Fig. 1

Workflow of the enrichment, isolation, and genomic characterization of anaerobic ureolytic bacteria from the rumen. Firstly, rumen microbiota samples were serially diluted and distributed into 96-well plates. After incubation, the wells with growth of ureolytic bacteria were identified by urease gene (ureC) specific PCR. Secondly, enriched ureolytic bacterial cells were serially diluted and embedded within agarose microspheres (aimed for one cell per microsphere). Thirdly, agarose microspheres were placed into dialysis bags that were placed into a rumen-simulating system for incubation. Finally, single strains in each microsphere were identified by 16S rRNA gene sequencing and subjected to whole-genome sequencing and analysis