Isolation, genome sequencing and functional analysis of two T7-like coliphages of avian pathogenic Escherichia coli

Chen, Mianmian; Xu, Juntian; Yao, Huochun; Chen, Lu; Zhang, Wei

doi:10.1016/j.gene.2016.01.049

Cited by 30 publications

(21 citation statements)

References 60 publications

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“…This is lower than the 56.3% found in the host, which is common amongst phages [ 29 ]. No close relatives of the Cluster A phages exist in the publicly available databases as determined by BLASTn, but the closest relative is the T7-like escherichia phage P694(KP090454) [ 30 ] which shares less than 1% sequence similarity with any Cluster A phages ( Figure 1 ). Cluster A phages are quite diverse, with the individual phages sharing only between 48.5% and 71.9% sequence similarity.…”

Section: Resultsmentioning

confidence: 99%

Unlocking the Potential of 46 New Bacteriophages for Biocontrol of Dickeya Solani

Carstens

Djurhuus

Kot

et al. 2018

Viruses

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Modern agriculture is expected to face an increasing global demand for food while also needing to comply with higher sustainability standards. Therefore, control of crop pathogens requires new, green alternatives to current methods. Potatoes are susceptible to several bacterial diseases, with infections by soft rot Enterobacteriaceae (SRE) being a significant contributor to the major annual losses. As there are currently no efficient ways of combating SRE, we sought to develop an approach that could easily be incorporated into the potato production pipeline. To this end, 46 phages infecting the emerging potato pathogen Dickeya solani were isolated and thoroughly characterized. The 46 isolated phages were grouped into three different groups based on DNA similarity, representing two distinct clusters and a singleton. One cluster showed similarity to phages previously used to successfully treat soft rot in potatoes, whereas the remaining phages were novel and showed only very limited similarity to previously isolated phages. We selected six diverse phages in order to create the hereto most complex phage cocktail against SRE. The cocktail was applied in a proof-of-principle experiment to treat soft rot in potatoes under simulated storage conditions. We show that the phage cocktail was able to significantly reduce the incidence of soft rot as well as disease severity after 5 days of storage post-infection with Dickeya solani. This confirms results from previous studies that phages represent promising biocontrol agents against SRE infection in potato.

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Section: Resultsmentioning

confidence: 99%

Unlocking the Potential of 46 New Bacteriophages for Biocontrol of Dickeya Solani

Carstens

Djurhuus

Kot

et al. 2018

Viruses

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“…Tailed bacteriophages constitute the most abundant and diverse group of dsDNA viruses, in which evolution is very complicated (Iranzo et al, 2016 ). Several results illustrated that phages may undergo genetic exchange by horizontal gene transfer from a large shared pool, and that horizontal gene transfer between phages is a component of evolution (Håggard-Ljungquist et al, 1992 ; Hendrix et al, 1999 ; Casjens, 2008 ; Dekel-Bird et al, 2013 ; Chen et al, 2016 ). In our study, both horizontal exchange and vertical gene order rearrangement may affect the organization of bacteriophage genomes and blur phylogenetic reconstructions in vB_EcoS-B2 and its relative phages.…”

Section: Resultsmentioning

confidence: 99%

Characterization and Genomic Study of Phage vB_EcoS-B2 Infecting Multidrug-Resistant Escherichia coli

et al. 2018

Front. Microbiol.

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The potential of bacteriophage as an alternative antibacterial agent has been reconsidered for control of pathogenic bacteria due to the widespread occurrence of multi-drug resistance bacteria. More and more lytic phages have been isolated recently. In the present study, we isolated a lytic phage named vB_EcoS-B2 from waste water. VB_EcoS-B2 has an icosahedral symmetry head and a long tail without a contractile sheath, indicating that it belongs to the family Siphoviridae. The complete genome of vB_EcoS-B2 is composed of a circular double stranded DNA of 44,283 bp in length, with 54.77% GC content. vB_EcoS-B2 is homologous to 14 relative phages (such as Escherichia phage SSL-2009a, Escherichia phage JL1, and Shigella phage EP23), but most of these phages exhibit different gene arrangement. Our results serve to extend our understanding toward phage evolution of family Siphoviridae of coliphages. Sixty-five putative open reading frames were predicted in the complete genome of vB_EcoS-B2. Twenty-one of proteins encoded by vB_EcoS-B2 were determined in phage particles by Mass Spectrometry. Bacteriophage genome and proteome analysis confirmed the lytic nature of vB_EcoS-B2, namely, the absence of toxin-coding genes, islands of pathogenicity, or genes through lysogeny or transduction. Furthermore, vB_EcoS-B2 significantly reduced the growth of E. coli MG1655 and also inhibited the growth of several multi-drug resistant clinical stains of E. coli. Phage vB_EcoS-B2 can kill some of the MRD E. coli entirely, strongly indicating us that it could be one of the components of phage cocktails to treat multi-drug resistant E. coli. This phage could be used to interrupt or reduce the spread of multi-drug resistant E. coli.

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“…The purified phage suspensions were stored at −80, 4, and 30 °C, respectively, and the phage titer was determined by double-agar overlay assay immediately and every week after storage 36 . For the thermal stability assay, equal volumes of phage (approximately 10 8 Pfu/ml) were added into the 1.5 ml tube and incubated at 28 °C, 37 °C, 50 °C, 60 °C, and 70 °C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a newly isolated phage infecting pathogenic Escherichia coli and analysis of its mosaic structural genes

Qin

Yuan

2018

Sci Rep

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The outbreak of multidrug-resistant pathogenic bacteria made the discovery of novel control strategies necessary. Phages have regained attention for their specific lytic activity against pathogenic bacterium. A newly isolated phage infecting the clinical Escherichia coli isolates, including several multidrug-resistant strains, was isolated, and this phage showed high control effects against the tested pathogenic E. coli strains. Host range analysis revealed that although the phage exhibited broad lytic spectrum against the tested E. coli strains, it could not lyse strains from the other species. Comparative genomic analysis showed that phages had undergone at least three genome recombination events during the evolutionary process at the position of the three phage tail genes, which was reported to be associated with the host range determination of the phage. The recombinant tail proteins contained functional domains that were highly similar with genes of the Salmonella phage and genes of Pseudomonas and Neisseria. The findings of this study not only provide resources for developing phage therapy against E. coli, but also showed the highly variable genome structure of the phage.

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Isolation, genome sequencing and functional analysis of two T7-like coliphages of avian pathogenic Escherichia coli

Cited by 30 publications

References 60 publications

Unlocking the Potential of 46 New Bacteriophages for Biocontrol of Dickeya Solani

Unlocking the Potential of 46 New Bacteriophages for Biocontrol of Dickeya Solani

Characterization and Genomic Study of Phage vB_EcoS-B2 Infecting Multidrug-Resistant Escherichia coli

Characterization of a newly isolated phage infecting pathogenic Escherichia coli and analysis of its mosaic structural genes

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