Evaluation of Biofilm Induced Urinary Infection Stone Formation in a Novel Laboratory Model System.

PURPOSE: Infection stones, which comprise approximately 15% of all urinary tract stones, are induced by infection with urease-positive pathogens. The bacteria in the stone matrix present significant treatment impediments compared to metabolic kidney stones. While much is known about how urinary composition regulates metabolic stone formation, there is a general lack of knowledge of which urinary factors regulate the rate of infection stone formation. Unfortunately more in-depth research into infection stones is limited by the lack of suitable models for real-time study of bacterial biofilm formation and stone formation under varying conditions. MATERIALS AND METHODS: We developed an in vitro model to study infection stone formation. The model closely represents the processes that occur in vivo, including the observed migration of ureolytic bacteria (our culture of Proteus mirabilis) from the bladder to the kidneys, followed by biofilm and stone formation in the kidney. We used scanning electron and confocal laser microscopy, x-ray diffraction, biological counts and dissolved chemical analyses to evaluate the model system. RESULTS: Crystals that formed in the system resembled clinically removed struvite stones in structure and composition. Results showed that the degree of ureolysis required to significantly change urine pH was minimal, bacterial communities inhabited the ureter, and upstream colonization and struvite formation required lag time. CONCLUSIONS: These results have implications for the detection and treatment of struvite stones. Currently this model is being used to study specific urinary factors that regulate struvite formation to identify treatment options, which combined with antibiotics would improve treatment of these stones and decrease recurrence.

Bacterially induced calcium carbonate precipitation and strontium coprecipitation in a porous media flow system.

Strontium-90 is a principal radionuclide contaminant in the subsurface at several Department of Energy sites in the Western U.S., causing a threat to groundwater quality in areas such as Hanford, WA. In this work, we used laboratory-scale porous media flow cells to examine a potential remediation strategy employing coprecipitation of strontium in carbonate minerals. CaCO(3) precipitation and strontium coprecipitation were induced via ureolysis by Sporosarcina pasteurii in two-dimensional porous media reactors. An injection strategy using pulsed injection of calcium mineralization medium was tested against a continuous injection strategy. The pulsed injection strategy involved periods of lowered calcite saturation index combined with short high fluid velocity flow periods of calcium mineralization medium followed by stagnation (no-flow) periods to promote homogeneous CaCO(3) precipitation. By alternating the addition of mineralization and growth media the pulsed strategy promoted CaCO(3) precipitation while sustaining the ureolytic culture over time. Both injection strategies achieved ureolysis with subsequent CaCO(3) precipitation and strontium coprecipitation. The pulsed injection strategy precipitated 71-85% of calcium and 59% of strontium, while the continuous injection was less efficient and precipitated 61% of calcium and 56% of strontium. Over the 60 day operation of the pulsed reactors, ureolysis was continually observed, suggesting that the balance between growth and precipitation phases allowed for continued cell viability. Our results support the pulsed injection strategy as a viable option for ureolysis-induced strontium coprecipitation because it may reduce the likelihood of injection well accumulation caused by localized mineral plugging while Sr coprecipitation efficiency is maintained in field-scale applications.

NMR measurement of hydrodynamic dispersion in porous media subject to biofilm mediated precipitation reactions.

Noninvasive measurements of hydrodynamic dispersion by nuclear magnetic resonance (NMR) are made in a model porous system before and after a biologically mediated precipitation reaction. Traditional magnetic resonance imaging (MRI) was unable to detect the small scale changes in pore structure visualized during light microscopy analysis after destructive sampling of the porous medium. However, pulse gradient spin echo nuclear magnetic resonance (PGSE NMR) measurements clearly indicated a change in hydrodynamics including increased pore scale mixing. These changes were detected through time-dependent measurement of the propagator by PGSE NMR. The dynamics indicate an increased pore scale mixing which alters the preasymptotic approach to asymptotic Gaussian dynamics governed by the advection diffusion equation. The methods described here can be used in the future to directly measure the transport of solutes in biomineral-affected porous media and contribute towards reactive transport models, which take into account the influence of pore scale changes in hydrodynamics.