title: Two Novel Myoviruses from the North of Iraq Reveal Insights into Clostridium difficile Phage Diversity and Biology.
abstract: Bacteriophages (phages) are increasingly being explored as therapeutic agents to combat bacterial diseases, includingClostridium difficileinfections. Therapeutic phages need to be able to efficiently target and kill a wide range of clinically relevant strains. While many phage groups have yet to be investigated in detail, those with new and useful properties can potentially be identified when phages from newly studied geographies are characterised. Here, we report the isolation ofC. difficilephages from soil samples from the north of Iraq. Two myoviruses, CDKM15 and CDKM9, were selected for detailed sequence analysis on the basis of their broad and potentially useful host range. CDKM9 infects 25/80 strains from 12/20C. difficileribotypes, and CDKM15 infects 20/80 strains from 9/20 ribotypes. Both phages can infect the clinically relevant ribotypes R027 and R001. Phylogenetic analysis based on whole genome sequencing revealed that the phages are genetically distinct from each other but closely related to other long-tailed myoviruses. A comparative genomic analysis revealed key differences in the genes predicted to encode for proteins involved in bacterial infection. Notably, CDKM15 carries a clustered regularly interspaced short palindromic repeat (CRISPR) array with spacers that are homologous to sequences in the CDKM9 genome and of phages from diverse localities. The findings presented suggest a possible shared evolutionary past for these phages and provides evidence of their widespread dispersal.
authors.affiliation: Department of Infection, Immunity and Inflammation, Medical Sciences Building, University of Leicester, University Road, Leicester, LE1 9HN, UK. saroa.rashid@gmail.com.
authors.forename: Srwa J
authors.initials: SJ
authors.lastname: Rashid
authors.affiliation: Department of Molecular Virology, Faculty of Biology, Adam Mickiewicz University in Poznan, Poznan 61-712, Poland. erytropoeta@gmail.com.
authors.forename: Jakub
authors.initials: J
authors.lastname: Barylski
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, OH 43201, USA. krhargr@gmail.com.
authors.forename: Katherine R
authors.initials: KR
authors.lastname: Hargreaves
authors.affiliation: Microbiology & Infection Unit, Warwick Medical School, University of Warwick, Coventry, CV4 7AL, UK. andrew.millard@warwick.ac.uk.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Millard
authors.affiliation: Department of Chemical Engineering, Loughborough University, Loughborough, LE11 3TU, UK. G.Vinner@lboro.ac.uk.
authors.forename: Gurinder K
authors.initials: GK
authors.lastname: Vinner
authors.affiliation: Department of Infection, Immunity and Inflammation, Medical Sciences Building, University of Leicester, University Road, Leicester, LE1 9HN, UK. mrjc1@le.ac.uk.
authors.forename: Martha R J
authors.initials: MR
authors.lastname: Clokie
mesh_terms: D001435:Bacteriophages / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D016360:Clostridioides difficile / Q000821:virology*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D016679:Genome, Viral; D058507:Host Specificity; D007493:Iraq; D046529:Microscopy, Electron, Transmission; D017900:Myoviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D014771:Virion / Q000648:ultrastructure
chemical_list: D004279:DNA, Viral
keywords: CRISPR/Cas system; Clostridium difficile; bacteriophage; endolysin; genome evolution; large terminase gene; phylogenetic analysis