3.2 The sewage community is rich in potent plasmid donors
In order to assess the potential of HI1A plasmid dissemination within sewage communities, we sorted and enriched R27 and Pb10 transconjugants from conjugation assays from Ellinge sewage community. Briefly, the enriched transconjugant sewage communities were grown in broth with plasmid selective antibiotics, and GFP expression was verified, to ensure plasmid maintenance and trackability. We subsequently quantified the plasmid transfer to a E. coli MG1655 recipient not encoding LacIq. This was furthermore compared to direct plasmid transfer from the E. coli donor to the E. coli recipient (figure 2b). The enriched transconjugant sewage community members harboring either R27 or pB10 and the E. coli donor were found to transfer their plasmids equally efficiently to the E. colirecipient (p > 0.05) (figure 2b). However, both the conjugation assays from E. coli to E. coli and enriched transconjugant sewage community to E. coli , revealed that R27 was transferred significantly less than pB10 (p<0.05) (figure 2b).