一株假单胞菌属低温噬菌体的分离及其特性研究

目的从纳帕海湿地土样中分离和培养低温菌及其噬菌体,并对其特性进行研究。方法分离纯化低温宿主菌,采用"双层平板法"分离获得噬菌体,通过16S rRNA基因分析对宿主菌进行初步鉴定,对噬菌体进行电镜形态观察,并进行噬菌体基因组酶切片段长度多态性分析及噬菌体生理特性研究。结果从纳帕海土样中分离获得一株裂解性低温噬菌体,命名为SV-12,其宿主菌S-12鉴定为恶臭假单胞菌(Pseudomonas putida)。噬菌体SV-12有囊膜,头部为典型的正六边形,立体对称结构,直径约68.7 nm;尾管长约122.5 nm,直径约25 nm;在4℃时具有侵染活性,4～25℃均能产生边缘清晰、透明的噬菌斑。最适感染温度约为10℃,pH耐受范围较广,在pH=10时具有最多出斑数。结论 SV-12属于肌尾噬菌体科,为裂解性噬菌体,对氯仿极度敏感。基因组为dsDNA,大小约为74 kb。     

一株红假单胞菌的分离及生理特性研究

光合细菌在水处理和水生态修复领域应用前景广阔,对其生理特性研究具有重要价值。从武汉东湖分离得到一株不产氧光合细菌PUF1,通过对菌落形态、细胞超微结构、特征吸收光谱以及系统发育等分析,初步确定为红假单胞菌属(Rhodopseudomonas sp.)紫色非硫光合细菌。PUF1细胞呈直的或稍弯曲的杆状,长3.05—10.06μm,直径0.32—0.68μm,具有片层膜结构。PUF1培养物呈深紫红色,主要色素为细菌叶绿素a(Bchl. a)和类胡萝卜素。初始pH 6.0—8.0,光照强度500—3000 lx,稳定期细菌生物量无显著差异,但培养液pH>8.0会显著抑制PUF1最大光量子产量(Fv/Fm)。试验进一步研究了细菌不同生长阶段粗蛋白含量、ATP酶(ATPase)活性及Fv/Fm的变化规律。结果表明, PUF1不同生长阶段其蛋白含量有所差异,以稳定期为最高(>60%),而ATPase活性随培养时间的延长而逐渐降低。此外, PUF1光合作用与其生长状态也有一定的关系,具体表现为细菌生长周期内Fv/Fm变化规律适用单峰高斯模型,且以对数期Fv/Fm为最高。研究结果可为光合细...     

几株沼泽红假单胞菌的分离和分类特性研究

分别从温州市郊区采集的4处淤泥中分离到4株光能异养型菌株W1,W2,W3,W4,对其形态特征和生理特性研究发现,4株菌基本相似,均为革兰氏阴性,弯曲杆状,出芽繁殖,在光照厌氧和黑暗好氧条件下均能生长.W1～W4菌株能利用许多有机物作为碳源且以乳酸钠、醋酸钠、丙酮酸、苹果酸、丁二酸为最适宜.酵母膏能显著刺激其生长,而谷氨酸钠和丙氨酸对其生长无刺激作用.在pH5.0～8.5的条件下4株菌均能生长,其最适pH均为7.0左右.根据上述结果,确证W1～W4均为沼泽红假单胞菌(Rhodopseudomonas palustris).     

铜绿假单胞菌噬菌体的分离鉴定及耐噬菌体突变频率测定

用铜绿假单胞菌为宿主菌自污水中分离到3株不同的铜绿假单胞菌噬菌体,命名为PaP1、PaP2及PaP3者均为DNA双链噬菌体,基因组大小分别约为47kb、34kb及24kb。3株噬菌体原液滴度（pfu）分别为109/mL、1011/mL和1011/mL。PaP1为裂菌性噬菌体,PaP2及PaP3为溶原性噬菌体。电镜观察,3株噬菌体头部均为多面体立体对称颗粒,直径分别约为70nm、55nm和65nm。PaP1属肌尾噬菌体科,PaP2和PaP3属短尾噬菌体科。研究中还发现了铜绿假单胞菌的耐噬菌体现象及耐受菌与敏感菌之间的“菌群交替”现象,经测定铜绿假单胞菌耐噬菌体的突变机率在1.4×10-7～7.9×10-7之间。     

一株荧光假单胞杆菌的分离鉴定与反硝化特性

【目的】从污水厂的活性污泥中获得一株高效反硝化细菌。【方法】采用低温驯化,进行初筛、复筛选取一株反硝化活性最高的菌株,命名为L2,通过形态学、生理生化特征及16S r RNA基因序列分析研究其分类地位,系统研究理化因素对该菌株反硝化性能的影响。【结果】菌株在低温条件下能够稳定高效地进行反硝化,鉴定该菌株为荧光假单胞杆菌(Pseudomonas fluorescens),其反硝化最适接种量为10%,温度为20°C,p H为7.0,盐浓度为0.5%,碳源为葡萄糖,C/N为5.0,能够耐受较高初始硝态氮浓度。【结论】菌株L2是一株耐低温、耐高浓度初始硝态氮、耐低C/N、兼性厌氧、高效反硝化的荧光假单胞杆菌。     

一株红假单胞菌分离及对幼鳖作用的初步研究

从露天养鳖池中分离到红假单胞菌CZ-2菌株。该菌具有生理需求简单、培养条件粗放、生长迅速的特点。在黑暗,好氧和光照/厌氧条件下均可良好生长,最适条件下培养6d,生物量可以达到1．57g/L。在温室幼鳖的饲料中添加CZ-2菌株培养液后幼鳖没有出现直接死亡,发病率也较对照池低50％左右。试验表明CZ-2菌株培养液对幼鳖生长有促进作用,比对照提高了8．9％。该文对CZ-2菌株培养特性及对幼鳖作用作了探讨。     

泡囊假单胞菌的分离及鉴定

从一例心包积液患者的心包液中,分离到一株氧化代谢、氧化酶阳性,具单极毛运动的革兰氏阴性杆菌。经形态、生理生化特性鉴定,DNA的G+C mol％测定和Biolog自动系列的验证,确认为泡囊假单胞菌(Pseudomonas vesieularis)。指出该菌可使免疫力低下患者发生感染。对该菌分类地位的变迁进行了讨论。     

杀虫遗传工程荧光假单胞菌IPP202部分生物学特性

对遗传工程荧光假单胞菌IPP202进行了质粒稳定性检 测、抑菌活性测定、在棉花根部和叶面定殖能力分析、杀虫蛋白抗紫外能力检测及田间杀虫 活性测定等试验。结果表明,工程菌与出发菌株P303相比,其抑菌、定殖等有益于植物的优 良特性未发生显著变化;经过连续培养和连续稀释培养后工程菌的质粒都非常稳定;广波长 紫外线照射2 h后,苏云金芽胞杆菌(Bacillus thuringiensis,简称Bt)由于裸露的伴 孢晶体杀虫蛋白受紫外线破坏因而杀虫活性大大下降,而荧光假单胞菌工程菌的活性变化不 大;IPP202田间杀虫活性与Bt野生菌株接近。工程菌有望解决Bt本身存在的杀虫蛋白多以裸 露晶体的形式存在而易受紫外线破坏的弱点,同时也发挥了P303菌株在多种植物上定殖能力 的优点,使其具有在植物周围大量繁殖而直到杀虫作用的优势。通过进一步研究,将有望构 建成更有实用价值的工程菌。     

沼泽红假单胞菌的研究进展

概述了沼泽红假单胞菌菌株的分离纯化,主要特性及应用研究方面的进展。     

301株铜绿假单胞菌的分布及耐药性分析

目的 通过对临床中分离的铜绿假单胞菌的分布及对临床常用13种抗生素的耐药性进行分析指导临床合理用药.方法 收集大连医科大学附属第二医院2010年1月至12月临床送检的标本,采用全自动细菌和药敏分析仪分离铜绿假单胞菌同时进行药敏试验.结果 铜绿假单胞菌在痰液标本中分离率高达79.06％;对头孢噻肟、头孢西丁、头孢唑啉3种药物的耐药率均大于50％;对头孢他啶,哌拉西林/他唑巴坦耐药率低于20％.结论 铜绿假单胞菌易产生多源耐药,加强耐药性监测,控制医院内感染,对临床医生选用有效的抗生素具有十分重要的意义.     

Complete Genome Sequence Analysis of the Cold-active Siphoviridae Bacteriophage from Pseudomonas fluorescens

Microbiology - Phages are considered the most abundant and diverse biological entities on Earth. Although many phages have been isolated, most of this diversity remains completely unexplored....     

Isolation and Characterization of a Bacteriophage Lytic for Desulfovibrio salexigens, a Salt-Requiring, Sulfate-Reducing Bacterium

A bacteriophage that lysed Desulfovibrio salexigens cells was isolated from marine sediments and preliminarily characterized by electron microscopy and electrophoretic analysis of structural proteins and genomic nucleic acid. The bacteriophage had an icosahedral head and a long flexible tail, and the buoyant density of the bacteriophage particles was 1.468 g/ml in cesium chloride. The particles consisted of a double-stranded DNA molecule about 33 kilobase pairs long and at least 11 structural proteins.     

Isolation and Characterization of a Pseudomonas fluorescens Strain Suppressive to Bipolaris maydis

Strain PJ0210, identified as Pseudomonas fluorescens, inhibited Bipolaris maydis both in vitro. The absence of detectable inhibitory substances assayed by a variety of methods indicated nutrient competition as the operative component of antagonism. This was further supported by observations of reduced inhibition by addition of glucose both in vitro and in vivo. Strain PJ0210 was adaptecd to the phyllosphere showing good survivability and colonizing ability both inplant house and field trials. Disease suppression was generally correlated with population sizes of strain PJ0210. Population sizes in the field were lower than those recorded in the plant house with a concomitant reduction in disease suppressive ability, of strain PJ0210.     

Isolation and Characterization of a Lytic Myoviridae Bacteriophage PAS-1 with Broad Infectivity in Aeromonas salmonicida

To search for candidate control agents against Aeromonas salmonicida subsp. salmonicida infections in aquaculture, one bacteriophage (phage), designated as PAS-1, was isolated from the sediment samples of the rainbow trout (Oncorhynchus mykiss) culture farm in Korea. The PAS-1 was morphologically classified as Myoviridae and possessed approximately 48 kb of double-strand genomic DNA. The phage showed broad host ranges to other subspecies of A. salmonicida as well as A. salmonicida subsp. salmonicida including antibiotic-resistant strains. Its latent period and burst size were estimated to be approximately 40 min and 116.7 PFU/cell, respectively. Furthermore, genomic and structural proteomic analysis of PAS-1 revealed that the phage was closely related to other Myoviridae phages infecting enterobacteria or Aeromonas species. The bacteriolytic activity of phage PAS-1 was evaluated using three subspecies of A. salmonicida strain at different doses of multiplicity of infection, and the results proved to be efficient for the reduction of bacterial growth. Based on these results, PAS-1 could be considered as a novel Aeromonas phage and might have potentiality to reduce the impacts of A. salmonicida infections in aquaculture.     

Characterization of Pseudomonas aeruginosa Bacteriophage 95

A new strain of bacteriophage, phage 95, specific to Pseudomonas aeruginosa and Ps. schuylkilliensis was isolated. It has been shown that the phage induces a lytic enzyme which hydrolyzes peptide bond between l-alanine and d-glutamic acid in the peptidoglycan of cell wall.

Here the characteristics of phage 95 are described. It possesses a hexagonal head of 650Å, a contractile tail of 1150Å and spike with fibers. Its DNA has a GC content of 48%, a density of 1.700 g · cm^?3 and Tm of 87.7°C. Photoreactivation was observed. Latent period is 20 min and burst size is 50. The phage is stable between pH 5 and 7.5, and unstable above 55°C. Culture condition and 300 liter scale cultivation are also presented.     

Isolation and Characterization of a Bacteriophage of Arthrobacter globiformis

A bacteriophage which reproduces on Arthrobacter globiformis ATCC 8010 was isolated from soil. This bacteriophage, designated phiAG8010, propagates either in soft agar or broth cultures of the host. Because of a slow adsorption rate, neither the latent period nor burst size was determined. The mature virion belongs to Bradley's group B and exhibits a hexagonal head measuring 69 nm (length) by 60 nm (width) attached to a sheathless tail 120 nm long. The buoyant density of the mature virion is 1.534 g/cm(3). The mature virion contains double-stranded DNA with a buoyant density of 1.722 g/cm(3) (equivalent to 63.3% G + C). Of 14 strains (representing 13 species) of Arthrobacter examined, including A. globiformis ATCC 4336, only A. globiformis ATCC 8010 supported replication of phiAG8010.     

Isolation and Characterization of a Bacteriophage for Vibrio fetus

Bacteriophages were isolated from 22 of 38 strains of Vibrio fetus by an enrichment process, utilizing the donor and host strains growing together in fluid thioglycollate medium. One phage, V-45, isolated by the conventional lawn-spot method, was characterized by stability in broth, growth kinetics, and morphology. It was sensitive to rapid thermal inactivation, chloroform, and pH values above 6.5. Calcium was required for phage replication and stability in broth. Magnesium provided the best protection against thermal inactivation at 50 C in the pH range of 6.5 to 7.5. The minimum latent period was 135 min, rise time was 75 min, and average burst size was 35 plaque-forming units per infected cell. Phage V-45 resembled Bradley's morphological group B, having a long tail without contractile sheath. Dimensions were: head, about 50 nm; tail, about 7 by 240 nm; and tail lumen, 2 to 3 nm.     

Isolation and Characterization of a Bacteriophage for Thiobacillus novellus

A DNA-containing bacteriophage for Thiobacillus novellus has been isolated from sewage and purified by ammonium sulfate precipitation, differential centrifugation, and cesium chloride equilibrium centrifugation. The buoyant density of this phage in CsCl is 1.51 g/cm(3). Electron microscopy studies have revealed a polyhedral head about 60 nm in diameter and a tail surrounded by a number of fine filaments. It has an adsorption rate constant of 1.1 x 10(-9) ml/min, a latent period of 45 min, and an average burst size of 150. The mole guanine and cytosine content in its DNA has been estimated to be 57 to 58%. Five structural proteins with molecular weights of 62,000, 42,500, 30,500, 17,750, and 13,500 have been detected by polyacrylamide gel electrophoresis techniques.     

Isolation of alginate-producing mutants of Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas mendocina

Spontaneous alginate-producing (muc) variants were isolated from strains of Pseudomonas fluorescens, P. putida and P. mendocina at a frequency of 1 in 10(8) by selecting for carbenicillin resistance. The infrared spectrum of the bacterial exopolysaccharide was typical of an acetylated alginate similar to that previously described in Azotobacter vinelandii and in mucoid variants of P. aeruginosa. Mucoid variants were not isolated from P. stutzeri, P. pseudoalcaligenes, P. testosteroni, P. diminuta, P. acidovorans, P. cepacia or P. maltophilia.