GROWTH OF PENICILLIN-RESISTANT AND PENICILLIN-SENSITIVE STRAINS OF STAPHYLOCOCCUS AUREUS

1963 ◽  
Vol 9 (2) ◽  
pp. 179-186
Author(s):  
Wendall E. Allen ◽  
Ilda McVeigh

Ten strains of naturally penicillin-resistant Staphylococcus aureus (obtained from patients), two in vitro derived resistant strains, and two sensitive strains, were grown at 37 C in Antibiotic Assay broth, and viable cell determinations were made at intervals. From these data, growth curves were plotted for each of the strains. The curves for the naturally penicillin-resistant and the sensitive strains are very similar. Little, if any, lag in growth of these strains occurred on transfer from maximum stationary-phase cultures to fresh medium. They grew at approximately the same rate during the logarithmic growth phase, which lasted for 3 to 4 hours; during the maximum stationary phase, about the same number of cells was present per milliliter in cultures of each of these strains. In contrast, the in vitro derived resistant strains underwent a lag of 2 to 6 hours on transfer to fresh medium and grew at a slower rate during the logarithmic growth phase. However, during the maximum stationary phase, which occurred after an incubation period of 24 to 32 hours, the cell titers were approximately the same as those of the naturally resistant and the sensitive strains. When grown in competition with either of the sensitive strains in Antibiotic Assay broth in the absence of penicillin, one of the naturally resistant strains persisted for 14 successive subcultures without any apparent change in ability to tolerate the antibiotic.

Author(s):  
B. L. Soloff ◽  
T. A. Rado

Mycobacteriophage R1 was originally isolated from a lysogenic culture of M. butyricum. The virus was propagated on a leucine-requiring derivative of M. smegmatis, 607 leu−, isolated by nitrosoguanidine mutagenesis of typestrain ATCC 607. Growth was accomplished in a minimal medium containing glycerol and glucose as carbon source and enriched by the addition of 80 μg/ ml L-leucine. Bacteria in early logarithmic growth phase were infected with virus at a multiplicity of 5, and incubated with aeration for 8 hours. The partially lysed suspension was diluted 1:10 in growth medium and incubated for a further 8 hours. This permitted stationary phase cells to re-enter logarithmic growth and resulted in complete lysis of the culture.


2002 ◽  
Vol 70 (6) ◽  
pp. 2758-2762 ◽  
Author(s):  
Christiane Wolz ◽  
Christiane Goerke ◽  
Regine Landmann ◽  
Werner Zimmerli ◽  
Ursula Fluckiger

ABSTRACT Staphylococcus aureus is one of the pathogens most frequently isolated in device-related infections. S. aureus is equipped with surface-associated proteins promoting specific binding to matrix molecules. Clumping factor A (ClfA, encoded by clfA) mediates adhesion to fibrinogen. Whereas the contribution of ClfA to pathogenicity is well documented, the influence of different growth and host parameters on gene activity is unclear. To elucidate this question, we investigated clfA transcript levels in an animal model of device-related infection and in planktonic and sessile bacteria grown in vitro. Specific mRNA from the S. aureus strains Newman, Reynolds, and RN6390 was quantified by LightCycler reverse transcription-PCR. In vitro, clfA transcript levels were low in the early logarithmic growth phase, but a clear increase was observed after the late logarithmic phase. Quantities of clfA transcripts were four to six times higher in the planktonic than in the sessile bacterial subpopulations grown to the stationary phase. During infection, in strains Newman and Reynolds levels of clfA transcripts in exudates accumulating in the infected devices were lower than those in the bacteria grown in vitro to stationary phase. clfA mRNA levels in the exudates increased during the initial phase of infection and remained constant after 96 h postinoculation. In contrast to the in vitro results, quantities of clfA transcripts in the unattached bacteria of the exudates never exceeded the level of clfA transcripts in the sessile bacteria attached to glass beads. However, a clear increase in clfA quantities in the sessile bacteria was observed late in infection after 144 h. In conclusion, maximal clfA transcript levels are reached late during growth in vitro and in vivo.


2012 ◽  
Vol 78 (7) ◽  
pp. 2120-2127 ◽  
Author(s):  
Lei Liu ◽  
Huichun Tong ◽  
Xiuzhu Dong

ABSTRACTComplex interspecies interactions occur constantly between oral commensals and the opportunistic pathogenStreptococcus mutansin dental plaque. Previously, we showed that oral commensalStreptococcus oligofermentanspossesses multiple enzymes for H2O2production, especially lactate oxidase (Lox), allowing it to out-competeS. mutans. In this study, through extensive biochemical and genetic studies, we identified a pyruvate oxidase (pox) gene inS. oligofermentans. Apoxdeletion mutant completely lost Pox activity, while ectopically expressedpoxrestored activity. Pox was determined to produce most of the H2O2in the earlier growth phase and log phase, while Lox mainly contributed to H2O2production in stationary phase. Bothpoxandloxwere expressed throughout the growth phase, while expression of theloxgene increased by about 2.5-fold when cells entered stationary phase. Since lactate accumulation occurred to a large degree in stationary phase, the differential Pox- and Lox-generated H2O2can be attributed to differential gene expression and substrate availability. Interestingly, inactivation ofpoxcauses a dramatic reduction in H2O2production from lactate, suggesting a synergistic action of the two oxidases in converting lactate into H2O2. In anin vitrotwo-species biofilm experiment, thepoxmutant ofS. oligofermentansfailed to inhibitS. mutanseven thoughloxwas active. In summary,S. oligofermentansdevelops a Pox-Lox synergy strategy to maximize its H2O2formation so as to win the interspecies competition.


1972 ◽  
Vol 18 (11) ◽  
pp. 1773-1775 ◽  
Author(s):  
M. M. Shahin

Protoplasts prepared from cells of different stages within the logarithmic growth phase and from transition phase showed different degrees of colony-forming ability. The cells yielding higher frequency of protoplast formation also gave protoplast with a high degree of colony-forming ability.


2005 ◽  
Vol 187 (6) ◽  
pp. 1951-1958 ◽  
Author(s):  
Toshiko Aiso ◽  
Hideji Yoshida ◽  
Akira Wada ◽  
Reiko Ohki

ABSTRACT The expression of ribosome modulation factor (RMF) is induced during stationary phase in Escherichia coli. RMF participates in the dimerization of 70S ribosomes to form the 100S ribosome, which is the translationally inactive form of the ribosome. To elucidate the involvement of the control of mRNA stability in growth-phase-specific rmf expression, we investigated rmf mRNA stability in stationary-phase cells and cells inoculated into fresh medium. The rmf mRNA was found to have an extremely long half-life during stationary phase, whereas destabilization of this mRNA took place after the culture was inoculated into fresh medium. RMF and 100S ribosomes disappeared from cells 1 min after inoculation. In addition to control by ppGpp-dependent transcription, these results indicate that the modulation of rmf mRNA stability is also involved in the regulation of growth-phase-specific rmf expression. Unexpectedly, the postinoculation degradation of rmf mRNA was suppressed by the addition of rifampin, suggesting that de novo RNA synthesis is necessary for degradation. This degradation was also suppressed in both a poly(A) polymerase-deficient and an rne-131 mutant strain. We cloned and sequenced the 3′-proximal regions of rmf mRNAs and found that most of these 3′ ends terminated at the ρ-independent terminator with the addition of a one- to five-A oligo(A) tail in either stationary-phase or inoculated cells. No difference was observed in the length of the poly(A) tail between stationary-phase and inoculated cells. These results suggest that a certain postinoculation-specific regulatory factor participates in the destabilization of rmf mRNA and is dependent on polyadenylation.


2019 ◽  
Vol 88 (1) ◽  
Author(s):  
Payal P. Balraadjsing ◽  
Lisbeth D. Lund ◽  
Yuri Souwer ◽  
Sebastian A. J. Zaat ◽  
Hanne Frøkiær ◽  
...  

ABSTRACT Staphylococcus aureus has evolved different strategies to evade the immune response, which play an important role in its pathogenesis. The bacteria express and shed various cell wall components and toxins during different stages of growth that may affect the protective T cell responses to extracellular and intracellular S. aureus. However, if and how the dendritic cell (DC)-mediated T cell response against S. aureus changes during growth of the bacterium remain elusive. In this study, we show that exponential-phase (EP) S. aureus bacteria were endocytosed very efficiently by human DCs, and these DCs strongly promoted production of the T cell polarizing factor interleukin-12 (IL-12). In contrast, stationary-phase (SP) S. aureus bacteria were endocytosed less efficiently by DCs, and these DCs produced small amounts of IL-12. The high level of IL-12 production induced by EP S. aureus led to the development of a T helper 1 (Th1) cell response, which was inhibited after neutralization of IL-12. Furthermore, preincubation with the staphylococcal cell wall component peptidoglycan (PGN), characteristically shed during the exponential growth phase, modulated the DC response to EP S. aureus. PGN preincubation appeared to inhibit IL-12p35 expression, leading to downregulation of IL-12 and an increase of IL-23 production by DCs, enhancing Th17 cell development. Taken together, our data indicate that exponential-phase S. aureus bacteria induce a stronger IL-12-dependent Th1 cell response than stationary-phase S. aureus and that this Th1 cell response shifted toward a Th17 cell response in the presence of PGN.


1970 ◽  
Vol 68 (1) ◽  
pp. 121-128 ◽  
Author(s):  
Barbara M. Hall

SummaryReasons for the accumulation of mercury-resistant strains of Staphylococcus aureus in hospital have been studied. A collection of paired strains, that is staphylococci similar in every respect except sensitivity to mercury salts, was made. Tests were made in an attempt to demonstrate a link between mercury resistance and some other factor which might aid survival, viz. resistance to drying and heat, production of bound coagulase, growth in the presence of sublethal amounts of tetracycline, survival in human blood at 37°C. and uptake by polymorphs at 30°0. and 37°C., development of resistance to antibiotics and competition in mixed cultures. It was not possible to demonstrate any consistent link between mercury resistance and any of these properties. Paper strips impregnated with the mercurial diuretic, Mersalyl, were shown to differentiate between mercury-resistant and -sensitive strains in vitro. Furthermore, development of resistance to mercury by passage in mercuric chloride-broth was demonstrated.It is proposed that mercury resistance has developed as a result of exposure to the mercury ion. Mercurial diuretics have been frequently used in medical and geriatric patients and it is among these that the higher carrier rates of mercury-resistant strains are found even when the local endemic strain is disregarded. In obstetric patients, where mercurials are seldom used, mercury-resistant strains are rare.Nasal carriage of factory workers exposed to mercury products showed that this group is likely to carry resistant or partially resistant strains.


2015 ◽  
Vol 59 (9) ◽  
pp. 5366-5376 ◽  
Author(s):  
Lukas Mechler ◽  
Alexander Herbig ◽  
Kerstin Paprotka ◽  
Martin Fraunholz ◽  
Kay Nieselt ◽  
...  

ABSTRACTRecalcitrance of genetically susceptible bacteria to antibiotic killing is a hallmark of bacterial drug tolerance. This phenomenon is prevalent in biofilms, persisters, and also planktonic cells and is associated with chronic or relapsing infections with pathogens such asStaphylococcus aureus. Here we report thein vitroevolution of anS. aureusstrain that exhibits a high degree of nonsusceptibility to daptomycin as a result of cyclic challenges with bactericidal concentrations of the drug. This phenotype was attributed to stationary growth phase-dependent drug tolerance and was clearly distinguished from resistance. The underlying genetic basis was revealed to be an adaptive point mutation in the putative inorganic phosphate (Pi) transporter genepitA. Drug tolerance caused by this allele, termedpitA6, was abrogated when the upstream genepitRwas inactivated. Enhanced tolerance toward daptomycin, as well as the acyldepsipeptide antibiotic ADEP4 and various combinations of other drugs, was accompanied by elevated intracellular concentrations of Piand polyphosphate, which may reversibly interfere with critical cellular functions. The evolved strain displayed increased rates of survival within human endothelial cells, demonstrating the correlation of intracellular persistence and drug tolerance. These findings will be useful for further investigations ofS. aureusdrug tolerance, toward the development of additional antipersister compounds and strategies.


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