Characterization of a new Bacillus megaterium bacteriophage,MJ-1, from tropical soil

A new Bacillus megaterium bacteriophage is characterized. It is a tailed phage with regular polyhedral head belonging to Bradley's group B. Head and tail dimensions are 56.4 and 300 nm, respectively. Lysis was restricted to strains of B. megaterium. No antigenic relationship with pumilus phage FP-1 or subtilis phage FS-1 was observed. The phage is sensitive to 60°C and moderately sensitive to chloroform. The nucleic acid is double-stranded linear DNA with a G-C mole % of 38.8 and a mol wt of (53±3)×10⁶.     

Physiological, morphological, and physicochemical characterization of a novel Escherichia coli bacteriophage, phage MM

A double-stranded DNA containing, T even-like, Escherichia coli bacteriophage, called MM, has been isolated from the local sewage and purified by polyethylene glycol precipitation followed by banding on a cesium chloride three-step gradient. It yields a burst size of 75 particles per infected cell, and has an adsorption coefficient of 3.3 x 10(-10) cm3/min and a latent period of 45 min. Electron microscopy of phage MM reveals an isometric icosahedral head, 92 nm long and 81 nm wide, and a 112-nm-long contractile tail with six pairs of 40-nm-long fibers attached to its baseplate. Phage MM appears similar to E. coli phage T4 or Salmonella phage O1. The density of phage MM in cesium chloride is 1.515 g/ml, and its total mass is 144 MDa. Gel electrophoresis of purified MM capsids displays two major capsid proteins in approximately equimolar amounts and with apparent molecular masses of 38 and 15 kDa. Similarly, purified MM tails yield two major polypeptides with apparent molecular masses of 55 and 16 kDa, most likely representing the major tail sheath and tail tube polypeptides. Its double-stranded DNA has a G-C content of 50%, a length of 131 kilobases (kb), and a mass of 89 MDa.     

Isolation and characterization of a new bacteriophage MMP17 from <Emphasis Type="Italic">Meiothermus</Emphasis>

Thermophiles and their viruses are extraordinarily important because of their roles in processes of evolution, biogeochemistry and ecology. Species of the genus Meiothermus share close relationship with genus Thermus, but no Meiothermus bacteriophage has been reported yet. In this work, a new thermophilic bacteriophage named MMP17 (Meiothermus Myoviridae phage 17) was isolated from a Meiothermus strain and was further characterized. MMP17 was a typical myovirus with an icosahedral head (42 nm in diameter) and a tail (120 nm in length and 17 nm in width). Its DNA was about 33.5–39.5 kb in size. MMP17 was very stable at 55–60°C and pH 6–7. According to the one-step growth curve, the latent period and the burst period were 60 and 30 min, respectively. An average of 15 phage particles was released from each infected cell. Four major bands were detected in purified virion preparation by SDS-PAGE. As MMP17 was a thermophilic bacteriophage with lower production temperature, its characterization and the relationship between MMP17 and Thermus bacteriophages deserved for further study.     

Isolation and characterization of a bacteriophage forBacillus licheniformis A5

The isolation of a virulent bacteriophage forBacillus licheniformis A5 is reported. This bacteriophage, designated NLP-1, has an icosahedral head 100 nm in diameter and a contractible tail with a maximum length of 130 nm. Its DNA has a density of 1.741 g/cm³ and aT _m of 78.4°C. Base composition analysis showed that thymine is absent and is replaced by hydroxymethyluracil. NLP-1 appears to belong to theBacillus group of bacteriophages that includes SP01 and SP82. It will infectB. cereus T andB. brevis 8185, but will not infectB. subtilis W23 or 168.     

Temperate bacteriophage ZF40 of Erwinia carotovora: phage particle structure and DNA restriction analysis

Structural organization of the temperate bacteriophage ZF40 of Erwinia carotovora was studied. Phage ZF40 proved to be a typical member of the Myoviridae family (morphotype A1). Phage particles consist of an isometric head 58.3 nm in diameter and a contractile 86.3-nm-long tail with a complex basal plate and short tail fibers (31.5 nm). Phage tail sheath, a truncated cone in shape, is characterized by specific packaging of structural subunits. The ZF40 phage genome is 45.8 kb in size, as determined by restriction analysis, and contains DNA cohesive ends. The ZF40 phage of Erwinia carotovora is assumed to be a new species of bacteriophages specific for enterobacteria.     

Tail-fiber attachement in bacteriophage T4D studied by quasielastic light scattering–band electrophoresis

Mixtures of bacteriophage T4D particles with up to six tail fibres attached were seprated and analyzed in a quasielastic light scattering-band electrophoresis apparatus. The electrophoretic mobilities and diffusion coefficients of the seprated bands were determined during the same experiment. Species differing in mobility by 0.05 × 10^?4 cm²/V sec were resolved. Henry's electropheresis theory for spheres indicates that each tail fiber contributes about 140 effective positive charges to the phage structure. Estimate of the charge using the permanent dipole moment and the electrophorectic mobility (Bontje et. al. (1977) Biopolymers 16 , 551–572) gives an effective charge of + 230 to ?250 per tail fiber. The charge distribution on the fiberless particles was estimated to be ?3380 on the head and ?290 on the tail if the dipole moment was assumed positive and ?1400 on the head and ?1000 on the tail for a negative dipole. Tail-fiber attachement does not proceed to completion in our invitro system, as substantial distributions as a function of tail-fiber input indicates the reaction is a random, noncooperative process.     

Temperate Bacteriophage ZF40 of Erwinia carotovora: Phage Particle Structure and DNA Restriction Analysis

Structural organization of the temperate bacteriophage ZF40 of Erwinia carotovora was studied. Phage ZF40 proved to be a typical member of the Myoviridae family (morphotype A1). Phage particles consist of an isometric head 58.3 nm in diameter and a contractile 86.3-nm-long tail with a complex basal plate and short tail fibers (31.5 nm). Phage tail sheath, a truncated cone in shape, is characterized by specific packaging of structural subunits. The ZF40 phage genome is 45.8 kb in size, as determined by restriction analysis, and contains DNA cohesive ends. The ZF40 phage ofErwinia carotovora is assumed to be a new species of bacteriophages specific for enterobacteria.     

Characterization of a lysin from deep-sea thermophilic bacteriophage GVE2

Thermostable enzymes from thermophiles have attracted extensive studies. However, little is known about thermophilic lysin of bacteriophage obtained from deep-sea hydrothermal vent. In this study, a lysin from deep-sea thermophilic bacteriophage Geobacillus virus E2 (GVE2) was characterized for the first time. It was found that the GVE2 lysin was highly homologous with N-acetylmuramoyl-L-alanine amidases. After expression in Escherichia coli, the recombinant GVE2 lysin was purified. The recombinant lysin was active over a range of temperature from 40 °C to 80 °C, with an optimum at 60 °C. Its optimal pH was 6.0, and it was stable over a wide range of pH from 4.0 to 10.0. The lysin was highly active when some enzyme inhibitors or detergents (phenylmethylsulfonyl fluoride, Tween 20, Triton X-100, and chaps) were used. However, it was strongly inhibited by sodium dodecyl sulfate and ethylene diamine tetraacetic acid. Its enzymatic activity could be slightly stimulated in the presence of Na⁺ and Li⁺. But the metal ions Mg²⁺, Ba²⁺, Zn²⁺, Fe³⁺, Ca²⁺, and Mn²⁺ at concentrations of 1 or 10 mM showed inhibitions to the lysin activity. Our study demonstrated the first characterization of lysin from deep-sea thermophilic bacteriophage.     

Characterization of a bacteriophage related to R-type pyocins.

A bacteriophage with a contractile tail which shows very similar features to R-type pyocins was isolated and characterized. This phage, named PS17,was purified by DEAE-cellulose chromatography and CsCl density gradient centrifugation. It was a DNA-containing phage, and the density of the purified particles in CsCl was found to be 1.468. DNA from this phage had a density of 1.720 in CsCl, indicating its guanine plus cytosine content to be 61.2%. The head was polyhedral, 69 nm in diameter, and the tail was 150 nm in length. This phage was neutralized by antiserum preparations against five R-type pyocins, and the antiserum against this phage was active in neutralizing R-type pyocins. The properties of this phage, PS17, were compared with another similar phage, PS3, which was previously reported.     

多重耐药肺炎克雷伯菌噬菌体KP002的生物学特性及基因组学研究

以上海某些医院临床分离到的多重耐药肺炎克雷伯菌为宿主菌,从不同环境的污水中分离获得1株肺炎克雷伯菌噬菌体KP002。电子显微镜显示其为有尾噬菌体,头部直径约70nm,尾长约80nm,尾宽约20nm。对其生物学特性进行研究,结果显示此株噬菌体在pH 3~9及4~50℃的环境中具有较高活性;6min吸附率达95%以上;潜伏期为10min,爆发期为50min;裂解量为172pfu/cell。结果表明,该噬菌体对pH值和温度适应范围较宽。对其全基因组进行测序分析,结果显示其基因组为环状双链DNA,全长47 173bp,GC含量为48%。本研究筛选获得1株对pH值和温度适应范围较宽的耐药肺炎克雷伯菌烈性噬菌体KP002,为建立耐药肺炎克雷伯菌的噬菌体库以用于治疗临床多重耐药菌感染提供了新的思路。     

BVPaP-3, a T7-Like Lytic Phage of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>: Its Isolation and Characterisation

The increasing emergence of antibiotic-resistant bacteria has produced a growing interest among scientists in bacteriophages as alternative antimicrobial agents. This article reports a lytic phage against an antibiotic-resistant strain of Pseudomonas aeruginosa. Phage BVPaP-3 is a member of the Podoviridae family and morphologically similar to the T7-like phage gh-1. The phage has a hexagonal head of 58–59 nm in diameter and a short tail of 10 × 8 nm. It is stable at a wide range of pH (6–10) and temperatures (4–40°C). Its optimal growth temperature is 37°C and the adsorption rate constant is 1.19 × 10⁻⁹. Latent and eclipse periods are 20 and 15 min, respectively, and the burst size is 44 after 35 min at 37°C. The phage has a DNA size of 41.31 kb and a proteome of 11 proteins. The major protein is 33 kDa in size.     

明永冰川融水中一株裂解性低温噬菌体的分离及特征

【目的】高山冰川是一类独特的生态系统,本研究探索从明永冰川地区分离和培养低温菌噬菌体,并对其特征进行研究。【方法】利用已分离的低温菌为宿主,采用"双层平板法"从明永冰川融水中分离纯化低温菌噬菌体;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长度多态性分析、衣壳蛋白组成分析及噬菌体生理特征研究。【结果】从明永冰川融水中分离获得一株裂解性低温噬菌体,命名为MYSP03(Mingyong Flavobacterium Siphoviridae Bacteriophage),其宿主菌MYB03鉴定为Flavobacterium菌株。噬菌体MYSP03为长尾型,无囊膜,头部具典型的正多面体立体对称结构,直径约72 nm;尾管长约240 nm,直径约10 nm;4℃时具侵染活性,在4℃-20℃范围内均可产生边缘清晰、透明的噬菌斑,最适感染温度约10℃,pH耐受范围较广,最适感染pH约9.4,对氯仿不敏感,基因组为双链DNA,大小约66 kb。     

Isolation and characterization of a temperate bacteriophage from the ruminal anaerobe Selenomonas ruminantium

A temperate bacteriophage was obtained from an isolate of the ruminal anaerobe Selenomonas ruminantium. Clear plaques that became turbid on further incubation occurred on a lawn of host bacteria. Cells picked from a turbid plaque produced healthy liquid cultures, but these often lysed on storage. Mid-log-phase liquid cultures incubated with the bacteriophage lysed and released infectious particles with a titer of up to 3 X 10(7) PFU/ml. A laboratory strain of S. ruminantium, HD-4, was also sensitive to this bacteriophage, which had an icosohedral head (diameter, 50 nm) and a flexible tail (length, 140 nm). The bacteriophage contained 30 kilobases of linear, double-stranded DNA, and a detailed restriction map was constructed. The lysogenic nature of infection was demonstrated by hybridization of bacteriophage DNA to specific restriction fragments of infected host genomic DNA and by identification of a bacteriophage genomic domain which may participate in integration of the bacteriophage DNA. Infection of S. ruminantium in vitro was demonstrated by two different methods of cell transformation with purified bacteriophage DNA.     

Isolation and characterization of a temperate bacteriophage from the ruminal anaerobe Selenomonas ruminantium.

A temperate bacteriophage was obtained from an isolate of the ruminal anaerobe Selenomonas ruminantium. Clear plaques that became turbid on further incubation occurred on a lawn of host bacteria. Cells picked from a turbid plaque produced healthy liquid cultures, but these often lysed on storage. Mid-log-phase liquid cultures incubated with the bacteriophage lysed and released infectious particles with a titer of up to 3 X 10(7) PFU/ml. A laboratory strain of S. ruminantium, HD-4, was also sensitive to this bacteriophage, which had an icosohedral head (diameter, 50 nm) and a flexible tail (length, 140 nm). The bacteriophage contained 30 kilobases of linear, double-stranded DNA, and a detailed restriction map was constructed. The lysogenic nature of infection was demonstrated by hybridization of bacteriophage DNA to specific restriction fragments of infected host genomic DNA and by identification of a bacteriophage genomic domain which may participate in integration of the bacteriophage DNA. Infection of S. ruminantium in vitro was demonstrated by two different methods of cell transformation with purified bacteriophage DNA.     

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.     

Characterization of a virulent Lactobacillus sake phage PWH2

A virulent bacteriophage PWH2 was isolated from fermented sausage. Out of 14 strains of Lactobacillus sake and 5 strains of L. curvatus tested as potential hosts, only L. sake Ls2 was sensitive. The plaques had a diameter of 0.5-1 mm. One-step growth kinetics of bacteriophage PWH2 revealed the following characteristics: a latent period of 1.5 h, a rise period of 1 h and a burst size of 110 (⁺ _– 10) phages per cell. From electron micrographs it was deduced that bacteriophage PWH2 has an icosahedral head, a long non-contractile tail and a complex base plate. The genome is linear and 35 kbp in length. The structural proteins consist of three major and two minor proteins. After an infection of L. sake Ls2, isolates were obtained that were resistant to PWH2. By treatment with mitomycin C, isolate R4a was found to be lysogenic. DNA/DNA hybridisation proved homology of phage PWH2 with the chromosomal DNA of strain R4a. Correspondence to: W. P. Hammes     

Isolation and characterization of a Lactobacillus plantarum bacteriophage isolated from a meat starter culture*

Morphological characterization of a bacteriophage isolated from the Lactobacillus plantarum portion of a commercial meat starter culture showed that the isolate, phage fri, belonged to the Bradley group A bacteriophages. It had a regular six-sided head (90 nm diameter), and a contractile tail (190 nm in length). Short tail fibres were observed at the distal end of the sheath. Fluorescent staining with acridine orange indicated that phage fri contained double-stranded DNA. The resistance to high concentrations of either chloroform or ether showed that its lipid content was negligible. Heat lability was demonstrated by inactivation of a phage fri population within 10 min at 60°C and within 5 min at 70°C. It tolerated pH levels of 3.0–8.0 and exhibited greater stability in the acid region than did its host strain. The latent and rise periods were both 75 min, and the average burst size 200 pfu/cell. Sensitivity was limited to the Lact. plantarum strain of only one manufacturer of the commercial meat starters investigated.     

Molecular evidence for a new bacteriophage of Borrelia burgdorferi

We have recovered a DNase-protected, chloroform-resistant molecule of DNA from the cell-free supernatant of a Borrelia burgdorferi culture. The DNA is a 32-kb double-stranded linear molecule that is derived from the 32-kb circular plasmids (cp32s) of the B. burgdorferi genome. Electron microscopy of samples from which the 32-kb DNA molecule was purified revealed bacteriophage particles. The bacteriophage has a polyhedral head with a diameter of 55 nm and appears to have a simple 100-nm-long tail. The phage is produced constitutively at low levels from growing cultures of some B. burgdorferi strains and is inducible to higher levels with 10 microg of 1-methyl-3-nitroso-nitroguanidine (MNNG) ml(-1). In addition, the prophage can be induced with MNNG from some Borrelia isolates that do not naturally produce phage. We have isolated and partially characterized the phage associated with B. burgdorferi CA-11.2A. To our knowledge, this is the first molecular characterization of a bacteriophage of B. burgdorferi.     

Isolation and characterization of a bacteriophage infectious to an extreme thermophile, Thermus thermophilus HB8.

A bacteriophage (phiYS40) infectious to an extreme thermophile, Thermus thermophilus HB8, was isolated and characterized. phiYS40 grows over the temperature range of 56 to 78 C, and the optimum growth temperature is about 65 C. The phage had a latent period of 80 min and a burst size of about 80 at 65 C. The phage has a hexagonal head 0.125 mum in diameter, a tail 0.178 mum long and 0.027 mum wide, a base plate and tail fibers. The phage is thermostable in broth but rather unstable in a buffer containing 10 mM Tris, 10 mM MgCl2, pH 7.5. The addition of Casamino Acids (1 percent), polypeptone (0.8 percent), yeast extract (0.4 percent), NaCl (0.1 M) or spermidine (1 mM) to the buffer restores the thermostability of phiYS40 to the same degree as in broth. The phage is also thermostable in water of the hot spring from which this phage was isolated. The nucleic acid of PhiYS40 is a double-stranded DNA and has a molecular weight of 1.36 X 10-8. The guanine plus cytosine content of the DNA was determined to be about 35 percent from chemical determinations, buoyant density (1.693 g/cm-3 in CsCl), and melting temperature (83.5 C in 0.15 M NaCl plus 0.015 M sodium citrate).     

Basic characterization of a Pseudomonas aeruginosa PAO1 bacteriophage

A bacteriophage of Pseudomonas aeruginosa PAO1 was characterized. Bacteriophage PIK was found to adsorb on the cell wall of the host organism. Electron microscopy of the phage PIK revealed that it had a bipyramidal hexagonal prismatic head of 110 nm in diameter, a tail which was 158 nm long and a tail plate of 47 nm width. This paper describes its basic characters, and a quantitative study was made of its adsorption to exponential phase cells of two different strains of P. aeruginosa. PIK was found to contain double stranded DNA and it appears to be virulent towards its host, P. aeruginosa PAO1. It was classified into the group of phages possessing a contractile tail.