Design ofVibrio 16S rRNA gene specific primers and their application in the analysis of seawaterVibrio community

The pathogenic species of genus Vibrio cause vibriosis, one of the most prevalent diseases of maricultured animals and seafood consumers. Monitoring their kinetics in the chain of seafood production, processing and consumption is of great importance for food and mariculture safety. In order to enrich Vibrio-representing 16S ribosomal RNA gene （rDNA） fragments and identify these bacteria further real-timely and synchronously among bacterial flora in the chain, a pair of primers that selectively amplify Vibrio 16S rDNA fragments were designed with their specificities and coverage testified in the analysis of seawater Vibrio community. The specificities and coverage of two primers, VF169 and VR744, were determined theoretically among bacterial 16S rDNAs available in GenBank by using BLAST program and practically by amplifying Vibrio 16S rDNA fragments from seawater DNA. More than 88.3% of sequences in GenBank, which showed identical matches with VR744, belong to Vibrio genus. A total of 33 clones were randomly selected and sequenced. All of the sequences showed their highest similarities to and clustered around those of diverse known Vibrio species. The primers designed are capable of retrieving a wide range of Vibrio 16S rDNA fragments specifically among bacterial flora in seawater, the most important natural environment of seafood cultivation.     

Marine bacterium strain screening and pyrethroid insecticide-degrading efficiency analysis

A pyrethroid insecticide-degrading bacterium, strain HS-24, was isolated from an offshore seawater environment. The strain, which can degrade cypermethrin(CYP) and deltamethrin(DEL), was identified as Methylophaga sp. The optimal culture and degradation conditions for CYP and DEL by strain HS-24 is pH 7 at 28°C. Under optimum culture conditions, strain HS-24 exhibited a broad degradation concentration range of 100, 200, 400, 600, and 800 mg/L for CYP and DEL. The metabolic intermediates were analyzed by NMR, which provided strong evidence that CYP and DEL removal occurred mainly because of a biological process. The toxicity of the degradation products of strain HS-24 was studied simultaneously by measuring the light output of the luminescence bacterium. This demonstrated that the biodegradation ability of strain HS-24 significantly decreased the toxicity of CYP- and DEL-contaminated aquaculture seawater. Finally, the findings of this paper indicate that strain HS-24 is thus revealed as a biological agent for the remediation of marine aquatic environments.     

铁细菌在北极特定海区沉积物中的分布

用平板法对取自北极加拿大海盆和楚科奇海10个沉积物岩芯中184个样品进行铁细菌检出率和含量分析.结果表明,4 ℃培养时铁细菌的检出率为10.33%,含量范围均为未检出～ 8.40×106 个/g(湿样,下同),平均含量为7.76×104 个/g.25 ℃培养时铁细菌的检出率为40.22%,含量范围均为未检出～1.00×108 个/g,平均含量为1.40×106 个/g.研究区沉积物中铁细菌含量高于太平洋深海沉积物中铁细菌含量;与前人在淡水沉积物中所作的调查相比,研究区的铁细菌含量落在前人调查结果范围内,但比细菌簇中铁细菌含量要低,这表明研究区海底沉积物的微生境较适合铁细菌的繁衍.对铁细菌在不同水深、经纬度以及沉积物深度中的分布进行了讨论,结果表明铁细菌检出率随沉积物深度增加而增大,含量变化呈现出浅层少、下层多之势.铁细菌的纬度分布范围较大,并显出有扩大的趋势.铁细菌含量与检出率随水深加大也呈跳跃式变化,水较深时铁细菌有较高的检出率和数量,但离陆距离的增加又会在物质供应上对其生长产生制约.培养温度对铁细菌分布有一定影响,温度提高不仅提高了铁细菌检出率,也增加了铁细菌的含量.这也许与部分铁细菌的最适宜生长温度较高有关,温度升高有利于某些铁细菌的生长发育.     

Distribution of general aerobic heterotrophic bacteria in sediment core taken from the Canadian basin and the Chukchi Sea

The occurrence percentage and abundance of General Aerobic Hetero- trophic Bacteria （GAB） were determined by using the method of MPN for 182 sub- samples from 10 sediment cores taken from the Canadian basin and the Chukchi Sea at two different culturing temperatures. The results showed that the general occurrence percentage of GAB was quite high, average abundanees of GAB at cultured temperatares of 4℃ and 25℃ were 4.46 ×10^7 and 5.47×10^7 cells·g^-1（wt）, respectively. The highest abundance of GAB occurred at 20 -22 cm section in the sediment. GAB abundances changed among the section of sediments, but there is a trend ： the a-bundances at the middle or lower sections were lower than those at upper section. Cultivation at 25℃ could improve the occurrence percentage and abundances of GAB, which suggests that the increasing of temperature may change the living circum-stances of GAB. The differences of GAB among the latitudes areas indicated that occurrence percentage and abundances of GAB in middle latitude areas were higher than those in the higher or lower latitude areas, and were more obvious at 4℃ than those at 25℃. The GAB abundances in sediment under the shallower water seemed to be low- er than those in sediments under the deeper water and this status was more obvious at 25℃ than that at 4℃.     

南极菲尔德斯半岛潮间带沉积物细菌的多样性及分离菌产酶测定

摘要:南极菲尔德斯半岛具有多种不同特征的生态地理微环境,如长城科考站、企鹅岛、生物湾与黄金湾流域、半岛南部、碧玉滩等。这些区域因水文地质、动植物分布、人类活动程度等不同而具有明显的生态地理差异。本研究从这些微环境海岸潮间带采集了7份代表性沉积物样品,采用16S rRNA基因高通量测序方法,对其细菌类群的多样性以及环境理化因子的影响进行了比较分析;同时采用常规可培养鉴定方法,对样品的分离菌株产酶状况进行了初步测定。结果表明：整体上看,所有样品的细菌类群主要分布在45门、104纲、442属当中,表现出比较高的多样性。其中,优势菌门Proteobacteria主要分布在半岛南部、碧玉滩和生物湾潮间带沉积物物中,优势菌门Bacteroidetes主要分布在企鹅岛两侧和黄金湾潮间带沉积物,长城站潮间带沉积物的优势门为Firmicutes。相似生态微环境的潮间带沉积物具有不同的优势菌门和纲;而不同生态微环境潮间带沉积物却具相同的优势菌门和纲。有机氮（TON）、NH4+-N、NO3--N、PO43--P、NO2--N含量与企鹅岛两侧、生物湾、黄金湾潮间带沉积物的菌群多样性具有相关性,其中TON对生物湾样品的影响最大,而有机碳（TOC）对所有样品的影响较小。分离菌株的产酶实验表明：在企鹅岛、生物湾、黄金湾等动物频繁出没的潮间带样品中,蕴藏着一批产淀粉酶、酪蛋白酶、脂肪酶、七叶苷酶、过氧化氢酶能力较强菌株,这为今后进一步的应用开发提供了宝贵的低温产酶菌株来源。 关键词: 菲尔德斯半岛; 潮间带沉积物; 细菌类群多样性; 产酶菌株     

海洋细菌在生态恢复中的作用

针对海洋生态环境日益恶化、需要积极治理、恢复的现象，本文论述了海洋细菌在海洋生态系统中占据的重要地位，并由此论述了海洋细菌在生态恢复过程中的重要作用。海洋细菌作为海洋生态系统的分解者和生产者，对海洋生态系统的稳定起到非常重要的作用；对于已经遭到破坏的海洋生态系统，充分认识和利用海洋细菌是获得稳定、可持续生态系统的根本途径。     

两株琼胶酶高产细菌的筛选和鉴定

从海洋环境中筛选得到两株琼胶酶的高产菌株，通过形态观察和生理生化反应，确定它们属于弧菌属，通过BIOLOG细菌鉴定系统鉴定，并同弧菌属标准菌株分析比较，确定上方宝剑两株菌都是塔式弧菌（Vibrio tubiashii)，这两株菌在碳源利用方面存在差别。     

大亚湾秋季小型底栖生物初步研究

2009年10月对大亚湾海域的小型底栖动物丰度和生物量进行了研究.结果表明,小型底栖动物的平均丰度为(593±265)ind·(10cm)?2,呈湾内向湾外递增趋势;平均生物量和生产量分别为(568.39±294.49)μg·(10cm)?2(干重)和(5115.53±2811.14)μg·(10cm)?2(干重).在分选的17个主要类群中,海洋线虫在丰度上占绝对优势达92.24%,其次是多毛类和桡足类,分别为2.05%和1.98%.在生物量上,海洋线虫(38.47%)和多毛类(29.97%)为绝对优势类群,其次是寡毛类(13.82%)和介形类(9.15%).在垂直分布上,占总量约53.02%的小型底栖动物和51.44%的海洋线虫分布于0—2cm表层.统计分析表明,本海域的小型底栖动物生物量与含水率、中值粒径和黏土+粉砂含量呈显著负相关(P<0.05),多毛类的生物量与小型底栖动物表现的显著相关性相同,桡足类的生物量与中值粒径值呈显著负相关(P<0.05),与沉积物的黏土+粉砂含量呈极显著负相关(P<0.01),而线虫生物量与环境因子无显著相关关系.由海洋线虫与桡足类的数量比(用 N/C 表示)与本海域环境状况的分析表明,该数值可以在一定尺度上为海洋环境检测提供参考.     

鳗弧菌(Vibrio anguillarum)M3菌株生长条件及其对蛋白酶产量的影响

采用体外测定细菌浓度、胞外产物(ECP)蛋白含量和蛋白酶活力的方法，进行了鳗弧菌M3菌株在2216E培养基中的培养条件研究。结果表明，该菌株用固体培养基培养至24h左右，可得到较高的菌体浓度、ECP蛋白含量和蛋白酶活力。采用响应面分析方法设计实验，用SAS统计软件分析数据，得到NaCl浓度、pH值和温度对菌体生长及蛋白酶产量影响的回归模型。在2216E培养基的基础上，添加不同氮源、碳源物质以及不同浓度蛋白胨进行生长研究。结果表明，胰大豆蛋白胨能促进菌体生长及ECP蛋白分泌；NH4Cl与酪蛋白水解物可抑制蛋白酶的产生；牙鲆肌肉匀浆对菌体、ECP蛋白产量和蛋白酶产生有不同程度的促进作用；培养基中蛋白胨浓度为4％时菌体量与ECP蛋白含量达最高值，在蛋白胨浓度为2％时蛋白酶分泌量已稳定；1％的葡萄糖、蔗糖、甘油均能显著地提高菌体及ECP蛋白产量，却抑制了蛋白酶的产生。