ABSTRACT
Orally administered phages to control zoonotic pathogens face important challenges, mainly related to the hostile conditions found in the gastrointestinal tract (GIT). These include temperature, salinity and primarily pH, which is exceptionally low in certain compartments. Phage survival under these conditions can be jeopardized and undermine treatment. Strategies like encapsulation have been attempted with relative success, but are typically complex and require several optimization steps. Here we report a simple and efficient alternative, consisting in the genetic engineering of phages to display lipids on their surfaces. Escherichia coli phage T7 was used as a model and the E. coli PhoE signal peptide was genetically fused to its major capsid protein (10 A), enabling phospholipid attachment to the phage capsid. The presence of phospholipids on the mutant phages was confirmed by High Performance Thin Layer Chromatography, Dynamic Light Scattering and phospholipase assays. The stability of phages was analysed in simulated GIT conditions, demonstrating improved stability of the mutant phages with survival rates 102-107 pfu.mL-1 higher than wild-type phages. Our work demonstrates that phage engineering can be a good strategy to improve phage tolerance to GIT conditions, having promising application for oral administration in veterinary medicine.
Subject(s)
Bacteriophage T7/genetics , Drug Resistance, Viral , Genetic Engineering , Administration, Oral , Animals , Bacteriophage T7/growth & development , Bacteriophage T7/metabolism , Capsid Proteins/genetics , Capsid Proteins/metabolism , Chromatography, Thin Layer , Dynamic Light Scattering , Escherichia coli/genetics , Escherichia coli/metabolism , Escherichia coli Proteins/chemistry , Escherichia coli Proteins/metabolism , Gastrointestinal Tract/virology , Hydrogen-Ion Concentration , Microscopy, Electron, Transmission , Phospholipids/analysis , Phospholipids/chemistry , Phospholipids/metabolism , Porins/chemistry , Porins/metabolism , Protein Sorting Signals/genetics , Temperature , Veterinary MedicineABSTRACT
New SHRIMP U-Pb zircon ages for the Portalegre and Alcáçovas orthogneisses document a complex pre- Variscan history for the Iberian basement in Portugal. The available geochemical and geochronological data for the Alcáçovas orthogneiss (ca. 540 Ma) tend to favor its involvement in a Cadomian orogenic event. This is consistent with the development of an active continental margin setting at the end of the Proterozoic and supports a Gondwanan provenance for the Iberian crust. On the other hand, the Ordovician emplacement age obtained for the magmatic precursors of the Portalegre orthogneisses (497 +/- 10 Ma) provides additional evidence for the occurrence of rift-related magmatic activity during the Lower Paleozoic.
ABSTRACT
We report on the synthesis of ZnO nanoparticles from Zn(CH3CO2)2 and NaOH in 2-propanol. Nucleation and growth are fast, and hence at longer times the particle size is controlled by coarsening. The coarsening kinetics are independent of the Zn(CH3CO2)2 concentration between 0.5 and 1.25 mM at a fixed [Zn(CH3CO2)2]:[NaOH] ratio of 0.625. The width of the size distribution was found to increase only slightly with aging time. In addition, at a fixed Zn(CH3CO2)2 concentration of 1 mM, the kinetics are independent of the [Zn(CH3CO2)2]:[NaOH] ratio between 0.476 and 0.625. The presence of water in the reaction mixture was found to only slightly affect the coarsening kinetics for water contents larger than about 20 mM. For lower water concentrations, the nucleation and growth of ZnO were very slow. It can be concluded that the synthesis method described provides a reliable source of ZnO nanoparticles due to its insensitivity to the reactant concentrations and the presence of water.
