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An Optimized Enrichment Technique for the Isolation of Arthrobacter Bacteriophage Species from Soil Sample Isolates
Published on: April 9, 2015
Isolation and molecular characterisation of Achromobacter phage phiAxp-3, an N4-like bacteriophage
Yanyan Ma1, Erna Li2, Zhizhen Qi3
1College of Food Science, Henan Institute of Science and Technology, Xinxiang, 453003, China.
Abstract:
Achromobacter xylosoxidans, an opportunistic pathogen, is responsible for various nosocomial and community-acquired infections. We isolated phiAxp-3, an N4-like bacteriophage that infects A. xylosoxidans, from hospital waste and studied its genomic and biological properties. Transmission electron microscopy revealed that, with a 67-nm diameter icosahedral head and a 20-nm non-contractile tail, phiAxp-3 has features characteristic of Podoviridae bacteriophages (order Caudovirales). With a burst size of 9000 plaque-forming units and a latent period of 80 min, phiAxp-3 had a host range limited to only four A. xylosoxidans strains of the 35 strains that were tested. The 72,825 bp phiAxp-3 DNA genome, with 416-bp terminal redundant ends, contains 80 predicted open reading frames, none of which are related to virulence or drug resistance. Genome sequence comparisons place phiAxp-3 more closely with JWAlpha and JWDelta Achromobacter phages than with other N4 viruses. Using proteomics, we identified 25 viral proteins from purified phiAxp-3 particles. Notably, investigation of the phage phiAxp-3 receptor on the surface of the host cell revealed that lipopolysaccharide serves as the receptor for the adsorption of phage phiAxp-3. Our findings advance current knowledge about A. xylosoxidans phages in an age where alternative therapies to combat antibiotic-resistant bacteria are urgently needed.
Insights
A novel bacteriophage, phiAxp-3, was isolated and characterized for its potential to infect Achromobacter xylosoxidans. This research provides insights into bacteriophage therapy against this opportunistic pathogen.
Area of Science:
- Microbiology
- Virology
- Bacteriophage Research
Background:
- Achromobacter xylosoxidans is an opportunistic pathogen causing nosocomial and community-acquired infections.
- There is an urgent need for alternative therapies against antibiotic-resistant bacteria.
Purpose of the Study:
- To isolate and characterize a bacteriophage infecting A. xylosoxidans.
- To investigate the genomic and biological properties of the isolated phage, named phiAxp-3.
- To explore the potential of phiAxp-3 as a therapeutic agent.
Main Methods:
- Isolation of bacteriophage phiAxp-3 from hospital waste.
- Transmission electron microscopy for morphological characterization.
- Determination of burst size, latent period, and host range.
- Whole-genome sequencing and bioinformatic analysis.
- Proteomic analysis to identify viral proteins.
- Investigation of the phage receptor on the host cell surface.
Main Results:
- phiAxp-3 is an N4-like bacteriophage belonging to the Podoviridae family, with an icosahedral head and non-contractile tail.
- The phage has a burst size of 9000 PFU and a latent period of 80 minutes, with a limited host range infecting 4 out of 35 A. xylosoxidans strains.
- The 72,825 bp genome contains 80 predicted open reading frames, with no identified virulence or drug resistance genes.
- Proteomics identified 25 viral proteins, and lipopolysaccharide was identified as the receptor for phage adsorption.
- Genome comparisons suggest a closer relationship to JWAlpha and JWDelta phages than other N4 viruses.
Conclusions:
- phiAxp-3 is a novel bacteriophage with potential for targeting A. xylosoxidans infections.
- Understanding its genomic and biological properties is crucial for developing phage-based therapies.
- The identification of lipopolysaccharide as the receptor provides a basis for further studies on phage-host interactions.
