Antibiotic resistance is increasing among Staphylococcus saprophyticus strains isolated from urinary tract infection. This necessitates alternative therapies. For this, a lytic phage (vB_SsapS-104) against S. saprophyticus, which formed round and clear plaques on bacterial culture plates, was isolated from hospital wastewater and characterized. Microscopy analysis showed that it had a small head (about 50 nm), tail (about 80 nm), and a collar (about 22 nm in length and 19 nm in width) indicating to be a phage within Siphoviridae family. Phage vB_SsapS-104 showed a large latency period of about 40 min, rapid adsorption rate that was significantly enhanced by MgCl2 and CaCl2, and high stability to a wide range of temperatures and pH values. Restriction analyses demonstrated that phage consists of a double-stranded DNA with an approximate genome size of 40 Kb. BLAST results did not show high similarity (megablast) with other previously identified phages. But, in Blastn, similarity with Staphylococcus phages was observed. Phage vB_SsapS-104 represented high anti-bacterial activity against S. saprophyticus isolates in vitro as it was able to lyse 8 of the 9 clinical isolates (%88.8) obtained from a hospital in Gorgan, Iran. It was a S. saprophyticus-specific phage because no lytic activity was observed on some other pathogenic bacteria tested. Therefore, phage vB_SsapS-104 can be considered as a specific virulent phage against of S. saprophyitcus isolated from urinary tract infection. This study provided the partial genomic characterization of S. saprophyticus phage and its application against urinary tract infection associated with S. saprophyticus. This phage also can be considered as a good candidate for a therapeutic alternative in the future.
- Klíčová slova
- Lytic bacteriophage, Siphoviridae, Staphylococcus saprophyticus, Urinary tract infection,
- MeSH
- antibakteriální látky farmakologie MeSH
- DNA virů MeSH
- fágová terapie MeSH
- genom virový MeSH
- hostitelská specificita MeSH
- infekce močového ústrojí mikrobiologie MeSH
- koncentrace vodíkových iontů MeSH
- latence viru MeSH
- lidé MeSH
- odpadní voda virologie MeSH
- sekvenční analýza DNA MeSH
- Siphoviridae genetika izolace a purifikace ultrastruktura MeSH
- stafylokokové bakteriofágy genetika MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus saprophyticus účinky léků virologie MeSH
- teplota MeSH
- transmisní elektronová mikroskopie MeSH
- virulence MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Írán MeSH
- Názvy látek
- antibakteriální látky MeSH
- DNA virů MeSH
- odpadní voda MeSH
In Staphylococcus aureus, generalized transduction mediated by temperate bacteriophages represents a highly efficient way of transferring antibiotic resistance genes between strains. In the present study, we identified and characterized in detail a new efficiently transducing bacteriophage of the family Siphoviridae, designated ϕJB, which resides as a prophage in the meticillin-resistant S. aureus (MRSA) strain Jevons B. Whole-genome sequencing followed by detailed in silico analysis uncovered a linear dsDNA genome consisting of 43 ,12 bp and comprising 70 ORFs, of which ∼40 encoded proteins with unknown function. A global genome alignment of ϕJB and other efficiently transducing phages ϕ11, ϕ53, ϕ80, ϕ80α and ϕNM4 showed a high degree of homology with ϕNM4 and substantial differences with regard to other phages. Using a model transduction system with a well-defined donor and recipient, ϕJB transferred the tetracycline resistance plasmid pT181 and a penicillinase plasmid with outstanding frequencies, beating most of the above-mentioned phages by an order of magnitude. Moreover, ϕJB demonstrated high frequencies of transferring antibiotic resistance plasmids even upon induction from a lysogenic donor strain. Considering such transducing potential, ϕJB and related bacteriophages may serve as a suitable tool for elucidating the nature of transduction and its contribution to the spread of antibiotic resistance genes in naturally occurring MRSA populations.
- MeSH
- aktivace viru MeSH
- bakteriální léková rezistence MeSH
- DNA virů chemie genetika MeSH
- fylogeneze MeSH
- genom virový MeSH
- lyzogenie MeSH
- methicilin rezistentní Staphylococcus aureus virologie MeSH
- molekulární sekvence - údaje MeSH
- otevřené čtecí rámce MeSH
- plazmidy MeSH
- pořadí genů MeSH
- přenos genů horizontální MeSH
- profágy genetika izolace a purifikace ultrastruktura MeSH
- sekvenční analýza DNA MeSH
- sekvenční homologie MeSH
- Siphoviridae genetika izolace a purifikace ultrastruktura MeSH
- syntenie MeSH
- transdukce genetická * MeSH
- transmisní elektronová mikroskopie MeSH
- výpočetní biologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA virů MeSH
In this study, we isolated and characterized a Siphoviridae phage isolated from the vicinity of a religious structure (Kaaba) in Makkah, Saudi Arabia. The phage was designated as φBM and characterized using transmission electron microscopy, restriction digestion of its DNA, and host range. Electron micrograph indicated that φBM phage has an icosahedral head with diameter of about 65 ± 5 nm and long, non-contractile tail with length of about 300 ± 10 nm and width of about 17 ± 2 nm, respectively. On the basis of the φBM phage morphology, we thus propose that φBM represents a member of Siphoviridae phages. The φBM phage was shown to be able to infect Bacillus megaterium and two other Bacillus species and has no effect on other tested bacteria. φBM was stable over the pH range of 5-9, chloroform resistant and stable at 4 °C. A one-step growth experiment showed a latent period of about 40 min and a burst size of approximately 65 per infected cell. The purified bacteriophage appeared to consist of ten proteins. The genome size was estimated to be ∼38 kb. To our knowledge, this is the first report on the isolation of a bacteriophage from Kaaba a heavily trafficked holy site in Saudi Arabia.
- MeSH
- Bacillus megaterium izolace a purifikace virologie MeSH
- chloroform toxicita MeSH
- DNA virů genetika metabolismus MeSH
- fágy bacilů klasifikace genetika izolace a purifikace ultrastruktura MeSH
- hostitelská specificita MeSH
- koncentrace vodíkových iontů MeSH
- mikrobiální viabilita účinky léků účinky záření MeSH
- mikrobiologie životního prostředí MeSH
- restrikční mapování MeSH
- Siphoviridae klasifikace genetika izolace a purifikace ultrastruktura MeSH
- teplota MeSH
- transmisní elektronová mikroskopie MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Saudská Arábie MeSH
- Názvy látek
- chloroform MeSH
- DNA virů MeSH
