Journal
MICROORGANISMS
Volume 9, Issue 3, Pages -Publisher
MDPI
DOI: 10.3390/microorganisms9030608
Keywords
bacterial surrogate; Escherichia coli; whole genome sequence; short reads; long reads; closed genome; high throughput sequencing
Categories
Funding
- USDA Agriculture and Food Research Initiative [2012-68003-30155]
- Texas Beef Council/Check-Off Program
- Beef Competitiveness Exceptional Item from the Texas legislature
- Texas A&M AgriLife Research
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This study involved sequencing the genomes of E. coli surrogates using two different platforms and found that combining long-read, short-read, and combined sequences can yield more accurate completed genomes. These data provide valuable insights for the adoption of WGS in the food industry and offer guidance for researchers and regulators in implementing related workflows.
In 2013, the U.S. Department of Agriculture Food Safety and Inspection Service (USDA-FSIS) began transitioning to whole genome sequencing (WGS) for foodborne disease outbreak- and recall-associated isolate identification of select bacterial species. While WGS offers greater precision, certain hurdles must be overcome before widespread application within the food industry is plausible. Challenges include diversity of sequencing platform outputs and lack of standardized bioinformatics workflows for data analyses. We sequenced DNA from USDA-FSIS approved, non-pathogenic E. coli surrogates and a derivative group of rifampicin-resistant mutants (rif(R)) via both Oxford Nanopore MinION and Illumina MiSeq platforms to generate and annotate complete genomes. Genome sequences from each clone were assembled separately so long-read, short-read, and combined sequence assemblies could be directly compared. The combined sequence data approach provides more accurate completed genomes. The genomes from these isolates were verified to lack functional key E. coli elements commonly associated with pathogenesis. Genetic alterations known to confer rif(R) were also identified. As the food industry adopts WGS within its food safety programs, these data provide completed genomes for commonly used surrogate strains, with a direct comparison of sequence platforms and assembly strategies relevant to research/testing workflows applicable for both processors and regulators.
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