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Isolation and molecular characterisation of Achromobacter phage phiAxp-3, an N4-like bacteriophage.

Ma Y, Li E, Qi Z, Li H, Wei X, Lin W, Zhao R, Jiang A, Yang H, Yin Z, Yuan J, Zhao X - Sci Rep (2016)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: College 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.

No MeSH data available.


Related in: MedlinePlus

Isolated Achromobacter phage phiAxp-3.(a) Plaque morphology of phage phiAxp-3. (b) Transmission electron micrographs of phiAxp-3. Arrows indicate the short noncontractile tails. Phage particles were negatively stained with 2% phosphotungstic acid. Scale bar, 100 nm. (c) One-step growth curves for phiAxp-3 with A. xylosoxidans strain A22732. Plaque-forming units per ml of A22732 culture at different time points. Each time point represents the mean value of three experiments.
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f1: Isolated Achromobacter phage phiAxp-3.(a) Plaque morphology of phage phiAxp-3. (b) Transmission electron micrographs of phiAxp-3. Arrows indicate the short noncontractile tails. Phage particles were negatively stained with 2% phosphotungstic acid. Scale bar, 100 nm. (c) One-step growth curves for phiAxp-3 with A. xylosoxidans strain A22732. Plaque-forming units per ml of A22732 culture at different time points. Each time point represents the mean value of three experiments.

Mentions: Phage phiAxp-3 was isolated from raw hospital sewage in China, using the A. xylosoxidans A22732 strain as the host; this bacterium produces OXA-114e and IMP-1 carbapenemases, which confer resistance to multiple β-lactam antibiotics including carbapenems16. Phage phiAxp-3 formed round plaques with transparent centres on double-layer plates (Fig. 1a). Transmission electron microscopy of the phiAxp-3 particles showed that phiAxp-3 possesses an isometric head with a diameter of about 67 nm and a short tail with an approximate length of 20 nm (Fig. 1b), thereby matching the typical morphological features of Podoviridae family viruses. Host range testing suggested that phiAxp-3 was able to successfully infect all A. xylosoxidans strains tested, unlike other species that were tested (Table 1). Besides the A22732 strain, which is reported to be multidrug-resistant16, all three of the other clinical A. xylosoxidans strains investigated here have been shown to be resistant to aztreonam and tobramycin15.


Isolation and molecular characterisation of Achromobacter phage phiAxp-3, an N4-like bacteriophage.

Ma Y, Li E, Qi Z, Li H, Wei X, Lin W, Zhao R, Jiang A, Yang H, Yin Z, Yuan J, Zhao X - Sci Rep (2016)

Isolated Achromobacter phage phiAxp-3.(a) Plaque morphology of phage phiAxp-3. (b) Transmission electron micrographs of phiAxp-3. Arrows indicate the short noncontractile tails. Phage particles were negatively stained with 2% phosphotungstic acid. Scale bar, 100 nm. (c) One-step growth curves for phiAxp-3 with A. xylosoxidans strain A22732. Plaque-forming units per ml of A22732 culture at different time points. Each time point represents the mean value of three experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837373&req=5

f1: Isolated Achromobacter phage phiAxp-3.(a) Plaque morphology of phage phiAxp-3. (b) Transmission electron micrographs of phiAxp-3. Arrows indicate the short noncontractile tails. Phage particles were negatively stained with 2% phosphotungstic acid. Scale bar, 100 nm. (c) One-step growth curves for phiAxp-3 with A. xylosoxidans strain A22732. Plaque-forming units per ml of A22732 culture at different time points. Each time point represents the mean value of three experiments.
Mentions: Phage phiAxp-3 was isolated from raw hospital sewage in China, using the A. xylosoxidans A22732 strain as the host; this bacterium produces OXA-114e and IMP-1 carbapenemases, which confer resistance to multiple β-lactam antibiotics including carbapenems16. Phage phiAxp-3 formed round plaques with transparent centres on double-layer plates (Fig. 1a). Transmission electron microscopy of the phiAxp-3 particles showed that phiAxp-3 possesses an isometric head with a diameter of about 67 nm and a short tail with an approximate length of 20 nm (Fig. 1b), thereby matching the typical morphological features of Podoviridae family viruses. Host range testing suggested that phiAxp-3 was able to successfully infect all A. xylosoxidans strains tested, unlike other species that were tested (Table 1). Besides the A22732 strain, which is reported to be multidrug-resistant16, all three of the other clinical A. xylosoxidans strains investigated here have been shown to be resistant to aztreonam and tobramycin15.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: College 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.

No MeSH data available.


Related in: MedlinePlus