Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i>
Extensive and multiple drug resistance in <i>P. aeruginosa</i> combined with the formation of biofilms is responsible for its high persistence in nosocomial infections. A sequential method to devise a suitable phage cocktail with a broad host range and high lytic efficiency against a bio...
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MDPI AG
2023-02-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/2/427 |
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| author | Medhavi Vashisth Anu Bala Jaglan Shikha Yashveer Priya Sharma Priyanka Bardajatya Nitin Virmani Bidhan Chand Bera Rajesh Kumar Vaid Taruna Anand |
| author_facet | Medhavi Vashisth Anu Bala Jaglan Shikha Yashveer Priya Sharma Priyanka Bardajatya Nitin Virmani Bidhan Chand Bera Rajesh Kumar Vaid Taruna Anand |
| author_sort | Medhavi Vashisth |
| collection | DOAJ |
| description | Extensive and multiple drug resistance in <i>P. aeruginosa</i> combined with the formation of biofilms is responsible for its high persistence in nosocomial infections. A sequential method to devise a suitable phage cocktail with a broad host range and high lytic efficiency against a biofilm forming XDR <i>P. aeruginosa</i> strain is presented here. Out of a total thirteen phages isolated against <i>P. aeruginosa</i>, five were selected on the basis of their high lytic spectra assessed using spot assay and productivity by efficiency of plating assay. Phages, after selection, were tested individually and in combinations of two-, three-, four-, and five-phage cocktails using liquid infection model. Out of total 22 combinations tested, the cocktail comprising four phages viz. φPA170, φPA172, φPA177, and φPA180 significantly inhibited the bacterial growth in liquid infection model (<i>p</i> < 0.0001). The minimal inhibitory dose of each phage in a cocktail was effectively reduced to >10 times than the individual dose in the inhibition of XDR <i>P. aeruginosa</i> host. Field emission-scanning electron microscopy was used to visualize phage cocktail mediated eradication of 4-day-old multi-layers of XDR <i>P. aeruginosa biofilms</i> from urinary catheters and glass cover slips, and was confirmed by absence of any viable cells. Differential bacterial inhibition was observed with different phage combinations where multiple phages were found to enhance the cocktail’s lytic range, but the addition of too many phages reduced the overall inhibition. This study elaborates an effective and sequential method for the preparation of a phage cocktail and evaluates its antimicrobial potential against biofilm forming XDR strains of <i>P. aeruginosa</i>. |
| first_indexed | 2024-03-11T08:01:17Z |
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| id | doaj.art-fbc900db7e0741bfbeea0f4be68d4f1b |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-11T08:01:17Z |
| publishDate | 2023-02-01 |
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| series | Viruses |
| spelling | doaj.art-fbc900db7e0741bfbeea0f4be68d4f1b2023-11-16T23:48:51ZengMDPI AGViruses1999-49152023-02-0115242710.3390/v15020427Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i>Medhavi Vashisth0Anu Bala Jaglan1Shikha Yashveer2Priya Sharma3Priyanka Bardajatya4Nitin Virmani5Bidhan Chand Bera6Rajesh Kumar Vaid7Taruna Anand8ICAR-National Research Centre on Equines, Hisar 125001, IndiaICAR-National Research Centre on Equines, Hisar 125001, IndiaDepartment of Molecular Biology and Biotechnology, College of Biotechnology, Chaudhary Charan Singh Haryana Agricultural University, Hisar 125004, IndiaICAR-National Research Centre on Equines, Hisar 125001, IndiaICAR-National Research Centre on Equines, Hisar 125001, IndiaICAR-National Research Centre on Equines, Hisar 125001, IndiaICAR-National Research Centre on Equines, Hisar 125001, IndiaICAR-National Research Centre on Equines, Hisar 125001, IndiaICAR-National Research Centre on Equines, Hisar 125001, IndiaExtensive and multiple drug resistance in <i>P. aeruginosa</i> combined with the formation of biofilms is responsible for its high persistence in nosocomial infections. A sequential method to devise a suitable phage cocktail with a broad host range and high lytic efficiency against a biofilm forming XDR <i>P. aeruginosa</i> strain is presented here. Out of a total thirteen phages isolated against <i>P. aeruginosa</i>, five were selected on the basis of their high lytic spectra assessed using spot assay and productivity by efficiency of plating assay. Phages, after selection, were tested individually and in combinations of two-, three-, four-, and five-phage cocktails using liquid infection model. Out of total 22 combinations tested, the cocktail comprising four phages viz. φPA170, φPA172, φPA177, and φPA180 significantly inhibited the bacterial growth in liquid infection model (<i>p</i> < 0.0001). The minimal inhibitory dose of each phage in a cocktail was effectively reduced to >10 times than the individual dose in the inhibition of XDR <i>P. aeruginosa</i> host. Field emission-scanning electron microscopy was used to visualize phage cocktail mediated eradication of 4-day-old multi-layers of XDR <i>P. aeruginosa biofilms</i> from urinary catheters and glass cover slips, and was confirmed by absence of any viable cells. Differential bacterial inhibition was observed with different phage combinations where multiple phages were found to enhance the cocktail’s lytic range, but the addition of too many phages reduced the overall inhibition. This study elaborates an effective and sequential method for the preparation of a phage cocktail and evaluates its antimicrobial potential against biofilm forming XDR strains of <i>P. aeruginosa</i>.https://www.mdpi.com/1999-4915/15/2/427bacteriophage<i>Pseudomonas aeruginosa</i>extensively drug resistant (XDR)biofilmphage cocktail |
| spellingShingle | Medhavi Vashisth Anu Bala Jaglan Shikha Yashveer Priya Sharma Priyanka Bardajatya Nitin Virmani Bidhan Chand Bera Rajesh Kumar Vaid Taruna Anand Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i> Viruses bacteriophage <i>Pseudomonas aeruginosa</i> extensively drug resistant (XDR) biofilm phage cocktail |
| title | Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i> |
| title_full | Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i> |
| title_fullStr | Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i> |
| title_full_unstemmed | Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i> |
| title_short | Development and Evaluation of Bacteriophage Cocktail to Eradicate Biofilms Formed by an Extensively Drug-Resistant (XDR) <i>Pseudomonas aeruginosa</i> |
| title_sort | development and evaluation of bacteriophage cocktail to eradicate biofilms formed by an extensively drug resistant xdr i pseudomonas aeruginosa i |
| topic | bacteriophage <i>Pseudomonas aeruginosa</i> extensively drug resistant (XDR) biofilm phage cocktail |
| url | https://www.mdpi.com/1999-4915/15/2/427 |
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