scholarly journals PHAGE FORMATION IN STAPHYLOCOCCUS MUSCAE CULTURES

1950 ◽  
Vol 34 (2) ◽  
pp. 231-250 ◽  
Author(s):  
Winston H. Price
Keyword(s):  
Aspartic Acid ◽  
Burst Size ◽  
Synthetic Medium ◽  
Growth Curves ◽  
Acid Synthesis ◽  
Virus Protein ◽  
Complete Medium ◽  
Multiplication Rate ◽  
One Step ◽  
Step Growth

1. Four strains of Staphylococcus muscae have been isolated which differ in their growth rates and phage syntheses in Fildes' synthetic medium. 2. Two of the strains when singly infected cannot release phage in Fildes' synthetic medium unless a substance present in certain acid-hydrolyzed proteins is added to the medium. One of these strains also requires other substance(s) present in acid-hydrolyzed proteins in order to grow in Fildes' medium. 3. The two strains which do not require the addition of the phage-stimulating factor have been found either to synthesize this substance, or one similar to it. One of these strains will not grow in Fildes' medium unless substance(s) present in acid-hydrolyzed proteins is added to the medium. 4. The purified acid-hydrolyzed protein factor necessary for virus liberation does not affect the multiplication rate of uninfected S. muscae cells in Fildes' synthetic medium. 5. The substance is not needed for the adsorption or the invasion of the host cell by the virus. In the absence of the factor, the virus is adsorbed to the cell and "kills" it. 6. An analysis carried out by means of the one-step growth curve technique has indicated that the substance is not concerned simply with the mechanism of virus release, but is necessary for some initial stage in virus synthesis. 7. With one bacterial strain not requiring the AHPF, aspartic acid had to be present at least during the minimum latent period for the cell to form virus. 8. In the absence of aspartic acid, the virus was adsorbed to the cell and killed it, but no virus was released from singly infected bacteria. 9. If the cells were grown in a medium containing aspartic acid and then resuspended in the medium minus aspartic acid, no virus was released, although such cells contained at least two times the amount of aspartic acid necessary for the burst size in the complete medium. 10. Aspartic acid, a constituent of the virus particle, appears from an analysis of one-step growth curves to take part in the initial phase of phage synthesis. 11. The effect of amino acids on virus formation is discussed in relation to the time sequence of virus protein and desoxyribonucleic acid synthesis.

scholarly journals PHAGE FORMATION IN STAPHYLOCOCCUS MUSCAE CULTURES

1950 ◽  
Vol 34 (2) ◽  
pp. 251-277 ◽  
Author(s):  
Winston H. Price
Keyword(s):  
Aspartic Acid ◽  
Phage Particle ◽  
Burst Size ◽  
Synthetic Medium ◽  
Acid Synthesis ◽  
Active Particle ◽  
Multiple Infection ◽  
Experimental Conditions ◽  
Virus Particles ◽  
Infected Cells

1. A strain of S. muscae which requires a substance present in certain acid-hydrolyzed proteins (AHPF) for virus liberation when singly infected in Fildes' synthetic medium no longer needs this substance when multiply infected. 2. In the absence of the AHPF under conditions of multiple infection the amount of phage released is approximately equal to the number of infecting particles between two to ten. Over ten particles per cell has no further effect on the yield of virus. 3. The experimental evidence indicates that it is the phage particle and not some other component in the lysate which can replace the AHPF. 4. The minimum latent period and rise period of cells singly infected in the presence of the AHPF and multiply infected in the absence of the AHPF are the same. 5. The desoxynucleic acid synthesis of cells, infected with a very few virus particles in the presence of excess AHPF and multiply infected with ten particles in the absence of the AHPF, occurs at approximately the same rate, with both infected samples synthesizing about the same amount of desoxynucleic acid and liberating the same yields of virus. 6. A strain of S. muscae which requires aspartic acid for virus synthesis when singly infected does not need this substance when multiply infected, the burst size under the latter conditions depending upon the multiplicity of infection between 3 to 12 particles per cell. 7. The data indicate that the virus released from multiply infected cells in the absence of added AHPF or aspartic acid is newly synthesized virus and not the original infecting particles. 8. The phage particle contains the AHPF and aspartic acid. 9. As a tentative working hypothesis, it is assumed that the AHPF and aspartic acid for phage formation under conditions of multiple infection, in the absence of added AHPF, or of aspartic acid, are contributed by the original infecting particles. 10. Ultraviolet-inactivated phage is adsorbed to the host cell and kills the cell although little virus is released under the experimental conditions. 11. Ultraviolet-inactivated phage particles, if added before the active particle is adsorbed, will greatly inhibit the liberation of new virus particles; but does not do so if added a few minutes after the active particle has been adsorbed. 12. Under the experimental conditions, reactivation of phage when present in multiply infected cells does not occur; and such ultraviolet-inactivated phage cannot serve as a source of the AHPF or aspartic acid, although the AHPF can be liberated from such inactivated particles by acid hydrolysis. 13. The results are discussed in relation to Luria's experiments with ultraviolet-treated phage and to his "gene pool" hypothesis of phage formation.

scholarly journals Aztreonam Lysine Increases the Activity of Phages E79 and phiKZ against Pseudomonas aeruginosa PA01

2021 ◽  
Vol 9 (1) ◽  
pp. 152
Author(s):  
Carly M. Davis ◽  
Jaclyn G. McCutcheon ◽  
Jonathan J. Dennis
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Morphological Changes ◽  
Burst Size ◽  
Type Iv Pili ◽  
Biofilm Growth ◽  
Promising Alternative ◽  
Type Iv ◽  
Alternative Treatment Option ◽  
One Step ◽  
Step Growth

Pseudomonas aeruginosa is a pernicious bacterial pathogen that is difficult to treat because of high levels of antibiotic resistance. A promising alternative treatment option for such bacteria is the application of bacteriophages; the correct combination of phages plus antibiotics can produce synergistic inhibitory effects. In this study, we describe morphological changes induced by sub-MIC levels of the antibiotic aztreonam lysine (AzLys) on P. aeruginosa PA01, which may in part explain the observed phage–antibiotic synergy (PAS). One-step growth curves for phage E79 showed increased adsorption rates, decreased infection latency, accelerated time to lysis and a minor reduction in burst size. Phage E79 plus AzLys PAS was also able to significantly reduce P. aeruginosa biofilm growth over 3-fold as compared to phage treatment alone. Sub-inhibitory AzLys-induced filamentation of P. aeruginosa cells resulted in loss of twitching motility and a reduction in swimming motility, likely due to a reduction in the number of polar Type IV pili and flagella, respectively, on the filamented cell surfaces. Phage phiKZ, which uses Type IV pili as a receptor, did not exhibit increased activity with AzLys at lower sub-inhibitory levels, but still produced phage–antibiotic synergistic killing with sub-inhibitory AzLys. A one-step growth curve indicates that phiKZ in the presence of AzLys also exhibits a decreased infection latency and moderately undergoes accelerated time to lysis. In contrast to prior PAS studies demonstrating that phages undergo delayed time to lysis with cell filamentation, these PAS results show that phages undergo accelerated time to lysis, which therefore suggests that PAS is dependent upon multiple factors, including the type of phages and antibiotics used, and the bacterial host being tested.

Fiber1, but not fiber2, is the essential fiber gene for fowl adenovirus 4 (FAdV-4)

2021 ◽  
Author(s):  
Xiaohui Zou ◽  
Yejing Rong ◽  
Xiaojuan Guo ◽  
Wenzhe Hou ◽  
Bingyu Yan ◽  
...  
Keyword(s):  
Reverse Genetics ◽  
Growth Curves ◽  
Dependent Manner ◽  
Helper Plasmid ◽  
Fowl Adenovirus ◽  
Virus Adsorption ◽  
One Step ◽  
Step Growth ◽  
Lmh Cells

Fibre is the viral protein that mediates the attachment and infection of adenovirus to the host cell. Fowl adenovirus 4 (FAdV-4) possesses two different fibre trimers on each penton capsomere, and roles of the separate fibres remain elusive. Here, we attempted to investigate the function of FAdV-4 fibres by using reverse genetics approaches. Adenoviral plasmids carrying fiber1 or fiber2 mutant genes were constructed and used to transfect chicken LMH cells. Fiber1-mutated recombinant virus could not be rescued. Such defective phenotype was complemented when a fiber1-bearing helper plasmid was included for co-transfection. The infection of fiber-intact FAdV-4 (FAdV4-GFP) to LMH cells could be blocked with purified fiber1 knob protein in a dose-dependent manner, while purifed fiber2 knob had no such function. On the contrary, fiber2-mutated FAdV-4, FAdV4XF2-GFP, was successfully rescued. The results of one-step growth curves showed that proliferative capacity of FAdV4XF2-GFP was 10 times lower than that of the control FAdV4-GFP. FAdV4XF2-GFP also caused fewer deaths of infected chicken embryos than FAdV4-GFP did, which resulted from poorer virus replication in vivo. These data illustrated that fiber1 mediated virus adsorption and was essential for FAdV-4, while fiber2 was dispensable although it significantly contributed to the virulence.

STUDIES OF INFECTIOUS BOVINE RHINOTRACHEITIS VIRUS: 1. PLAQUE ASSAY AND SOME CHARACTERISTICS IN BOVINE KIDNEY CELLS

1963 ◽  
Vol 9 (4) ◽  
pp. 567-576 ◽  
Author(s):  
Leslie R. Sabina ◽  
Raymond C. Parker
Keyword(s):  
Infectious Virus ◽  
Plaque Assay ◽  
Growth Curves ◽  
Kidney Cells ◽  
Infectious Bovine Rhinotracheitis ◽  
High Input ◽  
Bovine Kidney ◽  
One Step ◽  
Step Growth ◽  
Infectious Bovine Rhinotracheitis Virus

A reproducible plaquing procedure for infectious bovine rhinotracheitis virus (IBRV) in an established bovine kidney cell line is reported. The validity of this system for quantitative analysis has been established by conventional methods.After infection at different multiplicities, one-step growth curves have shown that the eclipse period for IBRV lasts approximately 4 hours and that the infectious virus increases at a logarithmic rate for 12 to 14 hours. The virus yield with the low and high input is 30 PFU and 210 PFU per cell, respectively. Only 1 to 9% of the total virus is released at 24 hours postinfection. The data presented indicate the half-life of IBRV at 37 °C and 42 °C to be 16 and 3.5 hours, respectively. A comparison of hyperimmune bovine and rabbit sera has shown that 92% of the infective particles are neutralized within 30 minutes.

scholarly journals External factors limiting the multiplication potential of Tetrahymena

1981 ◽  
Vol 50 (1) ◽  
pp. 407-418
Author(s):  
E. Hofmann ◽  
G. Cleffmann
Keyword(s):  
Cell Density ◽  
Synthetic Medium ◽  
Reference Conditions ◽  
Nitrogen Sources ◽  
Complete Medium ◽  
Multiplication Rate ◽  
Or Groups ◽  
Cell Densities ◽  
Culture Growth ◽  
O2 Supply

By variation of nutritional and other external conditions we have determined the factors that limit the multiplication rate and the culture growth in Tetrahymena thermophila. The enriched synthetic medium of Kidder & Dewey (1951), a culture temperature of 29 degrees C, and aeration by agitation were chosen as reference conditions. The final cell density is increased by and proportional to the amount of the complete set of nutrients. Testing single nutritional factors or groups of them revealed that only nitrogen sources yield higher cell densities. But none of them or any combination is as capable of increasing the cell density as the complete medium. Therefore, the medium has to be considered as well balanced. Ammonia, cell density, O2 supply, and pH have been excluded as factors limiting the capacity for multiplication. There are no known factors promoting or inhibiting culture growth.

scholarly journals Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1

2012 ◽  
Vol 194 (18) ◽  
pp. 5073-5079 ◽  
Author(s):  
Sherin Kannoly ◽  
Yongping Shao ◽  
Ing-Nang Wang
Keyword(s):  
Genome Organization ◽  
Cell Attachment ◽  
Phylogenetic Analyses ◽  
Bacterial Species ◽  
Growth Curves ◽  
Conjugative Plasmid ◽  
Content Type ◽  
Conjugative Plasmids ◽  
One Step ◽  
Step Growth

ABSTRACTWe have sequenced and characterized two R-plasmid-dependent single-stranded RNA bacteriophages (RPD ssRNA phages), C-1 and Hagl1. Phage C-1 requires a conjugative plasmid of the IncC group, while Hgal1 requires the IncH group. Both the adsorption rate constants and one-step growth curves are determined for both phages. We also empirically confirmed the lysis function of the predicted lysis genes. Genomic sequencing and phylogenetic analyses showed that both phages belong to theLevivirusgroup and are most closely related to another IncP-plasmid-dependent ssRNA phage, PRR1. Furthermore, our result strongly suggests that the stereotypical bauplans of genome organization found inLevivirusandAlloleviviruspredate phage specialization for conjugative plasmids, suggesting that the utilization of conjugative plasmids for cell attachment and entry comprises independent evolutionary events for these two main clades of ssRNA phages. Our result is also consistent with findings of a previous study, making theLevivirus-like genome organization ancestral and theAllolevivirus-like genome derived. To obtain a deeper insight into the evolution of ssRNA phages, more phages specializing for various conjugative plasmids and infecting different bacterial species would be needed.

One-step growth curves for vaccinia virus in cultures of monkey kidney cells

Virology ◽  
1959 ◽  
Vol 9 (3) ◽  
pp. 386-395 ◽  
Author(s):  
G. Furness ◽  
J.S. Youngner
Keyword(s):  
Vaccinia Virus ◽  
Growth Curves ◽  
Monkey Kidney ◽  
Kidney Cells ◽  
One Step ◽  
Step Growth
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