Abstract
Clostridium difficile is recognized as a problematic pathogen, causing severe enteric diseases including antibiotic-associated diarrhea and pseudomembranous colitis. The emergence of antibiotic resistant C. difficile has driven a search for alternative anti-infection modalities. A promising strategy for controlling bacterial infection includes the use of bacteriophages and their gene products. Currently, knowledge of phages active against C. difficile is still relatively limited by the fact that the isolation of phages for this organism is a technically demanding method since bacterial host themselves are difficult to culture. To isolate and characterize phages specific to C. difficile, a genotoxic agent, mitomycin C, was used to induce temperate phages from 12 clinical isolates of C. difficile. Five temperate phages consisting of ΦHR24, ΦHN10, ΦHN16-1, ΦHN16-2, and ΦHN50 were successfully induced and isolated. Spotting assays were performed against a panel of 92 C. difficile isolates to screen for susceptible bacterial hosts. The results revealed that all the C. difficile phages obtained in this work displayed a relatively narrow host range of 0–6.5% of the tested isolates. Electron microscopic characterization revealed that all isolated phages contained an icosahedral head connected to a long contractile tail, suggesting that they belonged to the Myoviridae family. Restriction enzyme analysis indicated that these phages possess unique double-stranded DNA genome. Further electron microscopic characterization revealed that the ΦHN10 absorbed to the bacterial surface via attachment to cell wall, potentially interacting with S-layer protein. Bacteriophages isolated from this study could lead to development of novel therapeutic agents and detection strategies for C. difficile.
Highlights
Antimicrobial resistance has become one of the most serious global healthcare problems
The PCR products were confirmed by DNA sequencing, which were further blasted against nucleotide database in National Center for Biotechnology Information (NCBI)
The results revealed that 66/73 clinical C. difficile isolates were lysogenics
Summary
Antimicrobial resistance has become one of the most serious global healthcare problems. Bacteriophages of Clostridium difficile and has been listed by the Centers for Disease Control and Prevention as an urgent threat. This anaerobic, toxin-producing bacterium is the leading cause of antibiotic-associated diarrhea in nosocomial setting (Rineh et al, 2014). The most recent option for recurrent CDI treatment is fecal microbiota transplantation (FMT), by using stool collected from healthy donors to treat CDI patients. This treatment succeeds to treat dysbiosis patients with remarkable high efficacy rate up to 81% (Drekonja et al, 2015). To mitigate the severity of CDI outbreaks and reduce the disease recurrence, alternative approaches for effective control of CDI are urgently needed (Joerger, 2003; Rea et al, 2013)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
