The distribution of phenylalkanes in the modern sediment associated with gas hydrate from the Gulf of Mexico

Phenylalkanes with carbon numbers between 16 and 19, characterized by the main carbon-18, have been identified in the modern sediments collected from gas hydrate area from the Gulf of Mexico. The structure of phenylalkanes with four isomers for every carbon number was determined by means of their mass spectra and previous studies. The distribution of the series characterized by a low molecular mass was similar to the distribution of n-alkane, alkylcyclohexanes and alkylbenzenes in each sample. There were differences in the distribution of the phenylalkane series between the S - 1, S - 4, S - 7, S - 9 samples and the S - 8, S - 10 and S - 11 samples. The phenylalkanes might be derived from Archaea associated with anaerobic oxidation of methane （AOM） processes in S - 1, S -4, S -7 and S -9 samples according to their distribution resembled with the distribution of the extract from a type of Archaea. The distribution of alkylcyclohexanes and alkylbenzenes in S - 1, S - 4, S - 7 and S - 9 sample was found to be similar to each other. The odd-over-even predominance of alkylcyclohexanes was seen as the input of some bac- terial.     

Antibacterial substance from mucus of a scleractinian coral, Symphyllia gigantea

Coral mucus covers the surface of coral and contains antibacterial substances as a first line of defense. Coral mucus not only enables the coral itself to resist disease, but also provides antibacterial agents for people. We collected mucus from a scleractinian coral (Symphyllia gigantea) at Sta. Sanya (China), then extracted the antibacterial substances using 10% glacial acetic acid with the help of antiprotease inhibitors, and tested the antibacterial activity by a terrestrial bacterium (Staphylococcus aureus) and a marine bacterium (Vibrio anguillarum). The result showed that, there were antibacterial agents in the mucus, and their antibacterial activities were lost by treatment of the sample at 90 ℃ water for 10 min.     

Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Karenia mikimotoi by fluorescence in situ hybridization

Harmful algal blooms recently have been under the spotlight throughout the world, because of their negative impact on the marine environment, aquaculture, fisheries as well as public health. The development of methods for rapid and precise identification and quantification of causative species is essential for the warning and monitoring of blooms, among which the techniques based on taxonomic probes are the most favored. In this study, two harmful algae, i.e., Prorocentrum minimum and Karenia mikimotoi were taken into consideration. The partial large subunit rDNA (D1-D2) of both species were firstly PCR-amplified, cloned and sequenced. The obtained sequences were then introduced to carry out alignment analysis for gene specific regions. Three respective candidate probes for each species were designed and used to screen the optimal probe by performing fluorescence in situ hybridization (FISH) tests. The results showed that the probes Pmin0443 and Kmik0602 displayed the best hybridization for P. minimum and K. mikimotoi, respectively. Both the specific (taxonomic) (Pmin0443 and Kmik0602) and the control probes (UniC0512 and UniR0499) were used for cross-reactivity tests with other microalgae in our laboratory. The probes Pmin0443 and Kmik0602 are specific and could be served as taxonomic probes introduced into the techniques targeting rRNA, such as FISH, sandwich hybridization, and DNA-microarray assay of P. minimum and K. mikimotoi in the future. Finally, FISH analyses with both probes were performed on the simulated field samples. The probes could hybridize exclusively with the target cells well, and no significant difference (p >0.05) was observed in the cell densities of the samples determined by FISH and light microscopy (LM). All suggest that the probes are specific and could be introduced into FISH for the monitoring of both harmful algae.     

Dynamic Behavior and Deformation Analysis of the Fish Cage System Using Mass-Spring Model

Fish cage systems are influenced by various oceanic conditions, and the movements and deformation of the system by the external forces can affect the safety of the system itself, as well as the species of fish being cultivated. Structural durability of the system against environmental factors has been major concern for the marine aquaculture system. In this research, a mathematical model and a simulation method were presented for analyzing the performance of the large-scale fish cage system influenced by current and waves. The cage system consisted of netting, mooring ropes, floats, sinkers and floating collar. All the elements were modeled by use of the mass-spring model. The structures were divided into finite elements and mass points were placed at the mid-point of each element, and mass points were connected by springs without mass. Each mass point was applied to external and internal forces, and total force was calculated in every integration step. The computation method was applied to the dynamic simulation of the actual fish cage systems rigged with synthetic fiber and copper wire simultaneously influenced by current and waves. Here, we also tried to find a relevant ratio between buoyancy and sinking force of the fish cages. The simulation results provide improved understanding of the behavior of the structure and valuable information concerning optimum ratio of the buoyancy to sinking force according to current speeds.     

Determination of cosmogenic 32P and 33P in environmental samples

Understanding phosphorus dynamics in marine environment is of great importance, and appropriate tracers for phosphorus cycling in oceans are invaluable. In this study, two methods were developed for extraction, purification, and determination of naturally occurring 32P and 33P in rainwater, marine plankton and sediments using both a low-level beta counter (LBC) and an ultra-low-level liquid scintillation spectrometer (LSS). Blanks, chemical yields and counting efficiencies were quantified for both methods. The chemical purification of 32P and 33P separated by both procedures was validated by their decay curves. The absorber thickness of aluminum for LBC was assessed as 39.2 mg/cm 2 . 32P and 33P specific activities in some rain samples were determined by both methods and showed good consistent results. The advantage of the LSS over the LBC is apparent in its high counting efficiency and in determining samples with high concentration of stable phosphorus. However, when measuring environmental samples with low concentration of stable phosphorus, such as rainwater, both methods can be used and each has its distinct advantage.     

基于Sentinel-3载荷OLCI和SRAL数据的内波同步探测研究

The ocean and land color instrument(OLCI) and synthetic aperture radar altimeter(SRAL) installed aboard the Sentinel-3 satellite have been in orbit for operational uses. In this study, data collected from Sentinel-3 are used to investigate internal waves in the South China Sea. An internal wave is detected using an OLCI image with a resolution of 300 m, and an analysis was performed with a quasi-synchronous moderate-resolution imaging spectroradiometer(MODIS) image. The opposite characteristics of OLCI and MODIS images of the same internal wave are explained by the critical angle in brightness reversals. The unique observational geometry of the OLCI image and its influence on observations of internal waves are discussed. The distribution of σ0 and sea surface height anomalies(SSHAs) induced by internal waves are studied using SRAL records. The σ0 records of SRAL occasionally show less sensitivity to the modulation of internal waves, which may be attributed to the observational geometry, while SSHAs show obvious variations. The synchronous pairing of OLCI images and SRAL records are analyzed to extract the three-dimensional sea surface signatures induced by internal waves. The analysis demonstrates that the profile of SSHAs in the surface shows an opposite phase to the profiles of internal waves in the ocean. The opposite phase relationship, observed in the remote sensing view, is also confirmed with a laboratory experiment.     

Development and validation of a TaqManTM fluorescent quantitative real-time PCR assay for the rapid detection of Edwardsiella tarda

Edwardsiella tarda is one of the most important emerging pathogens in the global aquaculture industries. As such, an accurate diagnosis and quantitative analytical methods are urgently needed for this bacterium. In this study, primers and a TaqMan probe specific to the conservative sequences of the 16S rRNA gene of E. tarda were designed. The concentration of primers and TaqMan probe were optimized to 200 nmol/L and 120 nmol/L, respectively. The detection sensitivity of the FQ-PCR assay was determined to be as low as five copies of the target sequence per reaction using the pGEM-16S rDNA recombinant plasmid as a template, which was 100 times more sensitive than conventional PCR. A standard curve by plotting the threshold cycle values （y） against the common logarithmic copies （log₁0 n_c as x; n_c is copy number） of pGEM-16S rDNA was generated. The results of intra-and inter-assay variability tests demonstrate that the established FQ-PCR method was highly reproducible. The assay was specific for E. tarda as it showed that there was no cross-reactivity to eight additional bacterial pathogen strains in aquaculture. Thus, the FQ-PCR assay has the potential for diagnostic purposes and for other applications, especially for the rapid detection and quantification of low-grade E. tarda infections.     

Multivariate analysis of fluorescence and source identification of dissolved organic matter in Jiaozhou Bay, China

Hierarchical clustering analysis and principal component analysis (PCA) methods were used to assess the similarities and dissimilarities of the entire Excitation-emission matrix spectroscopy (EEMs) data sets of samples collected from Jiaozhou Bay, China. The results demonstrate that multivariate analysis facilitates the complex data treatment and spectral sorting processes, and also enhances the probability to reveal otherwise hidden information concerning the chemical characteristics of the dissolved organic matter (DOM). The distribution of different water samples as revealed by multivariate results has been used to track the movement of DOM material in the study area, and the interpretation is supported by the results obtained from the numerical simulation model of substance tracing technique, which show that the substance discharged by Haibo River can be distributed in Jiaozhou Bay.     

THE EFFECT OF MINE TAILINGS ON THE PRODUCTION OF PLANKTON

Mine tailings at levels of ca 30 and 300 ppm were added to two plankton communities contained in 60 ton Controlled Experimental Ecosystems (CEE); a third CEE was used as a control. The mine tailings caused (1) a delay in the primary productivity and chlorophyll a maxima; (2) a shift in the size spectrum of primary producers towards autotrophic flagellates compared with diatoms in the control; (3) a suppression of heterotrophic microflagellates; and (4) an increased production of zooplankton compared with the control. Zooplankton were observed to have consumed tailings but no changes were observed in the structure of the zooplankton community as a result of tailings additions.     

可口革囊星虫Phasolosma esculenta体腔液的抗菌活性研究

可口革囊星虫Phasolosma esculenta为我国特有的一种可食用海洋经济动物。本文以可口革囊星虫为研究对象,利用琼脂径向扩散、涂板计数等方法研究其体腔液中物质的抗菌活性以及理化特性。研究结果表明,可口革囊星虫体腔液中存在抗菌活性物质,并且该物质对金黄色葡萄球菌Staphylococcus aureus等3种革兰氏阳性细菌具有明显的抗菌效果,但对大肠杆菌Escherichia coli等3种革兰氏阴性细菌以及葡萄牙假丝酵母Candida lusitaniae抗菌效果不明显。结果还发现:(1)不同提取方式出现明显不同的抗菌效果,与体腔液上清液和2%乙酸提取体腔液所得提取物的抗菌效果相比,超声破碎处理提取物的抗菌效果最为明显。(2)理化特性研究发现该抗菌物质抗菌效力持久,抗冻融,并且在5~30 ℃范围内最适抗菌温度约为25 ℃;该物质不耐高温,60 ℃处理10 min后该物质的抗菌作用完全消失;此外,该物质的抗菌作用不依赖金属离子的存在。(3)超滤管过滤截留琼脂扩散结果表明该抗菌物质的分子量在100 kDa以上。     

海洋球石藻(Emiliania huxleyi)活性物质分离鉴定及其抗菌抗氧化性能研究

以海洋球石藻Emiliania huxleyi为原料,提取分离生物活性物质,为该藻活性物质的研究与开发提供理论基础。通过对球石藻75%乙醇浸提物采用乙酸乙酯进行萃取,进一步利用薄层色谱、硅胶色谱、Sephadex LH-20凝胶色谱及高效液相色谱(HPLC)等方法,对化合物成分进行逐步分离纯化,获得纯度大于95%的单体化合物;通过~1HNMR、~(13)CNMR及HRMS等波谱分析技术鉴定化合物的结构,并对其抗菌、抗氧化性能进行初步分析。结果分离鉴定出3种单体化合物,分别为叶绿醇(C_(20)H_(40)O)、棕榈酸(C_(16)H_(32)O_2)和肉豆蔻酸(C_(14)H_(28)O_2),这些化合物均为首次从海洋球石藻中分离获得。其中叶绿醇具有较强的抗氧化能力,且对肝癌细胞Hep G2的增殖有较强的抑制作用,其他两种化合物对金黄色葡萄球菌、枯草芽孢杆菌及副溶血性弧菌均有不同程度的抗菌活性。另外,海洋球石藻中这些化合物除了具有一定的抗菌抗氧化性能外,可能在球石藻响应胁迫(如病毒感染)过程中也具有重要作用。     

腐殖酸抑制纳米银对菲律宾蛤仔鳃的氧化胁迫和遗传毒性

为了解DOM与AgNPs的相互作用及其对AgNPs毒性的影响,本文以菲律宾蛤仔为受试生物、以水环境广泛存在腐殖酸(Humic acid,HA)为代表性有机质。通过贝类体内抗氧化系统的生物标志物(过氧化氢酶(Catalase,CAT)、超氧化物歧化酶(Superoxide dismutase,SOD)、金属硫蛋白(Metallothionein,MT)和丙二醛(Malondialdehyde,MDA))的响应,研究了HA对AgNPs生物效应的影响。结果表明,HA显著增加了AgNPs的稳定性,抑制了AgNPs的聚集。AgNPs(20μg·L^-1)暴露3 d后,蛤仔鳃组织中Ag的蓄积显著上升,并随着时间的延长不断升高。AgNPs暴露第3、7天,不同浓度的HA条件下,鳃组织中Ag的蓄积无显著差异,但显著地低于无HA存在的AgNPs暴露组。CAT活性和MDA含量仅在无HA存在的AgNPs暴露组出现显著升高,其余各组之间无显著差异。表征DNA损伤程度的Olive尾矩(OTM)和尾长(TL)在无HA存在的AgNPs暴露组中随时间的延长不断上升。HA显著降低了AgNPs的对鳃组织细胞的遗传毒性。结果表明,HA通过增加AgNPs在介质中的稳定性而显著地抑制了鳃组织对Ag的蓄积,从而减弱了AgNPs对鳃组织的氧化胁迫和遗传毒性。结果证明了有机质含量等环境理化条件对纳米毒性存在显著影响。     

深度边缘光谱U-Net海水网箱养殖信息提取

网箱养殖是海水养殖的重要类型之一,传统网箱养殖目标光谱特征受近岸植被、水体影响较大,易出现噪声问题.新型深海网箱养殖目标离岸较远,但养殖目标海面框体部分较小,与自然水体光谱相似性较高,难以实现有效提取.本文提出深度边缘光谱U-Net模型对两种海水网箱养殖类型进行养殖信息提取.该模型通过Canny算子双边滤波算法去除波段...     

重组抗菌肽海藻酸钠微囊制备与体外释放特征研究

对重组表达的海洋生物抗菌肽对虾素3-2进行亲和层析纯化,以海藻酸钠为壁材,采用凝聚法制备了重组抗菌肽海藻酸钠微囊,以微囊的形态和包封率为指标优化制备工艺,对制备的微囊进行体外释放特征的初步研究。结果显示,在氯化钙浓度为1.5%,海藻酸钠浓度为2.0%时,制备的微囊为完整的球形,冷冻干燥后的直径约为1.1 mm,包封率为83.87%。微囊在模拟胃液(pH 2.0)中2 h左右释放量趋于稳定,释放量低于14%;微囊在模拟肠液(pH 7.8)中不断释放,5 h时释放量达98%,表明微囊具有良好的肠溶性而可以抵抗胃液的破坏,可以用作重组抗菌肽缓释/控释制剂,为抗菌肽在水产病害防治过程的口服给药提供实验基础。     

基于深度学习RCF模型的三都澳筏式养殖区提取研究

三都澳是中国重要的海水养殖海湾，在水产养殖中占有较高的经济地位。快速准确地获取养殖区的分布范围、数量以及面积等信息，对养殖区规划、产值估计、生态调查、风暴潮灾害预防等具有重要的意义。然而，随着养殖区域的扩大，海水背景状态愈发复杂多样，光谱特征差异较大，为养殖区提取带来困难。在本实验中，利用高分辨率遥感卫星GF-2图像，引入深度学习RCF（Richer Convolutional Features）网络模型对海湾内的筏式养殖区进行了提取。结果显示:该方法无需事先对区域进行水陆分离处理，且对水中泥沙较多的区域以及海浪较大的区域有很好的提取效果，提取精度达93%以上，适合进行大规模海水养殖区提取应用。     

泥蚶血红蛋白异源四聚体酶解多肽抗菌活性及其机理研究

采用凝胶层析技术从泥蚶血细胞裂解物中纯化得到两种血红蛋白Tg-HbⅡ(异源四聚体)和TgHbⅠ(同源二聚体),回收率分别为48.15%和24.91%。胰蛋白酶酶解Tg-HbⅡ,酶解液经反相C-18色谱柱分离纯化得7种血红蛋白源多肽F1~F7。通过琼脂扩散法检测酶解多肽抑菌活性,结果发现7种多肽对金黄色葡萄球菌、大肠杆菌、恶臭假单胞菌、枯草芽孢杆菌和坚强芽孢杆菌均无抑菌作用,但多肽F2~F7对副溶血弧菌、溶藻弧菌和哈维氏弧菌具有明显抗菌活性,且多肽F2~F7对副溶血弧菌、溶藻弧菌和哈维氏弧菌的MIC值在0.012~0.200mg/mL范围内。荧光显微镜观察酶解多肽杀菌效果发现多肽杀菌迅速,5min内弧菌细胞几乎全部死亡。利用透射电子显微镜观察正常的和经抗菌肽处理后的溶藻弧菌和哈维氏弧菌细胞的超微结构,结果显示,对照组正常细胞内部结构完整,而经抗菌肽处理过的细胞其细胞壁完整,但因缺乏细胞内容物支持导致局部区域内陷,细胞质膜明显收缩,细胞内物质外流形成许多空泡从而导致细胞死亡。研究结果表明血红蛋白源抗菌肽对泥蚶致病菌弧菌有明显的抗菌活性,其通过破坏细胞内部结构而起到杀菌作用,这些发现为血红蛋白抗菌机制的研究奠定了基础。     

太平洋双色鳗鲡(Anguilla bicolor pacifica)致病性类志贺邻单胞菌的鉴定和中药敏感性研究

从发病死亡的养殖太平洋双色鳗鲡(Anguilla bicolor pacifica)肝脏中分离到一株优势菌AMSH1,为研究其种属、毒力及中药敏感性,结合BIOLOG微生物鉴定系统和16S r DNA序列分析对该菌株进行鉴定,并通过人工回归感染试验确定其致病性。此外,还研究了10味中药单用、10种双联用和4种三联用对该菌株的体外抑菌作用。结果显示该菌株为类志贺邻单胞菌(Plesiomonas shigelloides)。经人工腹腔注射感染健康太平洋双色鳗鲡,证实该菌具有致病性,半致死量LD50为3.2×105CFU/g。药敏试验结果表明,10味中药均有较好的抑菌效果,其中五倍子(Galla chinensis)、丁香(Eugenia caryophllata)和生地榆(Sanguisorba officinalis)的效果最好,7种双联用复方和1种三联用复方均具有协同抑菌作用。本次引起太平洋双色鳗鲡发病的病原为类志贺邻单胞菌,该菌对太平洋双色鳗鲡的致病性及其中药药敏的研究属首次报道。建议选用含五倍子的中药双联用复方或三联用复方进行防治,皆绿色环保。     

基于ONDPI的海岸养殖池塘遥感影像提取研究

以Sentinel-2、Landsat-8卫星影像为数据源,以海岸养殖池塘为研究对象,针对养殖池塘的特点,提出了基于归一化差异池塘指数(NDPI)的面向对象结合法(ONDPI)。并以天津市某一海岸养殖区为例,与传统的水体指数结合直方图阈值分割法进行对比实验。实验表明ONDPI能够对遥感影像中的海岸养殖池塘进行有效提取,提取精度达到95%以上,优于其他方法,能够为海岸池塘养殖的规划管理提供参考数据。     

基于Sentinel-2影像的围海养殖信息提取

为了能够利用遥感图像快速准确地提取围海养殖矢量信息,本文选取养殖水体、堤坝及育苗室等交错分布的海参围海养殖区域作为研究区域,根据研究区域Sentinel-2遥感影像的光谱特征,选用归一化差异水体指数（Normalized Difference Water Index,NDWI）、改进归一化差异水体指数（Modified Normalized Difference Water Index,MNDWI）和增强水体指数（Enhanced Water Index,EWI）三类水体指数,分别进行提取实验,利用同时期高空间分辨率的高分二号卫星（GF-2）影像作为参考,验证不同方法的提取精度,精度评价结果表明：相较MNDWI和EWI两类水体指数,NDWI的分类精度更高,且利用NDWI提取研究区域的围海养殖信息的效果更好,所以该方法可在养殖区域的动态监测和规划管理中发挥数据支撑作用。