Rory Heffernan

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.

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.

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.