http://www.sciencedirect.com/science/article/pii/S1674987112001284

This study deals with the biogeography of Antarctic microflora （Antarctica acts as best model to study microbial biogeography） such as cyanobacteria and selected halophiles with special emphasis on Halomonas variabilis and Bacillus licheniformis.Halophiles are known to be resistant not only to salt stress,but also to extreme temperature,pressure,and aridity and they are capable of surviving in harsh environments such as polar regions,deep-sea habitats,and deserts.Many microbes are known to be resistant to hostile environmental conditions,and are capable of surviving in harsh environments.Our group has isolated 444 strains belonging to 28 genera of halophiles from various environments around the world.The 16S rRNA gene sequences revealed that many of the isolated strains from geographically distant habitats having different environmental conditions,were closely related to each other,with some strains possessing 100％ identical sequences.Organisms possessing survival mechanism such as spore formation are usually ubiquitous.The genus Halomonas is represented by potentially endemic strains and the ubiquitous H.variabilis,while spore-forming B.licheniformis showed cosmopolitan distribution.One potentially endemic （moderate endemicity that is regional and/or continental distribution） strain was reported from Syowa station,East Antarctica,and Mario Zucchelli station,West Antarctica,which are geographically separated by 3000 km.Moreover,15 strains having 100％ similarity with B.licheniformis were considered cosmopolitans.The results of this work provide support for the middle-ground model that some microbes have moderate endemicity and others have cosmopolitan distribution.These results will contribute to a greater understanding of microbial biogeography with special emphasis on Antarctica.     

宁波沿海陆源排污口拟杆菌(Bacteroidetes)分布的特点

采用高通量454焦磷酸测序方法对宁波沿海2个重点排污口、8个一般排污口的20个站位水样进行分析,得到2011年3月、5月、8月、10月份各排污口总体微生物以及拟杆菌纲的分布情况。通过对454测序结果分析,共成功鉴定24个门,主要包括变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和后壁菌门(Firmicutes)等。其中拟杆菌门的变化趋势与梭杆菌门(Fusobacteria)相似,均为随月份的增加而递增的趋势。在拟杆菌门下,共鉴定出3个纲3个目9个科27个属,4个采样月份大部分站点的优势属为黄杆菌纲(Flavobacteria),其次为拟杆菌纲(Bacteroidetes)、鞘脂杆菌纲(Sphingobacteria),通过聚类分析发现,拟杆菌门分布规律主要与排污口类型和季节有一定的关系。通过454测序检测到了极难培养的寡营养细菌鞘脂杆菌,也说明了454测序较传统微生物生态学研究方法的优势,同时针对普雷沃菌属(Prevotella sp.)及黄杆菌属(Flavobacterium sp.)两种具有代表性的属的分布情况进行了分析。     

侧孢短芽孢杆菌溶藻活性代谢产物部分特性的研究

研究了不同因素对侧孢短芽孢杆菌（Brevibaci Uuslaterosporus）无菌滤液溶藻效果的影响,并对无菌滤液进行了紫外光谱扫描分析,探讨侧孢短芽孢杆菌溶藻活性代谢产物的物质特性、探究物质的组成成分．结果显示：无菌滤液分别经不同温度、pH值、蛋白酶和紫外照射时间处理后,均仍具有极为显著的溶藻能力（P〈0．01）,溶藻实验7d后颤藻（Oscillatoria sp．）叶绿素a质量浓度的平均下降率分别为：58．65％、63．88％、61．50％和62．61％,藻体干重的平均下降率分别为：39．77％、39．30％、36．49％和37．02％,表明溶藻活性代谢物质具有耐酸、耐碱、耐高温、抗蛋白酶水解、抗紫外辐射的特性;同时,分别在4、28℃环境中贮存28d的无菌滤液,始终保持着极高的溶藻活性,溶藻实验7d后颤藻叶绿素a质量浓度的平均下降率分别为：50．84％和60．39％,藻体干重的平均下降率分别为：37．82％和37．54％,说明该物质的结构极其稳定,在环境中可长久存在;另外,加热与未加热处理的无菌滤液的紫外光谱最大吸收峰都在290～300nm之间．分析表明,溶藻活性代谢产物极大程度上属于具有极强稳定性的肽类物质．     

Enhancing the culturability of bacteria from the gastrointestinal tract of farmed adult turbot Scophthalmus maximus

Eighteen agar media were tested for the culture of gut-associated bacteria from farmed adult turbot(Scophthalmus maximus),including 16 agar media with or without 1% gastrointestinal(GI)supernatant,or with 2% or 4% GI supernatant.A total of 1 711 colonies were analyzed and 24 operational taxonomic units(OTUs)were identifi ed.The greatest bacterial diversity was isolated on Zobell 2216E/Zobell 2216E+ agar media,whereas MRS/MRS+ agar media produced a low diversity of colonies.Agar media with GI supernatant(1%,2%,or 4%)showed increased diversity and yielded different profi les of OTUs from the corresponding original media,suggesting that GI supernatant provides substances that enhance the culture effi ciency of bacteria from the turbot GI tract.The large majority of the colonies(82%)were γ-Proteobacteria,whereas 15.6% and 2.4% of colonies were Firmicutes and Actinobacteria,respectively.At the genus level,49.4% of all colonies were assigned to Vibrio.Other potential pathogens,including Pseudomonas,Photobacterium,and Enterobacter,and potential probiotics,including Bacillus,Paenibacillus,and Pseudomonas,were also isolated on agar media.Most cultured bacteria belonged to species that were fi rst described in the turbot GI tract.The impact of these species on turbot physiology and health should be investigated further.     

AreAcila divaricata and Acila mirabilis One Species or Two Distinct Species？ Evidence from COI Mitochondrial DNA

Acila divaricata （Hinds, 1843） and A. mirabilis （Adams and Reeve, 1850） are common benthic bivalves in China. A number of researchers have proposed that the latter species is a junior synonym of the former species. Because of morphological similarities, it is difficult to distinguish these two species based on visual examination only. For better understanding of their taxon-omy, the mitochondrial CO1 gene fragments of five individuals of A. divaricata from the East China Sea and six individuals of A. mirabilis from the Yellow Sea were sequenced in this study. The phylogenetic relationships of the obtained COI sequences, together with nineteen sequences of three species of the genus Nucula, were analyzed. The pairwise intra- and inter-specific distances for the CO1 sequences ranged from 0.002 to 0.017 and from 0.128 to 0.134, respectively, and no overlap was found. Phylogenetically, A. divaricata and A. mirabilis form distinct clades and cluster into a sister to all other Nucula species. The results indicated that A. di-varicata and A. mirabilis are two distinct species. The differences in the morphology and distribution between the two species were briefly discussed.     

芽孢杆菌(Bacillus)及其复合粘土对典型赤潮生物的去除作用研究

有害赤潮对海洋生态环境和沿海经济危害巨大,如何安全、有效地治理有害赤潮非常重要。文章考察了几种常见芽孢杆菌对近海典型赤潮生物——东海原甲藻(Prorocentrum donghaiense)的去除作用,发现解淀粉芽孢杆菌(Bacillus amyloliquefaciens)对其去除作用最强。进一步考察解淀粉芽孢杆菌对不同赤潮生物的去除作用,发现该芽孢杆菌对米氏凯伦藻(Karenia mikimotoi)和赤潮异弯藻(Heterosigma akashiwo)的去除作用高于东海原甲藻。在此基础上,将解淀粉芽孢杆菌与高岭土复合,研究了微生物复合粘土对典型赤潮生物的去除效果。结果表明,解淀粉芽孢杆菌与高岭土复合后,能够有效促进该菌的生长,进而提升了其对赤潮生物的去除能力。文章对芽孢杆菌及其复合粘土去除赤潮生物的机制进行了分析和探讨,为进一步发展和优化改性粘土治理赤潮技术体系提供了重要参考。     

菌株Alcaligenes sp.P156中一个新型烟酸羟化酶的克隆与功能验证

本研究预测naa cluster中的naaB基因负责催化烟酸到6-羟基烟酸的反应。通过对NaaB的蛋白序列进行分析,发现该酶含有三个亚基,分别由naaB_L1、naaB_S、naaB_L2基因编码,其中naaB_L1和naaB_L2分别编码2个含有钼辅因子结合结构的大亚基,naaB_S基因编码一个含有[2Fe-2S]簇的小亚基。本研究对naaB基因进行了克隆,将构建的重组质粒pME6032-naaB_L1SL2分别转化到无色杆菌(Achromobacter sp.)和大肠杆菌(Escherichia coli)中,通过HPLC和LC-MS检测验证了NaaB负责催化烟酸到6-羟基烟酸,并且在无色杆菌和大肠杆菌中NaaB均能够转化烟酸,这与之前报道烟酸羟化酶不能在大肠杆菌表达的结果不同,说明NaaB的基因编码和催化功能具有独特性。通过序列分析推测NaaB的这一特性可能跟该酶同时含有两个钼结合结构域的大亚基有关。     

铝土矿脱硅微生物的定向筛选及脱硅性能

利用铝硅酸盐矿物为能源的硅酸盐细菌选择性分离培养基,对从鄱阳湖地区不同性质土壤中筛选出了 18 株拟溶硅菌 种进行实验。通过对其产酸、产多糖、形成细菌-矿物复合体与溶硅能力的测定,定向筛选出P04与P17两株环状芽孢杆菌, 并以中国普通微生物菌种保藏中心环状芽孢杆菌 CGMCC1.0153(编号为 P19)(Bacillus circulans)为参照菌株,对其表型形态、 生理生化特性及脱硅效果进行了研究。结果表明:与其它菌种比较,这两株菌种具有较强的产酸、产多糖与溶硅能力,在 培养基中能形成明显的细菌-矿物复合体;通过与模式菌株的表型形态与生理生化特征及16S rRNA基因鉴定结果比较,可 认为 P04 与 P17 是环状芽孢杆菌;在实验条件下,P04,P17 与 P19 对铝土矿均有一定的脱硅能力,浸矿 12d 后,浸出上清液 中SiO2最大含量分别为54.26,42.78,47.98 mg/L,铝土矿的A/S从浸出前的5.17分别提高到10.99,7.61和8.90。表明具有 较强产酸、产多糖能力并能形成明显细菌 - 矿物复合体的环状芽孢杆菌的脱硅能力较强。     

大黄鱼(Pseudosciaena crocea)源美人鱼发光杆菌(Photobacterium damselae)的分离鉴定及致病性研究

于2010年从宁德患病大黄鱼中分离得到一株优势菌NPL1006,经鉴定该菌株为美人鱼发光杆菌。人工感染实验结果显示该菌对大黄鱼的半致死浓度(LD50)为7.96×103CFU/g体重。药敏性实验结果显示NPL1006对头孢唑啉、头孢噻吩和诺氟沙星等9种药物高度敏感。采用纸片法检测得到NPL1006的胞外产物具有蛋白酶、明胶酶和淀粉酶活性。胞外产物感染斑马鱼研究结果显示其LD50为1.20mg蛋白/kg体重。以上结果表明大黄鱼源美人鱼发光杆菌NPL1006胞外产物含有多种毒力因子,与该菌对大黄鱼的致病性密切相关。     

Cloning and characterization of a thermostable carboxylesterase from inshore hot spring thermophile Geobacillus sp. ZH1

The gene(741 bp) encoding carboxylesterase from the thermophilic bacterium Geobacillus sp.ZHl was cloned and overexpressed in Escherichia coli.The purified recombinant protein presented a molecular mass of about 40 kDa by SDS-PAGE analysis.Enzyme assays using p-nitrophenyl esters with different acyl chain lengths as the substrates confirmed its esterase activity,yielding highest specific activity with p-nitrophenyl acetate.Among the p-nitrophenyl esters tested,the carboxylesterase presented preference for p-nitrophenyl caprylate,but hydrolyzed p-nitrophenyl butyrate more efficiently.When p-nitrophenyl butyrate was used as a substrate,the recombinant carboxylesterase exhibited highest activity at pH 8.0 and 60℃.Almost no decrease in esterase activity was observed at 60℃for 3 h,and over 40% of activity was still maintained after incubation at 90℃for 3 h.These results indicate that Geobacillus sp.ZH1 recombinant esterase was thermostable.The enzymatic activity was inhibited by the addition of phenylmethylsulfonyl fluoride,indicating that it contains serine residue,which plays a key role in the catalytic mechanism.Except SDS and xylene,this esterase showed stability toward other tested detergents and organic solvents.Cloning,expression,and biochemical characterization of Geobacillus sp.ZH1 carboxylesterase lay a good foundation for its structural characterization and industrial application.