Isolation and genomic analysis of temperate phage 5W targeting multidrug-resistant Acinetobacter baumannii
Wenyi Peng1, Fei Zeng1, Zhiying Wu1
1Department of Public Health Laboratory Sciences, School of Public Health, Hengyang Medical School, University of South China, Hengyang, 421001, Hunan, China.
Abstract:
Temperate phages are potential therapeutic agents, but only a few temperate phages infecting multidrug-resistant Acinetobacter baumannii have been identified. In this study, we isolated 5W, a temperate phage that infects multidrug-resistant A. baumannii, from pond water using the enrichment method. A member of the Siphoviridae family, 5W has a narrow host range and infected only four of 19 A. baumannii clinical isolates. It exhibited rapid adsorption (> 90% in 6 min), a latency period of 20 min, and a burst size of ~ 180 plaque-forming units (PFU/cell). 5W contains a linear double-stranded DNA (dsDNA) genome of 43,032 bp with a GC content of 39.85%. The 5W genome contains 61 open reading frames, including lysogen-forming genes, but lacks any known virulence and antibiotic resistance genes. The lysin of 5W is an N-acetyl-β-D-muramidase belonging to the GH_108 family. The α-helical structure and highly positively charged amino acids in the C-terminal region indicate potential antibacterial activity against A. baumannii, and the M/S subunits of the restriction endonuclease are inserted into the lysogenic gene cluster. Comparative genome analysis revealed high similarity with two different prophages in A. baumannii ABCR01, suggesting that 5W may be derived from recombination of other prophages.
Insights
Researchers isolated temperate phage 5W, a potential therapeutic agent against multidrug-resistant Acinetobacter baumannii. This Siphoviridae family phage exhibits rapid adsorption and a distinct genome, offering new possibilities for combating resistant bacterial infections.
Area of Science:
- Microbiology
- Virology
- Genomics
Background:
- Temperate phages show therapeutic potential against multidrug-resistant bacteria.
- Few temperate phages infecting Acinetobacter baumannii have been identified, limiting therapeutic options.
Purpose of the Study:
- To isolate and characterize a novel temperate phage capable of infecting multidrug-resistant Acinetobacter baumannii.
- To evaluate the phage's biological and genomic properties for potential therapeutic applications.
Main Methods:
- Isolation of temperate phage 5W from pond water using enrichment techniques.
- Characterization of phage host range, adsorption rate, latency period, and burst size.
- Whole-genome sequencing and comparative genomic analysis of phage 5W.
Main Results:
- Phage 5W, belonging to the Siphoviridae family, was isolated and demonstrated a narrow host range against A. baumannii clinical isolates.
- Phage 5W exhibits efficient adsorption, a 20-minute latency period, and a burst size of approximately 180 PFU/cell.
- The phage possesses a 43,032 bp dsDNA genome with 61 ORFs, including lysogen-forming genes but lacking virulence and antibiotic resistance genes. Its lysin shows potential antibacterial activity.
Conclusions:
- Temperate phage 5W is a promising candidate for phage therapy against multidrug-resistant A. baumannii.
- The genomic features of 5W, including its lysin, suggest its potential utility in combating bacterial infections.
- Phage 5W's potential derivation from prophage recombination warrants further investigation for understanding its evolution and application.
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