腾冲热海一株嗜酸热硫化叶菌的分离与鉴定

从云南腾冲热海酸性温泉中分离纯化出一株极端嗜酸热菌株K4-1,并对其进行形态观察、生长特征、碳源和能源利用及16S rRNA基因分析.该菌株细胞形态为不规则球形,有单生鞭毛,严格好氧,兼性自养,能利用元素硫作为能源,也能利用酵母膏、精氨酸或核糖作为碳源和能源.其最适生长温度为75℃,最适pH为3.5.通过16S rRNA基因序列相似性对比对该菌株进行鉴定,结果表明该菌株与硫化叶茵属标准菌株的相似性介于86.6%～94.3%之间,与分离自腾冲热海的腾冲硫化叶菌Sulfolobus tengchongensis RT8-4菌株序列相似性最高,达到98.9%,可将菌株K4-1鉴定为硫化叶菌属菌株.菌株K4-1的16S rRNA基因序列号为EU729124.     

腾冲热海一株栖热菌裂解性噬菌体的分离及其特征

【目的】Thermus(栖热菌)属是嗜热细菌中比较古老的类群,本研究探索从腾冲热海热泉分离栖热菌噬菌体,并初步分析其特征。【方法】采用"双层平板法"从云南腾冲热海碱性热泉中分离纯化栖热菌噬菌体;对噬菌体及其宿主进行电镜形态观察,并进行噬菌体基因组限制性酶切片段多态性分析、噬菌体生理特征及蛋白组成分析。【结果】从腾冲热海热泉分离获得1株裂解性噬菌体,其宿主菌TC10通过16S rRNA基因序列分析鉴定为Thermus属菌株。此噬菌体为长尾型,头部直径67nm,尾管长837nm、宽10nm,最适感染温度为65℃-70℃,最适感染pH值为7.6,对氯仿不敏感,其形态与分离自俄罗斯勘察加半岛热泉的栖热菌噬菌体P23-45和P74-26有一定的差异,蛋白组成差异显著,为一株新的栖热菌噬菌体,命名为TTSP10(Tengchong Thermus Siphoviridae Bacteriophage)。     

浮霉菌门严格厌氧产氢细菌(Thermopirellula anaerolimosa)的分离及其生理特性

[目的]厌氧颗粒污泥中含有大量未知微生物资源,利用低浓度底物及添加抗生素的培养基进行厌氧发酵细菌的筛选,并对分离菌株进行生理生化特性研究.[方法]利用系列稀释法及亨盖特厌氧滚管技术从制糖废水厌氧处理反应器的颗粒污泥中分离到一株高温厌氧产氢细菌VM20-7T,通过16S rRNA基因序列同源性确定其系统发育地位.[结果]菌株VM20-7T为高温、严格厌氧、革兰氏阴性梨形细菌,细胞大小为(0.7-2.0)μm×(0.7-2.0) μm,不运动,不产芽胞.其生长温度范围为35℃-50℃(最适温度45℃),pH范围为6.0-8.3(最适pH7.0-7.5),NaCl耐受范围为0％-0.5％(w/v,最适浓度0％).菌株VM20-7T可利用葡萄糖、麦芽糖、核糖等多种糖类为唯一碳源生长,葡萄糖发酵终产物是乙酸和H2.该菌株不利用硝酸盐、硫酸盐等作为电子受体生长.G+C含量为60.9 mol％,16S rRNA基因序列同源性显示菌株属于浮霉菌门,但与已培养菌株的同源性较低,与梨形菌属一红小梨形菌属-芽殖小小梨形菌属(Pirellula-Rhodopirellul -Blastopirellula,PRB)分支的亲缘关系最近,但序列相似性也仅为82.7％-84.3％.[结论]利用低浓度糖类并添加抗生素分离厌氧颗粒污泥中的微生物,获得了浮霉菌门首例严格厌氧细菌VM20-7T.生理生化特性和系统发育分析显示,菌株VM20-7T为浮霉菌目的新属新种,命名为Thermopirellula anaerolimosa.该菌株的菌种保藏号为CGMCC 1.5169T=JCM 17478T=DSM 24165T.     

四株红树林促生菌的遗传分析鉴定及其促生能力

【目的】鉴定四株供试菌株的种属地位,了解菌株所具有的促进植物生长能力。【方法】运用nifH与16S rRNA基因序列对供试菌株进行遗传分析,采用钼锑抗比色法和乙炔还原法分别测定菌株的溶磷、固氮能力。通过接种试验验证菌株促进红树植物生长的能力。【结果】通过对菌株nifH与16S rRNA的同源性、系统发育树分析,HN011与需钠弧菌(Vibrio natriegens)的相似性最高,SZ7-1、SZ7-2与产酸克雷伯氏菌(Klebsiella oxytoca)的相似性最高。而SZ002在16S rRNA的系统发育分析中归属为类芽孢杆菌属(Paenibacillus sp.),却在nifH基因分析中与克雷伯氏菌(Klebsiella sp.)的相似性最高。供试菌株都具有较强的溶磷能力和高固氮酶活性。接种后植株有较好的生长表现,部分接种植株在干重、全氮、全磷含量等方面较对照有显著地增加(P0.05)。【结论】首次发现兼具溶磷-固氮两种能力的红树林植物促生菌,接种试验也表现菌株具有良好的促生能力,为红树林人工接种促生菌的应用提供了可靠的理论依据。     

一株弗兰西斯菌的鉴定及其微生物学特性研究

摘要:【目的】对广州市某空调冷却水中分离的一株L-半氨酸刺激生长性细菌08HL01032进行分类学鉴定及微生物学特性研究。【方法】通过培养基生长状况、菌株形态、生理生化特性、PCR鉴定、16r RNA基因序列分析、RNA聚合酶β亚单位（rpoB）基因序列分析、动物实验、药敏实验等多种表型与基因相结合的方法,探讨该菌株的分类进化地位及一些基本的生物学特性。【结果】该菌株一些生理生化特征与军团菌十分相似,为革兰氏阴性杆菌,氧化酶阴性,不还原硝酸盐,不分解尿素,血平板缓慢生长、易忽略,酵母活性炭缓冲（BCYE）琼脂48 h内生长良好,最初错误鉴定为军团菌。但16S rRNA基因（GenBank登陆号：FJ591095）、rpoB 基因（GenBank登陆号：FJ939309）系统发育进化显示,该菌株为弗兰西斯（Francisella）属细菌,与蜃楼弗兰西斯菌（F. philomiragia）最相近,相似度分别为95.3 %,87.3 %,菌落形态、表型特征亦与弗兰西斯菌属的相符合。其最适生长温度在25~28℃之间,42℃不生长;能耐受pH 2.2酸液10 min;万古霉素、青霉素、多粘霉素B等耐药,军团菌甘氨酸-万古霉素-多粘霉素B-放线菌酮（GVPC）选择性添加剂无生长抑制作用;菌体无芽孢、无鞭毛动力,透射电镜下观察到荚膜样结构,但动物实验初步显示对小鼠无致病性。【结论】菌株08HL01032为弗兰西斯菌属一潜在的新种,其典型特征为L-半胱氨酸刺激生长性,不同于军团菌的L-半胱氨酸依赖性生长特性。 关键词: L-半胱氨酸刺激生长性; 弗兰西斯菌; 鉴定; 微生物学特性     

瘤胃降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物的分离鉴定

【目的】本试验从瘤胃中分离鉴定降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物,为深入探究甲烷菌对厌氧真菌代谢途径的影响及相关调节机制奠定基础。【方法】利用厌氧滚管技术从荷斯坦奶牛瘤胃内容物中分离厌氧真菌与甲烷菌共培养物,通过形态学观察和DAPI染色以及甲烷菌16S rRNA基因序列分析方法分别对厌氧真菌及甲烷菌进行鉴定。【结果】从荷斯坦奶牛瘤胃中共分离到28株厌氧真菌与甲烷菌共培养物。共培养物中的厌氧真菌均为单中心菌株,分别属于Piromyces,Neocallimastix和Caeomyces属,所占百分比为53.57%,42.86%及3.57%。甲烷菌16S rRNA基因序列分析结果表明,共培养物中的甲烷菌均为甲烷短杆菌。本研究共获得四种不同的厌氧真菌与甲烷菌组合,分别为Piromyces/类Methanobrevibacter olleyae菌株,Neocallimastix/类Methanobrevibacter olleyae菌株,Neocallimastix/类Methanobrevibacter thaueri菌株及Caecomyces/类Methanobrevibacter olleyae菌株,分别占总数的53.57%,39.29%,3.57%及3.57%。【结论】分离得到的28株厌氧真菌和甲烷菌共培养物中,占优势的为具有丰富丝状假根的厌氧真菌Piromyces和Neocallimastix以及类Methanobrevibacter olleyae属的甲烷短杆菌。本研究为进一步研究瘤胃内厌氧真菌与甲烷菌相互代谢关系奠定基础。     

栖热菌属高温菌RH99-02菌株产类胡萝卜素的研究

从腾冲热海分离到最高生长温度达80℃的一株嗜热非芽孢菌RH99-02菌株,经鉴定属Thermus属,对其所产类胡萝卜素进行了吸收光谱扫描及薄层层析分析。发酵研究表明振荡培养时的菌体生物量和色素产量上均高于静置培养,培养至50h菌体生物量和色素产量达到最大值,菌体色素产量为762μg/g干重。     

8种贵州药用植物内生放线菌的分离及多样性研究

【背景】药用植物内生菌能产生多种活性较好的次级代谢产物,是新抗生素发现的重要来源。目前,药用植物内生菌的研究主要集中在内生真菌,内生放线菌研究相对较少,是一个尚未充分开发的新领域。【目的】研究贵州药用植物内生放线菌多样性,以期发现新种放线菌或产新活性物质的放线菌,为微生物药物研究奠定基础。【方法】药用植物样品表面消毒后用粉碎机粉碎,采用9种不同分离培养基分离放线菌;通过提取放线菌基因组DNA,PCR扩增16S rRNA基因序列,测定序列并提交EzBioCloud数据库进行序列比对,构建系统进化树,进行内生放线菌多样性分析。【结果】通过形态特征初步排重,从8种药用植物样品中共分离得到内生菌62株,测定其16S rRNA基因序列,序列比对结果表明其中的57株菌分布于放线菌域的5目8科16属,分别是短小杆菌属(Curtobacterium)、链霉菌属(Streptomyces)、微杆菌属(Microbacterium)、纤维单胞菌属(Cellulomonas)、植物杆菌属(Plantibacter)、原小单孢菌属(Promicromonospora)、动球菌属(Kineococcus)、壤球菌属(Agrococcus)、沼杆菌属(Patulibacter)、气微菌属(Aeromicrobium)、拉贝达氏菌属(Labedella)、叶居菌属(Frondihabitans)、冷杆菌属(Frigoribacterium)、节杆菌属(Arthrobacter)、动孢菌属(Kineosporia)、Amnibacterium属,其中短小杆菌属(Curtobacterium)为优势菌属,菌株M8JJ-5和M2KJ-4的16S rRNA基因序列分别与有效发表菌株AmnibacteriumkyonggienseKSL51201-037T(FJ527819)和Aeromicrobiumfastidiosum DSM10552T (Z78209)的相似性最高,相似性分别为97.29%和98.95%,可能为潜在新物种。【结论】贵州药用植物中蕴藏着多样性丰富的放线菌且具有从中发现新放线菌的潜力,能为新抗生素的发现提供菌种储备,也为今后对该地区放线菌资源的认识和开发利用提供理论依据。     

西伯利亚鲟(Acipenser baerii)致病性维氏气单胞菌的分离鉴定

摘要：【目的】本研究旨在寻找引起养殖西伯利亚鲟鱼(Acipenser baerii)病害的致病因子。【方法】从北京地区自然患病的西伯利亚鲟鱼体内分离到致病菌株X-1-06909,采用生理生化鉴定结合16S rRNA基因序列的系统发育学分析确定该菌株的系统发育地位。同时采用琼脂扩散法对抗菌类药物的敏感性进行测定。【结果】菌株X-1-06909与Aeromonas veronii ATCC 35624T的16S rRNA基因序列相似性达99.6％;结合形态特征与生理生化测定结果,革兰氏阴性杆菌,具极生单鞭毛     

一株高效降解木糖的耐酸、嗜热厌氧杆菌的生理特性及产物分析

【目的】分离高效降解木糖的嗜热厌氧杆菌菌株,用于发酵生产生物燃料乙醇,为后继的构建基因工程菌株及联合生物工艺提供材料。【方法】运用亨盖特厌氧操作技术从胜利油田油层采出液两年的富集样中分离到一株嗜热厌氧杆菌xyl-d。采用形态学观察、生理生化指标鉴定及基于16S rRNA的系统发育学分析确定其分类地位。【结果】菌株xyl-d为革兰氏阴性厌氧杆菌,菌体大小为(1.35-5.08)μm×(0.27-0.40)μm,单生、成对或成簇生长,芽胞圆形,端生。温度生长范围30-85℃(最适温度65℃);pH范围3.0-10.0(最适pH 7.5);NaCl浓度范围0%-4%(最适NaCl浓度2.0%)。发酵D-木糖的产物是乙醇、乙酸、CO2及少量的异丁醇、丙酸。菌株xyl-d的(G+C)mol%含量为45.6%,与热厌氧杆菌属模式菌株威吉利热厌氧杆菌(Thermoanaerobacter wiegelii)DSM10319T及嗜热乙醇杆菌(Thermoanaerobacter ethanolicus)DSM2246T的16S rRNA序列相似性均为99.3%。菌株利用D-木糖产乙醇的最佳初始pH为8.5;少量酵母粉能刺激生长并显著提高发酵D-木糖的产醇率,使乙醇成为主要的发酵产物;培养基中乙醇浓度达到7%(V/V)时菌体生长受到抑制,最佳生长条件下D-木糖的降解率可达91.37%,最佳产醇条件下发酵1摩尔D-木糖可产生1.29摩尔的乙醇。【结论】菌株xyl-d是从特殊生境(油藏)中分离到的一株高效降解D-木糖的耐酸、嗜热的厌氧杆菌,其为半纤维素降解产乙醇的联合生物工艺提供了菌源。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Chemokines and cytokines: axis and allies in asthma and allergy

Allergic asthma can be precipitated by many factors. For the atopic person, fungus, pollen, dust mites, cockroach antigens, and diesel exhaust are all agents that may trigger an allergic attack. Cytokines and chemokines are integral mediators of fungal asthma. From the earliest time points, they recruit and activate the cells required for the clearance of fungus as well as being critical factors involved in the immunopathology of this disease. In the final analysis, it is clear that these mediators can act to the benefit or the detriment of the host.     

Oxidation and nitration of ribonuclease and lysozyme by peroxynitrite and myeloperoxidase

In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier.