M. Tech. (Biotechnology, Department of Biosciences, Faculty of Applied and Computer Sciences), Vaal University of Technology. / Reverse osmosis (RO) membranes are widely used in water purification. The presence of biofilms in water and industrial water purification systems is prevalent. As a result, biofouling which is a biofilm problem causes adverse effects on reverse osmosis process, which include flux decline, shorter membrane lifetime and an increase in energy consumption The effect of biofouling on RO membranes was investigated at a water treatment facility at Sasol, Sasolburg by investigating the quality of water purified by the RO system and the extent of fouling that is attributed to biofouling. Chemical and microbiological data was averaged based on the results obtained from water analysis and samples from a fouled membrane. Bacteriological plate counts ranged between log 1.5 to 4 cfu/ml in water samples and log 3.9 to 4.5 cfu/cm2 on biofilm from the membrane surface. Water analysis indicated a high conductivity of 121 µS/cm in the feed and 81 ppm of the TDS, whereas in the permeate conductivity was found to be around 6 µS/cm and 3.8 ppm of the TDS. This indicated that components present in the feed were retained by the membrane. This was supported by membrane autopsy which showed that the bacteria and elements found in the feedwater were also present on the membrane surface, hence contributing to fouling. An average of 33% of cellular ATP was measured on the biofilm from membrane sample, showing that the fouling bacteria are metabolically active in situ. The results clearly indicated that an important biological activity occurred at the membrane surface.

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Bacterial biofilms represent a challenge to the healthcare system because of their resilience against antimicrobials and immune attack. Biofilms consist of bacterial aggregates embedded in an extracellular polymeric substance (EPS) composed of polysaccharides, nucleic acids and proteins. We hypothesised that carbohydrates could contribute to immune recognition of Pseudomonas aeruginosa biofilms by engaging C-type lectins. Here we show binding of Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin (DC-SIGN, CD209), mannose receptor (MR, CD206) and Dectin-2 to P. aeruginosa biofilms. We also demonstrate that DC-SIGN, unlike MR and Dectin-2, recognises planktonic P. aeruginosa cultures and this interaction depends on the presence of the common polysaccharide antigen. Within biofilms DC-SIGN, Dectin-2 and MR ligands appear as discrete clusters with dispersed DC-SIGN ligands also found among bacterial aggregates. DC-SIGN, MR and Dectin-2 bind to carbohydrates purified from P. aeruginosa biofilms, particularly the high molecular weight fraction (HMW; >132,000 Da), with Ks in the nM range. These HMW carbohydrates contain 74.9-80.9% mannose, display α-mannan segments, interfere with the endocytic activity of cell-associated DC-SIGN and MR and inhibit Dectin-2-mediated cellular activation. In addition, biofilm carbohydrates reduce the association of the DC-SIGN ligand Lewis, but not fucose, to human monocyte-derived dendritic cells (moDCs), and alter moDC morphology without affecting early cytokine production in response to lipopolysaccharide or P. aeruginosa cultures. This work identifies the presence of ligands for three important C-type lectins within P. aeruginosa biofilm structures and purified biofilm carbohydrates and highlights the potential for these receptors to impact immunity to P. aeruginosa infection.