Inactivating Host Bacteria for Characterization and Use of Phages
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 Anto...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-09-01
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| Series: | Biophysica |
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| Online Access: | https://www.mdpi.com/2673-4125/3/4/38 |
| _version_ | 1797381841161814016 |
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| author | James P. Chambers Elena T. Wright Barbara Hunter Philip Serwer |
| author_facet | James P. Chambers Elena T. Wright Barbara Hunter Philip Serwer |
| author_sort | James P. Chambers |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-08T20:58:16Z |
| format | Article |
| id | doaj.art-fbe79eec49e54c4f815c3aee25fcb6a1 |
| institution | Directory Open Access Journal |
| issn | 2673-4125 |
| language | English |
| last_indexed | 2024-03-08T20:58:16Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biophysica |
| spelling | doaj.art-fbe79eec49e54c4f815c3aee25fcb6a12023-12-22T13:56:11ZengMDPI AGBiophysica2673-41252023-09-013455856810.3390/biophysica3040038Inactivating Host Bacteria for Characterization and Use of PhagesJames P. Chambers0Elena T. Wright1Barbara Hunter2Philip Serwer3Department of Molecular Microbiology and Immunology, The University of Texas at San Antonio, San Antonio, TX 78249, USADepartment of Biochemistry and Structural Biology, UT Health, San Antonio, TX 78229, USADepartment of Pathology and Laboratory Medicine, UT Health, San Antonio, TX 78229, USADepartment of Biochemistry and Structural Biology, UT Health, San Antonio, TX 78229, USAPhage 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.https://www.mdpi.com/2673-4125/3/4/38electron microscopyphage characterizationmulti-drug resistant bacteriaphage therapythin sectioning |
| spellingShingle | James P. Chambers Elena T. Wright Barbara Hunter Philip Serwer Inactivating Host Bacteria for Characterization and Use of Phages Biophysica electron microscopy phage characterization multi-drug resistant bacteria phage therapy thin sectioning |
| title | Inactivating Host Bacteria for Characterization and Use of Phages |
| title_full | Inactivating Host Bacteria for Characterization and Use of Phages |
| title_fullStr | Inactivating Host Bacteria for Characterization and Use of Phages |
| title_full_unstemmed | Inactivating Host Bacteria for Characterization and Use of Phages |
| title_short | Inactivating Host Bacteria for Characterization and Use of Phages |
| title_sort | inactivating host bacteria for characterization and use of phages |
| topic | electron microscopy phage characterization multi-drug resistant bacteria phage therapy thin sectioning |
| url | https://www.mdpi.com/2673-4125/3/4/38 |
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