Inactivating Host Bacteria for Characterization and Use of Phages
dc.contributor.author | Chambers, James P. | |
dc.contributor.author | Wright, Elena T. | |
dc.contributor.author | Hunter, Barbara | |
dc.contributor.author | Serwer, Philip | |
dc.date.accessioned | 2024-06-28T15:02:45Z | |
dc.date.available | 2024-06-28T15:02:45Z | |
dc.date.issued | 2023-09-27 | |
dc.date.updated | 2024-06-28T15:02:45Z | |
dc.description.abstract | Phage characterization for research and therapy can involve newly isolated phages propagated in pathogenic bacteria. If so, characterization requires safety-managing the bacteria. In the current study, we adapt a common and inexpensive reagent, PrimeStore (Longhorn Vaccines and Diagnostics, San Antonio, TX, USA), to safety-manage bacteria in 20 min by selectively inactivating the bacteria. No bacterial survivors are observed among >10<sup>9</sup> bacteria per ml for a representative of both Gram-negative bacteria (<i>Escherichia coli</i>) and Gram-positive bacteria (<i>Bacillus thuringiensis</i>). This procedure causes no detected inactivation of podophage T3, myophage T4 and siphophage 0105phi7-2. Margins of safety for PrimeStore concentration exist for bacterial inactivation and phage non-inactivation. Thus, general applicability is expected. Subsequent dialysis is used to block long-term effects on phages. Nonetheless, comparable tests should be performed for each pathogenic bacterial strain/phage. Electron microscopy of thin sections reveals inactivation-altered bacterial cytoplasm and a non-disintegrated bacterial envelope (ghosts). Ghosting of <i>E. coli</i> includes re-arrangement of the cytoplasm and the release of endotoxin. The activity of the released endotoxin is >99% reduced after subsequent dialysis, which also removes PrimeStore components. Ghosting of <i>B. thuringiensis</i> includes apparent phase separation within <i>the</i> cytoplasm. The primary application envisaged is biophysical and other screening of phages for therapy of infectious disease. | |
dc.identifier | doi: 10.3390/biophysica3040038 | |
dc.identifier.citation | Biophysica 3 (4): 558-568 (2023) | |
dc.identifier.uri | https://hdl.handle.net/20.500.12588/6449 | |
dc.title | Inactivating Host Bacteria for Characterization and Use of Phages |