Characterization of a Thermophilic Bacteriophage for Bacillus stearothermophilus

Abstract

, AND L. LEON CAMPBELL. Characterization of a thermophilic bacteriophage for Bacillus stearothermophilus. J. Bacteriol. 91:340-348. 1965.-The biological and physical-chemical properties of the thermophilic bacteriophage TP-84 were investigated. TP-84 was shown to be lytic for 3 of 24 strains of Bacillus stearothermophilus tested over the temperature range of 43 to 76 C. The latent period of TP-84 on B. stearothermophilus strain 10 was 22 to 24 min. TP-84 has a hexagonal head, 53 m,u in diameter and 30 m,u on a side; its tail is 130 m,u long and 3 to 5 m,u wide. The phage has an S'20 of 436, and bands at a density of 1.508 g/cc in CsCl (pH 8.5). The diffusion coefficient of TP-84

Diffusion properties of bacteriophages through agarose gel membrane

“…In quantitative value, diffusion coefficients of phages were measured in previous studies to be 3 × 10 −12 m 2 /s – 3 × 10 −11 m 2 /s ( E. coli phages in water at 20°C),18, 29, 30 in agreement with the D app values we measured. Compared with the diffusion coefficients of antibiotics, the phages' diffusion coefficients are much smaller, agreeing with previous hypotheses that slow penetration rates might be a big bottleneck for treatment of biofilms with phage therapy.…”

Production and Purification of the Thermophilic Bacteriophage TP-84

“…Like the mesophilic phages, the thermophilic ones can be subdivided into two main groups: (i) virulent thermophilic phages (2-7, 10,12,13,16,18,19,27,28) and (ii) temperate phages (7, 17, 20-23, 25, 26). Unique in their extreme thermal stability (6,10,12,13,16,19,20,25,27,28), the thermophilic phages have attracted the attention of many investigators. Our interest in the thermophilic phage TP-84 derives from the assumption that thermophilic phages may represent the simplest form of thermophilic organisms and thus the mechanism(s) for their remarkable thermostability might be easier to elucidate.…”

“…The thermophilic bacteriophage used was isolated from greenhouse soil in 1952 by one of us (L. L. C.), using B. stearothermophilus strain 2184 as the host organism. Some of its properties have been reported (3,18,19).…”

“…Media and culture conditions. The media and growth conditions used in the early stages of this work have been described (19). 'Present address: Department of Bacteriology, The Hebrew University Hadassah Medical School, Jerusalem, Israel.…”

“…2. With either technique, the final purification of the phage was done by CsCl density gradient centrifugation as described by Saunders and Campbell (19). Solid CsCl (purity 99.9%, Penn Rare Metals Division) to a final density of 1.51 g/ml was added either directly to the dissolved magnesium carbonate-phage suspension or to the NaCl-polyethylene glycol-precipitated phage after it was suspended in TMC buffer containing 0.…”