Salmonella enterica serovar Kentucky is one of the food-borne zoonotic pathogens which is isolated in high frequency from poultry meat in the recent decades and is known for its multidrug resistance. The current study was aimed to isolate and characterize a bacteriophage against S. enterica serovar Kentucky isolate, 5925, which showed resistance to at least seven antibiotics and to study its efficiency to decontaminate S. Kentucky from chicken skin. The bacteriophage against S. enterica serovar Kentucky was isolated and was named vB_SenS_Ib_psk2 representing the place, source, and host. Electron microscopy revealed that the phage possesses isometric head and contractile tail, indicative of Siphoviridae family. Molecular detection of major capsid protein E gene yielded 511 bp, and NCBI blast analysis revealed that the phage belonged to the genus chivirus. The optimum temperature and pH for phage survival and multiplication were found to be - 20 to 42 °C and 6-10, respectively. One-step growth curve experiment of vB_SenS_Ib_psk2 revealed a latent period of 20 min and burst size of 253 phages/bacterial cell. The host susceptibility studies revealed that 83% of MDR isolates of S. enterica were susceptible to vB_SenS_Ib_psk2. Artificial spiking studies on chicken skin revealed that high multiplicity of infection (MOI) of phages of 106 pfu/mL is required for significant reduction (p ≤ 0.01) of bacterial concentration (0.14 ± 0.04) after 24-h incubation at 8 °C compared to group 1 (2.55 ± 0.89 cfu/mL).
- MeSH
- antibakteriální látky MeSH
- bakteriofágy * genetika MeSH
- Salmonella enterica * MeSH
- séroskupina MeSH
- Siphoviridae * genetika MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Kentucky MeSH
Alphaproteobacteria, which are the most abundant microorganisms of temperate oceans, produce phage-like particles called gene transfer agents (GTAs) that mediate lateral gene exchange. However, the mechanism by which GTAs deliver DNA into cells is unknown. Here we present the structure of the GTA of Rhodobacter capsulatus (RcGTA) and describe the conformational changes required for its DNA ejection. The structure of RcGTA resembles that of a tailed phage, but it has an oblate head shortened in the direction of the tail axis, which limits its packaging capacity to less than 4,500 base pairs of linear double-stranded DNA. The tail channel of RcGTA contains a trimer of proteins that possess features of both tape measure proteins of long-tailed phages from the family Siphoviridae and tail needle proteins of short-tailed phages from the family Podoviridae. The opening of a constriction within the RcGTA baseplate enables the ejection of DNA into bacterial periplasm.
- MeSH
- bakteriofágy genetika fyziologie ultrastruktura MeSH
- DNA bakterií genetika MeSH
- elektronová kryomikroskopie MeSH
- přenos genů horizontální MeSH
- regulace genové exprese u bakterií MeSH
- Rhodobacter capsulatus genetika virologie MeSH
- Siphoviridae genetika fyziologie ultrastruktura MeSH
- technika přenosu genů * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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.
- 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
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
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
Given the great biological importance and high diversity of temperate Staphylococcus aureus bacteriophages, a method is needed for the description of their genomic structure. Here we have updated a multiplex PCR strategy for the complex characterization of S. aureus phages of the family Siphoviridae. Based on the comparative genomic analysis of the available phage sequences, a multilocus PCR strategy for typing the major modules of the phage genome was designed. The genomic modules were classified on the basis of the genes for integrase (10 types), anti-repressor (five types), replication proteins polA, dnaC and dnaD (four types), dUTPase (four types), portal protein (eight types), tail appendices (four types) and endolysin (four types) corresponding to the integrase locus, lysogeny control region, and modules for DNA replication, transcription regulation, packaging, tail appendices and lysis respectively. The nine PCR assays designed for the above sequences were shown to be capable to identify the bacteriophage gene pool present both in the phage and bacterial genomes and their extensive mosaic structure. The established multiplex PCR-based multilocus diagnostic scheme is convenient for rapid and reliable phage and prophage classification and for the study of bacteriophage evolution.
- MeSH
- DNA virů genetika MeSH
- genom virový MeSH
- multilokusová sekvenční typizace MeSH
- polymerázová řetězová reakce metody MeSH
- profágy klasifikace genetika MeSH
- Siphoviridae klasifikace genetika MeSH
- srovnávací genomová hybridizace MeSH
- Staphylococcus aureus genetika virologie MeSH
- virové proteiny genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The entire double-stranded DNA genome of the Streptomyces aureofaciens phage mu1/6 was sequenced and analyzed. Its size is 38.194 kbp with an overall molar G+C content of 71.2 %. Fifty-two potential open reading frames (orfs) were identified, divided into two oppositely transcribed regions. In the left arm of the mu1/6 genome, an identified putative integrase and possible regulation proteins were identified. The rightwards transcribed region contains genes organized into apparently four functional units responsible for: (i) replication, (ii) DNA packaging and head assembly, (iii) tail morphogenesis, and (iv) lysis. Putative functions were assigned to twelve orfs based on bioinformatic analysis or experimental substantiation. Comparative analysis with three complete genomes of streptomycete phages revealed resemblance with respect to the organization of their genes into functional modules. Closer relationship was observed only between mu1/6 and S. venezuelae phage VWB.
- MeSH
- bakteriofágy genetika imunologie MeSH
- finanční podpora výzkumu jako téma MeSH
- genomová knihovna MeSH
- komponenty genomu genetika MeSH
- lyzogenie genetika MeSH
- otevřené čtecí rámce genetika MeSH
- Siphoviridae genetika izolace a purifikace MeSH
- Streptomyces aureofaciens genetika izolace a purifikace MeSH
- tetracyklin farmakologie izolace a purifikace MeSH
