Modulation of p53 gene expression in the intertidal copepod Tigriopus japonicus exposed to alkylphenols

The copepod, Tigriopus japonicus has been recognized as an excellent marine model species for ecotoxicological studies. The present study cloned and characterized the p53 gene of this copepod and studied its expression pattern. We discovered that p53 expression patterns varied among different developmental stages of the copepod, having the highest expression in the adult. Such variation was possibly associated with the molting cycle. By using real-time RT-PCR, we further investigated the modulatory pattern of the p53 gene in the copepod after exposure to three alkylphenols (i.e. nonylphenol, octylphenol, and bisphenol A) which are known as endocrine disruption chemicals (EDCs). The results showed that the three alkylphenols significantly induced p53 gene expression in the copepod, indicating the involvement of p53 in such stress-responses. Thus, the copepod p53 gene provides one of the stress-response biomarkers for exposure to EDC-like compounds.     

Ontogenetic habitat use by mangrove/seagrass-associated coral reef fishes shows flexibility in time and space

Tropical shallow-water habitats such as mangroves and seagrass beds are widely acknowledged as important juvenile habitats for various coral reef fish species, most of which are commercially important to fisheries. Spatio-temporal variability in ontogenetic habitat use by fish among these tropical coastal ecosystems has rarely been investigated, yet there are sufficient reasons to believe that this plays an important role. In the present study, we test the spatio-temporal variability in patterns of ontogenetic habitat use by some mangrove/seagrass-associated coral reef fishes (Lethrinus harak, Lethrinus lentjan, Lutjanus fulviflamma and Siganus sutor). Abundances of these four species were investigated during two years in Tanzanian coastal waters, using underwater visual census in mangrove, seagrass, shallow and deep mudflat, and shallow and deep coral reef habitats. The study covered four distinct seasons of the year and was done at two spatially separated (>40 km) locations. Averaged across locations, seasons and years, juveniles (≤10 cm length) of the four study species had significantly higher relative densities in shallow-water (mangroves and seagrass beds) than in deep-water habitats (deep mudflats or coral reefs), whereas the opposite pattern was found for the adults (>15 cm). These findings suggest a strong and general pattern of ontogenetic habitat shifts from shallow- to deep-water habitats. However, specific habitat-use patterns of juveniles as well as adults differed significantly in time and space. Various species showed subtle to considerable flexibility in juvenile as well as adult habitat use across seasons, years, or at different locations. Furthermore, for some species the data suggest presence of ontogenetic habitat shifts at one location but lack thereof at the other location. In summary, ontogenetic habitat use needs to be considered at various spatial and temporal scales for the interpretation of habitat utilization by fish during different life stages. This is important for conservation and management of these habitats, as essential habitats or seasons may be ignored or over-emphasized with respect to their importance for fish during different parts of their life cycle.     

Molecular cloning of superoxide dismutase (Cu/Zn-SOD) from aquatic molluscs

The potential of the first line of the active oxygen-scavenging system, partial cDNA encoding Cu/Zn superoxide dismutase (SOD) was isolated in three aquatic mollusc species: Ruditapes decussatus (marine clam), Dreissena polymorpha (continental water mussel) and Bathymodiolus azoricus (hydrothermal vent mussel). These SOD cDNA fragments were amplified by PCR with degenerate oligonucleotide primers derived from the amino acid sequence conserved in the Cu/Zn-SOD from several other organisms. A partial cDNA of CuZn-SOD was obtained for R. decussates (510 bp), D. polymorpha (510 bp) and B. azoricus (195 bp). The deduced amino acid sequence showed high similarity among the three mollusc species (57-63%) and among other species (50-65%). The residues involved in coordinating copper (His-47, 49, 64, 121) and zinc (His-64, 72, 81 and Asp-84) were well conserved among the three Cu/Zn-SOD sequences.     

扇贝外套腔内的共栖生纤毛虫 Ⅰ.蠕状康纤虫(Cohnilembus verminus)的形态学研究

研究扇贝外套腔内栖生的纤毛虫一蠕状康纤虫Cohnilembusverminus（Muller，1786）的形态、核器及纤毛图式。结果显示其口器结构为；M1呈单列结构，约占体长的1／2，M2由4组毛基粒构成，PM起自M2末端，Sc约由5个毛基粒构成。此外，还列表比较了不同作者报导的C．verminus种群及其它6种已知的康纤虫的主要性状特征。     

Identification of the multidrug resistance-associated protein (mrp) related gene in red mullet (Mullus barbatus)

Multixenobiotic resistance mechanism (MXR) in aquatic organisms is mediated by the activity of the P-glycoprotein (Pgp) transporter that binds and actively effluxes different chemicals out of cell. In addition to the Pgp, several other, non-Pgp transport proteins have been recently identified in different human and animal tissues. Given their characteristics and tissue distribution we hypothesized that members of the so-called multidrug resistance-associated protein (MRP) family may be expressed in aquatic organisms. This study attempted to identify MRP related genes in different tissues of several marine and freshwater bivalves (Mytilus galloprovincialis, Dreissena polymorpha, Anodonta cygnea) and fish species (Mullus barbatus, Cyprinus carpio, Salmo trutta). Following an alignment of known MRP1 and MRP2 human sequences, as well as the GenBank available mrp2 sequences from different animals, we determined highly conserved regions and used them to design three pairs of consensus primers. Total RNA was isolated, reverse transcribed to cDNA and the obtained cDNAs were PCR amplified with the corresponding primers. The amplified PCR products were sequenced and their homology compared with Pgp and MRP protein sequences from different species. The expression of MRP related mRNA was clearly identified only in liver tissue isolated from red mullet, with homologies at the protein level ranging from 75% to 76%. Described results clearly pointed at the possibility that at least in the red mullet MXR as a general defense mechanism may be mediated by the activities of at least two different types of transport proteins.     

流式细胞术比较研究4种双壳贝类血细胞的分群

应用流式细胞术对太平洋牡蛎(Crassostrea gigas)、海湾扇贝(Argopecten irradians)、虾夷扇贝(Patinopecten yessoensis)和毛蚶(Scapharca subcrenata)4种双壳贝类的血细胞分类进行了比较。根据其血细胞的前向角散射光(FSC)和侧向角散射光(SSC)特性的不同将血细胞分群,FSC和SSC二维图分析发现太平洋牡蛎、海湾扇贝和虾夷扇贝的血细胞都可分为3个亚群,即透明细胞、小颗粒细胞和大颗粒细胞,而毛蚶的血细胞只可区分为透明细胞和颗粒细胞2个亚群。同时还对这4种双壳贝类各种血细胞所占比例进行了比较研究,发现各亚群细胞所占比例差异很大。     

泉州湾洛阳江红树林自然保护区潮间带软体动物多样性及分布

为了解洛阳江红树林自然保护区不同生境的潮间带软体动物物种多样性、数量时空分布及群落结构现状,国家海洋局第三海洋研究所于2009年4月和11月在洛阳江红树林自然保护区内选择了光滩（Qch1）、屿头红树林区（Qch2）、增墁红树林区（Qch3）和米草区（Qch4））4条断面共3种生境,进行软体动物调查。共鉴定软体动物16科...     

Changes in Irish Sea Benthos: Possible Effects of 40 years of Dredging

From 1946 to 1951 Dr N. S. Jones sampled the benthos around the south of the Isle of Man from over 200 sites. Multivariate methods have been used here to compare subsets of this historical data with recent data from the same locations: of these locations some have been subject to heavy scallop dredging over the intervening 40 plus years and some to little dredging. Clear changes were apparent regardless of scallop dredging intensity. Some of the changes in the heavily dredged areas were those expected to result from extreme physical disturbance—an increased polychaete mollusc ratio, loss of some fragile species, and an increase in the predominance of scavenger/predator species. However, changes in the lightly dredged areas also included the loss of a number of species including some potentially fragile tube-dwellers. Reasons for these changes were not apparent.     

文蛤属(Meretrix)16S rRNA基因及ITS1序列的系统学分析

利用线粒体16SrRNA基因片段及核糖体DNA转录间隔区ITS1序列分析的方法,以近缘属的青蛤(Cyclinasinensis)为外群,对文蛤属的文蛤(M.meretrix)、丽文蛤(M.lusoria)、帘文蛤(M.lyrata)和斧文蛤(M.lamarckii)4种贝类进行了系统学研究。经ClustalX多重比对及DNAsp软件分析后,用PAUP4.0分析软件,计算出种间序列的碱基转换/颠换频率,利用邻接法NJ构建系统发育树。结果表明,帘文蛤与该属其他三种的分歧时间较早,两类序列与其他种类间的相对遗传距离分别在0.25866—0.28218和0.15644—0.20104之间。文蛤与丽文蛤间的16SrDNA及ITS1序列间的遗传距离仅为0.04805和0.04201,拓扑结构图显示关系很近,并且二者的分布区大面积重叠,其序列间的转换/颠换频率接近种内单元型(haplotype)间的序列变化,鉴于它们的贝壳形态有一定差别,应归为同一个种内的不同地理亚种比较恰当。     

合浦珠母贝26S蛋白酶体S4,S7亚基基因片段的分离与分析

26S蛋白酶体是真核生物中一种具有ATP依赖性的蛋白酶复合体,主要通过泛肽途径选择性降解细胞内与代谢调控、细胞周期有关的功能蛋白及异常蛋白,参与多种细胞活动的调控过程。26S蛋白酶体由具有催化活性的20S亚复合体和一个具有调节作用的19S亚复合体组成,其中19S亚复合体中的ATP酶亚基是调节26S蛋白酶体活性的重要组件。本通过简并引物PCR手段,从软体动物合浦珠母贝(Pinctada fucata)中扩增到参与构成19S亚复合体的S4和S7(MSS1)两个亚基的基因片段。这两个基因片段所编码的ATP酶组件包含有Gx4GKT,DEID,SAT和H／QRxGRxxR等26S蛋白酶体ATP酶亚基的共同功能基序。这是首次在软体动物中报道26S蛋白酶体的ATP酶亚基基因序列,为研究软体动物中26S蛋白酶体的结构与功能奠定了分子基础。     

Comparative salinity tolerances of four siphonariid limpets in relation to habitat restriction of the rare and endangered Siphonaria compressa

The salinity tolerances of four South African species of limpet in the genus Siphonaria were experimentally tested, in the context that one of them, S. compressa, is South Africa's most endangered marine mollusc and is restricted to two lagoonal localities that normally experience little variation in salinity. Its habitat is additionally restricted in that it occurs only on the eelgrass Zostera capensis. The other three species, S. capensis, S. concinna and S. serrata, are abundant and widespread on temperate rocky shores. We hypothesised that S. compressa would have limited tolerance to low salinities, contributing to its absence from otherwise suitable estuaries that experience regular fluctuations of salinity. The percentage survival of the limpets was recorded in salinities ranging from 2 to 35 over that period. Both salinity and species significantly affected the survival of the limpets over the 96 hours. Survival was highest in salinities close to those of normal seawater, and greatly reduced in the hypo saline waters. S. compressa had the lowest salinity tolerance of the four species, its time-to-50% mortality in hyposaline waters being significantly less than that for the other three species, which differed in a manner that could be correlated with their zonation patterns. Restriction to eelgrass, coupled with low salinity tolerance, probably explains why S. compressa is found at only two localities, both of which normally experience salinities close to that of seawater. Even there, mass mortalities of S. compressa have been recorded in association with die-backs of eelgrass after abnormal freshwater flooding or sedimentation—further emphasising the vulnerability of S. compressa to extinction.     

Unusual megafaunal assemblages on the continental slope off Cape Hatteras

Megafaunal assemblages were studied in August–September 1992 using a towed camera sled along seven cross-isobath transects on the continental slope off Cape Hatteras. A total of 20,722 megafaunal organisms were observed on 10,918 m² of the sea floor between the depths of 157 and 1 924 m. These data were compared with data previously collected off Cape Hatteras in 1985 and at other locations along the eastern U.S. coast between 1981 and 1987. Megafaunal populations on the upper and lower slopes off Cape Hatteras were fouond to be similar, in terms of density and species composition, to those observed at the other locations.In contrast, megafaunal abundances were found to be elevated (0.88 and 2.65 individuals per m² during 1985 and 1992, respectively) on the middle slope off Cape Hatteras when compared to most other slope locations (<0.5individuals per m²). These elevated abundances mainly reflect dense populations of three demersal fish, two eel pouts (Lysenchelys verrilli andLycodes atlanticus) and the witch flounderGlyptocephalus cynoglossus, and a large anemone (Actinauge verrilli). These four species dominated the megafauna off Cape Hatteras, whereas they represented only a minor component of megafaunal populations found at other slope locations. Additionally, numerous tubes of the foraminiferanBathysiphon filiformis were observed off Cape Hatteras, but not elsewhere. The high density of demersal fish found off Cape Hatteras appears to be related to the high densities of infaunal prey reported from this area. The high densities ofA. verrilli andB. fuliformis may be related to the same factors responsible for the high infaunal densities, namely enhanced nutrient inputs in the form of fine particles. Extreme patchiness also was observed in the distributions of the middle slope taxa off Cape Hatteras. This patchiness may reflect the habitat heterogeneity of this exceptionally rugged slope and the sedentary nature of the organisms inhabiting it.     

Further refutation of the primary-secondary settlement hypothesis for the brown mussel Perna perna

The primary-secondary settlement hypothesis, that mussels first settle in algae and then move to mussel beds, was rejected as the only mode of recruitment for the brown mussel Perna perna in a previous study at one location over one year on the south coast of South Africa. We investigated recruitment of P. perna into mussel beds and algal turfs over a five-year period (1995–2000) at 3–4 sites at each of four locations on the South and East coasts. Attachment to the two substrata was significantly temporally synchronous at two of the four locations. The disparity among locations could be due to differences in the algal species present or to differences in the importance of turf algae as a facilitator of recruitment. At Zululand, on the East Coast, attachment to mussel beds and algae was measured monthly and found not to be temporally synchronous. At this location, primary settlement into algae was significantly correlated with an increase in the density of juveniles in the mussel beds 2–3 months later, indicating that turf algae can act as irregular but important facultative facilitators of mussel recruitment, supplementing direct input into the mussel beds. However, as amounts of settlers and recruits in mussel beds exceeded those found in algal turfs most years and places, the primary-secondary settlement hypothesis is inapplicable to P. perna under most circumstances.     

长臂缨鲆核糖体RNA基因序列多态性特征分析

为了解鲽形目Pleuronectiformes鲆科Bothidae长臂缨鲆Crossorhombus kobensis (Jordan & Starks, 1906) 核糖体RNA基因的序列多态性特征, 本研究共获得该鱼类包括18S、5.8S、ITS1和ITS2全长及28S部分序列的128条克隆序列。经序列比对、聚类分析以及重组检测, 结果显示5.8S (158bp) 无长度变异, 而其他4个基因片段则表现出较高的长度多态性, 并可分为不同序列类型: 18S (1856~1893 bp) 有4种序列类型A、B、C和R; 28S (967~974bp) 和ITS1 (407~ 505bp) 均有3种类型A、B和R; ITS2 (423~447 bp)存在2种类型A和B。此外5个基因片段在碱基组成中均表现出GC偏倚, 并且ITS2 (71.14%)>ITS1 (65.37%)>28S (62.22%)>5.8S (57.67%)>18S (54.95%)。对具有不同序列类型的18S、28S和ITS进行真、假基因推断时, 通常的判别特征不足以提供有力依据, 因此增加了与4种鲆科近缘鱼类长冠羊舌鲆Arnoglossus macrolophus、青缨鲆Crossorhombus azureus、大鳞短额鲆Engyprosopon grandisquama以及冠毛鲆Lophonectes gallus相应基因片段的比对。各基因片段的插入/缺失以及特异性碱基差异位点比对结果显示: 18S和28S的短序列类型A与4种鲆科鱼类序列一致, 而其他序列类型则不同; ITS1序列类型A与4种鲆科鱼类在类型B的缺失位点均无缺失, 因此推测18S、28S和ITS1的A类型为真基因, 而其他类型为假基因。ITS2的A和B类型在差异位点上与4个鲆科鱼类不存在一致性, 没有足够的依据对两个类型做出真、假基因的推断。长臂缨鲆核糖体RNA基因中, 5.8S序列最为保守遵循协同进化的方式, 而其他4个基因片段为非协同进化的方式。     

Natural dispersion of mercury from Puhipuhi,northland, New Zealand

The mercury content of sediments and water in the Wairua (Wairoa) River, Northland, and of molluscs from the estuary of the river was determined to establish the extent of natural dispersion of mercury from deposits at the source of the river at Puhipuhi. The mercury content per gram wet weight of cockles, Chione stutchburyi, (0.032 ppm); rock oysters, Crassostrea glomerata, (0.081 ppm); pipi Paphies australe, (0.019 ppm); and green‐lipped mussels, Perna canaliculus, (0.017 ppm) was compared with that of specimens of the same species from other areas where presumably only background concentrations exist. Mercury could be detected in sediments at least 35 km from the deposits, but in water only up to about 8 km. Normal background levels were established for the soft parts and individual organs of the four species of mollusc investigated; of the molluscs found in the estuary of the Wairua River, only C. glomerata had anomalous amounts of mercury, but whether this indicates abnormally high mercury levels in the environment is unknown, because many other factors still need to be evaluated. Mercury levels of all molluscs were well below the generally accepted safety level of 0.5 ppm for fish for human consumption.     

威海近岸人工鱼礁布设对生物资源恢复效果

为了解人工鱼礁投放对区域内海洋生物资源恢复效果,于2013年3月至2014年1月在北黄海近岸人工鱼礁投放区进行调查。结果显示,该海区发现浮游动物53种,其中优势种为浮游桡足类。夏秋季底栖生物幼体成为季节性优势种。游泳动物共发现14种,主要为常见的礁区鱼类。底栖动物共12种,其中棘皮动物海燕为绝对优势种,为近岸养殖生物的主要敌害生物,主要的底栖资源优势种为虾蟹类与软体动物。投礁区的浮游动物与游泳动物物种数明显多于未投礁区,表明人工投礁有助于生物群落的恢复,提高生物多样性,改善区域生态系统。但该海域大型底栖藻类较少,还应进一步对大型底栖藻类增殖以提高海域初级生产。     

基于29次南极科考的普利兹湾鱼类多样性和分子种类鉴定

利用2013年29次南极科考对南极半岛普利兹湾的调查采集到的鱼类进行多样性和分类鉴定,所有站位的样品均由雪龙号采集。位于高纬度普利兹湾内的鱼类多样性和相关的评估尚未知晓。因此,有必要对该调查海域的鱼类多样性进行准确的评估。本航次共捕获鱼类样品99尾,为了克服形态鉴定不准确的缺点,我们结合DNA条形码（COI基因片段）技术对种类鉴定,同时下载NCBI基因库中的公开数据作为参考。利用NJ系统发育树和条形码间隙共准确鉴定出22种鱼类,其中与形态鉴定相对应的有13种,5种鱼类准确鉴定到属的水平,4种鉴定到其近缘种。公开发表的资料中显示南极鱼科鱼类在种类组成中属于优势种类,但是在本次调查中并未发现这一现象。受调查方法和站位数量的限制,本次调查鱼类表现出较低的多样性和较低生物量,这一结果可能导致对普利兹湾鱼类多样性认识的片面性。结果证明条形码技术可以有效的用于普利兹湾鱼类的种类鉴定,同时南极鱼类的种类鉴定和分布是了解南极鱼类物种多样性和生物地理学的一部分,因此,有必要对南极鱼类在大尺度上对其准确的种类鉴定。     

Reduction of flatfish habitat as a consequence of the proliferation of an invasive mollusc

Coastal bays provide habitats for juveniles and adults of many marine species. Mont Saint-Michel Bay (MSMB, France) hosts a highly diversified fish community and constitutes one of the most important nursery grounds for many commercially exploited marine species, such as sea bass, flatfish, clupeids and rays in the English Channel. Besides, MSMB also suffers from the massive invasion of an exotic mollusc, the American slipper-limpet (Crepidula fornicata, L.). This species arrived four decades ago and now represents the main filter-feeder biomass in the bay (150 Mt), an order of magnitude larger than local farmed and natural shellfishes. Recent analyses underlined the impact of this small gastropod on the trophic structure of this bay and its negative influence on juvenile sole densities in the nursery grounds. The present study uses a geostatistical approach to explore the effect of the extension of the slipper-limpet on flatfish (common sole Solea solea, L.; plaice Pleuronectes platessa, L.; brill Scophthalmus rhombus, L. and flounder Platichthys flesus, L.) spatial distribution. Data collected during survey of the MSMB at the end of the 1970s and three decades later have been used to build interpolated maps of (1) slipper-limpet and (2) flatfish spatial distributions. Slipper-limpets were concentrated in a small area, in the western part of the MSMB, in the 1970s while today they occupy half of the bay. This rapid proliferation led to the decrease of available surface for flatfishes, which previously occupied the whole bay and are now restricted to its eastern part. The present study highlighted that the negative influence on fish habitat in MSMB is apparently more related to changes in the substratum than to trophic interactions. This invasion has possible consequences on flatfish population renewal at a large scale and may also damage other benthic or demersal species, such as rays.