Genomic structure of the Aeromonas bacteriophage pAh6-C and its comparative genomic analysis

Jun, Jin Woo; Kim, Hyoun Joong; Yun, Sae Kil; Chai, Ji Young; Park, Taesung

doi:10.1007/s00705-014-2221-1

Cited by 13 publications

(8 citation statements)

References 9 publications

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“…Sequences of phages infecting bacteria other than gammaproteobacteria had to be excluded due to insufficient sequence similarity. The GJ1-specific primers were designed based on MCP sequences of members of the GJ1-like cluster [11] extended by Shewanella phage Spp001 [39] and Aeromonas phage pAh6-C [40]. The FO1 primer pair was designed based on the MCP sequences of the members of the Felix O1 cluster published by Grose and Casjens [11], except for FO1a, whose MCP sequence is identical to that of Felix O1.…”

Section: Methodsmentioning

confidence: 99%

A major-capsid-protein-based multiplex PCR assay for rapid identification of selected virulent bacteriophage types

et al. 2019

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Bacteriophages represent a promising alternative for controlling pathogenic bacteria. They are ubiquitous in the environment, and their isolation is usually simple and fast. However, not every phage is suitable for biocontrol applications. It must be virulent (i.e., strictly lytic), non-transducing, and safe. We have developed a method for identifying selected types of virulent phages at an early stage of the isolation process to simplify the search for suitable candidates. Using the major capsid protein (MCP) as a phylogenetic marker, we designed degenerate primers for the identification of Felix O1-, GJ1-, N4-, SP6-, T4-, T7-, and Vi1-like phages in multiplex PCR setups with single phage plaques as templates. Performance of the MCP PCR assay was evaluated with a set of 26 well-characterized phages. Neither false-positive nor false-negative results were obtained. In addition, 154 phages from enrichment cultures from various environmental samples were subjected to MCP PCR analysis. Eight of them, specific for Salmonella enterica , Escherichia coli , or Erwinia amylovora , belonged to one of the selected phage types. Their PCR-based identification was successfully confirmed by pulsed-field gel electrophoresis of the phage genomes, electron microscopy, and sequencing of the amplified mcp gene fragment. The MCP PCR assay was shown to be a simple method for preliminary assignment of new phages to a certain group and thus to identify candidates for biocontrol immediately after their isolation. Given that sufficient sequence data are available, this method can be extended to any phage group of interest. Electronic supplementary material The online version of this article (10.1007/s00705-019-04148-6) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

A major-capsid-protein-based multiplex PCR assay for rapid identification of selected virulent bacteriophage types

et al. 2019

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“…The earlier studies have analysed phenotypic and genotypic characterization and evaluated protective effect of phages against A. hydrophila , including Myoviridae pAh1‐C and pAh6‐C (Jun et al., 2013); Podoviridae Ahp1 (Wang et al., 2016); Myoviridae pAh‐1 (Easwaran et al., 2017); Myoviridae CT45P and TG25P (Hoang et al., 2019); Podoviridae MJG (Cao et al., 2020), Myoviridae AHP‐1 (Chandrarathna et al., 2020); Siphoviridae Akh‐2 (Akmal et al., 2020), Podoviridae LAh1‐LAh6, Siphoviridae LAh7 and Myoviridae LAh10 (Kabwe et al., 2020); Myoviridae PVN‐02 (Tu et al., 2020) and Myoviridae AhyVDH1 (Cheng et al., 2021). In previous studies, based on morphology and genomic data, the Aeromonas lytic phages pAh6‐C (Jun et al., 2013; Jun et al., 2015) and PVN02 (Dang et al., 2021; Tu et al., 2020) were initially classified as members of the Myoviridae family. However, phage pAh6‐C has recently been reclassified as Chaseviridae family, a new member of Caudovirales (https://talk.ictvonline.org/taxonomy/) (Adriaenssens et al., 2020).…”

Section: Introductionmentioning

confidence: 99%

Characterization and protective effects of lytic bacteriophage pAh6.2TG against a pathogenic multidrug‐resistant Aeromonas hydrophila in Nile tilapia ( Oreochromis niloticus )

Dien

Huy

et al. 2021

Transbounding Emerging Dis

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Bacteriophage (phage) is considered as one of the alternatives to antibiotics and an environmentally friendly approach to tackle antimicrobial resistance (AMR) in aquaculture. Here, we reported isolation, morphology and genomic characterizations of a newly isolated lytic phage, designated pAh6.2TG. Host range and stability of pAh6.2TG in different environmental conditions, and protective efficacy against a pathogenic multidrug‐resistant (MDR) Aeromonas hydrophila in Nile tilapia were subsequently evaluated. The results showed that pAh6.2TG is a member of the new family Chaseviridae which has genome size of 51,780 bp, encoding 65 putative open reading frames (ORFs) and is most closely related to Aeromonas phage PVN02 (99.33% nucleotide identity). The pAh6.2TG was highly specific to A. hydrophila and infected 83.3% tested strains of MDR A. hydrophila (10 out of 12) with relative stability at pH 7–9, temperature 0–40°C and salinity 0–40 ppt. In experimental challenge, pAh6.2TG treatments significantly improved survivability of Nile tilapia exposed to a lethal dose of the pathogenic MDR A. hydrophila, with relative per cent survival (RPS) of 73.3% and 50% for phage multiplicity of infection (MOI) 1.0 and 0.1, respectively. Phage treatment significantly reduced the concentration of A. hydrophila in both water and fish body. Interestingly, the surviving fish from A. hydrophila challenged groups provoked specific antibody (IgM) against this bacterium. In summary, the findings suggested that the lytic phage pAh6.2TG is an effective alternative to antibiotics to control MDR A. hydrophila in tilapia and possibly other freshwater fish.

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“…Lysogeny-associated, virulence-related and/or antibiotic-resistance genes should be absent in the genome of the bacteriophage, making genome sequencing essential for assessing the safety of a phage ( Jun et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producingEscherichia coliO157:H7

Amarillas

Cháidez

González‐Robles

et al. 2016

PeerJ

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BackgroundShiga toxin-producing Escherichia coli (STEC) is one of the most common and widely distributed foodborne pathogens that has been frequently implicated in gastrointestinal and urinary tract infections. Moreover, high rates of multiple antibiotic-resistant E. coli strains have been reported worldwide. Due to the emergence of antibiotic-resistant strains, bacteriophages are considered an attractive alternative to biocontrol pathogenic bacteria. Characterization is a preliminary step towards designing a phage for biocontrol.MethodsIn this study, we describe the characterization of a bacteriophage designated phiC119, which can infect and lyse several multidrug-resistant STEC strains and some Salmonella strains. The phage genome was screened to detect the stx-genes using PCR, morphological analysis, host range was determined, and genome sequencing were carried out, as well as an analysis of the cohesive ends and identification of the type of genetic material through enzymatic digestion of the genome.ResultsAnalysis of the bacteriophage particles by transmission electron microscopy showed that it had an icosahedral head and a long tail, characteristic of the family Siphoviridae. The phage exhibits broad host range against multidrug-resistant and highly virulent E. coli isolates. One-step growth experiments revealed that the phiC119 phage presented a large burst size (210 PFU/cell) and a latent period of 20 min. Based on genomic analysis, the phage contains a linear double-stranded DNA genome with a size of 47,319 bp. The phage encodes 75 putative proteins, but lysogeny and virulence genes were not found in the phiC119 genome.ConclusionThese results suggest that phage phiC119 may be a good biological control agent. However, further studies are required to ensure its control of STEC and to confirm the safety of phage use.

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Genomic structure of the Aeromonas bacteriophage pAh6-C and its comparative genomic analysis

Cited by 13 publications

References 9 publications

A major-capsid-protein-based multiplex PCR assay for rapid identification of selected virulent bacteriophage types

A major-capsid-protein-based multiplex PCR assay for rapid identification of selected virulent bacteriophage types

Characterization and protective effects of lytic bacteriophage pAh6.2TG against a pathogenic multidrug‐resistant Aeromonas hydrophila in Nile tilapia ( Oreochromis niloticus )

Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producingEscherichia coliO157:H7

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