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Evidence of In Vivo Prophage Induction during Clostridium difficile Infection.

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Title: Evidence of In Vivo Prophage Induction during Clostridium difficile Infection.
Authors: Meessen-Pinard, Mathieu1, Sekulovic, Ognjen1, Fortier, Louis-Charles1 louis-charles.fortier@usherbrooke.ca
Source: Applied & Environmental Microbiology. Nov2012, Vol. 78 Issue 21, p7662-7670. 9p.
Subjects: Clostridioides difficile, Bacterial diseases, Virulence of bacteria, Chromosomes, Bioinformatics, Ciprofloxacin, Moxifloxacin, Genetic transformation
Abstract: Prophages contribute to the evolution and virulence of most bacterial pathogens, but their role in Clostridium difficile is unclear. Here we describe the isolation of four Myoviridae phages, &phgr;MMP01, &phgr;MMP02, &phgr;MMP03, and &phgr;MMP04, that were recovered as free viral particles in the filter-sterilized stool supernatants of patients suffering from C. difficile infection (CDI). Furthermore, identical prophages were found in the chromosomes of C. difficile isolated from the corresponding fecal samples. We therefore provide, for the first time, evidence of in vivo prophage induction during CDI. We completely sequenced the genomes of &phgr;MMP02 and &phgr;MMP04, and bioinformatics analyses did not reveal the presence of virulence factors but underlined the unique character of &phgr;MMP04. We also studied the mobility of &phgr;MMP02 and &phgr;MMP04 prophages in vitro. Both prophages were spontaneously induced, with 4 to 5 log PFU/ml detected in the culture supernatants of the corresponding lysogens. When lyso-gens were grown in the presence of subinhibitory concentrations of ciprofloxacin, moxifloxacin, levofloxacin, or mitomycin C, the phage titers further increased, reaching 8 to 9 log PFU/ml in the case of &phgr;MMP04. In summary, our study highlights the extensive genetic diversity and mobility of C. difficile prophages. Moreover, antibiotics known to represent risk factors for CDI, such as quinolones, can stimulate prophage mobility in vitro and probably in vivo as well, which underscores their potential impact on phage-mediated horizontal gene transfer events and the evolution of C. difficile. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
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Abstract:Prophages contribute to the evolution and virulence of most bacterial pathogens, but their role in Clostridium difficile is unclear. Here we describe the isolation of four Myoviridae phages, &phgr;MMP01, &phgr;MMP02, &phgr;MMP03, and &phgr;MMP04, that were recovered as free viral particles in the filter-sterilized stool supernatants of patients suffering from C. difficile infection (CDI). Furthermore, identical prophages were found in the chromosomes of C. difficile isolated from the corresponding fecal samples. We therefore provide, for the first time, evidence of in vivo prophage induction during CDI. We completely sequenced the genomes of &phgr;MMP02 and &phgr;MMP04, and bioinformatics analyses did not reveal the presence of virulence factors but underlined the unique character of &phgr;MMP04. We also studied the mobility of &phgr;MMP02 and &phgr;MMP04 prophages in vitro. Both prophages were spontaneously induced, with 4 to 5 log PFU/ml detected in the culture supernatants of the corresponding lysogens. When lyso-gens were grown in the presence of subinhibitory concentrations of ciprofloxacin, moxifloxacin, levofloxacin, or mitomycin C, the phage titers further increased, reaching 8 to 9 log PFU/ml in the case of &phgr;MMP04. In summary, our study highlights the extensive genetic diversity and mobility of C. difficile prophages. Moreover, antibiotics known to represent risk factors for CDI, such as quinolones, can stimulate prophage mobility in vitro and probably in vivo as well, which underscores their potential impact on phage-mediated horizontal gene transfer events and the evolution of C. difficile. [ABSTRACT FROM AUTHOR]
ISSN:00992240
DOI:10.1128/AEM.02275-12