Skin and soft tissue infections (SSTIs) represent a significant healthcare challenge, particularly in the context of increasing antibiotic resistance. This study investigates the efficacy of a novel therapeutic approach combining bacteriophage (phage) therapy with a gum Karaya (GK)-based hydrogel delivery system in a porcine model of deep staphylococcal SSTIs. The study exploits the lytic activity and safety of the Staphylococcus phage 812K1/420 of the Kayvirus genus, which is active against methicillin-resistant Staphylococcus aureus (MRSA). The GK injectable hydrogels and hydrogel films, developed by our research group, serve as effective, non-toxic, and easy-to-apply delivery systems, supporting moist wound healing and re-epithelialization. In the porcine model, the combined treatment showed asynergistic effect, leading to a significant reduction in bacterial load (2.5 log CFU/gram of tissue) within one week. Local signs of inflammation were significantly reduced by day 8, with clear evidence of re-epithelialization and wound contraction. Importantly, no adverse effects of the GK-based delivery system were observed throughout the study. The results highlight the potential of this innovative therapeutic approach to effectively treat deep staphylococcal SSTIs, providing a promising avenue for further research and clinical application in the field of infections caused by antibiotic-resistant bacteria.
- Klíčová slova
- Bacteriophage, Gum Karaya, Hydrogel film, Injectable hydrogel, MRSA, Phage therapy, Staphylococcus aureus,
- MeSH
- fágová terapie * metody MeSH
- hojení ran účinky léků MeSH
- hydrogely * aplikace a dávkování chemie MeSH
- infekce v ráně * terapie mikrobiologie farmakoterapie MeSH
- methicilin rezistentní Staphylococcus aureus * účinky léků MeSH
- modely nemocí na zvířatech * MeSH
- prasata MeSH
- rostlinné gumy chemie MeSH
- stafylokokové bakteriofágy MeSH
- stafylokokové infekce * terapie farmakoterapie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- hydrogely * MeSH
- rostlinné gumy MeSH
AIM: Staphylococcus aureus strains are the cause of frightening hospital and community infections, especially when they are resistant to antimicrobials, have important pathogenicity factors, or have biofilm production ability. Looking for novel therapeutic options which would be effective against such strains is one of the highest priorities of medicine and medical research. The study aim was to describe the occurrence of S. aureus strains and proportion of methicillin resistant strains (MRSA) detected in laboratories of the Microbiological Institute, Faculty of Medicine, Masaryk University (FM MU) and St. Anne's University Hospital, Brno in 2011-2018. Selected strains of S. aureus were tested for biofilm production ability and susceptibility to antimicrobials and Stafal®, a phage therapeutic agent. A prerequisite was to develop a simple routine method suitable for phage susceptibility testing of bacteria. MATERIAL AND METHODS: Altogether 867 clinical isolates of S. aureus and 132 strains of other species of the genus Staphylococcus (isolated in 2011-2017) were tested for susceptibility to the phage therapy preparation Stafal® using the double-layer agar method. All strains of S. aureus were tested for biofilm production ability by the modified Christensen method with the use of titration microplates and for susceptibility to antistaphylococcal antibiotics by the disk diffusion test. For 95 S. aureus strains, the outcome of the double-layer agar method (DAM) was compared with that of our newly designed method (ODM) based on optical density decrease of the bacterial suspension. RESULTS: During the study period, the laboratories of the Faculty of Medicine, Masaryk University (FM MU) and St. Anne's University Hospital, Brno detected 2900 strains of S. aureus per year on average. The proportion of MRSA among S. aureus isolates from blood culture and venous catheters ranged between 8.8-15.2 %. S. aureus strains recovered from venous catheters and blood culture were confirmed as stronger biofilm producers than those from other clinical specimens. MRSA strains showed higher biofilm production than methicillin susceptible strains (MSSA). As many as 90.4 % of S. aureus strains tested susceptible to the Stafal® preparation. Even a higher proportion, i.e. 99.0 %, of MRSA strains were Stafal® susceptible. No relationship was found between Stafal® susceptibility and biofilm production ability. Although Stafal® targets primarily S. aureus, some susceptibility (26.5 %) was also found for other staphylococcal species. A novel simple method designed for routine testing of susceptibility to phage therapy preparations based on optical density decrease was comparably sensitive and reliable as the commonly used double-layer agar method (DAM) and, in addition to being easy and rapid to perform, after prolonged suspension culture and at higher measurement frequency, it has an extra advantage of providing the possibility for monitoring also phage action dynamics. CONCLUSIONS: The proportion of MRSA strains detected in this study is comparable to that reported for the whole Czech Republic, and the biofilm production data are consistent with scientific evidence. The host range of the Stafal® preparation is relatively wide and covers most strains of S. aureus and some coagulase negative staphylococci. The highest efficiency of Stafal® (99.4 %) was observed against MRSA strains with multiple types of antibiotic resistance. In vitro testing of 867 strains of S. aureus and 132 other staphylococcal species has shown the phage therapy preparation Stafal® to be a suitable candidate therapeutic option for the treatment of staphylococcal infections, especially in case of failure of conventional antibiotic therapy. Moreover, a simple method for routine phage susceptibility testing of clinical bacterial isolates has been designed, which is an essential tool to be used in phage therapy.
- Klíčová slova
- MRSA, Phage therapy, Stafal®, Staphylococcus, biofilm,
- MeSH
- antibakteriální látky terapeutické užití MeSH
- bakteriofágy * fyziologie MeSH
- lidé MeSH
- methicilin rezistentní Staphylococcus aureus virologie MeSH
- stafylokokové infekce * terapie virologie MeSH
- Staphylococcus * virologie MeSH
- techniky in vitro MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- antibakteriální látky MeSH
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
- Názvy látek
- auR protein, Staphylococcus aureus MeSH Prohlížeč
- fosfolipasy MeSH
- metaloendopeptidasy MeSH
Bacteriophages represent a simple viral model of basic research with many possibilities for practical application. Due to their ability to infect and kill bacteria, their potential in the treatment of bacterial infection has been examined since their discovery. With advances in molecular biology and gene engineering, the phage application spectrum has been expanded to various medical and biotechnological fields. The construction of bacteriophages with an extended host range or longer viability in the mammalian bloodstream enhances their potential as an alternative to conventional antibiotic treatment. Insertion of active depolymerase genes to their genomes can enforce the biofilm disposal. They can also be engineered to transfer various compounds to the eukaryotic organisms and the bacterial culture, applicable for the vaccine, drug or gene delivery. Phage recombinant lytic enzymes can be applied as enzybiotics in medicine as well as in biotechnology for pathogen detection or programmed cell death in bacterial expression strains. Besides, modified bacteriophages with high specificity can be applied as bioprobes in detection tools to estimate the presence of pathogens in food industry, or utilized in the control of food-borne pathogens as part of the constructed phage-based biosorbents.
- Klíčová slova
- bacteriophages, biopharmaceuticals, biotechnology, enzybiotics, genetically modified bacteriophages, pathogen detection, phage therapy,
- MeSH
- Bacteria účinky léků MeSH
- bakteriální infekce farmakoterapie MeSH
- bakteriofágy genetika MeSH
- biofilmy MeSH
- biologická terapie * MeSH
- biosenzitivní techniky MeSH
- biotechnologie metody MeSH
- genetické inženýrství MeSH
- lidé MeSH
- průmysl zpracování potravin MeSH
- průmyslová mikrobiologie * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
AIMS: The aim of this study was to assess the potential risk posed to the human population by the presence of Listeria monocytogenes serotype 1/2c in food based on the characterization of virulence factors of Listeria involved in the invasion of host cells and sensitivity to antimicrobial agents. METHODS AND RESULTS: In addition to sequencing of the inlA and inlB genes, the presence of genes lapB, aut, fbpA, ami, vip and llsX was tested. A premature stop codon (PMSC) in the inlA gene was detected in all tested strains of serotype 1/2c and, concurrently, two novel PMSC mutation types were identified. However, neither PMSC in the inlB gene nor deletion of the lapB, aut, fbpA, ami and vip genes were found in any of the strains. The presence of the llsX gene was not confirmed. Even though all L. monocytogenes strains showed sensitivity to the tested antimicrobials on the basis of their phenotype, sequencing revealed the presence of IS1542 insertion in the inlA gene, indicating the possibility of sharing of mobile genetic elements associated with antimicrobial resistance among strains. CONCLUSIONS: Other than the presence of PMSCs in the inlA gene, no PMSC in inlB or deletion of other factors linked to the invasiveness of listeria were detected. Tested strains showed sensitivity to antibiotics used in the therapy of listeriosis. SIGNIFICANCE AND IMPACT OF THE STUDY: Strains of L. monocytogenes serotype 1/2c typically carry a PMSC in the inlA gene, but these strains still represent a potential threat to public health. The possibility of transfer of IS1542, associated with resistance to vancomycin, between enterococci and Listeria spp. was revealed.
- Klíčová slova
- antibiotics, internalin A, internalin B, premature stop codon, sequencing,
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- bakteriální proteiny genetika metabolismus MeSH
- faktory virulence genetika metabolismus MeSH
- fenotyp MeSH
- lidé MeSH
- Listeria monocytogenes účinky léků genetika izolace a purifikace metabolismus MeSH
- nesmyslný kodon MeSH
- potravinářská mikrobiologie * MeSH
- séroskupina MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- bakteriální proteiny MeSH
- faktory virulence MeSH
- nesmyslný kodon MeSH
A prospective study (2007-2013) was undertaken to investigate clinical features and prognostic factors of necrotizing pneumonia caused by Staphylococcus aureus producing Panton-Valentine leukocidin (PVL) in the Czech Republic. Twelve cases of necrotizing pneumonia were detected in 12 patients (median age 25 years) without severe underlying disease. Eight cases occurred in December and January and the accumulation of cases in the winter months preceding the influenza season was statistically significant (P < 0·001). The course of pneumonia was very rapid, leading to early sepsis and/or septic shock in all but one patient. Seven patients died and mortality was fourfold higher in those patients presenting with primary pneumonia than with pneumonia complicating other staphylococcal/pyogenic infection elsewhere in the body. The S. aureus isolates displayed considerable genetic variability and were assigned to five lineages CC8 (n = 3), CC15 (n = 2), CC30 (n = 2), CC80 (n = 1), and CC121 (n = 3) and one was a singleton of ST154 (n = 1), all were reported to be associated with community-acquired infection. Four strains were methicillin resistant. The high case-fatality rate can only be reduced by improving the speed of diagnosis and a rapid test to detect S. aureus in the airways is needed.
- Klíčová slova
- Community-acquired pneumonia, Panton–Valentine leukocidin, Staphylococcus aureus, necrotizing pneumonia, septic shock,
- MeSH
- bakteriální pneumonie farmakoterapie mikrobiologie mortalita MeSH
- bakteriální toxiny biosyntéza MeSH
- dospělí MeSH
- exotoxiny biosyntéza MeSH
- genetická variace MeSH
- infekce získané v komunitě mikrobiologie MeSH
- kojenec MeSH
- leukocidiny biosyntéza MeSH
- lidé středního věku MeSH
- lidé MeSH
- methicilin rezistentní Staphylococcus aureus genetika metabolismus MeSH
- mladiství MeSH
- mladý dospělý MeSH
- nekróza mikrobiologie MeSH
- plíce patologie MeSH
- prognóza MeSH
- prospektivní studie MeSH
- roční období MeSH
- septický šok mikrobiologie MeSH
- Staphylococcus aureus MeSH
- streptokokové infekce farmakoterapie mikrobiologie mortalita MeSH
- Check Tag
- dospělí MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- bakteriální toxiny MeSH
- exotoxiny MeSH
- leukocidiny MeSH
- Panton-Valentine leukocidin MeSH Prohlížeč
In the last decade, there has been a rapid development in the use of molecular genetics methods in clinical microbiology. Novel technologies bring new knowledge and approaches to various disciplines of microbiology--taxonomy, identification of microbes, clinical diagnosis, epidemiology of infectious diseases and antibiotic resistance. This article summarizes the conclusions from the workshop of the Molecular Microbiology Working Group TIDE held during the Second Annual Meeting of the Society for Medical Microbiology of the J. E. Purkyne Czech Medical Association.
- MeSH
- Bacteria MeSH
- diagnostické techniky molekulární * MeSH
- DNA bakterií analýza MeSH
- infekce diagnóza MeSH
- lidé MeSH
- mikrobiologické techniky * MeSH
- molekulární biologie * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA bakterií MeSH
A solution to the problem of the increasing number of antibiotic-resistant bacterial strains can be the use of rational phage therapy. In the past, bacteriophages (phages) were often used for the treatment and prevention of infections and unlike antibiotic therapy, phage therapy caused almost no serious side effects. While previously several preparations containing whole phage particles were available for phage therapy, currently, the isolation of well characterised and purified phage components with antibacterial properties opens up new options for the management of intractable infections caused primarily by the bacterial genera Enterococcus, Escherichia, Klebsiella, Listeria, Proteus, Pseudomonas, Salmonella, Shigella, Staphylococcus and Streptococcus. In addition to human and veterinary medicine, the phage therapy principles also find use in the agriculture and food industry. Recent and former clinical studies as well as numerous animal model experiments have supported that phage therapy is an effective and safe alternative of antibiotic treatment of bacterial infections.
- MeSH
- antibiotická rezistence MeSH
- bakteriální infekce terapie MeSH
- bakteriofágy * MeSH
- lidé MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- přehledy MeSH
Mutations extended the host range of the polyvalent bacteriophage 812 of the family Myoviridae in up to 95 % of Staphylococcus aureus strains and 43 % of strains of different coagulase-positive and -negative Staphylococcus species. Mutational changes in the genome of several host-range mutants of phage 812 were identified. Host-range mutant 812F1 harbors a deletion in endolysin gene that arose together with intron excision. Four mutants (812i, 812b, 812p, 812F3) harbor deletion in the structural gene orf8 that results from a genome rearrangement associated with intron insertion. This rearrangement was also detected in the genome of the closely related phages U16 and phi131. Another intron was discovered in the recA812 gene in these four mutants. An insertion was found in a non-coding region of the restriction fragment PstI-O of three mutants (812b, 812F3, 812g) and phages U16 and phi131. The above results contribute to the explanation of genetic factors affecting the host range of polyvalent staphylococcal bacteriophages.
- MeSH
- bakteriofágy genetika MeSH
- endopeptidasy chemie genetika MeSH
- genom virový * MeSH
- molekulární sekvence - údaje MeSH
- mutace * MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- proteiny virových bičíků chemie genetika MeSH
- RNA virová chemie genetika MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- sekvenční seřazení MeSH
- Staphylococcus aureus virologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- endolysin MeSH Prohlížeč
- endopeptidasy MeSH
- proteiny virových bičíků MeSH
- RNA virová MeSH
This study describes the first molecular characterisation of clinical isolates of vancomycin-resistant enterococci (VRE) in the Czech Republic. Of 2647 patient isolates of Enterococcus spp. from 1997-2002, 121 (4.6%) were identified as VRE. The most common isolates were VanA+ Enterococcus faecium (78%) and VanB+ Enterococcus faecalis (10%). In addition, five VanA+ E. faecium isolates were obtained from environmental and staff sampling. Macrorestriction analysis of SmaI restriction fragment length polymorphism was performed for 54 VanA+ E. faecium clinical isolates and the five VanA+ E. faecium environmental isolates. Thirty-two unique restriction endonuclease patterns were identified, including two predominant clonal types represented by five or more isolates. Two environmental VanA+ E. faecium isolates were closely related to two patient isolates, which had an identical SmaI macrorestriction pattern. The results indicated potential survival of strains in the hospital environment and possible subsequent transmission to hospitalised patients.
- MeSH
- Enterococcus faecalis klasifikace izolace a purifikace MeSH
- Enterococcus faecium klasifikace genetika izolace a purifikace MeSH
- genotyp MeSH
- grampozitivní bakteriální infekce mikrobiologie MeSH
- hematologické nádory MeSH
- infekce spojené se zdravotní péčí mikrobiologie MeSH
- lidé MeSH
- nádory MeSH
- onkologická péče - zařízení * MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- rezistence na vankomycin genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
