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1.
筛选了六种抗冻保护剂(GLY[甘油],DMSO[二甲基亚砜],PG[丙二醇],EG[乙二醇],DMA[二甲基乙酰胺],MeOH[甲醇])及其五种不同浓度(6%,8%,10%,12%,14%,V/V)、三种稀释液(HBSS溶液,人工海水[ASW],过滤海水[FSW])、四种稀释比例(1︰1,1︰2,1︰4,1︰6)、三种添加剂(蔗糖,葡萄糖,海藻糖)、三个分步降温程序(程序A,程序B,程序C)对太平洋牡蛎(Crassostrea gigas)精液冷冻保存的影响,成功建立了太平洋牡蛎精液超低温保存方法:以10%DMSO为抗冻保护剂,HBSS溶液为稀释液,1:4的稀释比例,添加海藻糖,采用分布降温法冷冻保存太平洋牡蛎的精液,37°C水浴解冻后精液运动率达到71.27%±3.24%,显著高于其它实验组(P0.05),受精率和孵化率分别达到95.04%±1.99%、93.33%±1.33%,与鲜精(88.89%±15.16%,94.90%±0.95%)、程序冷冻前精液(95.96%±2.15%,92.67%±14.83%)差异不显著(P0.05),证明该太平洋牡蛎精液超低温保存方法可以应用于生产实践和科学研究中。  相似文献   

2.
采用分步降温法冷冻保存太平洋鳕(Gadus macrocephalus)精液,并用扫描电镜和透射电镜技术研究了精子的超微结构损伤。分析了添加剂(蛋黄)及五种抗冻剂(PG(丙二醇)、DMSO(二甲基亚砜)、EG(乙二醇)、GLY(甘油)、MeOH(甲醇))不同浓度(8%、10%、12%、14%、16%、18%)对太平洋鳕精子冷冻保存效果的影响。实验结果表明,蛋黄能够显著提高太平洋鳕冷冻保存效果(P0.05);12%PG中添加10%蛋黄保存的冻融精子运动率最高(85.87%±1.6%),显著高于其他冻存组(P0.05)。添加蛋黄的12%浓度的DMSO组解冻后精子运动速率较高,平均直线速度、平均曲线速度、平均路径速度分别达到了(120.39±20.78)μm/s、(120.39±20.78)μm/s、(155.64±12.02)μm/s,与其他各实验组差异显著(P0.05)。通过扫描电镜和透射电镜观察新鲜和冻融后的鳕鱼精子发现,鲜精中75.5%精子形态结构正常、24.5%精子形态结构异常;运动率最高的冻精中67%精子形态结构正常、33%精子形态结构异常。超低温保存对太平洋鳕精子结构产生了显著影响,细胞膜破裂、线粒体肿胀变形,鞭毛脱落、断裂。  相似文献   

3.
为防范浒苔绿潮次生灾害和降低海水养殖风险,文章采用人工授精的太平洋牡蛎胚胎、不同相对浓度的浒苔腐烂液以及基于发光细菌的生物毒性检测法,通过实验分析浒苔腐烂液的生物毒性及其对太平洋牡蛎胚胎发育的影响。研究结果表明:随着浒苔腐烂液相对浓度的提高,太平洋牡蛎D形幼虫相对孵化率逐渐降低至0,滞育和畸形等非正常胚胎增多,囊胚-原肠期和担轮幼虫期占比降至极低,易发生卵膜破裂和卵裂球分散,这些与太平洋牡蛎胚胎发育的阶段特点和浒苔腐烂液生物毒性的作用机制等有关;浒苔腐烂液相对浓度(0.25~10g/L)与生物毒性呈显著正相关,生物毒性(10%~97%)与相对孵化率呈显著负相关,且生物毒性达重毒水平时(由56%至70%)相对孵化率大幅下降(由66%至18%);建议在太平洋牡蛎胚胎发育的相对孵化率为50%时发出浒苔绿潮的渔业高风险预警,对应的浒苔腐烂液相对浓度为4.6g/L、生物毒性为55%。  相似文献   

4.
本文比较了胰酶消化法、网搓法和机械研磨法3种方法对获得游离日本蟳精子的效果。对获得的精子进行超低温冷冻保存,用曙红Y进行染色,检测精子的存活率。使用7种不同稀释液、不同浓度的二甲基亚砜(DMSO)、甘油(GLY)、1,2-丙二醇(PG)为抗冻保护剂,在6种不同的降温程序下,对日本蟳精子进行超低温冷冻保存,后在不同的温度下解冻复苏精子。实验结果表明:3种精子获取方法中以搓洗法的实验效果最好,胰酶消化法的效果次之,最差的是机械研磨法。其中搓洗法中以500目的筛绢效果最好,精子密度达2.56×10~(13)个/mL。日本蟳精子冷冻保存实验结果表明,稀释液以去钙人工海水、抗冻保护剂以5%DMSO、平衡时间20 min、降温程序P-1(0℃平衡20 min;-5℃/min降至-80℃;-80℃平衡5 min;-20℃/min降至-180℃;然后置于液氮中保存),水浴解冻37.5℃的效果最好,精子存活率最高。  相似文献   

5.
为探讨底物中钙赋存形态对牡蛎幼虫附着的诱导效应,本研究设计3因子随机区组实验,检验实验底物钙含量、钙赋存形态及海水中牡蛎幼虫丰度对实验底物上太平洋牡蛎(Crassostrea gigas)稚贝附着效果(密度和壳高)的影响。结果表明,在3个因子中,实验底物钙含量对附着稚贝密度和壳高的影响不显著(P0.05),而其它2个因子(钙赋存形态和海水中牡蛎幼虫丰度)均显著影响着实验底物上附着牡蛎稚贝的密度和壳高(P0.05)。在相同的牡蛎浮游幼虫丰度下,3种钙形态底物对牡蛎幼虫附着的诱导能力大小顺序为:有机钙碳酸钙=硫酸钙(P0.05);壳高的结果却恰恰相反。在同样的钙形态底物中,高牡蛎幼虫丰度下牡蛎稚贝附着量均显著高于低幼虫丰度的处理组(P0.05)。  相似文献   

6.
利用解剖法采集太平洋牡蛎精液 ,用自然海水做基础溶液 ,配制不同浓度的 DMSO(二甲亚砜 )作为超低温保护剂 ,在液氮 (L N2 )面上不同高度进行预处理 ,投入 LN2 内保存 ,升温解冻后 ,检测精子成活率。结果表明 ,DMSO处理浓度、预处理温度及时间和升温方式对超低温保存成活率均具有显著影响。其中 ,用 10 % DMSO在 LN2 面上 17cm预处理 6 min,采用 16~ 17℃流水不完全解冻 ,效果最好 ,超低温保存精子成活率可达 70 %。与新鲜精子相比 ,在 L N2 内保存一天的精子 ,授精能力没有明显变化。  相似文献   

7.
母源免疫因子能够通过卵细胞、母乳等方式转移给子代,在免疫系统发育成熟之前发挥重要的免疫保护作用,但贝类中的相关报道较少。为探索贝类非特异性免疫因子在子代发育早期的表达规律,本研究以魁蚶(Scapharca broughtonii)为研究对象,通过qRT-PCR和ELISA分析了大防御素在魁蚶胚胎及幼虫早期发育过程中mRNA和蛋白质的动态变化特征。结果显示,魁蚶卵子受精后发育至壳顶幼虫期,大防御素mRNA和蛋白质的表达变化趋势基本相同,即从受精卵期开始下降,至囊胚期最低,自担轮幼虫期表达量开始上升;与对照组相比,幼虫发育至担轮幼虫期,鳗弧菌(Vibrio anguillarum)处理组的大防御素升高更为显著,mRNA和蛋白的表达水平都基本高于对照组,说明鳗弧菌刺激能够上调担轮幼虫期后魁蚶幼体体内大防御素的表达量。总的来说,研究结果表明魁蚶母源性大防御素能够通过卵子传递给后代,从卵细胞受精后开始被消耗,当幼虫发育至担轮幼虫可能开始自身合成。  相似文献   

8.
用不同种类固着基及2种金属离子(K+、Cu2+)对香港巨牡蛎Crassostrea hongkongensis眼点幼虫固着变态的效果进行了研究。结果表明:黑色聚乙烯膜更有利于香港巨牡蛎眼点幼虫的固着,固着率达到49.85%,其次为黑色塑料板,固着率为19.33%,红色塑料板最低,固着率为5.14%;添加K+对牡蛎眼点幼虫的固着变态有诱导作用,当添加K+浓度为13.42mmol/L作用24h时,与对照组相比幼虫的固着提高率、变态提高率分别为42.39%、25.20%;而添加Cu2+对香港巨牡蛎眼点幼虫的固着变态没有诱导作用,随着Cu2+处理时间及处理浓度的增加,香港巨牡蛎眼点幼虫的死亡率明显提高,推测浓度过高的Cu2+对香港巨牡蛎眼点幼虫有毒性作用。  相似文献   

9.
多肽菌素作为一类具有抗菌活性的生物短肽,是一种可以替代抗生素的生物肽。为了推广多肽菌素在水产养殖中的应用,本研究以长牡蛎(Crassostreagigas)人工育苗为基础,以添加2×10_(–6) mol/L青霉素钠为对照组,以添加5个不同浓度(2×10_(–6)、4×10_(–6)、6×10_(–6)、8×10_(–6)、10×10_(–6) mol/L)多肽菌素为实验组,研究多肽菌素在长牡蛎人工育苗生产中的应用。结果显示:(1)在长牡蛎种贝培育方面,培育前8天各组差异不显著(P 0.05),培育后期添加6×10_(–6) mol/L多肽菌素组存活率均最高,对照组最低,且培育至32天时该实验组均显著高于其他组(P0.05)。(2)在长牡蛎幼虫培育方面,幼虫壳高日增长量在添加6×10_(–6) mol/L多肽菌素组中最大,为11.57μm/d,而对照组仅为9.61μm/d;幼虫存活率方面对照组始终最低,而添加6×10_(–6) mol/L与8×10_(–6) mol/L多肽菌素组均保持较高存活率,显著高于对照组(P 0.05)。(3)在长牡蛎幼虫附着变态方面,添加6×10_(–6) mol/L多肽菌素组附着率最高、附着时间最短,且均与对照组差异显著(P0.05)。(4)在长牡蛎稚贝培育方面,稚贝壳长日增长量实验组均高于对照组,且随添加浓度的上升,日增长量也随之增加;稚贝存活率在添加6×10_(–6) mol/L多肽菌素组达到最高,与对照组差异显著(P0.05)。结果表明,使用多肽菌素与使用青霉素钠相比,多肽菌素显著提高了整个牡蛎培育过程中的存活率、生长速度、及附着变态率,这也为多肽菌素在贝类育苗生产中的推广提供了基础依据。  相似文献   

10.
以中国南海常见的多毛类——华美盘管虫(Hydroides elegans)作为受试生物,分别研究了吡啶硫酮铜和吡啶硫酮锌对早期不同发育阶段的华美盘管虫的急性毒理效应。发现暴露于吡啶硫酮铜的华美盘管虫精子和卵子的受精半数抑制浓度(IC50)分别为19.49μg/L和88.44μg/L,担轮幼虫24 h半数致死浓度(24h-LC50)和48 h半数致死浓度(48h-LC50)分别为7.35μg/L和5.00μg/L,后担轮幼虫24h-LC50和48h-LC50分别为8.57μg/L和5.87μg/L;暴露于吡啶硫酮锌的华美盘管虫精子和卵子的受精IC50分别为36.74μg/L和159.59μg/L,担轮幼虫24 h-LC50和48 h-LC50分别为8.57μg/L和6.67μg/L,后担轮幼虫24 h-LC50和48 h-LC50分别为12.03μg/L和8.07μg/L。表明华美盘管虫早期发育的不同阶段对吡啶硫酮铜和吡啶硫酮锌的敏感性表现出一定差异,与配子受精率和后担轮幼虫的毒性反应相比,担轮幼虫对吡啶硫酮铜和吡啶硫酮锌的毒性表现出较高的敏感性;而且,对吡啶硫酮铜的敏感性普遍高于吡啶硫酮锌。  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.
海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明:土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。  相似文献   

13.
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   

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A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria ( 〉 99% ), but also between polar marine bacteria and bacteria from other aquatic environments ( ≥ 98.8% ), including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.  相似文献   

17.
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   

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《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   

20.
Sediment properties, such as water content and density, have been used to estimate the dry and wet weights, as well as the volume of sediment recovered and discharged, during benthic impact experiments conducted in the Pacific and Indian Oceans. The estimates show that the weights of dry (355-1332 t) and wet sediment (1651-4888 t) recovered during the experiments, are not only a function of the total duration and distance covered during the experiment, but also depend on water content and density of the sediment. Estimation of dry sediment and its volumetric ratio in the discharge, are key parameters for calculating the discharged volume. The estimated volume of wet sediment recovered (1427-4049 m 3 ) as well as discharged (2693-6951 m 3 ) during the experiments, provide important inputs to evaluate the sediment resettlement and migration. Using these estimates, the average depth of excavation on the seafloor, can also been calculated. However, the stages of sediment recovery and discharge are expressed differently in some of the experiments. In order to standardize these, different stages have been identified, and definitions of certain terms have been suggested, for use in the future. The methods of calculating different properties, as well as weight and volume of discharged sediment are described in the paper for use in other applications concerning deep-sea discharges.  相似文献   

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