Abstract P-46: Structure of A. Baumannii Phage Tapaz, Revealed with Cryo-Electron Microscopy

Background: Acinetobacter baumannii is an opportunistic pathogen and one of the six most important multidrug resistant microorganisms in hospitals worldwide. Some of its strains are resistant to most of the antibiotics, A. baumannii is included into the Priority 1 part of Global Priority List of Antibiotic-resistant Bacteria. Phage therapy is considered to be an alternative strategy to antibiotic treatments. Methods: A. baumannii strain NIPH601 cells were grown till OD6000.4 and infected with the phage at MOI 10:1. After complete lysis took place cell debris was spined down and phage particles were precipitated with the PEG6000 (final concentration 10% PEG 6000, 0.5 NaCl). Virus particles were collected by centrifugation, resuspended at SM buffer and applied on CsCl step gradient. Gradient was spinned down for 2 hours at 40000g and the fraction containing phage particles was collected and dialyzed against SM buffer. Purified phage particles were applied to Quantifoil 1.2/1.3 grids and plunge-froze in Vitrobot Mark IV (TFS) Micrographs were collected in HKU, Shenzhen campus with Titan Krios cryoelectron microscope (TFS), equipped with Gatan K3 direct electron detector. The micrographs were acquired with 1.06 Å pixel size and 1.5 um average defocus value in counting mode with 50 frames and 1.2 e/Å2/frame dose rate. All image processing was performed with Relion3.0 software, except for the particle picking step performed with cryolo. Results: Lytic A. baumannii phage TaPaz belongs to the family Myoviridae. BLAST search over NCBI “nr” (non-redundant) database revealed close homology with previously published sequences of Acinetobacter phage vB_AbaM_B9 and Acinetobacter phage BS46. However, no structural information about any homologous proteins was found among the PDB structures. The cryo-EM map was reconstructed with single particle analysis independently for the capsid, tail and baseplate regions. The capsid was reconstructed at 3.9 Å resolution with I3 symmetry applied (Fig. 1A). The baseplate region of the phage was reconstructed at 3.5 Å resolution with C3 symmetry (Fig. 1B). The tail region was reconstructed at 2.6 Å resolution with helical symmetry (Rise 36.4 Å, Twist 25.7 deg). Initial atomic model for the tail region was built from sequence with Deeptracer and was further refined in coot (Fig. 1C). Conclusion: We successfully obtained the near-atomic resolution structural map of phage TaPaz. The data obtained contribute to enhancing knowledge of structural diversity of bacterial viruses infecting A. baumannii

Experimental Study of Transient Flow Regimes in a Model Hydroturbine Draft Tube

Swirling flow with the formation of a precessing vortex core (PVC) in the draft tube model of a hydroturbine was studied. Experiments were performed on an aerodynamic setup under transient operating conditions of the hydroturbine. The turbine operating conditions were varied by continuously changing the flow rate at a constant runner speed. The transition from the partial load regime, when a precessing vortex core is formed, to the best efficiency point without a core is considered. Applied to this task, a comparison of the windowed Fourier transform with wavelet analysis is given. The dependence of the PVC lifetime in the transient regime correlates with the transient time. It is shown that the velocity profiles and the spectrum of pressure pulsations in transient regimes change quasistatically between part-load operation and the best efficiency point of the turbine. The phase-averaged velocity distributions in the transient regimes show that a transient regime is a sequence of quasisteady regimes

Laboratory Modeling of an Axial Flow Micro Hydraulic Turbine

This article is devoted to detailed experimental studies of the flow behind the impeller of an air model of a propeller-type microhydroturbine in a wide range of operating parameters. The measurements of two component distributions of averaged velocities and pulsations for conditions from part load to strong overload are conducted. It is shown that the flow at the impeller outlet becomes swirled when the hydraulic turbine operating mode shifts from the optimum one. The character of the behavior of the integral swirl number, which determines the state of the swirled flow, is revealed. Information about the flow peculiarities can be used when adjusting the hydraulic unit mode to optimal conditions and developing recommendations to expand the hydraulic turbine operation control range with preservation of high efficiency. This stage will significantly save time at the stage of equipment design for specific field conditions of water resource