Upon impact: the fate of adhering <i>Pseudomonas fluorescens</i> cells during Nanofiltration

Nanofiltration (NF) is a high-pressure membrane filtration process increasingly applied in drinking water treatment and water reuse processes. NF typically rejects divalent salts, organic matter, and micropollutants. However, the efficiency of NF is adversely affected by membrane biofouling, during which microorganisms adhere to the membrane and proliferate to create a biofilm. Here we show that adhered Pseudomonas fluorescens cells under high permeate flux conditions are met with high fluid shear and convective fluxes at the membrane-liquid interface, resulting in their structural damage and collapse. These results were confirmed by fluorescent staining, flow cytometry, and scanning electron microscopy. This present study offers a 'first-glimpse' of cell damage and death during the initial phases of bacterial adhesion to NF membranes and raises a key question about the role of this observed phenomena during early-stage biofilm formation under permeate flux and cross-flow conditions.European Research Council (ERC

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 NF270 membranes at 15 bar for permeate volumes of 0.5L, 2L, and 5L 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 minutes at 5 minutes 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 buildup of a homogenous fouling layer composed of both living and dead organisms (>10⁶cells.cm−2). Subsequent measurements of bacterial adhesion resulted in cell loadings of 0.2×10⁵, 1.0×10⁵cells×cm−2 and 2.6×10⁵ 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 proceduresand for the modelled feed solutions used in the experiment.Science Foundation IrelandEuropean Research Counci

Small secreted proteins enable biofilm development in the cyanobacterium Synechococcus elongatus

Small proteins characterized by a double-glycine (GG) secretion motif, typical of secreted bacterial antibiotics, are encoded by the genomes of diverse cyanobacteria, but their functions have not been investigated to date. Using a biofilm-forming mutant of Synechococcus elongatus PCC 7942 and a mutational approach, we demonstrate the involvement of four small secreted proteins and their GG-secretion motifs in biofilm development. These proteins are denoted EbfG1-4 (enable biofilm formation with a GG-motif). Furthermore, the conserved cysteine of the peptidase domain of the Synpcc7942_1133 gene product (dubbed PteB for peptidase transporter essential for biofilm) is crucial for biofilm development and is required for efficient secretion of the GG-motif containing proteins. Transcriptional profiling of ebfG1-4 indicated elevated transcript levels in the biofilm-forming mutant compared to wild type (WT). However, these transcripts decreased, acutely but transiently, when the mutant was cultured in extracellular fluids from a WT culture, and biofilm formation was inhibited. We propose that WT cells secrete inhibitor(s) that suppress transcription of ebfG1-4, whereas secretion of the inhibitor(s) is impaired in the biofilm-forming mutant, leading to synthesis and secretion of EbfG1-4 and supporting the formation of biofilms