Ashkan Safari

The importance of laboratory water quality for studying initial bacterial adhesion during NF filtration processes

Biofouling of nanofiltration (NF) and reverse osmosis (RO) membranes for water treatment has been the subject of increased research effort in recent years. A prerequisite for undertaking fundamental experimental investigation on NF and RO processes is a procedure called compaction. This involves an initial phase of clean water permeation at high pressures until a stable permeate flux is reached. However water quality used during the compaction process may vary from one laboratory to another. The aim of this study was to investigate the impact of laboratory water quality during compaction of NF membranes. A second objective was to investigate if the water quality used during compaction influences initial bacterial adhesion. Experiments were undertaken with NF 270 membranes at 15 bar for permeate volumes of 0.5 L, 2 L, and 5 L using MilliQ, deionized or tap water. Membrane autopsies were performed at each permeation point for membrane surface characterisation by contact angle measurements, profilometry, and scanning electron microscopy. The biological content of compacted membranes was assessed by direct epi-fluorescence observation following nucleic acid staining. The compacted membranes were also employed as substrata for monitoring the initial adhesion of Ps. fluorescens under dynamic flow conditions for 30 min at 5 min intervals. Compared to MilliQ water, membrane compaction using deionized and tap water led to decreases in permeate flux, increase in surface hydrophobicity and led to significant build-up of a homogeneous fouling layer composed of both living and dead organisms (>10(6) cells cm(-2)). Subsequent measurements of bacterial adhesion resulted in cell loadings of 0.2 × 10(5), 1.0 × 10(5) cells cm(-2) and 2.6 × 10(5) cells cm(-2) for deionized, tap water and MilliQ water, respectively. These differences in initial cell adhesion rates demonstrate that choice of laboratory water can significantly impact the results of bacterial adhesion on NF membranes. Standardized protocols are therefore needed for the fundamental studies of bacterial adhesion and biofouling formation on NF and RO membrane. This can be implemented by first employing pure water during all membrane compaction procedures and for the modelled feed solutions used in the experiment.

The significance of calcium ions on Pseudomonas fluorescens biofilms – a structural and mechanical study

The purpose of this study was to investigate the effects of calcium ions on the structural and mechanical properties of Pseudomonas fluorescens biofilms grown for 48 h. Advanced investigative techniques such as confocal laser scanning microscopy and atomic force spectroscopy were employed to characterize biofilm structure as well as biofilm mechanical properties following growth at different calcium concentrations. The presence of calcium during biofilm development led to higher surface coverage with distinct structural phenotypes in the form of a granular and heterogeneous surface, compared with the smoother and homogeneous biofilm surface in the absence of calcium. The presence of calcium also increased the adhesive nature of the biofilm, while reducing its elastic properties. These results suggest that calcium ions could have a functional role in biofilm development and have practical implications, for example, in analysis of biofouling in membrane-based water-treatment processes such as nanofiltration or reverse osmosis where elevated calcium concentrations may occur at the solid–liquid interface.

Mechanical properties of a mature biofilm from a wastewater system: from microscale to macroscale level.

A fundamental understanding of biofilm mechanical stability is critical in order to describe detachment and develop biofouling control strategies. It is thus important to characterise the elastic deformation and flow behaviour of the biofilm under different modes of applied force. In this study, the mechanical properties of a mature wastewater biofilm were investigated with methods including macroscale compression and microscale indentation using atomic force microscopy (AFM). The mature biofilm was found to be mechanically isotropic at the macroscale level as its mechanical properties did not depend on the scales and modes of loading. However, the biofilm showed a tendency for mechanical inhomogeneity at the microscale level as indentation progressed deeper into the matrix. Moreover, it was observed that the adhesion force had a significant influence on the elastic properties of the biofilm at the surface, subjected to microscale tensile loading. These results are expected to inform a damage-based model for biofilm detachment.

A physical impact of organic fouling layers on bacterial adhesion during nanofiltration

Organic conditioning films have been shown to alter properties of surfaces, such as hydrophobicity and surface free energy. Furthermore, initial bacterial adhesion has been shown to depend on the conditioning film surface properties as opposed to the properties of the virgin surface. For the particular case of nanofiltration membranes under permeate flux conditions, however, the conditioning film thickens to form a thin fouling layer. This study hence sought to determine if a thin fouling layer deposited on a nanofiltration membrane under permeate flux conditions governed bacterial adhesion in the same manner as a conditioning film on a surface. Thin fouling layers (less than 50 μm thick) of humic acid or alginic acid were formed on Dow Filmtec NF90 membranes and analysed using Atomic Force Microscopy (AFM), confocal microscopy and surface energy techniques. Fluorescent microscopy was then used to quantify adhesion of Pseudomonas fluorescens bacterial cells onto virgin or fouled membranes under filtration conditions. It was found that instead of adhering on or into the organic fouling layer, the bacterial cells penetrated the thin fouling layer and adhered directly to the membrane surface underneath. Contrary to what surface energy measurements of the fouling layer would indicate, bacteria adhered to a greater extent onto clean membranes (24 ± 3% surface coverage) than onto those fouled with humic acid (9.8 ± 4%) or alginic acid (7.5 ± 4%). These results were confirmed by AFM measurements which indicated that a considerable amount of energy (10(-7) J/μm) was dissipated when attempting to penetrate the fouling layers compared to adhering onto clean NF90 membranes (10(-15) J/μm). The added resistance of this fouling layer was thusly seen to reduce the number of bacterial cells which could reach the membrane surface under permeate conditions. This research has highlighted an important difference between fouling layers for the particular case of nanofiltration membranes under permeate flux conditions and surface conditioning films which should be considered when conducting adhesion experiments under filtration conditions. It has also shown AFM to be an integral tool for such experiments.

Interfacial separation of a mature biofilm from a glass surface - A combined experimental and cohesive zone modelling approach

A good understanding of the mechanical stability of biofilms is essential for biofouling management, particularly when mechanical forces are used. Previous biofilm studies lack a damage-based theoretical model to describe the biofilm separation from a surface. The purpose of the current study was to investigate the interfacial separation of a mature biofilm from a rigid glass substrate using a combined experimental and numerical modelling approach. In the current work, the biofilm-glass interfacial separation process was investigated under tensile and shear stresses at the macroscale level, known as modes I and II failure mechanisms respectively. The numerical simulations were performed using a Finite Volume (FV)-based simulation package (OpenFOAM®) to predict the separation initiation using the cohesive zone model (CZM). Atomic force microscopy (AFM)-based retraction curve was used to obtain the separation properties between the biofilm and glass colloid at microscale level, where the CZM parameters were estimated using the Johnson-Kendall-Roberts (JKR) model. In this study CZM is introduced as a reliable method for the investigation of interfacial separation between a biofilm and rigid substrate, in which a high local stress at the interface edge acts as an ultimate stress at the crack tip.This study demonstrated that the total interfacial failure energy measured at the macroscale, was significantly higher than the pure interfacial separation energy obtained by AFM at the microscale, indicating a highly ductile deformation behaviour within the bulk biofilm matrix. The results of this study can significantly contribute to the understanding of biofilm detachments.

Revealing region-specific biofilm viscoelastic properties by means of a microrheological approach

Particle-tracking microrheology is an in situ technique that allows quantification of biofilm material properties. It overcomes the limitations of alternative techniques such as bulk rheology or force spectroscopy by providing data on region specific material properties at any required biofilm location and can be combined with confocal microscopy and associated structural analysis. This article describes single particle tracking microrheology combined with confocal laser scanning microscopy to resolve the biofilm structure in 3 dimensions and calculate the creep compliances locally. Samples were analysed from Pseudomonas fluorescens biofilms that were cultivated over two timescales (24 h and 48 h) and alternate ionic conditions (with and without calcium chloride supplementation). The region-based creep compliance analysis showed that the creep compliance of biofilm void zones is the primary contributor to biofilm mechanical properties, contributing to the overall viscoelastic character.Analytics: A better look at biofilmsCombining two distinct techniques allows analysis of the structure and physical properties of bacterial biofilms in fine detail. Eoin Casey and colleagues, from University College Dublin, Ireland, and the University of Hong Kong, applied a combined non-destructive procedure to cultured biofilms, for determining biofilm slimy properties and structural features. Tracking the movement of tiny fluorescent beads embedded in the biofilms yielded insights into their localised material properties. The same samples were also analysed using a laser-based imaging technique called confocal microscopy, which added additional structural insight to the biofilm samples. This combination allowed an improved regional understanding of the biofilms’ varying material and mechanical properties, including viscosity and elasticity. In future, the researchers hope to add chemical analysis into the mix, producing a method to search for better ways to treat and eradicate biofilms.