Center for Biofilm Engineering (CBE)
Permanent URI for this communityhttps://scholarworks.montana.edu/handle/1/9334
At the Center for Biofilm Engineering (CBE), multidisciplinary research teams develop beneficial uses for microbial biofilms and find solutions to industrially relevant biofilm problems. The CBE was established at Montana State University, Bozeman, in 1990 as a National Science Foundation Engineering Research Center. As part of the MSU College of Engineering, the CBE gives students a chance to get a head start on their careers by working on research teams led by world-recognized leaders in the biofilm field.
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Item Hypoxia arising from concerted oxygen consumption by neutrophils and microorganisms in biofilms(2018-06) Wu, Yilin; Klapper, Isaac; Stewart, Philip S.Infections associated with microbial biofilms are often found to involve hypoxic or anoxic conditions within the biofilm or its vicinity. To shed light on the phenomenon of local oxygen depletion, mathematical reaction-diffusion models were derived that integrated the two principal oxygen sinks, microbial respiration and neutrophil consumption. Three simple one-dimensional problems were analyzed approximating biofilm near an air interface as in a dermal wound or mucus layer, biofilm on an implanted medical device, or biofilm aggregates dispersed in mucus or tissue. In all three geometries considered, hypoxia at the biofilm–neutrophil interface or within the biofilm was predicted for a subset of plausible parameter values. The finding that oxygen concentration at the biofilm–neutrophil juncture can be diminished to hypoxic levels is biologically relevant because oxygen depletion will reduce neutrophil killing ability. The finding that hypoxia can readily establish in the interior of the biofilm is biologically relevant because this change will alter microbial metabolism and persistence.Item Viscoelastic fluid description of bacterial biofilm material properties(2002-09) Klapper, Isaac; Rupp, Cory J.; Cargo, R.; Purevdorj, B.; Stoodley, PaulA mathematical model describing the constitutive properties of biofilms is required for predicting biofilm deformation, failure and detachment in response to mechanical forces. Laboratory observations indicate that biofilms are viscoelastic materials. Likewise, current knowledge of biofilm internal structure suggests modeling biofilms as associated polymer viscoelastic systems. Supporting experimental results and a system of viscoelastic fluid equations with a linear Jeffreys viscoelastic stress-strain law are presented here. This system of equations is based on elements of associated polymer physics and is also consistent with presented and previous experimental results. A number of predictions can be made. One particularly interesting result is the prediction of an elastic relaxation time on the order of a few minutes: biofilm disturbances on shorter time scales produce an elastic response, biofilm disturbances on longer time scales result in viscous flow, i.e., non-reversible biofilm deformation. Although not previously recognized, evidence of this phenomenon is in fact present in recent experimental results.Item Biofilm material properties as related to shear-induced deformation and detachment phenomena(2002-12) Stoodley, Paul; Cargo, R.; Rupp, Cory J.; Wilson, Suzanne; Klapper, IsaacBiofilms of various Pseudomonas aeruginosa strains were grown in glass flow cells under laminar and turbulent flows. By relating the physical deformation of biofilms to variations in fluid shear, we found that the biofilms were viscoelastic fluids which behaved like elastic solids over periods of a few seconds but like linear viscous fluids over longer times. These data can be explained using concepts of associated polymeric systems, suggesting that the extracellular polymeric slime matrix determines the cohesive strength. Biofilms grown under high shear tended to form filamentous streamers while those grown under low shear formed an isotropic pattern of mound-shaped microcolonies. In some cases, sustained creep and necking in response to elevated shear resulted in a time-dependent fracture failure of the "tail" of the streamer from the attached upstream "head." In addition to structural differences, our data suggest that biofilms grown under higher shear were more strongly attached and were cohesively stronger than those grown under lower shears.Item Commonality of elastic relaxation times in biofilms(2004-08) Shaw, T.; Winston, Matthew T.; Rupp, Cory J.; Klapper, Isaac; Stoodley, PaulBiofilms, sticky conglomerations of microorganisms and extracellular polymers, are among the Earth's most common life forms. One component for their survival is an ability to withstand external mechanical stress. Measurements indicate that biofilm elastic relaxation times are approximately the same (about 18 min) over a wide sample of biofilms though other material properties vary significantly. A possible survival significance of this time scale is that it is the shortest period over which a biofilm can mount a phenotypic response to transient mechanical stress.Item Adaptive responses to antimicrobial agents in biofilms(2005-08) Szomolay, Barbara; Klapper, Isaac; Dockery, Jack D.; Stewart, Philip S.Bacterial biofilms demonstrate adaptive resistance in response to antimicrobial stress more effectively than corresponding planktonic populations. We propose here that, in biofilms, reaction-diffusion limited penetration may result in only low levels of antimicrobial exposure to deeper regions of the biofilm. Sheltered cells are then able to enter an adapted resistant state if the local time scale for adaptation is faster than that for disinfection. This mechanism is not available to a planktonic population. A mathematical model is presented to illustrate. Results indicate that, for a sufficiently thick biofilm, cells in the biofilm implement adaptive responses more effectively than do freely suspended cells. Effective disinfection requires applied biocide concentration that increases quadratically or exponentially with biofilm thickness.Item A multidimensional multispecies continuum model for heterogeneous biofilm development(2007-01) Alpkvist, Erik; Klapper, IsaacWe propose a multidimensional continuum model for heterogeneous growth of biofilm systems with multiple species and multiple substrates. The new model provides a deterministic framework for the study of the interactions between several species and their effects on biofilm heterogeneity. It consists of a system of partial differential equations derived on the basis of conservation laws and reaction kinetics. The derivation and key assumptions are presented. The assumptions used are a combination of those used in the established one dimensional model, due to Wanner and Gujer, and for the viscous fluid model, of Dockery and Klapper. The work of Wanner and Gujer in particular has been extensively used through the years, and thus this new model is an extension to several spatial dimensions of an already proven working model. The model equations are solved using numerical techniques, for purposes of simulation and verification. The new model is applied to two different biofilm systems in several spatial dimensions, one of which is equivalent to a system originally studied by Wanner and Gujer. Dimensionless formulations for these two systems are given, and numerical simulation results with varying initial conditions are presented.Item Senescence can explain microbial persistence(2007-11) Klapper, Isaac; Gilbert, P.; Ayati, B. P.; Dockery, Jack D.; Stewart, Philip S.It has been known for many years that small fractions of persister cells resist killing in many bacterial colony–antimicrobial confrontations. These persisters are not believed to be mutants. Rather it has been hypothesized that they are phenotypic variants. Current models allow cells to switch in and out of the persister phenotype. Here, a different explanation is suggested for persistence, namely senescence. Using a mathematical model including age structure, it is shown that senescence provides a natural explanation for persistence-related phenomena, including the observations that the persister fraction depends on growth phase in batch culture and dilution rate in continuous culture.Item Description of mechanical response including detachment using a novel particle model of biofilm/flow interaction(2007-05) Alpkvist, Erik; Klapper, IsaacBacterial biofilms, while made up of microbial-scale objects, also function as meso- and macroscale materials. In particular, macro-scale material properties determine how biofilms respond to large-scale mechanical stresses, e.g. fluid shear. Viscoelastic and other constitutive properties influence biomass structure (through growth and fluid shear stresses) by erosion and sloughing detachment. In this paper, using the immersed boundary method, biofilm is modelled by a system of viscoelastic, breakable springs embedded in a fluid flow, evolving according to the basic physical laws of conservation of mass and momentum. We demonstrate in the context of computer simulation biofilm deformation and detachment under fluid shear stress.Item A multiscale model of biofilm as a senescence-structured fluid(2007-01) Ayati, B. P.; Klapper, IsaacWe derive a physiologically structured multiscale model for biofilm development. The model has components on two spatial scales, which induce different time scales into the problem. The macroscopic behavior of the system is modeled using growth-induced flow in a domain with a moving boundary. Cell-level processes are incorporated into the model using a so-called physiologically structured variable to represent cell senescence, which in turn affects cell division and mortality. We present computational results for our models which shed light on modeling the combined role senescence and the biofilm state play in the defense strategy of bacteria.Item Measurements of accumulation and displacement at the single cell cluster level in Pseudomonas aeruginosa biofilms(2008-09) Klayman, Benjamin J.; Klapper, Isaac; Stewart, Philip S.; Camper, Anne K.Quantitative descriptions of biofilm growth and dynamics at the individual cell level are largely missing from the literature. To fill this gap, research was done to describe growth, accumulation and displacement patterns in developing Pseudomonas aeruginosa biofilms. A parent strain of PAO1 was labelled with either a cyan or yellow fluorescent protein. These were then grown in a flow cell biofilm together so that pockets of dividing cells could be identified and their accumulation and displacement tracked. This analysis revealed a pattern of exponential accumulation for all clusters followed by a stationary accumulation phase. A background ‘carpet’ layer of cells uniformly colonizing the surface exhibited zero net accumulation of bio-volume. The individual clusters were found to have a mean accumulation rate of 0.34 h-1 with a range of 0.28–0.41 h-1. Cluster accumulation rates were negatively correlated with cluster size; larger clusters accumulated volume at a slower rate (P < 0.001). Pockets of cells on the inside of clusters initially accumulated at a comparable rate to the cluster within which they resided, but later invariably exhibited zero to slightly negative accumulation despite continued exponential (positive) accumulation of the cluster. Expanding clusters were able to displace neighbouring cells from the surface, and larger clusters displaced smaller clusters. This work provides a more detailed quantitative experimental observation of biofilm behaviour than has been described previously.