Molecular hydrogen as an energy source for Helicobacter pylori

The gastric pathogen Helicobacter pylori is known to be able to use molecular hydrogen as a respiratory substrate when grown in the laboratory. We found that hydrogen is available in the gastric mucosa of mice and that its use greatly increased the stomach colonization by H. pylori. Hydrogenase activity in H. pylori is constitutive but increased fivefold upon incubation with hydrogen. Hydrogen concentrations measured in the stomachs of live mice were found to be 10 to 50 times as high as the H. pylori affinity for hydrogen. A hydrogenase mutant strain is much less efficient in its colonization of mice. Therefore, hydrogen present in animals as a consequence of normal colonic flora is an energy-yielding substrate that can facilitate the maintenance of a pathogenic bacterium.     

溶藻细菌H5的分离、鉴定及溶藻特性研究

[目的]分离和鉴定溶藻细菌,研究其溶藻特性,为进一步研究溶藻细菌对水华的治理作用提供帮助。[方法]从香溪河春季水华集聚区水体中分离得到1株有高效溶藻效果的菌株(H5),采用16S rDNA序列相似性分析和Biolog微生物自动鉴定系统等对细菌进行鉴定。采用直接计数法,研究了其对汉斯冠盘藻(Stephanodiscus hantzschii)、倪氏拟多甲藻(Peridiniopsis niei)、具尾逗隐藻(Komma cau-data)的抑制效果,及对汉斯冠盘藻的溶藻作用方式。[结果]根据生理生化及16S rDNA序列分析鉴定,H5属于纺缍形赖氨酸芽孢杆菌(Lysinibacillus fusiformis)。该菌对汉斯冠盘藻、倪氏拟多甲藻、具尾逗隐藻的溶藻率最高为71.3%,最低为57.4%。培养滤液、热处理培养滤液对汉斯冠盘藻的生长具有明显抑制作用,而细菌无细胞提取物无溶藻能力。[结论]该菌对汉斯冠盘藻有较强的溶藻效果,且是通过分泌溶藻物质溶藻。     

Bacterial manganese reduction and growth with manganese oxide as the sole electron acceptor

Microbes that couple growth to the reduction of manganese could play an important role in the biogeochemistry of certain anaerobic environments. Such a bacterium, Alteromonas putrefaciens MR-1, couples its growth to the reduction of manganese oxides only under anaerobic conditions. The characteristics of this reduction are consistent with a biological, and not an indirect chemical, reduction of manganese, which suggest that this bacterium uses manganic oxide as a terminal electron acceptor. It can also utilize a large number of other compounds as terminal electron acceptors; this versatility could provide a distinct advantage in environments where electron-acceptor concentrations may vary.     

Antibody catalysis of the oxidation of water

Recently we reported that antibodies can generate hydrogen peroxide (H2O2) from singlet molecular oxygen (1O2*). We now show that this process is catalytic, and we identify the electron source for a quasi-unlimited generation of H2O2. Antibodies produce up to 500 mole equivalents of H2O2 from 1O2*, without a reduction in rate, and we have excluded metals or Cl- as the electron source. On the basis of isotope incorporation experiments and kinetic data, we propose that antibodies use H2O as an electron source, facilitating its addition to 1O2* to form H2O3 as the first intermediate in a reaction cascade that eventually leads to H2O2. X-ray crystallographic studies with xenon point to putative conserved oxygen binding sites within the antibody fold where this chemistry could be initiated. Our findings suggest a protective function of immunoglobulins against 1O2* and raise the question of whether the need to detoxify 1O2* has played a decisive role in the evolution of the immunoglobulin fold.     

烟草赤星病拮抗生防菌BS06-1的筛选

[目的]筛选对烟草赤星病具有明显拮抗作用的细菌。[方法]应用平板对峙培养法对从烟草根际分离到的细菌进行烟草赤星病拮抗性鉴定,对其中对烟草赤星病菌拮抗性较高的一株细菌利用细菌16S rRNA通用鉴定引物进行PCR扩增、测序和田间抗病性试验。[结果]共从烟草根际分离120份细菌,有6份对烟草赤星病病原菌具有一定拮抗作用,其中1份细菌拮抗作用明显,定名为BS06-1。将该细菌的16S rRNA测序结果登录NCBI网站进行Blastn序列比对,结果表明该菌株与枯草芽孢杆菌BSD-2同源性达到96%,与枯草芽孢杆菌BCRC 14718同源性达到95%,证明该菌属于枯草芽孢杆菌。田间试验证明该菌株BS06-1对烟草赤星病具有显著的抑菌效果。[结论]BS06-1细菌对烟草赤星病具有明显拮抗作用。     

抗MRSA活性菌株B2的理化特性和抑菌效果研究

[目的]确定既适合B2菌株生长又能产生较强活性物质的培养条件。[方法]在不同的温度、pH值、NaCl浓度和各种碳源培养条件下,进行菌体生长状况研究和抑菌活性检测。[结果]菌株B2在4～37℃、pH值2.0～11.0、NaCl浓度为1%～6%,以葡萄糖、麦芽糖、乳糖、蔗糖、淀粉为碳源时都能生长,其最适生长温度为28℃,最适pH值为7.0,最适NaCl浓度为1%,最适生长碳源为麦芽糖。[结论]当pH值为5.0、温度为28℃,以麦芽糖作唯一碳源时,产生的拮抗物质活性最强。     

A microbial arsenic cycle in a salt-saturated, extreme environment

Searles Lake is a salt-saturated, alkaline brine unusually rich in the toxic element arsenic. Arsenic speciation changed from arsenate [As(V)] to arsenite [As(III)] with sediment depth. Incubated anoxic sediment slurries displayed dissimilatory As(V)-reductase activity that was markedly stimulated by H2 or sulfide, whereas aerobic slurries had rapid As(III)-oxidase activity. An anaerobic, extremely haloalkaliphilic bacterium was isolated from the sediment that grew via As(V) respiration, using either lactate or sulfide as its electron donor. Hence, a full biogeochemical cycle of arsenic occurs in Searles Lake, driven in part by inorganic electron donors.     

黄瓜白粉病生防菌H_1、H_2的种类鉴定

通过形态特征、培养条件及生理生化指标,结合16SrDNA分子序列测定,对黄瓜白粉病生防菌H1、H2进行了鉴定.结果表明:生防菌H1、H2的形态特征及22项生理生化指标与枯草芽孢杆菌相符;生防菌H1、H216SrDNA序列与Genbank数据库中枯草芽孢杆菌的相似性高于99.5%,因此,H1、H2均为枯草芽孢杆菌亚种(Bacillus subtilis subsp.ubtilis).综合上述结果,最终确定生防菌H1、H2为枯草芽孢杆菌(Bacillus subtilis).     

萘降解菌NAP2-2的16S rRNA基因克隆和系统发育分析

筛选到1株萘降解菌NAP2-2,对其进行了表观及16S rRNA基因的系统发育研究。结果表明,NAP2-2能以萘作为唯一碳源和能源生长。16S rRNA基因同源性分析显示,该菌株为短芽孢杆菌属(Brevibacillus)的一个成员。对萘降解菌16S rRNA基因序列的系统发育学分析表明,降解菌主要分布在变形菌门和厚壁菌门。因此对NAP2-2菌株的分析揭示了短芽孢杆菌属新的生物学特性,为以后利用此类细菌处理多环芳烃污染提供了重要依据。     

香蕉枯萎病内生生防菌H4-3抑菌蛋白稳定性研究

铜绿假单胞菌H4-3菌株是福建省农科院农业生物资源研究所微生物研究中心从香蕉植株中分离到的内生菌,该菌株对香蕉枯萎病原菌具有很强的抑菌活性,其活性物质为大分子蛋白.研究测定了H4-3菌株抑菌蛋白的稳定性,结果表明：H4-3菌株的抑菌蛋白耐高温、强酸、强碱和高浓度蛋白酶,紫外线和反复冻融对H4-3菌株抑菌蛋白活性无显著影响.     

鸭源大肠杆菌O_(88)的分离鉴定及药敏试验

为了分离鸭大肠杆菌病病原,并筛选出敏感药物,从河北省安新县某大型鸭场30日龄病死雏鸭肝脏分离到一株革兰氏阴性杆菌,通过菌落形态观察、培养特性试验和生化试验鉴定以及药敏试验。确定为大肠杆菌。应用大肠埃希菌O抗原定型血清鉴定为O_(88)。动物试验结果显示,该分离菌株对小鼠有较强的致死性,为致病性大肠杆菌。药敏试验结果显示,分离菌株对庆大霉素、头孢曲松、头孢拉啶、呋喃妥因等药物敏感,对氨苄西林和链霉素等产生耐药。     

荧光假单孢细菌培养的无细胞滤液导致松萎蔫的病状观察

将松材线虫携带的致病细菌无细胞滤液、无菌松材线虫和致病细菌混合物、致病细菌鞭毛蛋白无细胞滤液、无菌松材线虫、无菌松材线虫和非致病细菌混合物接种黑松导致的松树萎蔫的病状与野生松材线虫导致松树萎蔫的病状进行观察和对比,结果为:(1)经培养的致病荧光假单孢菌GcM5-1A菌株的无细胞滤液、无菌松材线虫和致病细菌GcM5-1A混合物接种导致的黑松的病状变化趋势与松材线虫致死的黑松病状是一致的;无菌松材线虫、无菌松材线虫和非致病细菌AM2C混合物不能使黑松发病;(2)致病细菌GcM5-1A无细胞滤液和野生松材线虫导致的黑松病状和病程天数无显著差异;(3)接种无菌松材线虫和致病细菌GcM5-1A混合物的黑松病程显著长于接种野生松材线虫的黑松.结果可以用松萎蔫病是松材线虫和其携带的细菌的复合侵染病害的假说和松材线虫与其携带的细菌之间的互惠共生关系的论点来解释,说明松材线虫携带的致病细菌无细胞滤液是松萎蔫病致病的真正原因.     

Identification and Characterization of Lysobacterenzymogenes as a Biological Control Agent Against Some Fungal Pathogens

Strain OH11, a Gram-negative, nonspore forming, rod-shaped bacterium with powerful antagonistic activity, was isolated from rhizosphere of green pepper in Jiangsu Academy of Agricultural Sciences of China and characterized to determine its taxonomic position. 16S rRNA gene sequence analysis revealed that strain OH11 belongs to the Gammaproteobacteria and had the highest degree of sequence similarity to Lysobacter enzymogenes strain C3 （AY074793） （99%）, Lysobacter enzymogenes strain N4-7 （U89965） （99%）, Lysobacter antibioticus strain （AB019582） （97%）, and Lysobacter gummosus strain （AB16136） （97%）. Chemotaxonomic data revealed that strain OHI 1 possesses a quinine system with Q-8 as the predominant compound and C15：0 iso, C17：1 iso ω9c as the predominant iso-branched fatty acids, all of which corroborated the assignment of strain OH11 to the genus Lysobacter. Results of DNA-DNA hybridization and physiological and biochemical tests clearly showed that strain OH11 was classified as Lysobacter enzymogenes. Strain OH11 could produce protease, chitinase, and β-1,3-glucanase. It showed strong in vitro antifungal activity against Rhizoctonia solani, Sclerotinia scletotiorum, and several other phytopathogenic fungi. This is the first report of identification and characterization of Lysobacter enzymogenes as a biological control agent of plant diseases in China.     

高效降解菌Arthrobacter nicotianae ZAF-05对土霉素的减毒效应

  目的  明确1株土霉素(oxytetracycline, OTC)高效降解菌降低OTC生物毒性的效应,为菌株的实际应用提供初步理论依据。  方法  以前期获得的OTC高效降解菌Arthrobacter nicotianae ZAF-05为对象,利用高效液相色谱-电喷雾质谱联用技术(HPLC-Q-TOF-MS)分析了OTC的降解产物。以对OTC敏感的大肠埃希菌Escherichia coli、枯草芽孢杆菌Bacillus subtilis和斜生栅藻Scenedesmus obliquus作为指示生物,评估OTC降解产物的生物毒性。  结果  HPLC-Q-TOF-MS分析发现:经菌株ZAF-05作用后,体系中的高毒性OTC以及其自然水解产物差向异构土霉素(EOTC)含量显著降低,而低毒性的同分异构土霉素(ISO-OTC)含量略有上升。供试细菌和藻类在添加ZAF-05菌株的OTC体系中的生长情况明显好于OTC自然水解的处理和未添加降解菌的阴性对照,且差异极显著(P＜0.01)。与原药和自然水解物相比,经过ZAF-05菌株降解的OTC代谢产物生物毒性大大降低。菌株的降解作用缓解了OTC对斜生栅藻细胞超微结构的伤害,细胞叶绿体及细胞壁受损减轻。  结论  降解菌ZAF-05能降低OTC残留的环境生物毒性,而毒性的降低可能与体系中高毒性物质被转化成无毒或低毒的代谢产物有关。图7表1参26     

硬脂酸降解菌与嗜氢产甲烷杆菌共培养物的研究

本文介绍基本纯化的产氢乙酸的硬脂酸降解菌与嗜氢产甲烷杆菌的共培养物(LDB2-M2)在等当量的CaCl_2存在下,能降解C_1～C_18饱和脂肪酸;游离的硬脂酸能抑制M2的产甲烷活性;LDB 2短时间接触H_2,导致其活性暂时性抑制;当氢营养细菌数明显高于产氢产乙酸细菌数时,在分离过程中不必加其他氢营养菌作为伴生菌。     

DDT降解菌2D-1的分离鉴定与降解特性研究

从洪泽湖湿地淤泥中分离得到1株能高效降解DDT的革兰氏阳性细菌2D-1。通过生理生化分析,以及16SrDNA序列同源性比较分析,将2D-1鉴定为蜡状芽抱杆菌（Bacillus cereus）。在基础盐酵母培养基中,接种量5％,摇床转速150r／min,培养温度33℃,pH值6．5的条件下,2D-1菌株在8d内能对总量为53．46mg／L的DDT进行95．64％的降解,对p,p’-DDT、p,p＇-DDE、p,p＇-DDD和o,p＇-DDT的绝对降解浓度分别为5．32、22．98、6．66和16．17mg/L。结果表明,2D-1对4种异构体的适应性不同,p,p＇-DDE是其最适底物,且2D-1瞻解DDT的主娶产物有6种。     

一株生活垃圾除臭乳酸菌的筛选与鉴定

[目的]筛选出高效除臭菌株,为生活垃圾生物除臭提供菌种资源。[方法]采用初筛与复筛相结合的方法,分离高效除臭乳酸菌,并对其进行菌落及菌体形态学观察、生理生化特征及16S rDNA序列分析。[结果]从生活垃圾中分离获得一株高效除臭乳酸菌LDJ,经鉴定为干酪乳杆菌。[结论]该乳酸菌在实验室条件下24 h内对氨的去除率为90%,48 h内对硫化氢的去除率为46.8%,可较好地去除垃圾恶臭味。     

高产碱性β-葡萄糖苷酶的产纤维素酶菌株的筛选

为获得高产碱性β-葡萄糖苷酶的产纤维素酶菌株,利用碱性羧甲基纤维素钠平板筛选和β-葡萄糖苷酶平板筛选法相结合,从植物腐败堆积土壤中分离到1株高产碱性β-葡萄糖苷酶的产纤维素酶克雷伯氏杆菌,并对其进行初步酶学鉴定。结果表明:该菌胞外分泌液具有内切葡聚糖酶、外切葡聚糖酶及β-葡萄糖苷酶的活性,其中以β-葡萄糖苷酶活性最高,为48.9U/mL。其所产纤维素酶的最适反应pH值为10.0,最适反应温度为35℃,且Ca2+、Mg2+、Zn2+和Mn2+等金属离子对该酶具有较强的抑制作用。     

生姜根际姜瘟病拮抗菌的筛选及鉴定

从生姜根际土样分离获得了65个细菌分离物和42个放线菌分离物,通过离体拮抗试验,筛选出对姜瘟病有良好拮抗效果的细菌一株,放线菌两株,编号分别为H16-8、WF1和JW02-6。通过形态学观察、生理生化测定以及16S rDNA序列分析,初步确定:H16-8为短短芽孢杆菌(Brevibacillus brevis),WF1为弗雷德氏链霉菌(Streptomyces fradiae),JW02-6为劳伦链霉菌(Streptomyces laurentii)。H16-8、WF1和JW02-6的16S rDNA序列的GenBank登录号分别为:EU812754,FJ972686和FJ972687。     

嗜盐紫色硫细菌283-1的耐盐机制

菌株283-1是一株中度嗜盐紫色硫细菌,对其耐盐特性和机理进行了初步研究。结果表明,菌株283-1在改良的Pfennig紫色硫细菌培养基中最大能耐受2.3mol·L-1 NaCl,其生长对Na+有专一依赖性,对Cl-依赖性较弱;它主要通过在胞内积累相容性溶质甜菜碱来对抗胞外渗透压力,胞内甜菜碱的含量随培养基中NaCl浓度增加而增加,2.0mol·L-1 NaCl时浓度可达到156.4mg·g-1干重。外源添加甜菜碱可以明显提高该菌株耐盐生长的能力。