海洋烷烃降解菌Alcanivorax sp.A-11-3的分离鉴定及其降解酶基因研究

从马六岬海峡的表层海水中分离到一株石油降解菌A-11-3.经16S rDNA Blast结果表明其与Alcanivorax borkumensis SK2具有最高相似性为96%,该菌可能是食烷菌属的一个新种.该菌具有很强的石油降解能力.结果表明,A-11-3对烷烃的降解范围较宽,能以C8～C36的烷烃为唯一碳源和能源生长;在7d内该菌对柴油的降解率达到49.5%.此外,还从菌株中克隆到了大小为426bp的烷烃羟化酶基因alkB片段和843bp的P450 烷烃羟化酶基因的部分序列.序列分析表明,alkB片段编码的氨基酸序列与A. borkumensis SK2T的AlkB1和AlkB2的同源性分别为57.75%和40.14%;该菌的P450与SK2菌株的P450具有最高同源性,为87%.研究结果对于海洋石油降解微生物生态研究及石油污染生物修复技术开发有参考价值.     

海洋石油降解微生物的分离鉴定

以柴油为惟一碳源,从胜利油田黄河码头和厦门储油码头两个海水样品中富集得到两组柴油降解菌,共9株细菌,1株真菌,它们对柴油都有降解能力.16SrDNA鉴定结果表明这9株细菌中有3株摩加夫芽孢杆菌、1株施氏假单胞菌、1株腊样芽孢杆菌、1株反硝化产碱杆菌.1株阿氏葡萄球菌、1株食烷菌和1株类似很小海旋菌的未知新菌(M-5),其中B-5对柴油的降解能力最强,已报道的食烷菌属中其他种的同源性最高,为95.2%,表明它是该属中的一个新种.实验中获得的惟一一株真菌M-3属于假丝酵母,对柴油有较强的降解和乳化能力.实验中还从B5和施氏假单胞菌M-2中扩增到了烷烃降解的限速酶烷烃单加氧酶的基因片断,其中B5编码了一种新的烷烃单加氧酶.这些菌在石油污染的海水自净中起着重要作用,可用于海洋石油污染的生物修复.     

海洋石油烃降解细菌T1和T2的分子鉴定分类

从天津滨海潮间带被石油烃严重污染的沉积物（千样含油量0．2g／g）中，筛选分离出能够以柴油为唯一碳源生长的细菌，对其中生物量大、单株菌降解效率较高的两株细菌T1和T2进行16SrDNA克隆，通过测定和比较16SrDNA的部分序列对这两株细菌进行分子鉴定，以期用于石油污染的微生物修复中。结果表明，T1与深红红螺菌（Rhodospirillum rubrum）的同源性为89％，．r2与施氏甲单孢杆菌（Pseudomonas stutzeri）的同源性为99％。     

地芽孢杆菌属DYth03 DNA聚合酶基因的克隆、表达及性质分析

从东太平洋热液区E53站位的深海沉积物样品中分离出1株能在65℃生长的嗜热菌（DYth03）.该菌的16S rDNA序列与地芽孢杆菌属（Geobacillus）内各种之间的同源性为98%以上.克隆得到DYth03的DNA聚合酶基因（DYth-pol）,序列分析表明该基因全长为2 631 bp,G＋C含量为55.5%,推测编码为876个氨基酸,与BstDNApolI的同源性最高（达98%）.将该聚合酶基因克隆到pTTQ-h表达载体上,并在大肠杆菌DH1中进行表达.对纯化到的表达产物进行酶活性测定,结果表明该酶具有聚合酶活性和5’-3’外切酶活性.     

源于克雷伯氏菌Klebsiella sp.AC-11的3株噬菌体的生物学特性

从国家海洋局第三海洋研究所海水养殖池中分离得到一株细菌AC-11,16SrDNA序列分析表明其与肺炎克雷伯氏菌肺炎亚种(Klebsiella pneumoniae subsp.pneumoniae)具有最高的序列同源性达99.78%.以此菌株为宿主菌,于2008年11月采用双层平板法分离得到3株裂解性噬菌体KP1,KP...     

3种鲍16S rRNA基因片段序列的初步研究

本文对引自日本的盘鲍 (H aliotisdiscusdiscus)、皱纹盘鲍 (H.discushannai)和大鲍 (H.gi-gantiea) 3个自然群体的线粒体 DNA 1 6S r RNA基因片段进行了扩增和测序 ,分析了 52 8bp的碱基序列 ,结果显示 :3种鲍的基因序列中 A+T含量为 56.74%～ 57.1 2 %。 3个自然群体间碱基片段序列差异不显著 ,盘鲍与皱纹盘鲍、大鲍彼此均仅有 1处核苷酸检测到变异 ,皆为碱基转换 ,同源性为99.81 % ;皱纹盘鲍与大鲍有 2处碱基转换 ,同源性为 99.61 %。 1 6Sr RNA基因在鲍属内表现出很高的保守性。     

4株米氏凯伦藻(Karenia mikimotoi)藻际异养细菌的分离鉴定

采用分子生物学方法,对分离的米氏凯伦藻4株藻际异养细菌进行分子鉴定和功能分析。结果表明,经16S rRNA序列分析,4株藻际异养细菌分别隶属于Formosa、Erythrobacter、Shewanella和Marinobacter四个属,P1菌株与Formosa的16S rRNA核苷酸序列的同源性为95%,P2、P3和P4菌株与Erythrobacter、Shewanella和Marinobacter的同源性在99%以上。通过4株异养细菌及其同源性大的24株细菌构建进化树得出,28株细菌在系统发育树中主要分成4个分支,与同源性分析相符。     

海洋石油降解菌的筛选鉴定及其功能基因研究

以石油原油为唯一碳源,从中国海洋大学海洋微生物菌种库15株菌中筛选到5株石油降解菌。16SrRNA基因测序结果表明,WH072、WH069、WH303、ZXM008和ZXM183分别为耐盐芽孢杆菌(Bacillus halodurans)、除烃海杆状菌(Marinobacter hydrocarbonoclasticus)、弯曲芽孢杆菌(B.flexus)、普里兹湾假交替单胞菌(Pseudoalteromonas prydzensis)和嗜麦芽糖寡养单胞菌(Stenotrophomonas maltophilia)。其中对石油降解率最高的为WH072(高达36.4%),其次是WH069(20.0%)。对这5株菌检测已报道的石油烃降解酶基因,结果在WH072和WH069中均检测到了alkB、P450和almA3种功能基因。根据功能基因编码的氨基酸序列构建系统发育树并分析其各自的进化关系,发现P450基因在一定程度上具有种属特异性。对本研究首次报道的高效石油降解菌耐盐芽孢杆菌WH072,采用高效热不对称性交错PCR(hiTAIL-PCR)方法,成功获得了这3种功能基因的全长序列,其中alkB与已知同源基因序列相似性较低,其结构和功能有待进一步研究。     

养殖近江牡蛎致病弧菌的分离与鉴定

从广东省疫区患病近江牡蛎Crassostrea ariakensis消化盲囊及血液中分离纯化出10株细菌,人工注射感染实验发现其中3株细菌注射组死亡率显著高于对照组.3株细菌均为革兰氏阴性,短杆状,能运动.常规生理生化和Biolog 细菌鉴定系统测试结果表明,菌株A3G-2与溶藻弧菌Vibrio alginolyticus亲缘关系最近,菌株A2G-5B与解蛋白弧菌V. proteolyticus亲缘关系最近,菌株B1X-1与鲨鱼弧菌V. carchariae亲缘关系最近.为了进一步确定它们的分类学地位,分别测定了3株弧菌的16S rDNA序列,分析相关细菌相应序列的同源性,并构建系统进化树.结果表明, 菌株A3G-2与溶藻弧菌亲缘关系最近,菌株A2G-5B与解蛋白弧菌亲缘关系最近,菌株B1X-1与鲨鱼弧菌亲缘关系最近.因此菌株A3G-2、A2G-5B 和B1X-1分别被鉴定为溶藻弧菌、解蛋白弧菌和鲨鱼弧菌.组织病理观察发现患病牡蛎主要受损器官是消化盲囊,表现为腺泡管内壁吸收细胞崩解脱落,管腔扩大,进而管壁解体,消化盲囊出现大面积受损.     

一株深海嗜低温萘降解细菌Nah-1的分离及降解基因研究

以萘为唯一碳源和能源从南，中绳海槽深海沉积物中分离得到一株能降解萘的海洋嗜低温细菌Nah-1，测定了该茵的最适生长条件及生长曲线。16S rRNA基因（16SrDNA）序列同源性分析表明该菌属于解环菌属（Cyczocznsticus）。PCR扩增萘降解基因得到目标片段，比对结果表明，相似度最高的基因phnAl来自Cycloclasticus sp．A5，为99％，该基因编码的蛋白是萘双加氧酶大亚基。     

利用贝类废弃物制造水解动物蛋白

以扇贝加工废弃物—扇贝边为原料 ,采用枯草杆菌蛋白酶、胰蛋白酶、Flavourzyme对其进行深度水解 ;然后应用超滤技术对酶解液进行分离 ,除去未水解的蛋白质、细菌、微细的颗粒等物质 ,得到富含氨基酸和小分子肽的超滤透过液 ;再应用反渗透技术进行预浓缩 ,以除去其中过多水分 ;最后 ,对该浓缩液进行减压浓缩或喷雾干燥 ,即得到浓液状或粉状的水解动物蛋白产品。其中 ,粉状产品的粗蛋白含量为 6 6 .88% ,氨基酸态氮含量为 6 .96 % ,氨基酸态氮 /总氮 6 5 .0 5 % ,均优于国外同类产品     

5种南海海藻醇提取物活性初步研究

海藻是海洋生物代谢产物的主要来源之一,能产生丰富多样的生物活性物质.为了更好地利用中国南海的海藻资源,分别对采自中国湛江硇洲岛的1种绿藻基根硬毛藻Chaetomorpha basiretorsa Setchell和4种红藻刺状鱼栖苔Acantophora spicififera (Vahl) Boerg、三列凹顶藻Laurencia tristicha Tseng, Chang et Xia、紫杉状海门冬Asparagopsis taxiformis (Delile) Trevisan、鸡毛菜Pterocladia tenuis Okam进行了生物活性研究,分别对细胞毒活性(KB、Bel-7402、PC-3M、Ketr3和MCF-7)、Na~+,K~+-ATPase抑制活性以及犬血管平滑肌细胞增殖抑制活性进行了筛选,部分海藻提取物表现出一定的生理活性.     

Variability of the Kuroshio deduced from sea level data from the Izu Islands

Flow patterns and positions of the Kuroshio in the vicinity of the Izu Ridge are clarified by analyzing hydrographic observations and daily mean sea levels at Hachijo-jima, Miyake-jima and Oshima in the period from 1964 to 1981.Correlations are calculated between differences of dynamic depth anomaly at the surface refered to 1,000 db and differences of daily mean sea level between the two islands. The datum line of the tidal station at Hachijo-jima is about 90 cm higher than that at Miyake-jima, and about 20 cm higher than that at Oshima. A clear correlation is found between the cross-section transport of the Kuroshio and the mean sea-level difference between Hachijo-jima and Miyake-jima. The sea-level difference of the flow pattern without meander off Enshu-nada (type N) tends to be larger than that of the flow patterns with meander (type A and type B). This seems to indicate that the volume transport of the Kuroshio in the meandering period is smaller than that during the straight path period. Large sudden increases or decreases in the mean sea-level difference occur when the flow pattern changes and the Kuroshio axis shifts. The frequency of occurrence of quiet periods in the sea-level difference reflect the flow pattern of the Kuroshio.     

Lessons from inlet relocation: examples from Southern Portugal

Inlet relocation is a coastal management tool that, when applied to a migrating inlet, involves the artificial opening of a new tidal inlet along the historic migration path of the inlet. The old inlet is then artificially closed, or it is left open and will eventually close if the new inlet captures the entire tidal prism. Two inlets were relocated in such a manner in Southern Portugal and then were the subject of a monitoring program that included the acquisition of quantitative (topo-bathymetric surveys) data. Data was acquired for 4 years after the relocation at one of the inlets (Ancão Inlet) and for 2 years at the other (Fuzeta Inlet). The data obtained from the monitoring program were analysed together with the wave climate and then compared with historical information on the natural inlets, in order to assess the degree of success of the relocation actions. One of the relocations studied, Ancão Inlet, was considered to be successful even though an initial unexpected behaviour produced some material damage to property. On the contrary, the relocation of Fuzeta Inlet did not have the expected results, and the new inlet was affected by the same problems as the old one.It was found that the most important factor for a relocation action to succeed is the correct choice of the opening location. A theoretical procedure to enhance the possibilities of relocation success is suggested. (1) Hydrodynamic studies are needed in order to determine if the present conditions are similar to the historical ones. (2) The position for the inlet opening is chosen according to the hydrodynamic conditions, but there are other factors to be taken into account, i.e., the historical migration paths and typical inlet width of the natural inlet; the hydrodynamics of the backbarrier; the morphology of the backbarrier and, for multi-inlet barrier island systems, the proximity to adjacent inlets. (3) Once the position is chosen, environmental impact studies should be made in order to assess the risk of the relocation for the ecosystems of the area. Only if the environmental impact studies are favourable should a relocation action be performed.     

Biomarkers derived from heterolytic and homolytic cleavage of allylic hydroperoxides resulting from alkenone autoxidation

Laboratory incubation of alkenone mixtures with tert-butyl hydroperoxide and di-tert-butyl nitroxide (radical initiator) in hexane, as a means to simulate alkenone autoxidation processes, rapidly led to the formation of allylic hydroperoxides, whose presence was recently demonstrated in Emiliania huxleyi cells. After incubation in seawater and subsequent reduction with NaBH₄ (to reduce residual hydroperoxides before analysis), these reaction products quickly disappeared and were replaced by complex mixtures of n-alcohols, fatty acids, alkyldiols and hydroxyacids. Methyl alkenones produced saturated n-alkan-1-ols and fatty acids ranging from C₁₃ to C₁₆ and two series of C₁₃−C₁₆ (ω-1)-hydroxyacids and (1,ω-1)-diols. Ethyl alkenones also afforded C₁₃−C₁₆ saturated n-alkan-1-ols and fatty acids, accompanied by the production of C₁₄−C₁₇ (ω-2)-hydroxyacids and (1,ω-2)-diols. Deuterium labelling allowed us to show that most of the n-alkan-1-ols, hydroxyacids and alkyldiols resulted from the reduction during the NaBH₄ treatment of the corresponding aldehydes, ketoxyacids and ketoxyaldehydes formed from heterolytic or homolytic cleavages of allylic hydroperoxyl groups resulting from the oxidation of the double bonds of di- and triunsaturated alkenones. Amongst these products, the (ω-1)- and (ω-2)-hydroxyacids formed after NaBH₄ reduction of the (ω-1)- and (ω-2)-ketoxyacids were selected as potential biomarkers for alkenone autoxidation. Re-examination of lipid extracts of post-bloom seawater particulate matter samples from the DYFAMED station in the Ligurian Sea (where strong autoxidative alteration of the lipid distributions had previously been detected) showed the presence of significant amounts of 12-hydroxytetradecanoic, 13-hydroxytetradecanoic, 14-hydroxyhexadecanoic and 15-hydroxyhexadecanoic acids thus providing good evidence that these autoxidative processes occur in natural samples.     

南海冷泉区贻贝(Bathymodiolus platifrons)附生菌的分离培养与多样性分析

以采集后适应培养0~24 h以及添加甲烷和硫化钠培养24~240 h的南海冷泉区深海贻贝(Bathymodiolus platifrons)为材料,取其鳃部,分析附生菌的多样性与适应性变化状况。共分离鉴定出贻贝附生菌270株,对菌株的16S r DNA进行聚类分析,结果表明所分离菌株主要分布在4个门,21个属,其中变形菌的数量最多且多样性高。分析发现原位新采集的贻贝鳃部附生菌的多样性较高,6 h后附生菌的多样性明显降低。分别添加甲烷和硫化钠对深海贻贝进行培养,甲烷组与碳代谢有关的假单胞菌的数量逐渐增多,且在甲烷组240 h中发现2株食烷菌属菌株。硫化钠组的芽孢杆菌属所占比例升高。此外还发现4株潜在的新种。本研究实现了深海冷泉区贻贝的实验室培养,并对贻贝鳃部附生菌群结构进行了分析,丰富了海洋极端环境微生物资源库,并为深入解析贻贝与其附生菌之间的相互作用关系奠定了基础。     

蛇鲻胃蛋白酶的分离纯化和性质研究

以蛇鲻Saurida elongata内脏为原料,将蛇鲻胃经组织捣碎,硫酸铵分级沉淀,HitrapTMQ FF离子交换层析,Sephadex G-100凝胶过滤层析,得到胃蛋白酶纯品。经SDS-PAGE显示为单一条带,酶的分子量为37.7kDa,测得其最适温度为45℃,最适pH为2.2,酶的热稳定性很好,在40℃及以下温度处理3h酶活力没有任何变化,酶在pH 1—10以下都比较稳定,在强碱性条件下不稳定。EDTA和Trypsin inhibitor对该酶都没有明显的抑制作用,PMSF和Pepstatin A能完全抑制胃蛋白酶的活性。Ca2 、Mn2 、Mg2 、Ba2 对该酶影响不大,Hg2 对酶有较弱的抑制作用,而Zn2 、Cu2 、Fe3 对酶的抑制作用较强。     

The separation of Ceylon from Antarctica

The geometry of the junction between Ceylon and Antarctica during the Gondwanaland breakup is still under discussion. Analysis of the available geological-geophysical materials has allowed the peculiarities of Ceylon separation from Antarctica to be characterized, the new paleogeodynamical reconstruction to be elaborated, and a prognosis of the tectonic structure and mineral resources in the areas of Antarctic coast that were adjacent to Ceylon to be made.     

Criticism from former director of fisheries

In an article published in Marine Policy a year ago, (Lynne and Brian Chatterton, ‘How much political compromise can fisheries management stand’, Vol 5, No 1, April 1981), it was argued that management of South Australian coastal fisheries reflected the political ideology of the government in power, rather than the biological and socioeconomic imperatives of the resource. In the first article below, A.M. Olsen, former Director of Fisheries, criticizes the Chattertons' conclusions, maintaining that fisheries should be managed by professionals not politicians and that many of Brian Chatterton's problems during his tenure as Minister of Fisheries were of his own making. In the Chattertons' response, they point out that by allocating management of the resource between various vested interests (including government bureaucracy) a minister is making a political decision. In the third article, R.D. Walkerden, a former Permanent Secretary to Fisheries Ministers in South Australia, offers an administrator's view of the controversy. Detailed and parochial as some of the points at issue appear, the debate provides an illuminating view of some of the ideological and organizational conflicts that can hinder the effective management of a coastal resource.