Fig. 5: Gentamicin-resistant plasmid transfer study in Enterococcus.

a Schematic of the ex vivo study workflow and associated analyses, illustrating the study design within a 24-well plate format. b Antibiotic resistance gene profiling of donor and recipient strains. Colonies were individually picked from selective agar plates supplemented with the appropriate antibiotics and subjected to whole-genome sequencing using a combined long-read and short-read sequencing platform. A neighbour-joining tree was constructed to depict the relatedness between isolate genomes, aligned with plasmid replicons predicted within the genomes alongside their calculated genome sizes. The inset (top left) shows shotgun metagenomic contents (microbial taxa) of the infant faecal slurry utilised in the gut model experiment. Gene aac6-Ii is known to be a chromosomally encoded aminoglycoside acetyltransferase in Enterococcus spp. ARMA-59 and ARMA-73 are preterm-derived isolates (donor strains) whereas 64-3 (or, 64/3) is a plasmid-free laboratory strain. c Comparison of the computationally extracted antibiotic resistance gene aac-aph2 from the representative donor strain B1 and recipient (transformant) strains D1-1 and D3-1. d Comparison of complete contig sequences encoding the gentamicin resistance gene aac6-aph2, predicted to be either whole or partial sequences of potential mobile genetic elements, including plasmids.