Browsing by Autor "Rajal, Veronica B"

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Continuum heterogeneous biofilm model--a simple and accurate method for effectiveness factor determination.
(2012) Gonzo, Elio Emilio; Wuertz, Stefan; Rajal, Veronica B
We present a novel analytical approach to describe biofilm processes considering continuum variation of both biofilm density and substrate effective diffusivity. A simple perturbation and matching technique was used to quantify biofilm activity using the steady-state diffusion-reaction equation with continuum variable substrate effective diffusivity and biofilm density, along the coordinate normal to the biofilm surface. The procedure allows prediction of an effectiveness factor, η, defined as the ratio between the observed rate of substrate utilization (reaction rate with diffusion resistance) and the rate of substrate utilization without diffusion limitation. Main assumptions are that (i) the biofilm is a continuum, (ii) substrate is transferred by diffusion only and is consumed only by microorganisms at a rate according to Monod kinetics, (iii) biofilm density and substrate effective diffusivity change in the x direction, (iv) the substrate concentration above the biofilm surface is known, and (v) the substratum is impermeable. With this approach one can evaluate, in a fast and efficient way, the effect of different parameters that characterize a heterogeneous biofilm and the kinetics of the rate of substrate consumption on the behavior of the biological system. Based on a comparison of η profiles the activity of a homogeneous biofilm could be as much as 47.8% higher than that of a heterogeneous biofilm, under the given conditions. A comparison of η values estimated for first order kinetics and η values obtained by numerical techniques showed a maximum deviation of 1.75% in a narrow range of modified Thiele modulus values. When external mass transfer resistance, is also considered, a global effectiveness factor, η(0) , can be calculated. The main advantage of the approach lies in the analytical expression for the calculation of the intrinsic effectiveness factor η and its implementation in a computer program. For the test cases studied convergence was achieved quickly after four or five iterations. Therefore, the simulation and scale-up of heterogeneous biofilm reactors can be easily carried out.
Net growth rate of continuum heterogeneous biofilms with inhibition kinetics.
(2018) Gonzo, Elio Emilio; Wuertz, Stefan; Rajal, Veronica B
Biofilm systems can be modeled using a variety of analytical and numerical approaches, usually by making simplifying assumptions regarding biofilm heterogeneity and activity as well as effective diffusivity. Inhibition kinetics, albeit common in experimental systems, are rarely considered and analytical approaches are either lacking or consider effective diffusivity of the substrate and the biofilm density to remain constant. To address this obvious knowledge gap an analytical procedure to estimate the effectiveness factor (dimensionless substrate mass flux at the biofilm-fluid interface) was developed for a continuum heterogeneous biofilm with multiple limiting-substrate Monod kinetics to different types of inhibition kinetics. The simple perturbation technique, previously validated to quantify biofilm activity, was applied to systems where either the substrate or the inhibitor is the limiting component, and cases where the inhibitor is a reaction product or the substrate also acts as the inhibitor. Explicit analytical equations are presented for the effectiveness factor estimation and, therefore, the calculation of biomass growth rate or limiting substrate/inhibitor consumption rate, for a given biofilm thickness. The robustness of the new biofilm model was tested using kinetic parameters experimentally determined for the growth of Pseudomonas putida CCRC 14365 on phenol. Several additional cases have been analyzed, including examples where the effectiveness factor can reach values greater than unity, characteristic of systems with inhibition kinetics. Criteria to establish when the effectiveness factor can reach values greater than unity in each of the cases studied are also presented.
Simultaneous detection of four protozoan parasites on leafy greens using a novel multiplex PCR assay.
(2019) Shapiro, Karen; Kim, Minji; Rajal, Veronica B; Arrowood, Michael J; Packham, Andrea; Aguilar, Beatriz; Wuertz, Stefan
Pathogen contamination of fresh produce presents a health risk for consumers; however, the produce industry still lacks adequate tools for simultaneous detection of protozoan parasites. Here, a simple multiplex PCR (mPCR) assay was developed for detection of protozoan (oo)cysts and compared with previously published real-time PCR assays and microscopy methods. The assay was evaluated for simultaneous detection of Cryptosporidium, Giardia, Cyclospora cayetanensis, and Toxoplasma gondii followed by parasite differentiation via either a nested specific PCR or a restriction fragment length polymorphism (RFLP) assay. Spiking experiments using spinach as a model leafy green were performed for assay validation. Leaf-washing yielded higher recoveries and more consistent detection of parasites as compared with stomacher processing. Lowest limits of detection using the nested mPCR assay were 1-10 (oo)cysts/g spinach (in 10 g samples processed), and this method proved more sensitive than qPCR for parasite detection. Microscopy methods were more reliable for visual detection of parasites in lower spiking concentrations, but are more costly and laborious, require additional expertise, and lack molecular confirmation essential for accurate risk assessment. Overall, the nested mPCR assay provides a rapid (<24 h), inexpensive ($10 USD/sample), and simple approach for simultaneous detection of protozoan pathogens on fresh produce.
The continuum heterogeneous biofilm model with multiple limiting substrate Monod kinetics.
(2014) Gonzo, Elio Emilio; Wuertz, Stefan; Rajal, Veronica B
We describe a novel procedure to estimate the net growth rate of biofilms on multiple substrates. The approach is based on diffusion-reaction mass balances for chemical species in a continuum biofilm model with reaction kinetics corresponding to a Double-Monod expression. This analytical model considers a heterogeneous biofilm with variable distributions of biofilm density, activity, and effective diffusivity as a function of depth. We present the procedure to estimate the effectiveness factor analytically and compare the outcome with values obtained by the application of a rigorous numerical computational method using several theoretical examples and a test case. A comparison of the profiles of the effectiveness factor as a function of the Thiele modulus, φ, revealed that the activity of a homogeneous biofilm could be as much as 42% higher than that of a heterogeneous biofilm, under the given conditions. The maximum relative error between numerical and estimated effectiveness factor was 2.03% at φ near 0.7 (corresponding to a normalized Thiele modulus φ* = 1). For φ < 0.3 or φ > 1.4, the relative error was less than 0.5%. A biofilm containing aerobic ammonium oxidizers was chosen as a test case to illustrate the model's capability. We assumed a continuum heterogeneous biofilm model where the effective diffusivities of oxygen and ammonium change with biofilm position. Calculations were performed for two scenarios; Case I had low dissolved oxygen (DO) concentrations and Case II had high DO concentrations, with a concentration at the biofilm-fluid interface of 10 g O2 /m(3) . For Case II, ammonium was the limiting substrate for a biofilm surface concentration, CNs , ≤13.84 g of N/m(3) . At these concentrations ammonium was limiting inside the biofilm, and oxygen was fully penetrating. Conversely, for CNs > 13.84 g of N/m(3) , oxygen became the limiting substrate inside the biofilm and ammonium was fully penetrating. Finally, a generalized procedure to estimate the effectiveness factor for a system with multiple (n > 2) limiting substrates is given.