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.     

高通量测序技术揭示印度尼西亚北苏拉威西岛一株色素异常澄黄滨珊瑚特异的共生微生物群落

细菌性疾病对珊瑚的健康影响造成了很大影响,而致病菌与珊瑚疾病之间的对应性还不明朗。色素异常现象是野外澄黄滨珊瑚一种常见的不健康状态。我们采用Illumina 16s rRNA基因高通量测序技术比较分析了健康和色素异常澄黄滨珊瑚共生细菌的群落组成差异。色素异常澄黄滨珊瑚共生细菌的多样性明显高于健康澄黄滨珊瑚,同时两种珊瑚的共生细菌群落组成也有很大差异。一些特定的细菌类群在澄黄滨珊瑚共生细菌中的比例很高,推测这些细菌种类可能是导致澄黄滨珊瑚色素异常的“罪魁祸首”。     

海洋沉积物氨氧化菌群落结构的测序分析

本文以氨氧化细菌(ammonia-oxidizing bacteria,AOB)特征基因氨单加氧酶(ammonia monooxygenase,AMO)α亚基基因(amoA)为目标基因(～491bp),分别采用克隆文库、454高通量测序和illumina测序技术对其群落结构进行了比较研究。结果表明, 454高通量测序和illumina测序技术得到的菌群多样性及丰富度均高于克隆文库技术。在菌群组成方面,虽然3种测序技术检测到的优势类群均为亚硝化螺菌,但在菌群丰度方面存在差异。3种测序技术中,克隆文库技术在很大程度上可能高估物种丰度,且对部分低丰度类群的检测能力有限;illumina单端测序由于其测序片段较短,可能存在物种注释错误、不能准确区分各物种间序列差异等问题;比较而言, 454高通量测序技术能较为全面和准确地反映海洋沉积物中AOB的菌群结构特征。     

模拟增雨对荒漠土壤古菌多样性的影响

为探讨生长季增雨对荒漠土壤中古菌群落结构的影响,根据内蒙古磴口多年平均降水量,设计了模拟增雨试验,包括对照、2个增雨时段（生长季前期与后期）、每个时段2个增雨梯度（50%与100%）共计5种处理。增雨后提取荒漠土壤总DNA,对古菌群落编码16S rRNA基因的V4区进行Hiseq测序,分析古菌群落丰度、多样性及结构组成等。结果表明：该地区荒漠土壤中古菌群落均以奇古菌门（Thaumarchaeota）和广古菌门（Euryarchaeota）为优势菌群。NMDS和3种非相似性分析（MRPP、ANOSIM和Adonis）表明,生长季前期增雨未显著改变古菌的群落结构,生长季后期增雨显著改变了古菌的群落结构,当生长季后期增雨量达到100%时,古菌丰富度和多样性显著降低。与氨氧化过程密切相关的奇古菌门（Thaumarchaeota）相对丰度在生长季后期100%增雨条件下显著增加,将促进荒漠土壤的硝化过程。Mantel检验发现荒漠土壤古菌群落结构受土壤湿度、土壤温度、土壤硝态氮含量的影响。未来全球变化背景下降水格局变化将会对荒漠土壤古菌群落结构产生影响,并可能影响氮循环过程。     

On four closely related hypotrichous ciliates (Protozoa, Ciliophora, Spirotrichea): molecular characters, interspecific relationships and phylogeny defined with multigene sequence information

In order to clarify the phylogeny and relationships of the most confused hypotrichous ciliates, Holosticha-complex, four closely related holostichids (five populations), Holosticha bradburyae, H. diademata, Anteholosticha sp., and A. manca, were compared and analyzed using ITS2 secondary structures, ITS1-5.8S-ITS2 region and SSrRNA gene sequences. The ITS1-5.8S-ITS2 region sequences of these four species were first sequenced, and they shared sequence identities ranging from 68.0% to 90.1%, while two populations of Anteholosticha sp. differed in three nucleotides (sequence identity 99.8%). There were several minor differences among ITS2 secondary structures of these species, while two populations of Anteholosticha sp. had the identical secondary structure. Phylogenetic trees inferred from the ITS1-5.8S-ITS2 region sequences of stichotrichs using multiple algorithms (Neighbor-Joining, Maximum Parsimony and Bayesian) revealed similar topologies. The results show that:(1) Holosticha bradburyae and H. diademata firmly clustered together with strong bootstrap supports, forming a sister clade with Anteholosticha sp., (2) Anteholosticha appeared to be a paraphyletic assemblage, in which the morphotype A. manca was more closely related to Diaxonella trimarginata than to its congener Anteholosticha sp. Phylogenetic analyses based on the SSrRNA gene and the combined sequences of SSrRNA gene and ITS1-5.8S-ITS2 region revealed the similar relationships between Holosticha and Anteholosticha, nevertheless their positions within the subclass Stichotrichia differed from each other inferred from different genes.     

EM对SBR工艺处理生活污水的强化作用

生物强化技术对于改善现有污水处理工艺的效果具有重要作用 。实验研究以生活污水中添加有效微生物群(EM)和单以生活污水两种方式培养活性污泥,然 后分别将其引入SBR反应器,以考察EM对SBR工艺处理生活污水的强化作用。结果表明,在正 常生活污水浓度和最优工艺条件(污水pH6～8,曝气2h,静置沉淀0.5h)下,EM-SBR反应器对 污水CODcr、NH+4-N的平均去除率分别比普通SBR工艺高19.08%和23.17%;且具有较好 的稳定性。此外,EM强化的SBR工艺还具有极强的抗冲击负荷能力,当进水CODcr为2738mg．L-1时,处理3h后,出水CODcr即可达到《污水综合排放标准》的要求,去除率高于96 %,而普通SBR工艺即使处理6h也不能达标。     

鲈鱼生长催乳素的分离纯化与N-末端序列分析

生长催乳素(SL)是生长激素/催乳素家族的一种新的脑垂体蛋白质激素,本研究首次从鲈鱼脑垂体中分离得到.鲈鱼脑垂体在碱性条件下,经葡聚糖凝胶(SephadexG-100)过滤和反相高效液相色谱(rpHPLC)分离出纯化的鲈鱼生长催乳素,(sbSL),并与大麻哈鱼生长催乳素(sSL)抗体发生特异性的Western免疫印迹交叉反应证实.SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析,发现鲈鱼生长催乳素分别是由分子量为23.2×103和28.4×103u两种形式组成(后者可能是一种糖基化形式).根据Edman降解原理,测得鲈鱼生长催乳素的N-末端16个氨基酸的序列,它与已知鱼类生长催乳素的N-末端序列比较,具有较高的同源性;由此进一步证实,分离纯化得到的鲈鱼生长催乳素是正确的.     

Isolation and Identification of Lactic Acid Bacteria Isolated from a Traditional Jeotgal Product in Korea

Seventeen lactic acid bacterial strains (LAB) were isolated using MRS agar medium from Jeotgal, a Korean fermented food, purchased at the Jukdo market of Pohang. To identify the strains isolated, they were tested by examining their cell morphologies, gram-staining, catalase activity, arginine hydrolase activity, D-L lactate form and carbohydrate fermentation. According to the phenotypic characteristics, three strains were tent atively identified asLactobacillus spp., ten wereEnterococcus spp. (orStreptococcus spp., orPediococcus spp.) and the rest were Leuconostoc spp. (orWeissella spp.). Five strains among 17 were chosen by preliminary bacteriocin activity test. Four bacterial strains which inhibited both indicator microorganisms were identified by 16S rRNA sequencing. The results are as follows; Leuconostoc mesenteroides (HK 4),Leuconostoc mesenteroides (HK 5),Leuconostoc mesenteroides(HK 11),Streptococcus salivarius(HK 8). In order to check LAB which are showing a high survival rate in gut, we investigated three strains inhibiting both indicator microorganisms in artificial gastric acid and bile juice -all except HK8. The three strains mentioned above grew in extreme low acid conditions.     

基于低压静电场及高通量测序技术对中华管鞭虾(Solenocera crassicornis)保鲜过程中微生物群落影响分析

中华管鞭虾(Solenocera crassicornis)隶属于十足目、管鞭虾科、管鞭虾属, 主要分布于我国东南沿海。其富含多种必需氨基酸和矿物质, 高蛋白低脂肪, 但易受微生物污染而发生腐败变质, 需引用基于16S rRNA的高通量测序技术准确测定其微生物群落组成成分变化。以中华管鞭虾为试验材料, 采用高通量测序技术对虾肉的微生物群落碱基进行基因测序, 将产生的碱基序列通过OTU聚类、多样性及相关性分析手段研究了贮藏过程中电场处理后影响中华管鞭虾品质的微生物发育信息和群落组成变化规律。分组为新鲜样本(G0), 2 kV、3 kV及对照组贮藏中期(G1、G2、G3), 2 kV、3 kV、对照组贮藏末期(G4、G5、G6), 通过物种韦恩图、Alpha和Beta多样性以及物种组成分析, 研究不同强度电场环境对中华管鞭虾微生物群落组成的影响。结果表明, OTU聚类分析中优势菌群从属水平看为假交替单胞菌属(Pseudoalteromonas)、假单胞菌属(Pseudomonas)、嗜冷杆菌属(Psychrobacter)和热杀索丝菌属(Brochothrix)等, 其中假交替单胞菌属能够适应高盐浓度环境, 是中华管鞭虾微生物腐败主要致腐菌。贮藏中期微生物物种多样性顺序为3 kV组>2 kV组>对照组(0 kV), 电场能够有效抑制优势菌的生长; 贮藏末期3组物种结构相近, 此时电场对虾肉菌群结构无明显影响。PCoA主成分分析图中发现, 在坐标系中贮藏中期的G1、G2、G3组距离较为接近, 贮藏末期的G4、G5、G6组距离较为接近, 说明在同一贮藏期间的各组微生物种群结构类似。综上, 贮藏前中期低压静电场能够抑制优势菌的生长, 维持微生物物种间平衡并使其物种多样性较高, 且3kV更佳, 贮藏末期时由于优势菌更适应环境, 热杀索丝菌属和肉食杆菌属占比增加, 电场对物种多样性的影响不大。研究结果可为深入探讨影响中华管鞭虾贮藏保鲜效果的微生物群落组成和基础发育信息提供有效依据。     

A Preliminary Genetic Investigation of Rastrelliger Kanagurta Based on Random Amplified Polymorphic DNA and Mitochondrial ND2 Gene

In a preliminary investigation, Random Amplified Polymorphie DNA (RAPD) analysis and partial mitochon-drial ND2 gene sequencing were conducted to study the genetic variation of the Indian mackerel, Rastrelliger kanagurta along a 450 km stretch of its distribution on the west coast of Peninsular Malaysia. A total of 53 individuals from 6 popu-lations were analyzed using 4 RAPD primers and a sub-sample of 15 individuals was chosen for sequencing of partial ND2 gene. Comparison between the 2 markers revealed genetic structuring in the RAPD results but genetic homogeneity for ND2 gene. Based on the former there may be at least 2 genetically differentiated groups of Rastrelliger kanagurta a-long this stretch.