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1.
海湾扇贝样品不同保存条件下DNA的提取及RAPD扩增比较   总被引:17,自引:4,他引:17  
以海湾扇贝为材料,研究了鲜活样品以及采用冷冻保存、70%乙醇固定和10%福尔马林固定等方法保存的闭壳肌样品用于DNA提取的不同效果,并分析了不同保存条件下所提得DNA用于RAPD扩增的结果,研究表明,鲜活样品以及采肜冷冻保存和70%乙醇固定的样品可以得到很好的DNA,其质量和得率没有显著差异,利用上述DNA模板进行RAPD分析,可以获得一致性很好的扩增结果。10%福尔马林休固定样品则没有得到可用的DNA模板。采用70%乙醇固定保存海洋生物组织,是用于DNA提取及相关的分子生物学研究的快速、简易和有效的方法。  相似文献   

2.
分离草虾杆状病毒包涵体的一种新方法   总被引:2,自引:0,他引:2  
杨丰  林斌 《台湾海峡》1995,14(1):80-82
本文报道了一种纯化草虾杆状病毒(MBV)包涵体的方法。取出受MBV感染的虾肝胰组织,在TESP溶液中(50mmol/dm^3Tris-HC1 pH8.0,50mmol/dm^3EDTA,0.1mol/dm^3NaC1,0.5mmol/dm^3PMSF)破碎,差速离心,再经30%~70%蔗糖浓度梯度高速离心,可得到含包涵体的区带。电镜下观察到较纯的包涵体,经蛋白酶K降解后,PCR扩增,经琼脂糖凝胶电泳可见清晰的DNA区带。  相似文献   

3.
三唑磷对四种水生生物的毒性及安全评价研究   总被引:21,自引:0,他引:21  
本文研究了三唑磷对小球藻、卤虫、南美白对虾、泥蚶四种水生生物的急性毒性作用,结果显示,三唑磷对小球藻的48h的EC50为24μg/dm^3,72h的EC50为15μg/km^3;对卤虫的24hLC50为1.64mg/dm^3,48hLC50为0.8mg/dm^3;对南美白对虾仔虾的48hLG0为3.2μg/dm^3,96hLC50为1.1μg/dm^3。;对泥蚶的48hLC50为21.0mg/dm^3,96hLC50为10.2mg/dm^3。三唑磷对小球藻、卤虫、南美白对虾为高毒农药,泥蚶对三唑磷有很强的耐受力。  相似文献   

4.
用氯化锂法从眼点拟微球藻(Nannochloropisis oculata)中提取DNA并对其提取条件进行了优化。新鲜藻细胞在提取液(0.6mol/L LiCl,0.8%Sakosyl,20mmol/L EDTA(pH8.0),0.4%PVP,10mmol/L Tris-HCl(pH8.0),5%β—巯基乙醇)中经过55℃温育30min,每mg鲜藻可提取20—40mg DNA,提取的DNA分子量在23kb左右,可用于PCR、限制性酶切等。  相似文献   

5.
黑鱾血液的生理生化指标研究   总被引:2,自引:0,他引:2  
孙敏  徐善良  唐道军 《台湾海峡》2009,28(4):482-487
本文采用F-820半自动血细胞分析仪和全自动生化分析仪,对黑纪的血液常规指标及血清中的17项生化指标进行了分析测定,并用Wright’s染色法制作血涂片对白细胞进行分类计数.结果表明:(1)黑纪的血红细胞数量为(2.49±0.47)×10^12个/dm^3;白细胞数量为(1.08±0.21)×10^11个/dm^3;在白细胞中以嗜中性粒细胞数量最多,占52.42%±4.67%,其次为淋巴细胞,占25.72%±2.88%;单核细胞体积最大,占总数的3.27%±0.62%.(2)黑纪红细胞脆性大,抗性差,在0.65%~0.70%的NaCl溶液中开始溶血,在0.50%~0.55%的NaCl溶液中完全溶血.(3)黑纪的血红蛋白(HGB)为109.11±15.72g/dm^3,高于一般鱼类,与其较强的耐低氧生理能力相适应.(4)血清中血糖浓度为4.72±1.35mmol/dm^3,明显低于凶猛肉食性鱼类,与其岛礁性生态习性和杂食性相吻合.(5)雌雄鱼的乳酸脱氢酶分别为332.67±188.48U/dm^3和905.66±257.85U/dm^3,雌鱼显著高于雄鱼,可能与繁殖活动相关.  相似文献   

6.
用室内培养的塔玛亚历山大藻(Alezandrium tamarense ATHK藻株)作为实验对象,分别选择了5%福尔马林固定后常温保存、95%乙醇固定后常温保存、-20℃冷冻保存、鲁哥氏液(Lugol’s solution)固定后常温保存等方法,在5,15,30,60d后对保存的样品进行单细胞PCR反应,并与新鲜样品进行比较。实验结果表明,用95%乙醇保存、-20℃冷冻保存、鲁哥氏液保存的样品在60d后仍可扩增出目标条带,与新鲜样品没有差别,但是样品中的藻细胞形态有一些改变;而以5%福尔马林固定保存的样品只能在5d后扩增出目标条带,但藻细胞形态比较完好。因此,对用于单细胞PCR实验的微藻样品,短期内(不多于5d)可以采用福尔马林保存,而需要长期保存的样品,应当采用乙醇或鲁哥氏液固定,或者采用-20℃冷冻保存的方法。  相似文献   

7.
不同保存条件下中华哲水蚤基因组DNA提取的比较   总被引:4,自引:2,他引:4       下载免费PDF全文
以采自厦门港的中华哲水蚤(Calanus sinicus)为对象,研究多种保存条件对中华哲水蚤基因组DNA提取的影响。结果表明,除5%福尔马林保存的样品没有提取到DNA之外,其余样品均能成功提取出基因组DNA;以该DNA为模板通过相应引物成功扩增出线粒体DNACOI基因片段。其中无水乙醇保存样品提取可靠的基因组DNA方法的建立,为野外样品保存工作提供了一个简便,经济的途径。  相似文献   

8.
2006~2008年间对进入福建沿海4个主要港口的12艘国际航船所携带的压舱水进行浮游植物种类组成和丰度分布的调查.此调查共检测出浮游植物7个门86属239种(含变种和变型),77μm孔径网滤样和20μm孔径网滤样的平均藻类丰度分别为1.2×10^2cells/dm^3(变动在0~9.1×10^2cells/dm^3间)和3.4×10^3cells/dm^3(变动在0~3.0×10^4cells/dm^3间).同时对其中6艘船舶的压舱水样品用f/2培养基培养,共有13种硅藻和1种甲藻被成功培养.文中还分析了压舱水中浮游植物丰度与盐度、水龄的关系,并结合历史资料讨论了压舱水里各类浮游植物的分布.  相似文献   

9.
单条固定线虫基因组DNA提取及18S rRNA基因PCR扩增   总被引:2,自引:0,他引:2  
根据线虫18S核糖体RM基因PCB扩增效果比较了丙酮、乙醇、乙醇 0.05mol/L FDTA(pH8.0)和5%海水福尔马林4种固定剂,碱裂解和蛋白酶K处理2种单条线虫基因组DNA提取方法的优劣。用乙醇固定的样品最适合制备RR模板DNA,而5%海水福尔马林固定的样品能最完整地保持样品形态。蛋白酶K处理获得的DNA较碱裂解获得的更适合PCR扩增。结果有助于分子生物学方法在海洋线虫分类、多样性和生态学研究中的应用。  相似文献   

10.
高密度轮虫对点带石斑鱼早期仔鱼培育影响的研究   总被引:4,自引:1,他引:4  
曾庆民 《台湾海峡》2004,23(1):69-75
本试验在水体为1000dm^3、仔鱼密度为15ind./dm^3条件下,研究不同轮虫密度(10、25、40、70、100ind./cm^3)对点带石斑鱼早期仔鱼摄食和生长的影响.至仔鱼12日龄的培育试验结果表明,在点带石斑鱼早期仔鱼的培育过程中,投放轮虫对水质有影响,特别是在2~6日龄间,DO值和pH值都有一降低过程,但随后即回升并相对稳定;轮虫密度越高则pH值越低,pH值最低出现在4~6日龄,4日龄时10ind./cm^3组的pH值比100ind./cm^3组高0.32.在10~100ind./cm^3的轮虫密度范围内,较低的轮虫密度对仔鱼的生长有利,12日龄10ind./cm。组仔鱼的全长比100ind./cm^3组高31.83%,但100ind./cm^3组仔鱼的成活率比10ind./cm^3组的高出近一倍;不同轮虫密度下仔鱼摄食轮虫数量的差异并不明显;但9日龄以后,过高的轮虫密度100ind./cm^3下仔鱼生长速度显著降低.  相似文献   

11.
The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths.  相似文献   

12.
Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone.  相似文献   

13.
The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics.  相似文献   

14.
15.
《海洋学报(英文版)》2014,(9):F0003-F0003
<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology,  相似文献   

16.
The relationship between the flux of exotic benthic foraminiferal tests (i.e. tests which are supplied from open-sea sources alone) in a tidal inlet and that of bulk sediment was analysed, which can be expressed as two first-order linear equations. According to this relationship, in order to determine net sediment transport directions in the entrance, the test concentration in surficial sediments of the tidal basin can be compared against a ‘ critical level ’. The critical level is determined for the conditions that no net transport of bulk sediment is present within the entrance. If the observed concentration (averaged over the tidal basin) is higher than the simulated critical level, then the net sediment transport is directed to landward. This method is applied to the analysis of net sand transport at Christchurch Harbour, a tidal inlet system located in southern England. In this investigation, concentrations of exotic foraminiferal tests in the surficial sediments of the tidal basin and ebb tidal delta area were obtained from the analysis of sea-bed sediment samples. A series of probable critical levels were calculated based upon the data sets with regard to: (1) sediment discharge from the rivers; (2) magnitude of sediment discharge within the entrance during the ebb; (3) the test concentration outside the harbour; (4) the thickness of the moving layer; and (5) two parameters associated with dispersive processes. The results show that the concentration in the tidal basin sediment is higher than a number of simulated critical concentrations for representative cases. Consequently, the high level of the concentration of exotic benthic foraminiferal tests within the harbour should be explained as a result of landward net transport of sands within the entrance.  相似文献   

17.
18.
Observations on seafloor features from subbottom profiling and seabed photography data show that the undisturbed and natural conditions before the benthic disturbance undergo a substantial change after operation of the disturber. The effects can be seen in the form of deep trenches and grooves, formed by the action of the sleds and the pumping of the sediment along its path, as well as sediment piles on either side of the track. Biological traces such as fecal coils, casts, tubes, burrows, and trails get obliterated in the areas of disturbance and resedimentation. The smooth, uniform nature of the seafloor observed in predisturbance phase shows microtopographic changes in and around the disturbed area as a result of sediment excavation and resettlement. Vertical mixing as well as lateral transport of sediment alter the geological, biological, physical, and chemical conditions on the seafloor, which need to be monitored over time to assess the process and the time taken for restoration of environmental conditions.  相似文献   

19.
Abstract. Long-term changes in the distributional patterns of commercial sponges ( Spongia spp. and Hippospongia spp.) within the West Indian Region indicates that: 1) commercial sponges had a widespread distribution throughout the whole West Indian Region and were ubiquitous in very shallow water until about the first half of the present century; 2) they were fished commercially not only in the traditional northern Caribbean sites (Florida, Gulf of Mexico, Bahamas) but also in the Greater ( e. g. , Hispaniola, Jamaica) and Lesser Antilles; and 3) they became extinct throughout most of the Lesser Antilles ( e. g. , Puerto Rico, Vieques, St. Thomas) sometime during the first half of this century. Mortalities of spongiids within the Antilles were found to differ from other marine mortalities in that: 1) species disappeared from a large region; 2) species vanished from different habitats and depths; and 3) natural populations never recovered. Species richness patterns suggest that commercial sponge genera ( Spongia and Hippospongia ) evolved under slightly cooler elimatic conditions than those found at present, and that these extinctions occurred as a direct or indirect effect of positive thermal anomalies in sea surface and atmospheric temperatures between 1900–1950. The concept that species diversity is stable on a regional scale is questioned.  相似文献   

20.
The behaviour of an axially pre-loaded cylindrical member of an offshore structure, hit by a supply ship, has been investigated. The effects of axial pre-loading on the dynamic properties of members, the extent of damage and propagation of dynamic instability in the tubular members have also been investigated. Numerical models have been validated using available experimental data from the literature although these are mostly static with no pre-loading. Axial pre-loading was found to change the dynamic characteristics of some cylindrical members quite dramatically while it had no significant effect on others. The study examined whether there was any change in the behaviour between local and global modes when the loading was dynamic rather than static. The effect of damping on the dynamic instability of axially pre-loaded tubes under lateral dynamic loads was also studied. It was observed that for some geometries the quasi-static response can be used to define the boundary between bounded and unbounded dynamic responses. The main contribution of this study, compared with previous investigations reported in the literature, is that both pre-loading and dynamic effects have been included.  相似文献   

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