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Isolation and Genomic Characterization of an Acinetobacter johnsonii Bacteriophage AJO2 From Bulking Activated Sludge.

ABSTRACT: A novel Podoviridae lytic phage AJO2, specifically infecting Acinetobacter johnsonii, was isolated from bulking activated sludge. The one-step growth experiment showed that the latent period and burst size of AJO2 were estimated to be 30 min and 78.1 phages per infected cell, respectively. The viability test indicated that neutral conditions (pH 6-8) were table for AJO2 survival, while it was sensitive to high temperature (≥60°C) and ultraviolet treatment (254 nm). Genomic sequencing revealed that the AJO2 had a linearly permuted, double-stranded (ds) DNA consisting of 38,124 bp, with the G+C content of 41 mol%. A total of 58 putative open reading frames (ORFs), 11 pairs of repeats and 11 promoters were identified. The AJO2 genome had a modular gene structure which shared some similarities to those of A. baumanii phages. However, genomic comparative analysis revealed many differences among them, and novel genes were identified in the AJO2 genome. These results contribute to subsequent researches on the interaction between bacteriophages and hosts in wastewater treatment, especially during the bulking period. Additionally, the newly isolated phage could be a good candidate as a therapeutic agent to control nosocomial infections caused by A. johnsonii.

SUBMITTER: Fan N

PROVIDER: S-EPMC6401600 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND: Acinetobacter baumannii is an emerging nosocomial pathogen worldwide with increasing prevalence of multi-drug and pan-drug resistance. A. baumannii exists widely in natural environment, especially in health care settings, and has been shown difficult to be eradicated. Bacteriophages are often considered alternative agent for controlling bacterial infection and contamination. In this study, we described the isolation and characterization of one virulent bacteriophage AB1 capable of specifically infecting A. baumannii. RESULTS: A virulent bacteriophage AB1, specific for infecting a clinical strain A. baumannii KD311, was first isolated from marine sediment sample. Restriction analysis indicated that phage AB1 was a dsDNA virus with an approximate genome size of 45.2 kb to 46.9 kb. Transmission electron microscopy showed that phage AB1 had an icosahedral head with a non-contractile tail and collar or whisker structures, and might be tentatively classified as a member of the Siphoviridae family. Proteomic pattern of phage AB1, generated by SDS-PAGE using purified phage particles, revealed five major bands and six minor bands with molecular weight ranging from 14 to 80 kilo-dalton. Also determined was the adsorption rate of phage AB1 to the host bacterium, which was significantly enhanced by addition of 10 mM CaCl2. In a single step growth test, phage AB1 was shown having a latent period of 18 minutes and a burst size of 409. Moreover, pH and thermal stability of phage AB1 were also investigated. At the optimal pH 6.0, 73.2% of phages survived after 60 min incubation at 50 degrees C. When phage AB1 was used to infect four additional clinical isolates of A. baumannii, one clinical isolate of Stenotrophomonas maltophilia, and Pseudomonas aeruginosa lab strains PAK and PAO1, none of the tested strains was found susceptible, indicating a relatively narrow host range for phage AB1. CONCLUSION: Phage AB1 was capable of eliciting efficient lysis of A. baumannii, revealing its potential as a non-toxic sanitizer for controlling A. baumannii infection and contamination in both hospital and other public environments.

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Isolation and Genomic Characterization of an Acinetobacter johnsonii Bacteriophage AJO2 From Bulking Activated Sludge.

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