2. Abstract
Over the last 35 years in the UK the burden of Shiga toxin-producing Escherichia coli (STEC) O157:H7 infection has, during different periods of time, been associated with five different sub-lineages (1983-1995: Ia, I/IIa and I/IIb, 1996-2014: Ic and 2015-2018: IIb). The acquisition of a stx2a-encoding bacteriophage by these five sub-lineages appears to have coincided with their respective emergences. Oxford Nanopore Technology (ONT) was used to sequence, characterise and compare the stx-encoding prophage harboured by each sub-lineage to investigate the integration of this key virulence factor. The stx2a-encoding prophage from each of the lineages causing clinical disease in the UK were all different, including the two UK sub-lineages (Ia and I/IIa) circulating concurrently and causing severe disease in the early 1980s. Comparisons between the stx2a-encoding prophage in sub-lineages I/IIb and IIb revealed similarity to the prophage commonly found to encode stx2c, and the same site of bacteriophage integration (sbcB) as stx2c encoding prophage. These data suggest independent acquisition of previously unobserved stx2a-encoding phage is more likely to have contributed to the emergence of STEC O157:H7 sub-lineages in the UK than intra-UK lineage to lineage phage transmission. In contrast, the stx2c-encoding prophage showed a high level of similarity across lineage and time, consistent with the model of stx2c being present in the common ancestor to extant STEC O157:H7 and maintained by vertical inheritance in the majority of the population. Studying the nature of the stx-encoding bacteriophage contributes to our understanding of the emergence of highly pathogenic strains of STEC O157:H7.
Impact statement The application of ONT technology to sequence UK epidemic strains of STEC O157:H7 revealed stx2a-encoding prophage exhibit a high level of diversity. There was little evidence of geographical or temporal patterns of relatedness, or of intra-UK transmission of stx2a-encoding prophage between indigenous strains. The stx2a-encoding prophage in the UK lineages associated with severe disease appear to be acquired independently and most likely from different geographical and/or environmental sources. These data provide supporting evidence for the existence of a dynamic environmental reservoir of stx2a-encoding prophage that pose a threat public health due to their potential for integration into competent, indigenous sub-lineages of STEC O157:H7. We also provide further evidence that stx2c-encoding prophage exhibit a high level of similarity across lineage, geographical region and time, and have likely been maintained and inherited vertically.
Data summary All FASTQ files and assemblies of samples sequenced in this project were submitted to the National Centre for Biotechnology Information (NCBI). All data can be found under BioProject: PRJNA315192 - https://www.ncbi.nlm.nih.gov/bioproject/?term=PRJNA315192. Strain specific details can be found in the methods section under data deposition.
Publicly available data used in this project can be found via Table 1 and data bibliography.