%和81.28%±4.53%、3.04%±0.49%,而南方均在16L:8D条件下达到最大值,分别为89.52%±0.88%、3.18%±0.30%和88.66%±7.09%、3.22%±0.26%;(5)经过20 d的培养观察,我国北方萱藻丝状体发育的适宜光周期范围为8L:16D~10L:14D,而南方的适宜光周期范围为10L:14D~12L:12D。     

不同光照条件对小新月菱形藻和等鞭金藻8701生长及生化成分的影响

研究了小新月菱形藻(Nitzschiaclosteriumf.minutissima)(MACC/B228)和等鞭金藻8701(IsochrysisgalbanaParke8701)(MACC/H060)在4种光照条件下的生长情况,以及不同光强下、不同时期收获的藻细胞中总脂和碳水化合物含量的变化。结果表明,小新月菱形藻在光强70μmol·s-1·m-2时细胞分裂频率(μ)最大,最适光强70～140μmol·s-1·m-2;260μmol·s-1·m-2光强能导致细胞分裂频率变慢,指数期缩短。等鞭金藻8701在光强140μmol·s-1·m-2时μ值最大,260μmol·s-1·m-2下略有降低。2种藻均在低光下脂肪含量多,小新月和等鞭金藻脂肪含量分别占干重的25.5%～35.3%和28.3%～37.5%;而碳水化合物含量少,分别占干重的5.5%～18.2%、3.9%～11.4%;在高光强下相反。不考虑光照条件的影响,2种藻细胞内碳水化合物的含量均在静止期达到最大,分别占干重的18.98%和23.32%。小新月菱形藻在70μmol·s-1·m-2光强下的指数期细胞有最大的脂肪含量,等鞭金藻的最高脂肪含量出现于光强20μmol·s-1·m-2下指数末期的细胞中。二者在光强140μmol·s-1·m-2下生长且进入静止期时均可获得较大生物量,同时细胞的脂肪和碳水化合物的含量也处于较高水平。     

不同光强处理对长茎葡萄蕨藻叶绿素荧光特性的影响

通过调制叶绿素荧光仪(MINI-PAM),对长茎葡萄蕨藻(Caulerpa lentillifera)在10μmol/(m~2·s)、120μmol/(m~2·s)、360μmol/(m~2·s)光强下的叶绿素荧光特性进行了研究。结果表明:随着光照强度从10μmol/(m~2·s)上升到360μmol/(m~2·s),长茎葡萄蕨藻最大光量子产量(F_v/F_m)、实际光量子产量、相对电子传递速率(rETR)、快速光曲线初始斜率均呈下降趋势;用120μmol/(m~2·s)和360μmol/(m~2·s)光强处理的样品光化学猝灭(qP)较高,而非光化学猝灭(NPQ)较低;不同光强处理4 h和8 h时,与10μmol/(m~2·s)处理组相比,120μmol/(m~2·s)处理组最大相对电子传递速率(rETR_(max))、半饱和光强(I_k)均显著上升,360μmol/(m~2·s)处理组I_k显著上升,rETR_(max)明显降低;在360μmol/(m~2·s)光强下处理12 h与处理8 h相比,样品rETR_(max)、I_k均显著降低。上述结果说明长茎葡萄蕨藻偏向适应10μmol/(m~2·s)的低光条件,在120μmol/(m~2·s)和360μmol/(m~2·s)的高光条件下能通过不同的方式提高对光强的耐受能力,但是在360μmol/(m~2·s)光强下处理12 h会引起光氧化对光合结构造成破坏。     

生态因子对萱藻生长发育影响的研究现状及展望

主要阐述了国内外有关于生态因子对萱藻(Scytosiphon lomentaria)生长发育影响的研究现状,并对有关于萱藻的进一步研究提出了展望。温度、光照、盐度、营养液及抗生素等生态因子均会对萱藻生长发育造成不同程度的影响,温度和光照变化对萱藻生长发育的影响最为明显。萱藻配子体阶段的适宜温度较低,为6～12℃,而孢子体阶段在5～23℃范围内均可生长,生长速度随温度上升而增加;萱藻不同生长阶段的最适光照强度也有所不同,萱藻配子体阶段的最适光照范围为36～54μmol·m~(-2)·s~(-1),较低光强6～30μmol·m~(-2)·s~(-1)是萱藻孢子体生长发育的适宜范围,而高光强54μmol·m~(-2)·s~(-1)有利于萱藻孢子的附着;盐度、pH对萱藻生长发育影响显著,过高或过低均不利于萱藻生长;不同营养盐、维生素及抗生素对萱藻的生长发育产生的影响不同,在萱藻不同的生长阶段,应根据生长状况及时给予适宜浓度的营养盐或药物添加剂以促进萱藻生长。     

巨藻幼苗光合、呼吸作用的初步研究

为了研究巨藻幼苗发育及生理生态适应性规律,以巨藻配子体为材料,研究光照周期(10 L∶14 D、12 L∶12 D、14 L∶10 D、16 L∶8 D)对配子体采苗的影响;以光照强度、温度、营养盐浓度3种环境因子设置单因素实验,研究其对巨藻幼苗(0.5~1 cm)的表观光合速率(Pn)、暗呼吸速率(Rd)及相对生长速率(relative growth rate,RGR)的影响,并进行上述3因素对RGR的正交实验。结果显示:(1)10 d后显微观察发现,长光照周期(14 L∶10 D、16 L∶8 D)下采苗效果较好,配子体大量发育为孢子体。(2)光强为70μmol/(m2·s)、温度为13℃和氮营养盐浓度为2 mg/L(N/P=10∶1,下同)的条件下幼苗的RGR较大,表观光合速率较强。90μmol/(m2·s)光强组的幼苗生长受到抑制,其RGR显著小于其他实验组。10℃组的RGR显著小于13℃(P0.05)。氮营养盐浓度上升到8 mg/L,RGR显著降低。(3)随着营养盐浓度的增加,幼苗的表观光合速率、暗呼吸速率整体呈现上升趋势,0.2 mg/L组的Pn显著小于6 mg/L、8 mg/L组,且Rd与其他组差异性显著(P0.05)。(4)相同光强下,温度升高,Pn随之降低,在40~80μmol/(m2·s),10℃、13℃组与16℃、19℃组的Pn差异性显著。经正交实验验证,适宜巨藻幼苗培养条件为光照强度40~70μmol/(m2·s)、温度13℃、氮营养盐浓度2 mg/L。     

多肋藻雌配子体种质保存和扩培的温光条件研究

跟踪观察了温度和光照强度双因子对多肋藻雌配子体生长的影响,旨在探明其适宜的种质保存与扩增培养的温光条件。试验设置3个温度,分别为10、15和20℃,每个温度下设5个光照强度,分别为10、20、40、60和80μmol/(m~2·s),实验进行了4个月。结果表明,配子体在10℃,光强≤10μmol/(m~2·s)时适宜种质保存,15～20℃、20μmol/(m~2·s)时适宜短期扩增培养,15℃、20μmol/(m~2·s)时适宜长期扩增培养。     

二氧化锗对共培养萱藻丝状体和硅藻生长的影响

为抑制萱藻丝状体保存和扩增过程中出现的小伪菱形藻与碎片菱形藻的生长,本实验应用实验生态学方法,分别建立了丝状体与小伪菱形藻、丝状体与碎片菱形藻、丝状体与小伪菱形藻和碎片菱形藻的共培养体系,研究了1.00～4.00μg/mL二氧化锗(GeO_2)对共培养条件下丝状体生长发育及附生硅藻生长的影响。结果显示:(1)处理萱藻丝状体和硅藻共培养体系的适宜GeO_2浓度为1.00～2.50μg/mL,各实验组14 d的硅藻抑制率均高于67.33%±5.18%,且丝状体生长发育良好,2.00μg/mL为最适浓度,此浓度下丝状体日均增长率最高,在各培养体系中均大于11.00%,且诱导后孢子囊枝比例和孢子囊直径分别为57.47%±5.31%和(24.55±1.01)μm,与对照组差异不显著;(2) 3.50和4.00μg/mL GeO_2虽对硅藻抑制效果更佳,但同时也会抑制丝状体生长和后期孢子囊的形成与发育,其中4.00μg/mL GeO_2可导致丝状体死亡;(3)碎片菱形藻较小伪菱形藻对GeO_2更敏感。实验14 d,各浓度GeO_2对碎片菱形藻的抑制率为(82.10%±2.40%)～(96.35%±0.79%),均高于同浓度GeO_2对小伪菱形藻的抑制率;同时在丝状体与小伪菱形藻和碎片菱形藻的共培养体系中,碎片菱形藻占硅藻比例随GeO_2浓度升高而相应下降。     

盐度-光照强度-温度对角毛藻生长及高不饱和脂肪酸含量的影响

为获知盐度—光照强度—温度对半咸水硅藻——角毛藻SHOU-B98生长及高不饱和脂肪酸含量的影响,实验采用L_9(3~4)正交设计探讨其在不同盐度(6、12和18)、光照强度[10、30和50μmol/(m~2·s)]及温度(10、20和30℃)组合条件下的生长及藻细胞ARA、EPA和DHA含量的变化。结果显示,盐度、光照强度、温度三因素对角毛藻SHOUB98的生长及高不饱和脂肪酸含量均有显著影响,且存在因素间的交互作用。角毛藻SHOU-B98在盐度12、光照强度30μmol/(m~2·s)和温度30℃及盐度18、光照强度50μmol/(m~2·s)和温度20℃的组合条件下生长最快,第4天后这2组做藻的生物量显著高于其他组(P0.05)。角毛藻细胞积累不同高不饱和脂肪酸的最佳培养条件也不同,细胞积累ARA的最优培养组合条件为盐度6、光照强度50μmol/(m~2·s)和温度30℃;细胞积累EPA和DHA的最优培养组合条件为盐度18、光照强度50μmol/(m~2·s)和温度20℃。研究表明,角毛藻SHOU-B98生长快且细胞脂肪酸营养价值高的培养条件为盐度18、光照强度50μmol/(m~2·s)和温度20℃的组合。     

水流和光照条件对舌状蜈蚣藻Grateloupia livida(Harv)Yamad果孢子萌发生长的影响

在不同水流和不同光照条件下,分别对舌状蜈蚣藻Grateloupia livida(Harv)Yamad果孢子萌发情况展开研究,以期为其苗种生产奠定基础。舌状蜈蚣藻果孢子在附着1 d、15 d后,置于不同水流速度(0、1、1.5、4、7、10、14 cm/s;1.5 cm/s流速由充气控制,其他流速由流水装置控制)培养20 d;舌状蜈蚣藻果孢子附着1 d后,在不同光照周期(6L∶18D、8L∶16D、10L∶14D、12L∶12D、14L∶10D、16L∶8D)和不同光照强度(35、85、135μmol photons/m~2·s)交叉条件下培养70 d。结果显示,附着1 d的果孢子萌发体在静止条件下的萌发生长明显优于其他水流速度下的(P0.05);附着15 d的果孢子萌发体在1.5 cm/s流速下的萌发生长最好(P0.05),在静止条件下的萌发生长明显慢于其他流速下的各组(P0.05)。光照周期和光照强度对果孢子萌发生长的影响显著(P0.05),这二者对盘状体和孢苗生长的影响具有交互作用(P0.05),对盘状体萌发出直立体的影响没有交互作用(P0.05)。光照时间为8–10 h,盘状体萌发直立体的百分率以及孢苗的高度明显高于其他光照时间下的各组。光照强度为35μmol photons/m~2·s时,盘状体萌发直立体的百分率以及孢苗的高度明显比其他光照强度下的高。研究表明,在舌状蜈蚣藻育苗生产中,可采取孢子刚附着时静水培养,附苗后15 d充气培养,并控制光照时间为8–10 h和光照强度为35μmol photons/m~2·s。     

Antifreeze proteins in the urine of marine fish

Several species of marine teleosts have evolved blood plasma antifreeze polypeptides which enable them to survive in ice-laden seawater. Four distinct antifreeze protein classes differing in carbohydrate content, amino acid composition, protein sequence and secondary structure are currently known. Although all of these antifreezes are relatively small (2.6–33 kd) it was generally thought that they were excluded from the urine by a variety of glomerular mechanisms. In the present study antifreeze polypeptides were found in the bladder urine of winter flounder (Pseudopleuronectes americanus), sea raven (Hemitripterus americanus), ocean pout (Macrozoarces americanus) and Atlantic cod (Gadus morhua). Since the plasma of each of these fish contains a different antifreeze class it would appear that all four classes of antifreeze can enter the urine. The major antifreeze components in the urine of winter flounder were found to be identical to the major plasma components in terms of high performance liquid chromatography retention times and amino acid composition. It is concluded that plasma antifreeze peptides need not be chemically modified before they can enter the urine.     

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.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.     

东太平洋公海茎柔鱼种群遗传结构初步研究

根据2009年7月到10月我国大型鱿钓渔船在东太平洋(81.9°～94.9°W,8.9°N～11.3°S)秘鲁和哥斯达黎加外海作业期间采集的样本,利用线粒体DNA细胞色素b基因的724 bp部分序列,分析了采自秘鲁7个采样点和哥斯达黎加3个采样点的155个个体的序列多样性与种群遗传结构.结果显示,724 bp片段中发现了16个变异位点,155个个体出现43个单倍型.序列多样性分析结果揭示,155个个体的平均单倍型多样性指数为0.873,核苷酸多样性指数为0.003 69.群体间共享6个单倍型,秘鲁外海茎柔鱼群体享有最多特有单倍型(20个).分子方差分析揭示,82.70％的遗传变异性出现在种群内；群体间的FST分析揭示部分秘鲁群体与哥斯达黎加群体,部分秘鲁群体间存在着显著的遗传分化；而哥斯达黎加群体内部不存在显著的遗传分化.     

Bioaugmentation in the growth and water quality of livebearing ornamental fishes

Bacillus subtilis isolated from the intestine of Cirrhinus mrigala (Hamilton) was incorporated into the rearing water of Poecilia reticulata (Peters), Poecilia sphenops (Valenciennes), Xiphophorus helleri (Heckel) and Xiphophorus maculatus (Gunther) at four different concentrations (5 × 10⁸ cells ml⁻¹, 5 × 10⁷ cells ml⁻¹, 5 × 10⁶ cells ml⁻¹ and 5 × 10⁵ cells ml⁻¹) and its effect on fish growth performance and survival, water quality parameters and bacterial population of water were assessed. The results showed that the addition of bacterial cells in the rearing water resulted in greater survival and a faster growth rate and, hence, greater length and weight increments of the livebearers. The use of a bioaugmentor in the rearing water of the livebearing fishes resulted in significantly lower (P < 0.05) concentrations of dissolved organic matter and total ammonium nitrogen. The counts of motile aeromonads and total coliforms recorded in the water of bioaugmented tanks were also lower than that in the control tank. Bioaugmentation between 10⁶ and 10⁸ cells ml⁻¹ in the rearing water is sufficient in establishing a bioaugmentor and the use of a higher concentration of bacterial cells did not always lead to significantly better results.     

青岛斋堂岛附近海域薛氏海龙繁殖生物学

根据2015年3-5月在青岛斋堂岛附近海域使用张网采集的1250尾薛氏海龙(Syngnathus schlegeli)样本,分析了该水域薛氏海龙的性比、副性征、性腺指数、性腺成熟度与卵径、育儿袋中胚胎等繁殖生物学特征。结果表明,青岛斋堂岛附近海域薛氏海龙春季繁殖种群雌雄比例分别为63.28%和36.24%,未成年个体占0.48%,雌雄比为1.75:1,明显偏离1:1的性别比例;繁殖群体中雄性个体的性腺指数始终保持在较低水平,为0.01%~4.85%,雌性个体相对较高,为0.05%~128.99%,二者差异极显著(P0.01);组织学观察发现,性腺在3—5月份迅速发育成熟,雄性排精后精巢腔中仍剩余较多精细胞,表明雄性个体为多次排精;雌性Ⅲ~Ⅳ期卵巢中均分布有不同时相的卵子,且卵径分布呈现两个峰值,表明雌性个体为分批产卵;根据雌性个体V期卵巢中成熟卵子的卵粒数与雄性个体育儿袋中的平均怀卵数基本相同,育儿袋中胚胎的发育阶段也基本一致,推测其交配模式可能为一雄一雌制,抑或是雄性在同一天内与多个雌性交配的一雄多雌制。斋堂岛附近海域薛氏海龙的交配模式还有待室内行为观察并结合微卫星标记等手段进一步确认。     

Use of Microbacterium sp. and Exiguobacterium mexicanum to improve the survival and development of Artemia under xenic conditions

The effect of Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N was evaluated in the diet of Artemia under xenic conditions. Viable cultures of bacteria were provided to xenic cultures of Artemia in combination with Sacharomyces cerevisiae, cornflour or Spirulina, and the effect on the survival and growth was recorded. The use of these bacterial strains improves significantly the survival of Artemia independently of the used food (P < 0.05), and variable results were observed in the growth.     

Use of ongrownArtemia in nursery culturing of the tiger shrimp

Juvenile and adultArtemia produced in a semi flow-through culture system were used as food for postlarval shrimp. The growth performance of shrimp reared on such ongrownArtemia live prey is identical to the growth obtained when feeding newly hatchedArtemia. However, a significantly better stress resistance is obtained when the postlarvae are exposed to a low salinity in a stress test. Besides nutritional and energetic advantages, the use ofArtemia biomass for feeding postlarval shrimp also results in improved economics as expenses for cysts and weaning diets can be reduced.     

基于线粒体COⅠ和Cytb基因序列的北太平洋柔鱼种群遗传结构研究

为准确掌握柔鱼的种群遗传结构,拟通过线粒体DNA的COI和Cytb基因序列分析方法对柔鱼不同产卵季节群体的遗传多样性和遗传结构进行研究。经PCR扩增与测序分别获得600 bp COI与481 bp Cytb基因序列。基于COI基因序列分析得到的单倍型数、单倍型多样性指数、核苷酸多样性指数及平均核苷酸差异数分别为24、(0.729 ±0.033)、(0.005 70 ± 0.003 25)和3.421。基于Cytb基因序列分析得到的单倍型数、单倍型多样性指数、核苷酸多样性指数及平均核苷酸差异数分别为28、(0.852 ± 0.016)、(0.006 45 ± 0.003 73)和3.101。分析认为, 北太平洋柔鱼2个产卵季节群体均具有较高的单倍型多样性指数和较低的核苷酸多样性指数。单倍型邻接树、两两群体间的F_st值以及AMOVA分析结果均表明, 北太平洋柔鱼2个产卵季节群体间的遗传差异不显著, 不存在显著的群体遗传结构。初步认为, 该海域因缺乏地理上的障碍, 加之北太平洋海流的作用以及柔鱼个体较强的游泳能力, 使得群体之间具有较强的基因流。