Staphylococcus aureus may be a highly virulent human pathogen, especially when it is able to form a biofilm, and it is resistant to antibiotic. Infections caused by these bacteria significantly affect morbidity and mortality, primarily in hospitalized patients. Treatment becomes more expensive, more toxic, and prolonged. This is the reason why research on alternative therapies should be one of the main priorities of medicine and biotechnology. A promising alternative treatment approach is bacteriophage therapy. The effect of the anti-staphylococcal bacteriophage preparation Stafal® on biofilm reduction was assessed on nine S. aureus strains using both sonication with subsequent quantification of surviving cells on the catheter surface and evaluation of biofilm reduction in microtiter plates. It was demonstrated that the bacteriophages destroy planktonic cells very effectively. However, to destroy cells embedded in the biofilm effectively requires a concentration at least ten times higher than that provided by the commercial preparation. The catheter disc method (CDM) allowed easier comparison of the effect on planktonic cells and cells in a biofilm than the microtiter plate (MTP) method.
- MeSH
- antiinfekční látky * MeSH
- bakteriologické techniky MeSH
- biofilmy * MeSH
- lidé MeSH
- methicilin rezistentní Staphylococcus aureus růst a vývoj izolace a purifikace virologie MeSH
- mikrobiální viabilita MeSH
- počet mikrobiálních kolonií MeSH
- stafylokokové bakteriofágy fyziologie MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus aureus růst a vývoj izolace a purifikace virologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The bacteriophage K1/420 is a member of genus Kayvirus that was extensively studied as an alternative treatment to combat bacterial infections caused by antibiotic-resistant Staphylococcus aureus strains. Despite the promise of phage therapy, the development of clinical applications of phages is facing regulatory and technical hurdles before it can receive acceptance in the Western World. Suitable simple and accurate diagnostic techniques to control the quality of the phage, which would satisfy the requirements of regulatory authorities are still being discussed. Here, we present the conditions for the simultaneous separation and detection of phage K1/420 and S. aureus by CZE and by CIEF were found, and the phage isoelectric point was determined to be 3.6. After removing the cell debris, the phage was successfully purified from the crude phage lysate and pre-concentrated by preparative isoelectric focusing. Its zone was localized by the positions of colored pI markers in the cellulose bed. The phage from the harvested zone had a decreased ability to infect its host. However, it was suitable for its separation, detection and identification by capillary electrophoretic methods, MALDI-TOF MS and electron microscopy.
Staphylococcus sciuri is a bacterial pathogen associated with infections in animals and humans, and represents a reservoir for the mecA gene encoding methicillin-resistance in staphylococci. No S. sciuri siphophages were known. Here the identification and characterization of two temperate S. sciuri phages from the Siphoviridae family designated ϕ575 and ϕ879 are presented. The phages have icosahedral heads and flexible noncontractile tails that end with a tail spike. The genomes of the phages are 42,160 and 41,448 bp long and encode 58 and 55 ORFs, respectively, arranged in functional modules. Their head-tail morphogenesis modules are similar to those of Staphylococcus aureus ϕ13-like serogroup F phages, suggesting their common evolutionary origin. The genome of phage ϕ575 harbours genes for staphylokinase and phospholipase that might enhance the virulence of the bacterial hosts. In addition both of the phages package a homologue of the mecA gene, which is a requirement for its lateral transfer. Phage ϕ879 transduces tetracycline and aminoglycoside pSTS7-like resistance plasmids from its host to other S. sciuri strains and to S. aureus. Furthermore, both of the phages efficiently adsorb to numerous staphylococcal species, indicating that they may contribute to interspecies horizontal gene transfer.
- MeSH
- bakteriální geny * MeSH
- fosfolipasy metabolismus MeSH
- genom virový MeSH
- genomika metody MeSH
- hostitelská specificita MeSH
- metaloendopeptidasy metabolismus MeSH
- plazmidy genetika MeSH
- přenos genů horizontální MeSH
- přichycení viru MeSH
- stafylokokové bakteriofágy fyziologie ultrastruktura MeSH
- Staphylococcus virologie MeSH
- transdukce genetická * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The transduction mediated by bacteriophages is considered to be one of the primary driving forces in horizontal gene transfer in staphylococci, which is crucial to their adaptation and successful evolution. For a transduction to be effective, it is generally accepted that the recipient strain should be susceptible to the transducing phage. In this study, we demonstrate that the plasmid DNAs are effectively transduced into the recipient Staphylococcus aureus strains in spite of their insensitivity to the lytic action of the transducing phage, provided that these phages adsorb effectively to the bacterial cells. The tetracycline and penicillinase plasmids were transduced to insensitive laboratory and clinical strains by bacteriophages ϕ29, ϕ52A and ϕ80α as well as by prophage ϕ53 and naturally occurring prophages induced from donor lysogenic strains. Comparable frequencies of transduction were achieved in both phage-sensitive and phage-insensitive recipient strains. We have demonstrated that such mechanisms as the restriction of DNA and lysogenic immunity which are responsible for insensitivity of cells to phages may not be a barrier to the transfer, maintenance and effective spread of plasmids to a wider range of potential recipients in the staphylococcal population.
- MeSH
- antibakteriální látky farmakologie MeSH
- lyzogenie MeSH
- mnohočetná bakteriální léková rezistence MeSH
- penicilinasa genetika MeSH
- plazmidy * MeSH
- profágy genetika fyziologie MeSH
- stafylokokové bakteriofágy genetika fyziologie MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus aureus genetika virologie MeSH
- tetracyklin farmakologie MeSH
- transdukce genetická * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
