A culture-dependent survey of thermophilic bacteria from hot springs in Xiamen area in China

Microbes are believed to play important roles in ecosystem function in many environments. The hot springs of Xiamen Island are close to the Xiamen Sea, and may have some characteristics different from those of inland hot springs. Microbes living in the hot springs of Xiamen may have new characteristics. However, little is known about microbial communities of hot springs close to the Xiamen Sea. A cuhure-dependent survey of microbial population in the Xiamen hot springs was pcrformed by using an approach combining total cellular protein profile identification and 16S rRNA gene sequencing. A total of 328 isolates of bacteria were obtained from liquid and sediment samples from the Xiamen hot springs, including neutrophilie thermophilic bacteria and moderately thermophilic acidophiles. Neutrophilic thermophilic bacteria, which grow at a temperature range of 55-90℃ including Rhodothermus marinus (Strain 1) , Thermus thermophilus (Strain 2), Thermus thiopara (Strain 3) , Geobacillus stearothermophilus(Strain 4) , Geobacillus thermoleovorans (Strain 5) , and Pseudomonas pseudoal-caligenes (Strain 6), were recovered by 2216E plates. Moderately thermophilic acidophiles, which can grow at temperatures above 50℃ and a pH range of 1. 8-3.5 such as Alicyclobacillus acidoterrestris (Strain 8) , Sul-fobacillus acidophilus (Strain 9), and Sulfobacillus thermosulfidooxidans (Strain 10), were isolated on selective solid medium containing sulfur and Fe2+. Among these strains, Rhodothermus marinus, Thermus thermophilus and Geobacillus stearothermophilus are not only thermophilcs, but also halophiles. One bacterium strain (Strain 6) shared 99% nucleotide sequence homology with Pseudomonas pseudoalcaligenes on the 16S rRNA gene se-quence, but was quite different from Pseudomonas pseudoalcaligenes in biological characteristics, suggesting that it may represent a novel thermophilic species. Results indicated that various species of neutrophilic thermophiles and moderately thermophilic acidophiles were widely distributed in the Xiamen hot springs and that Rhodother-mus marinus and Thermus thermophilus dominated the cultivable microbial community.     

Complete genome sequence and proteomic analysis of a thermophilic bacteriophage BV1

Viruses of thermophiles are of great interest due to their roles in gene transfer, global geochemical cycle and evolution of life on earth. However, the thermophilic bacteriophages have not been studied extensively. In this investigation, a typical bacteriophage BV1 was obtained from a thermophilic bacterium Geobacillus sp. 6k512, which was isolated from an inshore hot spring in Xiamen of China. The BV1 contained a double-stranded linear DNA of 35 055 bp, which encodes 54 open reading frames (ORFs). Interestingly, eight of the 54 BV1 ORFs shared sequence similarities to genes from human disease-relevant bacteria. Seven proteins of the purified BV1 virions were identified by proteomic analysis. Determination of BV1 functional genomics would facilitate the better understanding of the mechanism for virus-thermophile interaction.     

印尼热泉中产嗜热碱性蛋白酶菌株筛选及酶学性质研究

采用MSM寡营养培养基从印度尼西亚Pantai cermin,Kalianda和Banyuwedang三个地区的热泉水样、泥样以及沉积物样品中分离获得细菌菌株,通过检测蛋白酶产生透明圈和福林酚蛋白酶活性测定相结合的方法,从中筛选出1株产嗜热蛋白酶的菌株PBI,该菌株经初步鉴定为短杆菌属(Brevibacillus),并对其酶学性质进行了研究。结果表明,菌株PBI产蛋白酶的最适酶活温度为60℃,最适pH值为8.0～9.0,具有较好的热稳定性和pH稳定性,60℃时酶的半衰期为30min,70℃条件下20min仍保持46.1%的酶活,该酶在pH值为5.0～9.0范围的缓冲液中酶活相对稳定。其产嗜热蛋白酶的酶活力最高可达到60.53U/mL,在100℃条件下仍能保留26.37%的相对酶活。Fe2+,Fe3+,Cu2+和Zn2+对嗜热蛋白酶活性具有明显的抑制作用。该嗜热蛋白酶对EDTA敏感,苯甲基磺酰氟(PMSF)、亮抑酶肽(Leupeptin)、苄眯(Benzamidine)和胃蛋白酶抑制剂(Pepstain A)对该嗜热蛋白酶都有一定的抑制作用,说明其酶活性受到丝氨酸、半胱氨酸残基的影响。结果表明,该菌株是1株具有进一步改造利用价值的产蛋白酶菌株。     

海洋微生物对甲胺磷农药的降解作用

作者从沿岸海域分离了38株有机磷农药的耐药菌,用分批培养法进行富集培养,得到有机磷农药的降解菌,并着重研究了其中两株菌对甲胺磷农药的降解情况,其结果表明,在10d内,降解菌株1对甲胺磷的降解率为59．1％,降解菌株2的降解率为53．3％,在各自的安磷培养液中,降解菌株1的数量大于降解菌株2;在甲胺降解过程中,其毒性逐渐减弱,降解菌株1所引起的毒性下降幅度大于降解菌株2。     

印度尼西亚卡利安达岛近岸热泉微生物和氢酶基因的多样性

研究对2011年7月采自印度尼西亚卡利安达岛近岸热泉的样品,进行了细菌和古菌16S rRNA基因和细菌氢酶基因克隆文库的构建、序列测定和多样性分析。结果表明:热泉系统的细菌和古菌由16个门组成,Proteobacteria为热液流体和沉积物的优势类群,丰度分别为59.5%和73.3%,Cyanobacteria为菌苔的优势类群(丰度,56.5%);具有氨氧化作用的Crenarchaeota为古菌的优势类群。热泉系统中,70%以上的种类与海洋细菌的亲缘关系最近,说明该系统具有海洋特性;约20%的种类与陆地热泉中嗜热或中等嗜热细菌的亲缘关系最近,这些种类可能来源于高温地层内部,并参与了铁氧化还原、氢氧化、硫氧化和硝酸还原等过程。热泉系统中,NAD(P)-关联的双向NiFe-氢酶基因和FeFe-氢酶基因的组成分布也受到温度和盐度分布变化的影响。研究为揭示浅海热液系统提供理论参考。     

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.     

厦门海区潮间带海洋动植物共附生微生物的抗菌活性

从厦门海区潮间带石纯、浒苔、江蓠、海兔、海葵及鲨鱼肠道分离得２８９株共附生微生物,以８种敏感菌为指示菌,从中获得拮抗活性的幼苗２８株,放线菌１７株,真菌８株。拮抗细菌以假单胞菌（３９％）、弧菌（２５％）和芽直菌（１４％）为主,主要为抗革兰氏阳性细菌,抗真菌的菌株比例较低。拮抗放线菌分属于链霉菌和小单孢菌,其抗革兰氏阳性菌、阴性菌和真菌的菌株比例分别为６５％、４７％和２３％。拮抗真菌分青霉属和芽枝霉     

来自大黄鱼(Pseudosciaena crocea)肠道的弧菌拮抗菌的筛选与鉴定

采用点种法并以副溶血弧菌(Vp1.2164)和溶藻弧菌(Va-Y)为弧菌指示菌,对从健康的大黄鱼肠道内分离的115株细菌进行体外拮抗试验。并对筛选出的拮抗菌进行抗菌谱测定,最后利用VITEK-32细菌快速鉴定系统和16SrRNA基因序列对拮抗菌进行鉴定。结果表明,从中筛选到3株有较强拮抗作用的菌株,编号分别为X1402-1、X1108和X3914,且菌株X1402-1的抗菌作用最强,抗菌谱最广,21株病原指示菌中,对16株均能产生较强的抑制作用,其中对Vp-1.2164的抑菌直径最大,达到28.5mm,对Va-Y的抑菌直径为8.7mm;其次为菌株X1108和X3914,对两种指示弧菌的抑菌直径分别为:12.0mm、9.0mm和0mm、7.2mm。经VITEK-32细菌快速鉴定系统鉴定,3株拮抗菌分别与铜绿假单胞菌,恶臭假单胞菌,地衣芽孢杆菌的相似性均为99%。通过16SrRNA基因序列分析表明,3株拮抗菌分别与类产碱假单胞菌Pseudomonas pseudoalcaligenes(EU419918)、恶臭假单胞菌Pseudomon asputida(FJ440105)和地衣芽孢杆菌Bacillus licheniformis(DQ988136)的16SrRNA基因同源性最高,分别为100.0%、97.5%、99.0%。综合分析试验结果,X1402-1、X1108和X3914分别被确定为类产碱假单胞菌、恶臭假单胞菌和地衣芽孢杆菌。     

基于低压变频电场技术对带鱼(Trichiurus lepturus)保鲜过程中微生物群落影响分析

带鱼(Trichiurus lepturus)在低温条件运输和贮藏过程中,易受微生物和内源性蛋白酶的影响而产生胺类和醛类物质导致腐败。为了研究低压变频电场对微冻带鱼在贮藏过程中的微生物群落特征,采用Miseq基因组测序技术,对贮藏至40d的舟山带鱼的微生物群落进行碱基信息测序。将产生的碱基序列通过OTU聚类、多样性分析、相关性分析等手段分析了影响带鱼保鲜效果的微生物群落组成和发育信息。在此基础上,研究了不同强度电场(0,2,2.5,3 kV/m)处理对带鱼微生物群落组成的影响。根据菌属相对丰度结果,推测希瓦氏菌属(Shewanella)、假单胞菌属(Pseudomonas)、紫色杆菌属(Janthinobacterium)是带鱼微生物腐败主要致腐菌。结果表明,40 d后,经3 kV/m处理的样品其OTU6嗜冷菌属和OTU13假单胞菌属c的相对丰度均小于1%,而且OTU7、OTU14、OTU8、OTU9等菌属未被OTU聚类。结果表明,经3kV/m电场强度的低压静电场处理的抑菌保鲜效果最好,表现为在贮藏过程中杂菌种类最少,样本微生物群落物种丰度和多样性均最低,腐败程度最低。     

南极菲尔德斯半岛可培养土壤微生物多样性及理化性质鉴定

为了探索南极可培养土壤微生物的多样性,本研究对中国第31次南极科学考察采集自南极菲尔德斯半岛的5份土壤样品进行了细菌、真菌的分离培养。选择形态差异较大的细菌和真菌进行了16S rDNA和ITS鉴定及系统发育分析。最终共鉴定得到20个属的33株细菌和6个属的8株真菌,其中8株细菌,10株真菌序列相似性较低,可能是新种。该结果表明南极菲尔德斯半岛地区具有丰富的微生物多样性,其中假单胞菌属的细菌居多。对分离得到的细菌和真菌分别进行理化性质和胞外酶活性鉴定,实验结果显示,分离获得的细菌和真菌绝大多数可产生水解酶类并同化多种碳源。初步认定这些微生物在参与南极物质代谢、适应南极极端环境方面发挥作用。本研究丰富了对南极菲尔德斯半岛可培养土壤微生物多样性的认识,并筛选获得了一些产低温酶特性的菌株,这为极地微生物资源的利用研究奠定了基础。     

Contribution of thermogenic organic matter to the formation of biogenic gas hydrate: Evidence from geochemical and microbial characteristics of hydrate-containing sediments in the Taixinan Basin,South China Sea

High-saturation (40–100%), microbial gas hydrates have been acquired by expedition GMGS2 from the Taixinan Basin. In this study, geochemical and microbial features of hydrate-containing sediments from the drilling cores (GMGS2-09 and GMGS2-16) were characterized to explore their relationships with gas hydrate formation. Results showed that the average TOC content of GMGS2-09 and GMGS2-16 were 0.45% and 0.63%, respectively. They could meet the threshold for in situ gas hydrate formation, but were not available for the formation of high-saturation gas hydrates. The dominant members of Bacteria at the class taxonomic level were Alphaproteobacteria, Bacilli, Bacteroidia, Epsilonproteobacteria and Gammaproteobacteria, and those in Archaea were Marine_Benthic_Group_B (MBGB), Miscellaneous_Crenarchaeotic_Group (MCG), Group C3, Methanomicrobia and Methanobacteria. Indicators of microbes associated with thermogenic organic matter were measured. These include: (1) most of the dominant microbes had been found dominant in other gas hydrates bearing sediments, mud volcanos as well as oil/coal deposits; (2) hydrogenotrophic methanogens and an oilfield-origin thermophilic, methylotrophic methanogen were found dominant the methanogen community; (3) hydrocarbon-assimilating bacteria and other hyperthermophiles were frequently detected. Therefore, thermogenic signatures were inferred existed in the sediments. This deduction is consistent with the interpretation from the seismic reflection profiles. Owing to the inconsistency between low TOC content and gas hydrates with high saturation, secondary microbial methane generated from the bioconversion of thermogenic organic matters (oil or coal) was speculated to serve as enhanced gas flux for the formation of high-saturation gas hydrates. A preliminary formation model of high-saturation biogenic gas hydrates was proposed, in which diagenesis processes, tectonic movements and microbial activities were all emphasized regarding to their contribution to gas hydrates formation. In short, this research helps explain how microbial act and what kind of organic matter they use in forming biogenic gas hydrates with high saturations.     

泥螺消化道内的细菌学分析

泥螺(Ｂｕｌｌａｃｔａｅｘａｒａｔａ)隶属于软体动物门,腹足纲 ,后鳃亚纲,头盾目 ,阿地螺科。其味道鲜美,具有较高的营养价值和经济价值 ,是浙江沿海重要的滩涂养殖品种,至２０００年全省养殖面积已达约１１３００ｈａ。泥螺的生长是以不断吞食周围滩涂中的涂泥和底栖藻类等为营养基础 ,已知滩涂沉积物及动植物碎屑中所含有的微生物从事着大量的转化和分解工作［４］,它们在泥螺体内又会怎样呢 ?为探索泥螺的消化生理功能及与养殖、病害和防治的关系,本文从泥螺消化道内的细菌菌群组成与功能研究入手作初步分析。关于贝类体内的细菌学调查…     

季风环流影响下的南海海洋细菌多样性特征初探

季风驱动下的南海海洋动力过程有着季节性的多尺度变化特征, 显著影响南海海洋生态系统的演化进程。海洋细菌作为海洋生态系统中物质循环和能量流动的重要组成部分, 对环境变化和海洋多尺度动力过程有着积极的响应。将微生物生态过程与海洋动力过程相结合, 研究微生物的群落结构、动态变化及其与海洋环境过程的耦合, 是目前国际海洋科学多学科交叉研究的热点之一。文章以国家自然科学基金重点项目“季风环流影响下的南海海洋细菌多样性特征及其生物海洋学意义”的主要研究成果为基础, 从南海北部上升流、海洋锋面、中尺度涡旋、次中尺度过程及其对海洋细菌多样性和生态系统可能的影响等几个方面, 探讨当前的研究进展和初步研究成果。     

青岛即墨温泉盆地热水沉积指示物及其标识性意义

经野外取样和XRD测试技术,在即墨温泉盆地发现硅质沉淀物、盐华、石膏、自然硫等温泉热水沉积指示物.根据热水沉积指示物的空间分布和含量的变化,阐述温泉热水沉积指示物的标识性意义：（1） 热水沉积指示物是划分第四纪沉积物层系的依据;（2） 是温泉地下热储温度的指示剂;（3） 依据热水沉积指示物百分含量的空间变化,可圈定温泉地热中心,指导地热资源开发;（4） 是寻找地下盲温泉和古地热水活动的标志;（5） 温泉热水沉积模式对现代海底热水沉积作用、成矿作用环境的模拟研究具有重要参考作用.     

Analysis of cultivable aerobic bacterial community composition and screening for facultative sulfate-reducing bacteria in marine corrosive steel

Anaerobic, aerobic, and facultative bacteria are all present in corrosive environments. However, as previous studies to address corrosion in the marine environment have largely focused on anaerobic bacteria, limited attention has been paid to the composition and function of aerobic and facultative bacteria in this process. For analysis in this study, ten samples were collected from rust layers on steel plates that had been immersed in seawater for diff erent periods (i.e., six months and eight years) at Sanya and Xiamen, China. The cultivable aerobic bacterial community structure as well as the number of sulfate-reducing bacteria (SRB) were analyzed in both cases, while the proportion of facultative SRB among the isolated aerobic bacteria in each sample was also evaluated using a novel approach. Bacterial abundance results show that the proportions are related to sea location and immersion time;abundances of culturable aerobic bacteria (CAB) and SRB from Sanya were greater in most corrosion samples than those from Xiamen, and abundances of both bacterial groups were greater in samples immersed for six months than for eight years. A total of 213 isolates were obtained from all samples in terms of CAB community composition, and a phylogenetic analysis revealed that the taxa comprised four phyla and 31 genera. Bacterial species composition is related to marine location;the results show that Firmicutes and Proteobacteria were the dominant phyla, accounting for 98.13% of the total, while Bacillus and Vibrio were the dominant genera, accounting for 53.06% of the total. An additional sixfacultative SRB strains were also screened from the isolates obtained and were found to encompass the genus Vibrio (four strains), Staphylococcus (one strain), and Photobacterium (one strain). It is noteworthy that mentions of Photobacterium species have so far been absent from the literature, both in terms of its membership of the SRB group and its relationship to corrosion.     

海洋光合细菌的分离鉴定及其生长条件的研究

用红色硫细菌培养基从青岛沿海潮间带海泥中分离出8株光合细菌,经鉴定菌株HD_1属最小着色菌(Chromatium minutissimum),HD_2、HD_4为细着色菌(Chromatium gracile);菌株SL_1、SL_1x、PL_1和PL_2为嗜酸红假单胞菌(Rhodopseudomonas acidophila),菌株HCx为沼泽红假单胞菌(Rhodopseudomonaspalustris)。分离菌株均具运动性,不需生长因子。菌株HD_1、HD_2和HD_4在厌氧光照条件下利用硫化物,且在细胞内贮存硫粒。菌株HD_1、HD_2的最适盐度为4.0％,最适pH为7.0,8.0。当培养基中硫化物含量为0.01—0.04％时,菌株HD_1、HD_4生长良好。菌株PL_2在初始pH为6时生长最好。酵母膏对菌株HCx具有明显的促生长作用。     

中国对虾成虾肠道微生物区系

对野生健康中国对虾成虾肠道微生物区系进行了研究.从其肠道中分离出47株菌,它们分别属于弧菌属、发光杆菌属、不动杆菌属、假单胞菌属、黄杆菌属、气单胞菌属、屈挠杆菌属和色杆菌属8个属.其中弧菌属和发光杆菌属在整个肠道中为优势菌属,不动杆菌属和假单胞菌属为次优势菌属,黄杆菌属、气单胞菌属、屈挠杆菌属和色杆菌属为非优势菌属.在对虾的整个肠道中,前肠和中肠的优势菌属为弧菌属,而后肠的优势菌属为发光杆菌属.在弧菌属中溶藻胶弧菌、漂浮弧菌和坎贝氏弧菌为优势菌,哈唯氏弧菌为非优势菌.前肠、中肠和后肠的菌量分别为1.3×105、2.8×105和1.1×104cfu/虾体.     

异硫氰酸苄酯(BITC)防污损效果研究──对3种海洋细菌的抑菌作用

异硫氰酸苄酯（ＢＩＴＣ）的抑菌试验表明：ＢＩＴＣ对３种海洋细菌（假单胞菌Ｐｓｅｕｄｏｍｏｎａｓｓｐ．Ｔ１０８株,假单胞菌Ｐｓｅｕｄｏｍｏｎａｓ．ｓｐ．Ｅ２０４株和异单胞菌Ａｌｔｅｒｍｏｎａｓ．ｓｐ．Ｅ１１３株）的完全抑制浓度分别为４．０,１．０和２．０ｍｇ／Ｌ,５０％抑制该３种海洋细菌生长（相对生长率）的浓度分别为２．６７,０．３７和０．４４ｍｇ／Ｌ。     

Isolation and diversity analysis of heterotrophic bacteria associated with sea anemones

A study was undertaken to investigate the heterotrophic bacterial flora associated with the sea anemones. Samples of the sea anemones Anthopleura midori were collected from the coast of Weihai and bacteria were isolated from these samples. Additionally, high numbers of viable bacteria were obtained from the celom wall and surface of anemone, the community of cultivable bacteria was very diverse. As a result of this isolation, 60 strains were obtained, 56 of them were selected for identification and characterization by 16S rRNA gene sequence analysis and limited phenotypic testing. Among these isolates, 16 strains were phylogenetically related to members of the genus Pseudoalteromonas and neighboring taxa. Other isolates included members of the genera Colwellia, Vibrio, Acinetobacter, Pseudomonas, Endozoicomonas, Roseovarius, Paracoccus, Loktanella, Leisingera, Sulfitobacter, Bacillus, Staphylococcus, Plantibacter, Microbacterium, Micrococcus, Joostella, Psychroserpens, Cellulophaga, Krokinobacter, Polaribacter and Psychrobacter. Seven potential novel species were found. Among 60 strains, 17 of them can produce proteolytic exoenzyme, 20 can produce lipolytic exoenzyme. Strain NQ8 has strong antagonistic effects on some Vibrio strains. This study demonstrates that the culturable fraction of bacteria from the sea anemones Anthopleura midori is diverse and appears to possess much potential as a source for the discovery of novel bioactive materials.     

不同碳源富集的石油烃降解菌群结构的分析

以原油和正十六烷为唯一碳源,从长期受石油污染的环境中富集和分离具有不同功能的石油烃降解菌,并利用平板法和PCR-DGGE法时不同碳源富集到的菌群结构进行分析.结果表明,利用2216E平板从以原油和正十六烷为碳源的富集液中各得到两株菌,分另11为TJ-1、TJ-2和TJL-1和TJL-2,分子生物学方法鉴定结果表明,这4株菌分别为短小芽孢杆菌(Bacillus pumilus)、威尼斯不动杆菌(Acinetobacter venetianus)、Oceanobacillus picturaeP和短小芽孢杆菌(Bacillus pumilus).PCR-DGGE分析结果表明,以原油和正十六烷为碳源的富集液中优势菌分别有5种和2种,且2种富集液中的优势菌明显不同.对比PCR-DGGE和平板法分析结果,可以看出PCR-DGGE法能够提供更全面的菌群结构信息.