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In this research, tetracycline resistant (tet R ) and tetracycline susceptible (tet S ) Escherichia coli isolates were retrieved from dairy manure and the influence of tetracycline resistance on the transport of E. coli in saturated porous media was investigated through laboratory column transport experiments. Experimental results showed that tet R E. coli strains had higher mobility than the tet S strains in saturated porous media. Measurements of cell surface properties suggested that tet R E. coli strains exhibited lower zeta potentials than the tet S strains. Because the surface of clean quartz sands is negatively charged, the repulsive electrostatic double layer (EDL) interaction between the tet R cells and the surface of sands was stronger and thus facilitated the transport of the tet R cells. Although no difference was observed in surface acidity, cell size, lipopolysaccharides (LPS) sugar content and cell-bound protein levels between the tet R and tet S strains, they displayed distinct outer membrane protein (OMP) profiles. It was likely that the difference in OMPs, some potentially related to drug efflux pumps, between the tet R and tet S strains led to alteration in cell surface properties which in turn affected cell transport in saturated porous media. Findings from this research suggested that manure-derived tet R E. coli could spread more widely in the groundwater system and pose serious public health risks.

Enterococcus was selected by US EPA as a Gram-positive indicator microorganism for groundwater fecal contamination. It was recently reported that enterococcal surface protein (esp) was more prevalent in Enterococcus from human sources than in Enterococcus from nonhuman sources and esp could potentially be used as a source tracking tool for fecal contamination (Scott et al., 2005). In this research, we performed laboratory column transport experiments to investigate the transport of Enterococcus faecium within saturated quartz sands. Particularly, we used a wild type strain (E1162) and a mutant (E1162Δesp) to examine the influence of esp on the transport behavior of E. faecium. Our results showed that esp could significantly enhance the attachment of E. faecium cells onto the surface of silica sands and thus lower the mobility of E. faecium within sand packs. Cell surface properties (e.g., zeta potential) were determined and the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory was applied to explain the effects of esp on the retention of E. faecium. Overall, our results suggested that E. faecium strains with esp could display lower mobility within saturated sand packs than E. faecium strains without esp. The disparity in the transport behavior of E. faecium with and without esp could limit the effectiveness of esp as a source tracking tool within the groundwater system.

It was recently reported that tetracycline could enhance the mobility of manure-derived Escherichia coli within saturated porous media (Walczak et al. (Water Research 45:1681–1690, 2011)). It was also shown, however, that E. coli from various sources could display marked variation in their mobility (Bolster et al. (Journal of Environmental Quality 35:1018–1025, 2009)). The focus of this research was to examine if the observed difference in the mobility of manure-derived tetracycline-resistant (tetR) and tetracycline-susceptible (tetS) E. coli strains was source-dependent. Specifically, E. coli were isolated from Lake Michigan, and the influence of tetracycline resistance on Lake Michigan-derived E. coli was investigated through column transport experiments. Additionally, a variety of cell morphology and surface properties were determined and related to the observed bacterial transport behavior. Our experimental results showed that, consistent with previous observations, the deposition rate coefficients of the tetR E. coli strain was ~20–100% higher than those of the tetS E. coli strain. The zeta potential of the tetR E. coli cells was ~25 mV more negative than the tetS E. coli cells. Because the surfaces of the E. coli cells and the quartz sands were negatively charged, the repulsive electrostatic double-layer interaction between the tetR E. coli cells and the quartz sands was stronger, and the mobility of the tetR E. coli cells in the sand packs was thus higher. The tetR E. coli cells were also more hydrophilic than the tetS E. coli cells. Results from migration to hydrocarbon phase (MATH) tests showed that about ~35% more tetS E. coli cells partitioned to the hydrocarbon phase. As it was previously shown that cell hydrophobicity could enhance the attachment of bacterial cells to quartz sand, the difference in cell hydrophobicity could also have contributed to the observed higher mobility of the tetR E. coli cells. The size of the tetR and tetS E. coli cells were similar, suggesting that the observed difference in their mobility was not size-related. Characterization of cell surface properties also showed that tetR and tetS E. coli cells differed slightly in cell-bound lipopolysaccharide contents and had distinct outer membrane protein profiles. Such difference could alter cell surface properties which in turn led to changes in cell mobility.

The outer membrane protein (OMP) TolC is the cell surface component of several drug efflux pumps that are responsible for bacterial resistance against a variety of antibiotics. In this research, we investigated the effects of OMP TolC on E. coli transport within saturated sands through column experiments using a wide type E. coli K12 strain (with OMP TolC), as well as the corresponding transposon mutant (tolC∷kan) and the markerless deletion mutant (ΔtolC). Our results showed OMP TolC could significantly enhance the transport of E. coli when the ionic strength was 20 mM NaCl or higher. The deposition rate coefficients for the wild type E. coli strain (with OMP TolC) was usually >50% lower than those of the tolC-negative mutants. The measurements of contact angles using three probe liquids suggested that TolC altered the surface tension components of E. coli cells and lead to lower Hamaker constants for the cell-water-sand system. The interaction energy calculations using the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory suggested that the deposition of the E. coli cell primarily occurred at the secondary energy minimum. The depth of the secondary energy minimum increased with ionic strength, and was greater for the TolC-deletion strains under high ionic strength conditions. Overall, the transport behavior of three E. coli strain within saturated sands could be explained by the XDLVO calculations. Results from this research suggested that antibiotic resistant bacteria expressing OMP TolC could spread more widely within sandy aquifers.