Characterisation of three bacteriophages infecting serovars of salmonella enterica

A collection of three newly isolated Salmonella bacteriophages, vB_SenS-Ent1, vB_SenS-Ent2 and vB_SenS-Ent3 was established. These bacteriophages were characterised by electron microscopy, host range, sensitivity to restriction enzymes and profiles of structural proteins on SDS-PAGE gels. The complete genome sequences of each bacteriophage were established to greater than 30x coverage and bioinformatics analysis identified the functions of a number of coding sequences. Since the last update of virus taxonomy by the ICTV a number of additional genome sequences for bacteriophages infecting the genus Salmonella have been reported in the literature. To date, all but one of the Siphoviridae comprising the Salmonella bacteriophages with fully sequenced genomes remain unclassified by the ICTV. Comparative genomic analysis reveals that a number of these phages form a coherent group within the Siphoviridae and supports the establishment of a new genus, the “Setp3likeviruses”. The proposed genus includes 5 bacteriophages infecting Salmonella; SETP3, vB_SenS-Ent1, SE2, wksl3 and SS3e, and 5 infecting Escherichia; K1G, K1H, K1ind1, K1ind2 and K1ind3. This group share identical virion morphology, have terminally redundant, circularly permuted genomes ranging between 42-45 kb in size and are characterised by high nucleotide sequence similarity, shared homologous proteins and conservation of gene order.Bioluminescent bacterial reporters, transformed to express the luxCDABE operon of Photorhabdus luminescens, were used to establish the activity of the vB_SenS-Ent bacteriophages in microtitre broth lysis assays, efficacy as biological control agents for the removal of Salmonella in contaminated foods and for spatial measurements of plaque expansion in agar overlays

CoreGenes3.5: A webserver for the determination of core genes from sets of viral and small bacterial genomes

Background: CoreGenes3.5 is a webserver that determines sets of core genes from viral and small bacterial genomes as an automated batch process. Previous versions of CoreGenes have been used to classify bacteriophage genomes and mine data from pathogen genomes. Findings. CoreGenes3.5 accepts as input GenBank accession numbers of genomes and performs iterative BLASTP analyses to output a set of core genes. After completion of the program run, the results can be either displayed in a new window for one pair of reference and query genomes or emailed to the user for multiple pairs of small genomes in tabular format. Conclusions: With the number of genomes sequenced increasing daily and interest in determining phylogenetic relationships, CoreGenes3.5 provides a user-friendly web interface for wet-bench biologists to process multiple small genomes for core gene determinations. CoreGenes3.5 is available at. © 2013 Turner et al.; licensee BioMed Central Ltd

From orphan phage to a proposed new family–the diversity of N4-like viruses

Escherichia phage N4 was isolated in 1966 in Italy and has remained a genomic orphan for a long time. It encodes an extremely large virion-associated RNA polymerase unique for bacterial viruses that became characteristic for this group. In recent years, due to new and relatively inexpensive sequencing techniques the number of publicly available phage genome sequences expanded rapidly. This revealed new members of the N4-like phage group, from 33 members in 2015 to 115 N4-like viruses in 2020. Using new technologies and methods for classification, the Bacterial and Archaeal Viruses Subcommittee of the International Committee on Taxonomy of Viruses (ICTV) has moved the classification and taxonomy of bacterial viruses from mere morphological approaches to genomic and proteomic methods. The analysis of 115 N4-like genomes resulted in a huge reassessment of this group and the proposal of a new family “Schitoviridae”, including eight subfamilies and numerous new genera

Genomic analysis of two novel bacteriophages infecting Acinetobacter beijerinckii and halotolerans species

Bacteriophages are the most diverse genetic entities on Earth. In this study, two novel bacteriophages, nACB1 (Podoviridae morphotype) and nACB2 (Myoviridae morphotype), which infect Acinetobacter beijerinckii and Acinetobacter halotolerans, respectively, were isolated from sewage samples. The genome sequences of nACB1 and nACB2 revealed that their genome sizes were 80,310 bp and 136,560 bp, respectively. Comparative analysis showed that both genomes are novel members of the Schitoviridae and the Ackermannviridae families, sharing ≤ 40% overall nucleotide identities with any other phages. Interestingly, among other genetic features, nACB1 encoded a very large RNA polymerase, while nACB2 displayed three putative depolymerases (two capsular depolymerases and one capsular esterase) encoded in tandem. This is the first report of phages infecting A. halotolerans and beijerinckii human pathogenic species. The findings regarding these two phages will allow us to further explore phage—Acinetobacter interactions and the genetic evolution for this group of phages.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit, and by LABBELS—Associate Laboratory in Biotechnology, Bioengineering and Microelectromechnical Systems, LA/P/0029/2020.info:eu-repo/semantics/publishedVersio

A roadmap for genome-based phage taxonomy

Bacteriophage (phage) taxonomy has been in flux since its inception over four decades ago. Genome sequencing has put pressure on the classification system and recent years have seen significant changes to phage taxonomy. Here, we reflect on the state of phage taxonomy and provide a roadmap for the future, including the abolition of the order Caudovirales and the families Myoviridae, Podoviridae, and Siphoviridae. Furthermore, we specify guidelines for the demarcation of species, genus, subfamily and family-level ranks of tailed phage taxonomy

Genome sequence of vB_AbaS_TRS1, a viable prophage isolated from Acinetobacter baumannii strain A118

© 2016 Turner et al. A novel temperate phage, vB_AbaS_TRS1, was isolated from cultures of Acinetobacter baumannii strain A118 that had been exposed to mitomycin C. Phage TRS1 belongs to the Siphoviridae family of bacteriophages and encapsulates a 40,749-bp genome encoding 70 coding sequences and a single tRNA

Pectobacterium atrosepticum phage vB_PatP_CB5: A member of the proposed genus ‘Phimunavirus’

© 2018 by the authors. Licensee MDPI, Basel, Switzerland. Pectobacterium atrosepticum is a phytopathogen of economic importance as it is the causative agent of potato blackleg and soft rot. Here we describe the Pectobacterium phage vB_PatP_CB5 (abbreviated as CB5), which specifically infects the bacterium. The bacteriophage is characterized in detail and TEM micrographs indicate that it belongs to the Podoviridae family. CB5 shares significant pairwise nucleotide identity (≥80%) with P. atrosepticum phages 'M1, Peat1, and PP90 and also shares common genome organization. Phylograms constructed using conserved proteins and whole-genome comparison-based amino acid sequences show that these phages form a distinct clade within the Autographivirinae. They also possess conserved RNA polymerase recognition and specificity loop sequences. Their lysis cassette resembles that of KP34virus, containing in sequential order a U-spanin, a holin, and a signal–arrest–release (SAR) endolysin. However, they share low pairwise nucleotide identity with the type phage of the KP34virus genus, Klebsiella phage KP34. In addition, phage KP34 does not possess several conserved proteins associated with these P. atrosepticum phages. As such, we propose the allocation of phages CB5, Peat1, 'M1, and PP90 to a separate new genus designated Phimunavirus

Abolishment of morphology‑based taxa and change to binomial species names: 2022 taxonomy update of the ICTV bacterial viruses subcommittee

This article summarises the activities of the Bacterial Viruses Subcommittee of the International Committee on Taxonomy of Viruses for the period of March 2021−March 2022. We provide an overview of the new taxa proposed in 2021, approved by the Executive Committee, and ratifed by vote in 2022. Signifcant changes to the taxonomy of bacterial viruses were introduced: the paraphyletic morphological families Podoviridae, Siphoviridae, and Myoviridae as well as the order Caudovirales were abolished, and a binomial system of nomenclature for species was established. In addition, one order, 22 families, 30 subfamilies, 321 genera, and 862 species were newly created, promoted, or moved

Comparative analysis of 37 Acinetobacter bacteriophages

© 2017 by the authors. Licensee MDPI, Basel, Switzerland. Members of the genus Acinetobacter are ubiquitous in the environment and the multipledrug resistant species A. baumannii is of significant clinical concern. This clinical relevance is currently driving research on bacterial viruses infecting A. baumannii, in an effort to implement phage therapy and phage-derived antimicrobials. Initially, a total of 42 Acinetobacter phage genome sequences were available in the international nucleotide sequence databases, corresponding to a total of 2.87 Mbp of sequence information and representing all three families of the order Caudovirales and a single member of the Leviviridae. A comparative bioinformatics analysis of 37 Acinetobacter phages revealed that they form six discrete clusters and two singletons based on genomic organisation and nucleotide sequence identity. The assignment of these phages to clusters was further supported by proteomic relationships established using OrthoMCL. The 4067 proteins encoded by the 37 phage genomes formed 737 groups and 974 orphans. Notably, over half of the proteins encoded by the Acinetobacter phages are of unknown function. The comparative analysis and clustering presented enables an updated taxonomic framing of these clades

Greywater irrigation as a source of organic micro-pollutants to shallow groundwater and nearby surface water

Increased water demands due to population growth and increased urbanisation have driven adoption of various water reuse practices. The irrigation of greywater (water from all household uses, except toilets) has been proposed as one potential sustainable practice. Research has clearly identified environmental harm from the presence of micro-pollutants in soils, groundwater and surface water. Greywater contains a range of micro pollutants yet very little is known about their potential environmental fate when greywater is irrigated to soil. Therefore, this study assessed whether organic micro-pollutants in irrigated greywater were transferred to shallow groundwater and an adjacent surface waterway. A total of 22 organic micro-pollutants were detected in greywater. Six of these (acesulfame, caffeine, DEET, paracetamol, salicylic acid and triclosan) were selected as potential tracers of greywater contamination. Three of these chemicals (acesulfame, caffeine, DEET) were detected in the groundwater, while salicylic acid was also detected in adjacent surface water. Caffeine and DEET in surface water were directly attributable to greywater irrigation. Thus the practice of greywater irrigation can act as a source of organic micro-pollutants to shallow groundwater and nearby surface water. The full list of micro-pollutants that could be introduced via greywater and the risk they pose to aquatic ecosystems is not yet known