Park M, Lee JH, Shin H, Kim M, Choi J, Kang DH et al.. Characterization and comparative genomic analysis of a novel bacteriophage, SFP10, simultaneously inhibiting both Salmonella enterica and Escherichia coli O157:H7. Appl Environ Microbiol 78: 58-69

Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Research Institute for Agriculture and Life Sciences, and Center for Agricultural Biomaterials, Seoul National University, Seoul, South Korea.
Applied and Environmental Microbiology (Impact Factor: 3.67). 01/2012; 78(1):58-69. DOI: 10.1128/AEM.06231-11
Source: PubMed


Salmonella enterica and Escherichia coli O157:H7 are major food-borne pathogens causing serious illness. Phage SFP10, which revealed effective infection of both S. enterica and E. coli O157:H7, was isolated and characterized. SFP10 contains a 158-kb double-stranded DNA genome belonging to the Vi01 phage-like
family Myoviridae. In vitro adsorption assays showed that the adsorption constant rates to both Salmonella enterica serovar Typhimurium and E. coli O157:H7 were 2.50 × 10−8 ml/min and 1.91 × 10−8 ml/min, respectively. One-step growth analysis revealed that SFP10 has a shorter latent period (25 min) and a larger burst
size (>200 PFU) than ordinary Myoviridae phages, suggesting effective host infection and lytic activity. However, differential development of resistance to SFP10
in S. Typhimurium and E. coli O157:H7 was observed; bacteriophage-insensitive mutant (BIM) frequencies of 1.19 × 10−2 CFU/ml for S. Typhimurium and 4.58 × 10−5 CFU/ml for E. coli O157:H7 were found, indicating that SFP10 should be active and stable for control of E. coli O157:H7 with minimal emergence of SFP10-resistant pathogens but may not be for S. Typhimurium. Specific mutation of rfaL in S. Typhimurium and E. coli O157:H7 revealed the O antigen as an SFP10 receptor for both bacteria. Genome sequence analysis of SFP10 and its comparative
analysis with homologous Salmonella Vi01 and Shigella phiSboM-AG3 phages revealed that their tail fiber and tail spike genes share low sequence identity, implying that the genes
are major host specificity determinants. This is the first report identifying specific infection and inhibition of Salmonella Typhimurium and E. coli O157:H7 by a single bacteriophage.

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    • "Although future experimental work is needed, a role of the C-terminal variable regions in phage host specificity is supported by the observation that one of the tailspikes of the Salmonella Typhi specific phage Vi1 has a conserved acetyl esterase domain that recognizes the Typhi capsule as receptor [38]. Phage SFP10, which was reported to infect both Salmonella and E. coli O157:H7, also carries tailspikes that presented similarity to tailspikes found in both E. coli and Salmonella phages [39], suggesting a possible role of these regions in host specificity. "
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    ABSTRACT: Salmonella is a widely distributed foodborne pathogen that causes tens of millions of salmonellosis cases globally every year. While the genomic diversity of Salmonella is increasingly well studied, our knowledge of Salmonella phage genomic diversity is still rather limited, despite the contributions of both lysogenic and lytic phages to Salmonella virulence, diversity and ecology (e.g., through horizontal gene transfer and Salmonella lysis). To gain a better understanding of phage diversity in a specific ecological niche, we sequenced 22 Salmonella phages isolated from a number of dairy farms from New York State (United States) and analyzed them using a comparative genomics approach. Classification of the 22 phages according to the presence/absence of orthologous genes allowed for classification into 8 well supported clusters. In addition to two phage clusters that represent novel virulent Salmonella phages, we also identified four phage clusters that each contained previously characterized phages from multiple continents. Our analyses also identified two clusters of phages that carry putative virulence (e.g., adhesins) and antimicrobial resistance (tellurite and bicyclomycin) genes as well as virulent and temperate transducing phages. Insights into phage evolution from our analyses include (i) identification of DNA metabolism genes that may facilitate nucleotide synthesis in phages with a G+C % distinct from Salmonella, and (ii) evidence of Salmonella phage tailspike and fiber diversity due to both single nucleotide polymorphisms and major re-arrangements, which may affect the host specificity of Salmonella phages. Genomics-based characterization of 22 Salmonella phages isolated from dairy farms allowed for identification of a number of novel Salmonella phages. While the comparative genomics analyses of these phages provide a number of new insights in the evolution and diversity of Salmonella phages, they only represent a first glimpse into the diversity of Salmonella phages that is likely to be discovered when phages from different environments are characterized.
    BMC Genomics 07/2013; 14(1):481. DOI:10.1186/1471-2164-14-481 · 3.99 Impact Factor
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    • "Burst size of PhaxI is about 420 particles per cell. Burst sizes of AR1, CEV1, CEV2, AKFV33, CBA120 and SFP10 are 34, 150, 350, 350, 440 and 100 p.f.u. per cell, respectively (Kutter et al., 2011; Niu et al., 2012; Park et al., 2012; Raya et al., 2006, 2011; Ronner & Cliver, 1990). "
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    ABSTRACT: Bacteriophages are considered as promising biological agents for the control of infectious diseases. Sequencing of their genomes can ascertain the absence of antibiotic resistance, toxin or virulence genes. The anti-O157:H7 coliphage, PhaxI, was isolated from a sewage sample in Iran. Morphological studies by TEM showed that it has an icosahedral capsid of 85-86 nm and a contractile tail of 115 x 15 nm. PhaxI contains double-stranded DNA composed of 156,628 nucleotides with a G+C content of 44.5% that encodes 209 putative proteins. In MS analysis of phage particles altogether 92 structural proteins were identified. PhaxI lyses E. coli O157:H7 in LB medium and milk, has an eclipse period of 20 minutes, a latent period of 40 minutes, and a burst size of about 420 particles per cell. PhaxI is a member of the genus "Viunalikevirus" of the Myoviridae family and is specific for Escherichia coli O157:H7.
    Microbiology 05/2013; 159(Pt_8). DOI:10.1099/mic.0.063776-0 · 2.56 Impact Factor
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    • "The scientific literature lists over fifty phages described as being E.coli O157-specific. These include sixteen phages (V1-V16) comprising part of a phage typing scheme for this bacterium [6] plus phages 38, 39, 41, 42, ECB7 and ECA1 [7]; AR1 [8,9]; Bo-21, Av-05, Av-06, and Av-08 [10]; CA933P, CA911 MFA933P and MFA45D [11]; CEV1 and CEV2 [12,13]; CSLO157 [14]; DC22 [15], e4/1c and e11/2 [16]; ECML-4, ECML-117, and ECML-134 [17]; JK06; KH1, KH4 and KH5 [18]; LG1 [19]; φV10 [20,21]; PP01 [22]; SFP10 [23]; SH1 [24]; SP15, SP21, and SP22 [25]; vB_EcoM_CBA120 [5]; vB_EcoS_AKFV33 [4]; and, vB_EcoS_Rogue1 [26]. However, relatively little or consistent information on morphology and taxonomic position, host range, receptor specificity, genome size and characterization is available for many of these viruses. "
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