Magnetic resonance imaging of biofilm mass transport processes with gadolinium tracers

Lead Research Organisation: University of Glasgow
Department Name: School of Geographical & Earth Sciences

Abstract

Biofilms are dense cities of bacteria which adhere together by excreting a slimy, glue-like substance. Significantly, these slimy communities offer huge potential in an array of important biotechnological applications, such as sewage treatment, biofuel production and the generation of electricity in microbial fuel cells. They also play an important role in controlling the chemistry of the natural environment. For a biofilm to function, however, reactants (e.g. the sewage in sewage treatment plants) must be efficiently transported through the biofilm where they are processed by bacteria. Significantly, the rate at which the biofilm can operate is controlled by the rate at which these reactants move through the biofilm. Consequently, it is vital for engineers and microbiologists to be able to measure the rate of reactant supply. Critically, this data is essential to our understanding of the way biofilms work and our ability to enhance biofilm performance. Whilst tools for measuring transport in biofilms exist, they cannot measure all the parameters needed (for example, some are restricted to either high or low molecular mass reactants) and some are invasive, potentially damaging the biofilm, altering results. Magnetic resonance imaging (MRI), however, has tremendous potential to bridge this technology gap. MRI is non-invasive and so it quite literally enables us to look inside the biofilm and measure the movement of reactants without harming the biofilm in any way. Problematically, while MRI can measure the movement of water in biofilms (which can be used as a proxy for the movement of other low molecular mass compounds), measurement of high molecular mass molecules is difficult. This, however, can change. By labeling these molecules with a paramagnetic ion (in this case gadolinium), the molecule suddenly becomes easily visible with MRI. This technology is already applied in clinical research, where gadolinium is used to make molecules readily visible in human and other mammalian tissues. Here, we aim to demonstrate that paramagnetically labeled molecules can be used to track mass transport within biofilms. In this investigation, we will image the transport of a range of commercially available gadolinium labeled molecules in biofilms from laboratory wastewater treatment bioreactors and from natural systems. A calibration protocol will used to convert MRI data into actual gadolinium concentrations, enabling us to determine the concentration of Gd in each image pixel at each time interval. From this, diffusion coefficients for each gadolinium labeled molecule in each biofilm will be calculated. A 3D model will also be used the generate maps of diffusion coefficients throughout each biofilm. If successful, this technology would be an invaluable tool providing microbiologists and engineers alike with essential transport data needed to harness the full power of these complex biological communities.

Publications

10 25 50
 
Description Biofilms are dense cities of bacteria which adhere together by excreting a slimy, glue-like substance. Significantly, these slimy communities offer huge potential in an array of important biotechnological applications, such as sewage treatment, biofuel production and the generation of electricity in microbial fuel cells. For a biofilm to function, however, reactants (e.g. the sewage in sewage treatment plants) must be efficiently transported through the biofilm where they are processed by the bacteria. Significantly, the rate at which the biofilm can operate is controlled by the rate at which these reactants move through the biofilm. Consequently, it is vital for engineers and microbiologists to be able to measure the rate of movement of these reactants through the biofilm. Critically, this data is essential to our understanding of the way biofilms work and our ability to enhance biofilm performance.



In repose to this, this proposal set out to show how we could image the movement of molecules (reactants) through biofilms using MRI. Molecules become readily visible to MRI when labelled with a paramagnetic tag. Consequently, MRI can be used to image their transport through porous media. In this study, we demonstrated that this method could be applied to imaging movement of molecules through biofilms.



In the first instance the transport of Gd-DTPA, a commercially available paramagnetic molecule, was imaged in both agar (as a homogeneous test system) and in a relatively thick biofilm (1 cm thick). A calibration protocol was applied to convert these images into concentration maps, thus revealing the spatially resolved concentration of this tracer at different time intervals. Comparing the data obtained from the agar and the phototrophic biofilm experiments with diffusion models revealed that mass transport of Gd-DTPA in agar was purely via diffusion with a diffusion coefficient of 7.2x10-10 m2 s-1 while its was transported inside the Phormidium biofilm by both diffusion and advection.



To advance upon this, we built a bespoke RF coil with a smaller diameter than used previously (5 mm compared to 35 mm used in the above experiment). This enabled us to obtain better image resolution and thus work with smaller biofilms (2 mm thick). This size is of particular relevance to granular biofilms used in wastewater treatment. Again, we were able to image the transport of GD-DTPA through both agar and a thin biofilm, 2 mm in thickness. Transport models were again applied to determine diffusion coefficients for the molecule in agar and the biofilm.



This technology can be used to explore further mass transport processes in biofilms, either using the wide range of commercially available paramagnetically tagged molecules and nanoparticles, or by using bespoke tagged molecule designed to mimic specific pollutants. This would enable researchers to collect key transport data (such as diffusion coefficients) for large molecular pollutants and particulates. This data is key to underpinning better models of how biofilms work, and could thus lead to better design of biofilms in, for example, waste water treatment systems and fuel cells.
Exploitation Route Can help to imporve design of sewage treatment systems, making them more efficient and cost effective. Biofilm performance in bioreactors such as waste water treatment systems is heavily infuenced by the rate of transport of reactant molecules (such as the organic waste in waste water) through the biofilm. Knowing the transport rates of these molecules helps to develop better models of biofilm function from which to design optimal biofilm based bioreactors.



We expect our appraoch (using MRI to image molecular transport) to be of primary interest in the first instants to academia. However, secondary beneficiaries are those in the industrial sector utilizing large scale bioreactors such as sewage treatment systems. Although any biofilm based technology has potential to benefit from this method, waste water treatment is by far the largest sector and thus has the potential to benefit the most. Our MRI approach can be used to determine the transport behaviour of a wide range of large organic molecules and particulates which could potentially be used by the industrial sector to optimize waste waterreatment systems
Sectors Environment

 
Description New collaboration with hydrogeologists at University of Birmingham 
Organisation University of Birmingham
Country United Kingdom 
Sector Academic/University 
PI Contribution We will now be applying this novel MRI method to image nanoparticle transport through rock. In collaboration with hydrogeologists at the University of Birmingham, we will use this unique data to develop better transport models of nanoparticle pollutants through groundwater systems. These are much needed for better environmental risk assessment. Discussion lead to a succesful EPSRC grant application which is detailed elsewhere in this ROS.
Start Year 2012
 
Title Application of paramagnetic tracers for imaging mass transport in biofilms 
Description This work provided a new technique for imaging mass transport of molecules through biofilms. Here we demonstrated that paramagnetically tagged moecules (commonly used in biomedical sciences) can be used to image molecular transport through biofilms 
Type Of Technology New/Improved Technique/Technology 
Year Produced 2010 
Impact
 
Description Report in The Engineer magazine (UK based engineering magazine) 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact I was interviewed by The Engineer Magazine (I believe they saw the grant had just been funded on the EPSRC website) and a short report on the project was published in April 2009.

these are unknown
Year(s) Of Engagement Activity 2009