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
Bacteriophages of the significant veterinary pathogen Staphylococcus pseudintermedius are rarely described morphologically and genomically in detail, and mostly include phages of the Siphoviridae family. There is currently no taxonomical classification for phages of this bacterial species. Here we describe a new phage designated vB_SpsS_QT1, which is related to phage 2638A originally described as a Staphylococcus aureus phage. Propagating strain S. aureus 2854 of the latter was reclassified by rpoB gene sequencing as S. pseudintermedius 2854 in this work. Both phages have a narrow but different host range determined on 54 strains. Morphologically, both of them belong to the family Siphoviridae, share the B1 morphotype, and differ from other staphylococcal phage genera by a single long fibre at the terminus of the tail. The complete genome of phage vB_SpsS_QT1 was sequenced with the IonTorrent platform and expertly annotated. Its linear genome with cohesive ends is 43,029 bp long and encodes 60 predicted genes with the typical modular structure of staphylococcal siphophages. A global alignment found the genomes of vB_SpsS_QT1 and 2638A to share 84% nucleotide identity, but they have no significant similarity of nucleotide sequences with other phage genomes available in public databases. Based on the morphological, phylogenetic, and genomic analyses, a novel genus Fibralongavirus in the family Siphoviridae is described with phage species vB_SpsS_QT1 and 2638A.
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
Critical behaviour pervades scientific disciplines as diverse as geology, economy or sociology. The critical behaviour of cell control systems is an open issue whose role has not yet been fully explored. The control of the expression of lambda phage DNA in the host cell can be classified as a system with critical behaviour. Lambda phage is a virus that infects Escherichia coli. Its core genes maintain one of two states; lysogeny or lysis. Current knowledge of the lambda phage genetic network allows to build a computational model of transcriptional control of the genes involved in the lytic-lysogenic switch and to simulate the temporal changes of their expression. Here, we focused on the computational simulation of these gene expressions to demonstrate critical behaviour of the system.
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
This study presents synthesis of paramagnetic particles formed by nanomaghemite (γ-Fe2O3) core, whose surface was modified with LaCl3 and entire composite is able to specifically bind the bacteriophage λ. In this study we described the structure of bacteriophage λ, its biochemical properties and role in the therapy, biotechnology and nanomedicine. In the next part we summarized accessible information about paramagnetic particles, their attributes and specificity of binding the bacteriophage λ. Paramagnetic particles show the ability to immobilize bacteriophage λ for subsequent analysis using ion-exchange chromatography with Vis detection. The presence of bacteriophage λ on the surface of paramagnetic beads was demonstrated by sodium dodecyl sulphate gel electrophoresis (SDS-PAGE). It can be stated that paramagnetic beads show significant binding ability towards bacteriophage λ. This phenomenon can be employed for magnetic separation of bacteriophages prior their possible modifications and subsequent application as a cytostatic nanotransporters or antimicrobial agents usable in hard-to-heal bacterial infections.
- Klíčová slova
- superparamagnetic iron oxide nanoparticles,
- MeSH
- bakteriofág lambda * chemie MeSH
- bakteriofágy MeSH
- chromatografie iontoměničová MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- magnetické nanočástice * MeSH
- nanomedicína MeSH
- nanotechnologie MeSH
- oxidy MeSH
- železnaté sloučeniny MeSH
- železo MeSH
- Publikační typ
- 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
Protein phosphorylation is the most abundant and best studied protein posttranslational modification, dedicated to the regulation of protein function and subcellular localization as well as to protein-protein interactions. Identification and quantitation of the dynamic, conditional protein phosphorylation can be achieved by either metabolic labeling of the protein of interest with (32)P-labeled ATP followed by autoradiographic analysis, the use of specific monoclonal or polyclonal antibodies against the phosphorylated protein species and finally by phosphoproteome delineation using mass spectrometry.Hereby we present a fourth alternative which relies on the enforced-affinity-based-electrophoretic separation of phosphorylated from non-phosphorylated protein species by standard SDS-PAGE systems co-polymerized with Phos-Tag™ and Mn(2+) or Zn(2+) cations. Phosphate groups of phosphorylated Ser, Thr, and Tyr residues form complexes with Mn(2+) and Zn(2+) cations with polyacrylamide immobilized Phos-Tag™. Following appropriate treatment of the gels, separated proteins can be quantitatively transferred to PVDF or nitrocellulose membranes and probed with common-not phosphorylation state specific-antibodies and delineate the occurrence of a certain phosphoprotein species against its non-phosphorylated counterpart.
- MeSH
- akrylamid chemie MeSH
- Arabidopsis enzymologie růst a vývoj MeSH
- bakteriofág lambda enzymologie MeSH
- elektroforéza v polyakrylamidovém gelu metody MeSH
- fenol chemie MeSH
- fosfatasy metabolismus MeSH
- fosfoproteiny izolace a purifikace metabolismus MeSH
- fosforylace MeSH
- kultivační techniky MeSH
- membrány umělé MeSH
- mitogenem aktivované proteinkinasy izolace a purifikace metabolismus MeSH
- polyvinyly chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this study, we discussed the use of bacteriophage λ as a nanocarrier of anthracycline drug doxorubicin. This drug is used in treatment of cancer, which is the cause of death of every fourth patient in developed countries. Doxorubicin has many severe side effects for patients' healthy cells, which lowers their well-being. These can be eliminated by encapsulation of doxorubicin into suitable nanocarrier, such as bacteriophage λ capsid. In this work, infusion method was used for this encapsulation and proven by absorbance and fluorescence measurement of whole bacteriophage. It was concluded, that bacteriophage both intercalates into DNA and binds to capsid proteins of bacteriophage λ.
- Klíčová slova
- enkapsulace léčiv,
- MeSH
- bakteriofág lambda * MeSH
- DNA MeSH
- doxorubicin * MeSH
- fluorescenční spektrometrie statistika a číselné údaje MeSH
- lékové transportní systémy * MeSH
- nanočástice MeSH
- virové plášťové proteiny MeSH
- virům podobné částice * MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
