Effect of dissolved oxygen on two bacterial pathogens examined using ATR-FTIR spectroscopy, microelectrophoresis, and potentiometric titration.

The effects of dissolved oxygen tension during bacterial growth and acclimation on the cell surface properties and biochemical composition of the bacterial pathogens Escherichia coli O157:H7 and Yersinia enterocolitica are characterized. Three experimental techniques are used in an effort to understand the influence of bacterial growth and acclimation conditions on cell surface charge and the composition of the bacterial cell: (i) electrophoretic mobility measurements; (ii) potentiometric titration; and (iii) ATR-FTIR spectroscopy. Potentiometric titration data analyzed using chemical speciation software are related to measured electrophoretic mobilities at the pH of interest. Titration of bacterial cells is used to identify the major proton-active functional groups and the overall concentration of these cell surface ligands at the cell membrane. Analysis of titration data shows notable differences between strains and conditions, confirming the appropriateness of this tool for an overall charge characterization. ATR-FTIR spectroscopy of whole cells is used to further characterize the bacterial biochemical composition and macromolecular structures that might be involved in the development of the net surficial charge of the organisms examined. The evaluation of the integrated intensities of HPO(2)(-) and carbohydrate absorption bands in the IR spectra reveals clear differences between growth protocols. Taken together, the three techniques seem to indicate that the dissolved oxygen tension during cell growth or acclimation can noticeably influence the expression of cell surface molecules and the measurable cell surface charge, though in a strain-dependent fashion.

Transport of selected bacterial pathogens in agricultural soil and quartz sand.

The protection of groundwater supplies from microbial contamination necessitates a solid understanding of the key factors controlling the migration and retention of pathogenic organisms through the subsurface environment. The transport behavior of five waterborne pathogens was examined using laboratory-scale columns packed with clean quartz at two solution ionic strengths (10 mM and 30 mM). Escherichia coli O157:H7 and Yersinia enterocolitica were selected as representative Gram-negative pathogens, Enterococcus faecalis was selected as a representative Gram-positive organism, and two cyanobacteria (Microcystis aeruginosa and Anabaena flos-aquae) were also studied. The five organisms exhibit differing attachment efficiencies to the quartz sand. The surface (zeta) potential of the microorganisms was characterized over a broad range of pH values (2-8) at two ionic strengths (10 mM and 30 mM). These measurements are used to evaluate the observed attachment behavior within the context of the DLVO theory of colloidal stability. To better understand the possible link between bacterial transport in model quartz sand systems and natural soil matrices, additional experiments were conducted with two of the selected organisms using columns packed with loamy sand obtained from an agricultural field. This investigation highlights the need for further characterization of waterborne pathogen surface properties and transport behavior over a broader range of environmentally relevant conditions.

Role of oxygen tension on the transport and retention of two pathogenic bacteria in saturated porous media.

To examine the influence of variations in the dissolved oxygen (DO) concentration on pathogen mobility, laboratory-scale filtration experiments were performed using the enterohemorrhagic strain Escherichia coli O157:H7 and the enteroinvasive organism Yersinia enterocolitica. Cells were incubated either in the absence (anaerobic) or in the presence (aerobic) of oxygen to understand how these two growth conditions may affect bacterial transport and retention in water-saturated granular porous media. The influence of DO during growth is found to be organism dependent, whereby E. coli O157:H7 exhibits decreased transport potential when grown in the presence of 02 and Y. enterocolitica exhibits greater transport when grown aerobically. To understand the influence of DO changes during cell acclimation and transport, bacteria were resuspended and acclimated in either oxygen-depleted (low DO) or oxygen-rich (saturated DO) electrolytes prior to conduction of filtration experiments. The effect of DO on bacterial transport and retention is shown to be dependent on the antecedent growth conditions and on the organism studied. Measurements of the cell surface charge, shape, and size reveal some variability when the oxygen tension is changed during bacterial growth or acclimation and are linked to the observed bacterial transport behavior.

Relevance of nontoxigenic strains as surrogates for Escherichia coli O157:H7 in groundwater contamination potential: role of temperature and cell acclimation time.

Nontoxigenic bacteria are commonly used as indicators for predicting the contamination potential of pathogens in natural or engineered aqueous environments. In this study, column transport experiments were used to examine the relevance of two nontoxigenic strains of Escherichia coli O157:H7 as potential surrogates for the well-known pathogen. Experiments conducted at 11 degrees C indicate that only one of the nontoxigenic strains may be an appropriate surrogate for predicting the migration potential of the pathogen at low solution ionic strengths. Results of various bacterial characterization methods indicate that differences in cell attachment could qualitatively, but convincingly, be related to differences in cell surface charge. Additional experiments conducted at 22 degrees C reveal the influence of temperature on bacterial cell surface charge and cell attachmentto sand. The role of cell acclimation time to an artificial groundwater solution is also examined, showing little change in the degree of cell attachment over a period of several weeks.