Enterococcus faecalis is a commensal bacterium, which upon acquisition of virulence factors on mobile genetic elements can cause sepsis, urinary tract infections and endocarditis. E. faecalis isolates can be multi-drug resistant and have been implicated in the dissemination of antibiotic resistance genes to other genera. Although the host range of pheromone inducible conjugative plasmids is restricted to Enterococci, they often carry transposons, which are capable of transposing into the chromosome of other genera. The plasmid pCF10 contains the antibiotic resistance gene tetM on a conjugative transposon Tn925. Tn925 is a Tn916-like plasmid and is capable of pCF10-independent conjugative transfer to multiple bacterial species at low levels. Biofilms are communities of bacteria growing within a matrix. In biofilms, bacteria are more difficult to kill because of their lower susceptibility to antibiotics. In hospital settings, biofilms can grow on medically implanted devices, catheters or even human tissue. In mixed species biofilms, antibiotic resistances are able to be transferred through horizontal gene transfer from E. faecalis to other bacterial species. In mixed species biofilms, it has been show that Tn925 can transpose into S. aureus at rates of 10-8 by Ella Massie Schuh. Using static mixed species biofilms, the transfer of tetM from E. faecalis to S. aureus was studied, hoping to better understand the underlying mechanisms. The goal of these studies was to determine if residence on pCF10 increased the transfer frequency of Tn925 in mixed species biofilms. Mixed species biofilms containing E. faecalis (pCF10) and S. aureus (pALC2073aPSM) were established and pCF10 conjugation was induced with pheromone cCF10. Transfer of Tn925 / Biomedical Sciences

Indicator organisms like coliforms, E. coli and enterococci are normally associated with the intestinal tract of warm-blooded animals, and can thus be used for indicating fecal contamination of water. This study investigated the removal efficiency of indicator organisms in a free water surface constructed wetland used for additional treatment of municipal wastewater. The species composition of enterococci and the prevalence of the tetracycline resistance gene tetM were also investigated. Samplings were done at five different sites: at the inlet, within and at the outlet of the wetland, as well as upstream and downstream of the wetland outlet into the adjoining river. A total of 86 enterococci isolates were characterized as E. faecalis, E. faecium or other Enterococcus spp., and the two former were also screened for the tetM gene. The investigated wetland showed a high removal rate of indicator organisms, with a removal of approximately 98%. E. faecium was identified as the predominant species in the wetland at a mean of 55.8%. In comparison, only 9.3% were E. faecalis, while other Enterococcus spp. were found to comprise 34.9%. Of the 56 isolates identified as E. feacalis or E. faecium 10.7% were positive for the tetM gene. The high removal of bacteria shown by this investigation provides further evidence of the benefits of constructed wetlands on wastewater treatment. It also offers a first indication of the Enterococcus spp. composition in Ekeby wetland, Eskilstuna, Sweden, and shows that the prevalence of the tetM gene is relatively low at this point in time.