Building next-generation sequencing (NGS) capacity for One Health and antimicrobial stewardship projects An on-going case study of multidrug-resistant Salmonella in zoo mammals - SUMMARY The overall goal of this project is to build next-generation sequencing (NGS) capacity that will support One Health and antimicrobial stewardship. Currently, the NDSU VDL along with multiple partners representing North Dakota zoos, human health lab, CDC, FDA, Vet-LIRN, NVSL, and USDA are investigating a multidrug-resistant (MDR) Salmonella serovar Newport in zoo mammals that clusters with a known human pathogenic strain, REPJJP01, making it a One Health issue. Implementation of the MIRO CANVAS would allow the NDSU VDL to more rapidly and cost effectively prepare NGS libraries for sequencing rather than waiting to batch samples. Additionally, the MIRO CANVAS coupled with a laptop, small sequencer, portable power supply, pipettes, minimal supplies, and small cooler bag could provide a SLAB, or Sequencing Lab-in- A-Bag, that could be extremely versatile and deployable in the field or point-of-care site. A zoological facility has agreed to help in the proof-of-concept of the SLAB. This equipment proposal has two specific aims. 1) Purchase and implement a MIRO CANVAS digital microfluidics platform for walk-away next-generation sequencing (NGS) library preparation to sequence bacterial and viral pathogens, especially Salmonella isolates. 2) Prepare a portable sequencing lab-in-a-bag or “SLAB” that can be deployed to the field or point-of-care site to detect and identify potential pathogens relatively rapidly and on-site.