Nanofiltration and seawater reverse osmosis desalination are still considered energy-intensive processes. Seawater desalination can be 25 times more energy-intensive compared to conventional water treatment processes. Biofouling is a significant problem in achieving sustainable desalination, as it increases the energy demands and the overall water cost. Limiting the biodegradable substrate concentration in the feed water is proposed as a suitable approach to control biofouling in desalination membranes. Until now, nutrient manipulation studies have not fully elucidated to which extent this technique affects biofilm morphology and if the manipulated biofilms are easier to control and remove with a chemical-free approach. The main objective of this Ph.D. study is to provide a comprehensive assessment of the effect of nutrient manipulation on the physical properties of the developed biofilm to decrease the impact of biofouling on system performance and enhance the cleanability of biofilms in membrane systems. The aspects of the study included biofilm development and related system performance under varying feed water biodegradable carbon and phosphorous concentrations and the impact of permeation. The results of this study indicate that lowering the assimilable organic carbon and phosphorus concentration in the feed water controls biofilm formation and prolongs membrane system performance. A strategy of enhancing the hydraulic cleanability of biofilms in RO systems could involve avoiding the increase of the phosphorus concentration by eliminating the use of phosphonate-based antiscalants. The higher detachment for biofilms grown at a lower phosphorus concentration was explained by more soluble polymers in the EPS, resulting in a lower biofilm cohesive and adhesive strength. We demonstrated that the phosphorus concentration in the feed water affected the microbial and EPS composition. A homogenous bacterial community composition was found over the biofilm height. Permeation played a role in shaping biofilm localization, and therefore, the observed impact on the system performance parameters. This Ph.D. dissertation represents an exciting advance towards greener desalination by controlling and enhancing the cleanability of biofilms through feed water nutrient manipulation.

The aim of this thesis was to develop stable-isotope steady-state metabolic flux analysis (MFA) based on ¹³C labeling to quantify intracellular fluxes of central carbon metabolism in plants. The experiments focus on the analysis of a heterotrophic cell suspension culture of Arabidopsis thaliana (L) Heynh. (ecotype Landsberg erecta). The first objective was to develop a robust methodology based on combining high quality steady-state stable labeling data, metabolic modeling and computational analysis. A comprehensive analysis of the factors that influence the outcome of MFA was undertaken and best practice established. This allowed a critical analysis of the subcellular compartmentation of carbohydrate oxidation in the cell culture. The second objective was to apply the methodology to nutritional perturbations of the cell suspension. A comparison of growth on different nitrogen sources revealed that transfer to an ammonium-free medium: (i) increased flux through the oxidative pentose phosphate pathway (oxPPP) by 10% relative to glucose utilisation; (ii) caused a substantial decrease in entry of carbon into the tricarboxylic acid cycle (TCA); and (iii) increased the carbon conversion efficiency from 55% to 69%. Although growth on nitrate alone might be expected to increase the demand for reductant, the cells responded by decreasing the assimilation of inorganic N. Cells were also grown in media containing different levels of inorganic phosphate (Pi). Comparison of the flux maps showed that decreasing Pi availability: (i) decreased flux through the oxPPP; (ii) increased the proportion of substrate fully oxidised by the TCA cycle; and (iii) decreased carbon conversion efficiency. These changes are consistent with redirection of metabolism away from biosynthesis towards cell maintenance as Pi is depleted. Although published genome-wide transcriptomic and metabolomic studies suggest that Pi starvation leads to the restructuring of carbon and nitrogen metabolism, the current analysis suggests that the impact on metabolic organisation is much less extreme.