中国对虾育苗池水中哈维氏弧菌的检测

１９９６年４月～５月,在山东莱州市大华育苗池、山东即墨市丰城育苗场发病育苗池、环境水体及活体饵料中共采集８０份样品,用间接ＥＬＩＳＡ技术对其进行了苗期对虾病原菌－哈维氏弧菌的检测。结果表明,发病育苗池中哈维氏弧菌的检出率（５８．３％）明显高于正常育苗池（２０．４％）,证实丰城对虾育苗场暴发的流行病与育苗水体中的哈维氏弧菌有关;大华育苗场育苗水中也检测出哈维氏弧菌,但大华育苗场并未发病,说明哈维氏弧菌似乎是条件致病菌。在丰城育苗场及大华育苗场的蓄水池中均检测到哈维氏弧菌,而活性饵料中未检出,表明育苗水中的哈维氏弧菌来源于外海水。     

海洋水环境中藻毒素的检测技术及分布研究进展

海洋藻毒素是海洋有害藻产生的次级代谢产物,具有种类多、毒性强和生物蓄积性等特点,对海洋生态系统、海产养殖业以及人类健康会造成严重威胁。近年来,近海水环境中的海洋藻毒素已引起人们的广泛关注。通过对近海水环境中海洋藻毒素的富集技术、仪器分析方法和分布特征方面的研究进展进行归纳和总结发现:被动固相吸附和固相萃取技术是海水中海洋藻毒素的主要富集技术,结合液相色谱-质谱联用技术进行检测,是目前近海水环境中各类海洋藻毒素检测的最有效方法;多种海洋藻毒素在全球近海水环境中有分布,且欧洲近海海水中藻毒素种类最丰富、浓度也相对较高;在中国近海水环境中多种脂溶性藻毒素广泛分布。在全球气候变化以及近海污染不断加剧的背景下,海洋藻毒素对近海水环境的影响不容忽视,今后的研究方向将主要包括:重要养殖区海水中藻毒素的长期连续监测;海水中典型藻毒素的现场快速检测技术;近海水环境中海洋藻毒素的来源及其影响因素;典型海洋藻毒素的迁移转化规律及其生物可利用性等。     

海水悬浮物浓度原位测量技术进展

海水悬浮物指海水中的悬浮颗粒物和浮游微生物等，对海水悬浮物浓度进行准确的测量具有重要的意义。海水悬浮物浓度测量技术，无论是传统方法（即现场采水），还是现代方法（即间接测量方式，包括光学法和声学法等）都有一定的不足。本文主要综述了几种海水悬浮物浓度间接测量方法、国内外有关海水原位过滤测量的技术研发进展及深海原位微孔过滤技术。最后，对海水悬浮物原位过滤技术的发展方向进行了探讨。     

共沉淀富集法测定海水中稀土元素含量方法研究

稀土元素已越来越多地应用于示踪各类地球化学体系的物质来源与岩石矿物形成等演化过程。但在进行海水样品的稀土元素检测时，由于样品的高盐特征和稀土元素的痕量特征又使得分析具有相当的挑战性。本研究旨在从高盐的海水基质中有效分离并富集痕量的稀土元素，同时结合电感耦合等离子体质谱（Inductively Coupled Plasma Mass Spectrometry, ICP-MS）技术对海水中的稀土进行定量分析。通过对海水样品稀土元素的定量限分析、加标回收实验和梯度加标回收实验，验证了方法的准确性，评估方法的分离富集效率。通过共沉淀过程中的pH比较、氢氧化钠与氨水作为共沉淀剂的比较，确定了富集过程中共沉淀剂用量、种类等影响因素。通过稀土元素配分曲线的绘制与类似样品结果进行比对，以及标准海水的检测结果，验证了方法在实际样品中的可行性。本研究探索了共沉淀富集法在分离富集海水中稀土元素的实验条件，实现高盐背景下痕量稀土的准确定量，并将其用于近岸海水与孔隙水的实际检测。     

海水中溶解二氧化碳监测技术跟踪

综述了研究监测海水中二氧化碳含量的目的及意义和二氧化碳在海水中的存在形式,以及测量海水中溶解二氧化碳浓度的监测方法,着重介绍了直接测量的几种原理和目前主要采用的基于红外分析原理的研制方法,并对该领域的现状和发展进行了分析.     

基于电子鼻和HS-SPME-GC-MS方法研究 海水中七氯的快速检测技术

为了建立海水中有机氯农药的快速检测技术, 本文基于电子鼻和顶空固相微萃取气质联用法(HS-SPME-GC-MS)对海水中的七氯进行检测。结果表明, 电子鼻可以识别海水中的挥发性七氯成分及浓度, 最低检测限0.01mg/L。利用线性判别分析方法(LDA)的累计判别贡献率和欧氏距离、判别函数法、相关性对未知样品的识别率均达到了80%以上, 具有良好的区分效果; PLS模型预测七氯浓度跟实际值之间的相对误差在0.49%—16%之间。HS-SPME-GC-MS检测浓度小于0.10mg/L时, 相对误差在10%左右; 而大于此浓度时, 相对误差随之增大。检测到的主要挥发性物质基本呈现规律性的变化, 与电子鼻确认结果一致。对比两种检测技术, 发现电子鼻检测方法比HS-SPME-GC-MS更迅速、灵敏, 检测限更低, 相对误差也更小, 仪器便携、投资相对较小。     

动态式抽取、气相色谱法测定海水中的甲烷

本文报道了用动态（ｄｙｎａｍｉｃ）技术，经抽取、气相色谱法测定海水中溶存甲烷浓度的方法．详细论述了溶存甲烷经起泡、收集（吸附）、解吸和检测四步完成，并逐步讨论了起泡时间、起泡气体流速以及收集温度等有关因子对测定的影响。结果表明：该方法分析速度快、精密度好、灵敏度高，可广泛应用于海上现场测定海水中溶存的甲烷。     

应用双特异分子探针技术定性定量检测圆海链藻(Thalassiosira rotula)

设计了一套圆海链藻(Thalassiosira rotula)特异性探针,运用双特异分子探针技术,对圆海链藻(Thalassiosira rotula)进行了定性定量检测.结果表明,本实验中设计的一套探针与其它十几种藻无交叉反应,具有种特异性;细胞裂解液直接杂交检测优于提纯RNA样品检测;对自然样品做了初步检测,发现天然海水中的其它浮游生物对该检测方法影响很小.     

基于分光光度法的多量程海水营养盐原位传感器检测系统设计

针对传统海水营养盐检测方法不能满足海水营养盐长期原位监测需求的问题,研制了一种基于分光光度法的多量程海水营养盐原位传感器检测系统,通过对系统的高度集成及对多量程检测、低功耗技术、漏液保护技术的应用,实现了对海水5项营养盐参数快速、宽范围、高精度的原位测量。经过实验室和青岛中苑码头现场测试,表明本营养盐传感器检测系统具有低功耗、高可靠性能,可满足对5项营养盐参数的快速精确测量要求,实现了对海水营养盐参数的原位监测,为相关部门及时了解海洋生态环境和水体富营养化程度提供了数据支持,具有重大现实意义。     

多波束测深数据处理及成图

针对多波束测深系统测量的特点,分别分析声速改正技术和潮位改正技术。从声速在海水中的传播出发,阐述海水中声速的测量,对声速的较正方法进行探讨,随后针对潮汐效应的影响,对多波束测深数据进行潮位改正,并利用海上试验实测的多波束测深数据,将处理后的数据绘制成海底地形数字地图。     

Characterization of bacterial communities associated with seasonal water masses from Tongyoung in South Sea of Korea

Understanding the patterns of microbial diversity and their composition in seawater is necessary to assess the impacts of microbes on marine ecosystem. Although the potential roles of microbes in the ocean have been studied, their diversity and relationship with environmental factors remain unclear. In this study, we adopted a 16S rRNA gene tag-pyrosequencing technique to investigate the bacterial communities associated with two different water masses from Tongyoung in the South Sea of Korea. Our results revealed highly diverse bacterial communities up to 900 Operational Taxonomic Units (OTUs) estimated from each seawater which was collected in the month of March and May, when the environmental conditions including temperature differed significantly: 7.2°C and 17.6°C in March and May, respectively. Altogether, 13 bacterial phyla were recovered from the seawater, of which Proteobacteria was the most dominant group. In addition, the value of the Shannon index, which measures the evenness of the distribution of individuals among OTUs, in May is higher than that in March, indicating that it displays a wider diversity of bacteria. Interestingly, the proportion of pathogenic bacteria was significantly increased in the month of May compared to March, suggesting that pathogenic bacteria were increasingly emerging in May. In particular, Pseudoalteromonas and Vibrio spp. were determined as major pathogenic bacteria from both water masses, of which Vibrio spp. were dominant.     

海洋放线菌不同分离方法的比较研究

采用平板涂布法,以青岛海区潮间带沉积物为对象进行海洋放线菌的分离方法研究.具体分析了不同样品预处理方式、稀释液、海水浓度和培养基种类等对分离效果的影响.结果表明,55℃预处理样品6 min能有效减少细菌数量,利于潮间带沉积物中放线菌的分离;以1/4林格氏溶液稀释样品、纯海水配置培养基,可以分离得到较多的放线菌;不同培养基对沉积物中放线菌的分离效果差别很大,本实验设置的M1 ～ M9培养基中,M1、M6、M7和M8培养基的分离效果优于其他5种.     

Starvation effects on pathogenic Vibrio alginolyticus in natural seawater

To get a better understanding of the starvation survival strategy of pathogenic Vibrio alginolyticus, log-phase cells were inoculated into sterile natural seawater for starvation studies. The results showed that all of total bacteria number, viable bacteria number and CFU number of V. alginolyticus increased remarkably at the initial starvation stage; after reaching their peaks at 5 d, both total bacteria number and viable bacteria number of V. alginolyticus fell slowly, while the CFU number fell more quickly after reaching its peak at 10 d; V. alginolyticus elongated their cells at the prophase of starvation, and then shrunk their volume and turned their shapes into ovals from rods at the anaphase of starvation; starved cells showed more sensitivity to heating and UV; starved cells showed no significant difference from unstarved ones at the lowest detection limit determined by indirect enzyme-linked immu- nosorbent assay （ELISA） ; starved cells＇ ability to adhere to the skin mucus of large yellow croaker （ Pseudosciaena crocea） showed a sharp decline as the starvation time increases; the cellular protein of V. alginolyticus increased remarkably at the ariaphase of starvation. The results indicated that pathogenic V. alginolyticus could survive in starvation for relatively long periods of time （ ≥2 months） in 28℃ natural seawater due to the morphological and physiological changes; however, starved V. alginolyticus cells showed less virulence and higher sensitivity under environmental stresses.     

光谱法测定海水中溶解态无机氮的研究进展

溶解态无机氮（dissolved inorganic nitrogen, DIN）主要由亚硝酸盐-氮（NO^-₂-N）、硝酸盐-氮（NO^-₃-N）和铵氮（NH⁺₄-N）组成,它们在海洋的生物地球化学循环过程中起重要作用。但人类活动向海洋输入了大量无机氮,导致一系列环境问题。为了更好地开展海洋氮循环研究和环境污染管理,需对海水中的DIN进行测定。在众多分析方法中,光谱法因其通用性好、适用范围广、所需设备简单,成为测定海水DIN的首选。本文总结了近10年来基于光谱法测定海水DIN的研究进展,包括紫外分光光度法测定NO^-₃-N、萘乙二胺分光光度法测定NO^-₂-N和NO^-₃-N、次溴酸盐氧化-分光光度法测定NH⁺₄-N、靛酚蓝分光光度法测定NH⁺₄-N、酸碱指示剂-分光光度法测定NH⁺₄-N、荧光法和化学发光法测定DIN等,比较了各分析方法的特点,并展望了光谱法测定海水DIN的发展趋势。总的来说,在分析方法上,新试剂的使用以及一些新合成材料的出现,丰富了DIN的分析手段;在分析仪器上,以流动分析技术为基础的分析仪器在DIN的实验室及现场分析中得到了广泛应用。DIN的分析方法均朝着简单便捷、全自动化、分析速度快、精确度高、可适用范围广的方向发展。     

Studies on the determination of inorganic iodine in seawater

Existing methods for the determination of the various forms of iodine present in seawater are laborious or unwieldy in use. This paper describes modified forms of the iodate and total iodine methods described by Barkley and Thompson (1960), a spectrophotometric procedure for iodate determination derived from Johannesson's (1958) work, and an automatic method for total iodine determination. Procedures for iodate and total-iodine determination which are suitable for use aboard ship, are recommended. Both procedures for iodate determination returned a standard deviation close to 0.5 μg/l when several replicate samples of a seawater containing approximately 30 μg/l of iodate-iodine were analysed. The automatic method for total iodine determination yielded a standard deviation of 0.8 μg/l by the repeated analysis of a seawater containing a total of approximately 50 μg/l of iodine. Suitable methods for the filtration and storage of seawater are also described.     

南麂岛海域沉积物中海洋放线菌的分离研究

采用稀释涂板法研究了南麂岛海域沉积物中海洋放线菌的分离技术,分析了真空干燥处理、热处理、海水浓度、培养基种类等因素对分离效果的影响;首次将羧甲基纤维素钠(CMC)溶液应用于海洋沉积物样品的预处理,探讨了不同浓度的CMC对海洋沉积物样品中放线菌分离结果的影响。结果表明:海洋沉积物经干燥处理、50℃热处理20 min均能有效地减少细菌数量,利于海洋沉积物中放线菌的分离;60%的海水配制的培养基对海洋放线菌的分离效果优于纯海水所配培养基,并且培养基中添加海泥浸出液能有效地增加海洋放线菌的出菌率;以质量浓度为2 g/L的CMC为分散剂时,不仅能良好地分散沉积物样品中的放线菌孢子,而且能明显地增加海洋放线菌的检出量及稀有放线菌的数量。     

海水中硼的测定方法

介绍了海水中硼的测定主要方法,包括容量法,分光光度法、极谱法、光谱法、离子选择电极法等及其研究进展。     

A new method to detect viable bacteria in natural seawater using 16SrRNA oligonucleotide probe

Fluorescent oligonucleotide probes were used to detectVibrio cholerae directly under epifluorescence microscope. The probe which is complementary to the specific 16SrRNA sequence ofVibrio cholerae was labelled with Texas-Red, whereas the universal probe was labelled with Fluorescein. These probes allowed the distinctionVibrio cholerae from other eubacteria under the same microscopic field. In order to detect and enumerate specific bacteria in natural seawater, this method was combined with the direct viable count (DVC) technique. The combined method increased intracellular rRNA levels in the sample, and made it possible to detect the target bacteria with the specific gene probe. The applicability of this new method was confirmed both for the laboratory mixed culture system and natural seawater.     

Filter-feeder macroinvertebrates as key players in culturable bacteria biodiversity control: a case of study with Sabella spallanzanii (Polychaeta: Sabellidae)

The present study investigates the effect of the filtering activity of Sabella spallanzanii on the culturable heterotrophic bacterial community through the comparison of the bacterial diversity in transplanted polychaetes and the surrounding seawater. For isolation of culturable heterotrophic bacteria, seawater samples as well as polychaete homogenates were plated in triplicates onto Bacto Marine Agar 2216 (Difco). All the colonies grown were isolated, subcultured and identified by several morphological, biochemical and cultural methods. Some bacterial genera showed higher average abundances in polychaetes than in seawater (i.e. Lucibacterium and Photobacterium). Aeromonas represented a conspicuous component of the bacterial community both in S. spallanzanii and seawater. The presence of Cytophaga and Pseudomonas was also relevant in the examined seawater samples. The selective concentrations of some bacterial genera inside S. spallanzanii either by grazing on bacteria or their capability as bacterial reservoirs, provides evidence for the role of macrobenthic invertebrates as key determinants for microbial diversity.     

中国重点海水浴场中粪肠球菌和主要肠道病毒的调查研究

本文调查了中国十大典型海水浴场,旨在监测人类肠道病毒在海水浴场的发生情况,并且找出表面海水中肠道病毒的存在与传统的粪便污染指示物一粪大肠菌群是否存在关系,如果存在,是什么关系。分别采用常规的平板计数法和RT-PCR技术对粪大肠菌群和人类致病性的主要肠道病毒（甲肝病毒、轮状病毒、脊髓灰质炎病毒）进行了检测。结果显示,在2...