大肠杆菌O157:H7的活的非可培养状态

将大肠杆菌 O157:H7培养于低温贫养条件下 ,以涂布平板法 (PC)和最大近似值法(MPN)检测可培养细菌数 ,95～ 115d后表明可培养菌数下降为零。吖啶橙荧光显微镜直接计数法(AODC)检测细菌总数 ,表明细菌总数始终变化不大 ,而活菌直接镜检计数 (DVC)检测到的活菌数保持在 10 6个 / m L。实验证明了大肠杆菌 O157:H7在一定的条件下可进入活的非可培养状态(VBNC)。     

环丙沙星作DNA抑制剂应用于活菌直接计数

参照Isabel方法用不同浓度的环丙沙星(Ciprofl oxacin),处理两株革兰氏阳性菌;北京棒状杆菌(Corynebacterium pakinense);枯草芽 孢杆菌(Bacillus subtilis)和两株革兰氏阴性菌：霍乱弧菌(Vibrio cholerae)和大肠杆 菌 O157：H7(Escherichia coli O157:H7)。结果发现经乳酸环丙沙星处理后的细菌形 态均变长变大,计数结果表明环丙沙星处理的细菌的活菌直接计数与用萘啶酮酸(Nalidixic acid)处理的活菌直接计数法(DVC)结果相似,高于平板菌落计数结果,而且对部分革兰氏 阳性菌也可应用此法进行活菌直接计数。     

渤海湾近岸海域的细菌数量分析

于2004年2月和5月应用AODC、DVC、HPC和TC法测定了渤海湾近岸海水的细菌丰度。其中AODC值变化范围为(5.3~192.0)×108/L个,DVC值为(3.8~122.0)×108/L个,HPC值为(2.00×104~4.95×107)/L,TC值为(0~>11 000).L-1个。海河口附近细菌丰度高于其它海域,说明该海域细菌分布受海河排污影响很大。同一海水样品的各细菌计数结果之间存在显著的相关性(r>0.935,P<0.01)。研究海域细菌丰度高于国内其它海区,海水受到的污染依然严重。     

副溶血弧菌菌株1211U的活的非可培养状态

将副溶血弧菌（Ｖｉｂｒｉｏ ｐａｒａｈａｅｍｏｌｙｔｉｃｕｓ）菌株１２１１培养于低温贫养条件下,以平板菌落计数法（ＰＣ）和最大近似数法（ＭＰＮ）检测可培养细菌数,５０￣６０ｄ后表明可培养数下降为零。吖啶橙荧光显微镜直接计数法（ＡＯＤＣ）检测细菌总数,表明细菌总数始终变化不大,而活菌直接计数法（ＤＶＣ）检测到的活菌数保持在１０＾５个／ｍｌ。本实验证明了副溶血弧菌１２１１Ｕ株在一定的条件下或进入活的非     

黄海冷水团可培养细菌的多样性研究

本文研究了2008年7月黄海冷水团海域可培养细菌的多样性。AODC和DVC计数总菌数和活菌数分别为(1.3~4.8)×105/mL和(0.6~1.6)×105/mL;2216E平板以菌落计数法可培养细菌浓度为(0.56~2.2)×103cfu/mL。从分离到的475株中选取163株进行16S rDNA扩增,并用HhaⅠ酶进行ARDRA(扩增性rDNA限制酶切片段分析)多态性分析,取114株不同带型测序。结果显示,冷水团细菌归为4个细菌类群:变形菌门(Proteobacteria)、放线菌门(Acti-nobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes),共24个属,其中包括α-,β-,γ-变形菌纲;非冷水团细菌也归为这4个门类,共15个属,但未分离到β-变形菌纲。γ-变形菌纲在冷水团和非冷水团区域的不同深度都是优势菌群,不同的是冷水团区域α-变形菌纲比例较高(26.8%),而非冷水团区域α-变形菌纲比例相对较低(15.6%)。这表明黄海冷水团和非冷水团区域细菌多样性都很丰富,但其群落组成和优势菌群有所不同。此外,16S rDNA测序结果表明,6株细菌可能为海洋细菌新种。     

基于高通量测序方法分析荣成月湖潮间带趋磁细菌多样性

荣成月湖是一个典型天然潟湖,潮间带沉积物中存在多种形态趋磁细菌。通过Roche454高通量测序平台,对沉积物样品(B_S)和磁收样品(B_M)进行16SrRNA基因高通量测序,认识荣成潮间带沉积物的细菌群落结构,并了解趋磁细菌多样性及系统进化地位。研究结果表明沉积物样品中主要的细菌类群为δ-变形菌纲,占总细菌数的26.4%,其次是γ-变形菌纲和α-变形菌纲;而磁收样品中细菌多样性和种类明显降低,以α-变形菌纲占绝对优势,相对比例达72.6%。在磁收样品和沉积物样品分别发现了1 612条和186条reads与趋磁细菌相关,分别占细菌总数的5.76%和0.85%,磁收样品中趋磁细菌数是沉积物样品的6.8倍。对两个样品中获得的趋磁细菌序列进行系统进化分析,发现这些序列多数属于变形菌门的α-变形菌纲,以趋磁球菌占绝对优势,少数属于δ-变形菌纲,与多细胞趋磁原核生物亲缘关系最近。海洋趋磁螺菌属仅在B_M样品中检出,趋磁弧菌属在B_M样品优势度高于B_S样品,而多细胞趋磁原核生物和趋磁螺菌属在B_S样品中优势度更高。通过分析样品间的差异OTUs,认为荣成潮间带沉积物中可能存在大量未知的趋磁细菌新类群。研究结果为下一步培养和开发趋磁细菌这一功能菌群,发现趋磁细菌新类群及趋磁细菌生态功能提供了基础资料。     

中国对虾血淋巴抗菌活力的测定

采用溶藻胶弧菌（Ｖｉｂｒｉｏ ａｌｇｉｎｏｌｙｔｉｃｕｓ）、副溶血弧菌（Ｖ． ｐａｒａｈａｅｍ ｏｌｙｔｉｃｕｓ）、哈维氏弧菌（Ｖ．ｈａｒｖｅｙｉ）、嗜水气单胞菌（Ａｅｒｏｍ ｏｎａｓｈｙｄｒｏｐｈｉｌａ）对中国对虾（Ｐｅｎａｅｕｓ ｃｈｉｎｅｎｓｉｓ）血浆样品、血细胞溶解物（ＨＬＳ）样品的抗菌活力进行了测定。表明中国对虾血浆样品对四种细菌均表现出一定的抗菌活力。其中,三种弧菌活细胞数在孵育１ｈ 时呈现最低;嗜水气单胞菌活细胞数在孵育４ｈ 时最少。在三种弧菌中,血浆样品对哈维氏弧菌表现出的抗菌活力最强,而对副溶血弧菌表现出的抗菌活力最弱。血细胞溶解物样品对三种弧菌几乎没有表现出抗菌活力;嗜水气单胞菌在与血细胞溶解物样品孵育２ｈ 时,其活细胞数呈现最低,说明血细胞溶解物样品对该菌表现出了一定的抗菌活力。由此显示,抗菌活力在中国对虾血浆样品中表现得较强,而在血细胞溶解物样品中表现得较弱,血淋巴抗菌因子可以在短时间内对细菌表现出其最强的活力,但对不同种细菌其抗菌作用强弱不同。     

大洋锰结核区细菌的初步研究

本文叙述了1985年12月—1986年4月在北太平洋(北纬7°—13°,西经178°—165°)锰结核调查区所进行的细菌研究工作。采用落射荧光显微镜直接计数法,分别对19个表层沉积样品和12个锰结核样品进行了总菌计数。结果表明,其范围在10~4—10~7个/克;细菌种群密度结核样普遍高于泥样。对现场分离得到的异养细菌及锰氧化细菌进行了初步鉴定分类及部分生理生化特性试验,发现锰氧化细菌在沉积物细菌区系中占相当比例(12％),其中革兰氏阴性假单胞菌属Pseudomonas占50％。     

乳山湾外海溶氧低值区细菌多样性初步研究

采用化学裂解法从乳山湾外海溶氧低值区不同溶解氧质量浓度(3.7～7.0mg.L-1)的6个站位海水样品中提取了环境DNA样品。以试剂盒纯化后的DNA样品为模板扩增其16SrRNA基因V3区,通过变性梯度凝胶电泳、分子文库构建及DNA测序对溶氧低值区海水中的细菌群落结构进行研究。结果表明,乳山湾外海溶氧低值区不同溶解氧质量浓度的6个站位底层海水样品中的细菌群落结构是相似的,它们均由隶属于Alteromonas(交替单胞菌属)、Salegentibacter(需盐杆菌属)等10个属的18种细菌组成。系统发育分析发现这些细菌分别属于α变形菌纲(2种)、γ变形菌纲(12种)和黄杆菌纲(3种)三个大类。在乳山湾外海溶氧低值区的海水样品中细菌多样性最高的类群是γ变形菌纲。     

TiCl4水解制备金红石型TiO2纳米粉

实验以TiCl4为主要原料，采用直接水解法制备出了TiO2样品。经差热-热重(DTA-TG)、X-射线衍射(XRD)、透射电子显微镜（TEM）和比表面(BET)分析，得出样品TiO2的晶型为金红石型，其粒子近似呈球形，各个晶面所对应的粒径基本一致，分布在7.0～12.0nm之间，平均粒径为10.5nm，对应的比表面积和孔容积分别为166m2/g和0.12mL/g。     

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.     

Heterotrophic bacteria and their relationship with plankton in a New Zealand freshwater lake

The viable heterotrophic bacteria of Lake Grasmere in inland Canterbury (43° 05 S, 171° 45 E) were studied. Numbers of bacteria in water above a spring were positively correlated with rainfall in the 7 days before sampling, but bacteria in open water and from over the macrophyte Elodea canadensis were not. There was some increase in the bacterial population in the water over E. canadensis in the autumn to winter months. Numbers of bacteria in the open water ranged from 55 to 1020 cells per millilitre, and were outnumbered by algae by a factor of between 4 and 34. The bacterial population tended to increase after blooms of zooplankton, but there was little or no response to changes in phytoplankton populations. Benthic bacteria rarely increased in numbers after the deposition of organic matter.     

霍乱弧菌的越冬方式—活的非可培养状态及其检测方法

霍乱弧菌是河口与海洋环境中自然微生物区系的成员。霍乱弧菌在诸如低温、低盐和低营养物浓度的不良环境条件下,可以变成非可培养状态。利用活菌直接镜检技术和免疫萤光抗体染色技术证明,霍乱弧菌是活的非可培养状态。实验还证明霍乱弧菌的越冬是通过活的非可培养状态实现的。     

苗期中国对虾幼体异养细菌区系及其变化与病害发生的关系

研究了2个育苗场不同发育期的中国对虾(Penaeus chinensis)幼体的异养菌和弧菌种群变化的动态过程。以典型特征法、BIOLOGGN法和数值分类法对菌株进行鉴定。结果表明，对虾幼体样品中所分离的细菌，大多是革兰氏阴性杆菌。鉴定为假单胞菌属(Pseudomonas)、气单胞菌属(Aeromonas)和弧菌属(Vibrio)。弧菌属主要为溶藻弧菌(Vibrioal-ginozyticus)和哈维氏弧菌(Vibrio harveyi)。在对虾幼体发育早期，假单胞菌(Pseudomonas)和气单胞菌(Aeromonas)占优势，随着对虾幼体的发育，弧菌(Vibrio)渐成为优势，其中溶藻弧菌(Vibrio alginolyticus)和哈维氏弧菌(Vibrio harveyi)优势明显。二者的消长与苗期病害的发生相关，溶藻弧菌总是在虾幼体健康时出现或成为优势，而哈维氏弧菌成为优势时，苗期病害容易发生。     

Bacteria on petroleum globules in the Philippine sea in January, 1973

About 10⁶ of total bacteria (direct microscopic count) or heterotrophic bacteria (colony count with Medium 2216E) and about 10⁴ of petroleumlytic bacteria (oil-oxidizing bacteria) were enumerated per square centimeter of the surface of petroleum globules suspended in the topmost 10 meters of water from a station (22°00.2N, 125°51.9E) in the western North Pacific central water, where about 10⁷ of total bacteria, about 10³ of heterotrophic bacteria and about 10 of petroleumlytic bacteria were enumerated per liter of seawater.     

长江口及邻近海域表层海水细菌多样性及群落结构

从长江口及邻近海域10 个站点采集表层海水, 通过DAPI 染色计数和克隆文库构建的方法, 对细菌的多样性及群落结构进行初步研究。DAPI 染色计数结果显示长江口及邻近海域表层海水中 细菌丰度总体较高, 生物量变化较大, 从1.16×10⁵(A04 站点)到 1.48×10⁶ cells/mL (B03 站点); 总体 趋势从长江口向外海增加。细菌克隆文库的系统发育分析表明: 细菌序列以变形菌门为主(占总文库 的60.61%), 其中α-变形菌纲是绝对优势类群(50.62%); 其次是拟杆菌门(15.18%)、放线菌门(14.79%) 和蓝细菌门(4.61%), 还有少量厚壁菌门、绿弯菌门、酸杆菌门、浮霉菌门、纤维杆菌门、疣微菌门 等; 另外柔膜菌门和SAR 类群也有发现。文库多样性分析结果显示长江口及邻近海域部分站点表层 海水中细菌多样性显著, 与研究区海域水产养殖活动有关, 并受到区域洋流和海洋化学环境(如低氧 带)的影响。     

Short-term bacterial reactivity of nitrogen-enriched seawater of a eutrophic lagoon

We carried out, in September, a 24 h experiment to determine the potential reactivity of bacterial communities found in the sea water in Arcachon Basin (France). To create eutrophic systems, batches of seawater were enriched with 30 mgl^?1 of either ammonium chloride or amino acids. Samples were taken every 3 h. Quantitative measurements were made of direct counts (AODC), biomass and ATP content. The heterotrophic bacterial communities were defined in terms of their catabolic potentialities and ‘specific’ diversity. Bacterial heterotrophic activity was established by measuring heterotrophic uptake and the mineralization percentage of labelled glucose or amino acids. From these data, ratios of AMP/ATP and P/B were calculated where P is the uptake of labelled substrate and B is the biomass.After the nitrogen enrichment an increase of the respiration was seen at first. After 6 h, a biomass peak appeared associated with a continuous increase in the number of cells. Bacterial growth was concurrent with a qualitative modification of the community: UAI increased, and diversity dropped. A less diversified community resulted suggesting an immature ecological state (zymogenous bacterial). Low values of respiration percentage (20%) characterized high growth yields.     

北黄海微表层细菌丰度与可培养细菌群落结构分析

为分析海洋微表层这一特殊生境中的细菌类群,于2010—2011年4个航次对北黄海微表层和次表层海水中的总菌丰度、可培养细菌丰度和群落结构进行了分析。采用流式细胞仪测定总菌丰度,平板计数法测定可培养细菌丰度,PCR-16SrDNA分析可培养细菌的群落结构。结果表明,调查海域微表层海水总菌均值为1.76×106 cell/mL,次表层海水总菌均值为1.07×106 cell/mL。可培养细菌丰度范围是1.00×102~1.70×106 CFU/mL,微表层和次表层可培养细菌所占总菌数量的百分比分别为13.05%和0.45%。微表层对总菌的富集因子(EF)均值为2.02,可培养细菌的EF均值为74.16。PCR-16SrDNA序列分析结果表明,该海域可培养细菌分属变形菌门(Proteobacteria)(94.34%)、厚壁菌门(Firmicutes)(1.89%)、拟杆菌门(Bacteroidetes)(1.89%)和放线菌门(Actinobacteria)(1.89%)4个类群。本研究初步发现,微表层对细菌具有较强的聚集作用,尤其对可培养细菌聚集作用更为明显。微表层中可培养细菌的群落结构与次表层有所不同,其种类丰富,来源和功能多样。由此可见,微表层独特的生境成就了其独特的微生物类群,其生态功能有待于进一步研究。     

Limno-tolerant bacteria govern nitrate concentration in Mandovi Estuary,India

The spatial and temporal abundances of limno-tolerant and halo-tolerant bacteria were investigated in the tide-dominated Mandovi estuary along the west coast of India. These investigations were carried out in relation to various environmental parameters on a monthly basis at three fixed stations for a year. On an annual basis, the estuary showed an average salinity of 28.2, 17.4, and 12.6 at the mouth, midstream and upstream region. Halo-tolerant retrievable count (HTRC) and limno-tolerant retrievable count (LTRC) of bacteria were in the order of 10⁶ L⁻¹. Among the environmental parameters, a strong negative relationship between salinity and nitrate (r = −0.806; p < 0.001) suggested that 64% of the variation could be due to fresh water influence in the estuary. The limno-tolerant retrievable count (LTRC) brought about 23% variations in nitrate concentration. This influence was maximum during the monsoon (r = 0.522; p < 0.05) especially in the surface waters (r = 0.624; p < 0.001) suggesting nitrate reduction by LTRC. Measurements of nitrate reducing activity (NRA) in whole-water samples along the salinity gradient in the estuary also revealed higher reduction rates at lower salinity upstream. This was further confirmed by culture experiments where the limno-tolerant bacteria showed higher NRA than halo-tolerant forms. It is therefore suggested that LTRC is more actively involved in the variation of nitrate that enters the Mandovi estuary particularly during the monsoon.