Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i>
Johne’s disease (JD), a chronic infectious enteritis of ruminants, causes major economic losses in the dairy industry globally. This enteritis is caused by <i>Mycobacterium avium</i> subsp. <i>Paratuberculosis</i> (MAP). Currently there is no cure for JD and test-based cullin...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-12-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/16/1/20 |
| _version_ | 1797342451465191424 |
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| author | Victoria K. Harman-McKenna Jeroen De Buck |
| author_facet | Victoria K. Harman-McKenna Jeroen De Buck |
| author_sort | Victoria K. Harman-McKenna |
| collection | DOAJ |
| description | Johne’s disease (JD), a chronic infectious enteritis of ruminants, causes major economic losses in the dairy industry globally. This enteritis is caused by <i>Mycobacterium avium</i> subsp. <i>Paratuberculosis</i> (MAP). Currently there is no cure for JD and test-based culling has proved ineffective at preventing the spread. To isolate new mycobacteriophages (mbps) that can potentially be used to control JD transmission and infection on dairy farms, we optimized an isolation protocol by fecal spiking and the testing of different isolation solution compositions. Using this protocol, we successfully enhanced the yield of mbps from spiked fecal samples, elevating it from less than 1% to 59%. With this method, we isolated 14 mbps from 475 environmental samples collected from MAP-positive dairy farms, after in-sample enrichment with MAP and the fast-growing <i>M. smegmatis</i>. The sample sources included soil, manure pits, lactation barns, feces, milk, and drain water. After fingerprinting these mbps by restriction enzyme profiling, we concluded that 12 were distinct and novel. Further characterization of their host range revealed that eight were capable of lysing multiple MAP strains. We also studied the cross-resistance, lysogeny, the effect of pH and their antimycobacterial properties in milk replacer. Each novel mbp showed limited cross-resistance and prophage immunity and showed no reduction in the titer in a range of pHs after 4 h. The novel phages were also able to reduce the mycobacterial counts to zero after 8 h in milk replacer. In conclusion, these novel mbps could be considered to be used in the control strategies of JD on farms. |
| first_indexed | 2024-03-08T10:33:28Z |
| format | Article |
| id | doaj.art-74b8daad340d46999ab16cca251f783c |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-08T10:33:28Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-74b8daad340d46999ab16cca251f783c2024-01-26T18:46:50ZengMDPI AGViruses1999-49152023-12-011612010.3390/v16010020Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i>Victoria K. Harman-McKenna0Jeroen De Buck1Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 1N4, CanadaFaculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 1N4, CanadaJohne’s disease (JD), a chronic infectious enteritis of ruminants, causes major economic losses in the dairy industry globally. This enteritis is caused by <i>Mycobacterium avium</i> subsp. <i>Paratuberculosis</i> (MAP). Currently there is no cure for JD and test-based culling has proved ineffective at preventing the spread. To isolate new mycobacteriophages (mbps) that can potentially be used to control JD transmission and infection on dairy farms, we optimized an isolation protocol by fecal spiking and the testing of different isolation solution compositions. Using this protocol, we successfully enhanced the yield of mbps from spiked fecal samples, elevating it from less than 1% to 59%. With this method, we isolated 14 mbps from 475 environmental samples collected from MAP-positive dairy farms, after in-sample enrichment with MAP and the fast-growing <i>M. smegmatis</i>. The sample sources included soil, manure pits, lactation barns, feces, milk, and drain water. After fingerprinting these mbps by restriction enzyme profiling, we concluded that 12 were distinct and novel. Further characterization of their host range revealed that eight were capable of lysing multiple MAP strains. We also studied the cross-resistance, lysogeny, the effect of pH and their antimycobacterial properties in milk replacer. Each novel mbp showed limited cross-resistance and prophage immunity and showed no reduction in the titer in a range of pHs after 4 h. The novel phages were also able to reduce the mycobacterial counts to zero after 8 h in milk replacer. In conclusion, these novel mbps could be considered to be used in the control strategies of JD on farms.https://www.mdpi.com/1999-4915/16/1/20<i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i>bacteriophage therapyphage cocktail |
| spellingShingle | Victoria K. Harman-McKenna Jeroen De Buck Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> Viruses <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> bacteriophage therapy phage cocktail |
| title | Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> |
| title_full | Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> |
| title_fullStr | Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> |
| title_full_unstemmed | Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> |
| title_short | Effective Isolation and Characterization of Mycobacteriophages with the Ability to Lyse <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> |
| title_sort | effective isolation and characterization of mycobacteriophages with the ability to lyse i mycobacterium avium i subsp i paratuberculosis i |
| topic | <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> bacteriophage therapy phage cocktail |
| url | https://www.mdpi.com/1999-4915/16/1/20 |
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