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Habimana, Olivier
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Habimana, Olivier
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Habimana, Olivier
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Now showing 1 - 10 of 17
- PublicationCharacterisitics of Streptomyces griseus biofilms in continuous flow tubular reactorsThe purpose of this study was to investigate the feasibility of cultivating the biotechnologically important bacterium Streptomyces griseus in single-species and mixed- species biofilms using a Tubular Biofilm Reactor (TBR). Streptomyces griseus biofilm development was found to be cyclical, starting with the initial adhesion and subsequent development of a visible biofilm after 24 hours growth, followed by the complete detachment of the biofilm as a single mass, and ending with the re-colonization of the tube. Fluorescence microscopy revealed that the filamentous structure of the biofilm was lost upon treatment with protease, but not DNase or metaperiodate, indicating that the extracellular polymeric substance is predominantly protein. When the biofilm was cultivated in conjunction with Bacillus amyloliquefaciens, no detachment was observed after 96 h, although once subjected to flow detachment occurred. Electron microscopy confirmed the presence of both bacteria in the biofilm and revealed a network of fimbriae-like structures that were much less apparent in single-species biofilm, and are likely to increase mechanical stability when developing in a TBR. This study presents the very first attempt in engineering Streptomyces griseus biofilms for continuous bioprocess applications.
880Scopus© Citations 15 - PublicationBacterial adhesion onto nanofiltration and reverse osmosis membranes: Effect of permeate fluxThe influence of permeate flux on bacterial adhesion to NF and RO membranes was examined using two model Pseudomonas species, namely Pseudomonas fluorescens and Pseudomonas putida. To better understand the initial biofouling profile during NF/RO processes, deposition experiments were conducted in cross flow under permeate flux varying from 0.5 up to 120 L/(h m2), using six NF and RO membranes each having different surface properties. All experiments were performed at a Reynolds number of 579. Complementary adhesion experiments were performed using Pseudomonas cells grown to early-, mid- and late-exponential growth phases to evaluate the effect of bacterial cell surface properties during cell adhesion under permeate flux conditions. Results from this study show that initial bacterial adhesion is strongly dependent on the permeate flux conditions, where increased adhesion was obtained with increased permeate flux, until a maximum of 40% coverage was reached. Membrane surface properties or bacterial growth stages was further found to have little impact on bacterial adhesion to NF and RO membrane surfaces under the conditions tested. These results emphasise the importance of conducting adhesion and biofouling experiments under realistic permeate flux conditions, and raises questions about the efficacy of the methods for the evaluation of antifouling membranes in which bacterial adhesion is commonly assessed under zero-flux or low flux conditions, unrepresentative of full-scale NF/RO processes.
488Scopus© Citations 20 - PublicationThe importance of laboratory water quality for studying initial bacterial adhesion during NF filtration processes(Elsevier, 2013-05-15)
; ; ; ; ; 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.979Scopus© Citations 24 - PublicationUpon impact: the fate of adhering Pseudomonas fluorescens cells during Nanofiltration(American Chemical Society, 2014-07-29)
; ; 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.281Scopus© Citations 9 - PublicationThe significance of calcium ions on Pseudomonas fluorescens biofilms – a structural and mechanical studyThe 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.
366Scopus© Citations 32 - PublicationNanofiltration and reverse osmosis surface topographical heterogeneities: do they matter for initial bacterial adhesion?(Elsevier, 2015-07-15)
; ; ; ; ; The role of the physicochemical and surface properties of NF/RO membranes influencing bacterial adhesion has been widely studied. However, there exists a poor understanding of the potential role membrane topographical heterogeneities can have on bacterial adhesion. Heterogeneities on material surfaces have been shown to influence bacterial adhesion and biofilm development. The purpose of this study was therefore to investigate whether the presence of membrane topographical heterogeneities had a significant role during bacterial adhesion as this could significantly impact on how biofouling develops on membranes during NF/RO operation. An extensive study was devised in which surface topographical heterogeneities from two commercial membranes, NF270 and BW30, were assessed for their role in the adhesion of two model organisms of different geometrical shapes, Pseudomonas fluorescens and Staphylococcus epidermidis. The influence of cross-flow velocity and permeate flux was also tested, as well as the angle to which bacteria adhered compared to the flow direction. Bacterial adhesion onto the membranes and in their surface topographical heterogeneities was assessed using Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), fluorescence microscopy and image analysis. Results showed that up to 30% of total adhered cells were found in membrane defect areas when defect areas only covered up to 13% of the membrane surface area. This suggests that topographical heterogeneities may play a significant role in establishing environmental niches during the early stages of biofilm development. Furthermore, no noticeable difference between the angle of cell attachment in defect areas compared to the rest of the membrane surface was found.384Scopus© Citations 16 - PublicationComparison of biomass detachment from two different Pseudomonas spp. biofilms under constant shear conditionsIn the context of biofilm development, detachment is of practical importance when placed in a biofilm management perspective. The objective of the present study was to examine biofilm structure and biofilm detachment under controlled conditions for two distinct microorganisms grown under constant shear conditions. Detached biofilm biomass was regularly collected and analysed over the course of 72 h biofilm growth by Pseudomonas putida and Pseudomonas fluorescens cells, and biofilm structural development assessed using confocal microscopy. The two Pseudomonas spp., which had very similar specific growth rates in planktonic culture, presented notably different characteristics in terms of biofilm morphology but their detachment behaviours over time were very similar. These findings underline the intrinsic complexity of the detachment phenomenon.
270Scopus© Citations 4 - PublicationA physical impact of organic fouling layers on bacterial adhesion during nanofiltration(Elsevier, 2014-12-15)
; ; ; ; ; 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.329Scopus© Citations 20 - PublicationAntifouling activity of enzyme-functionalized silica nanobeadsThe amelioration of biofouling in industrial processing equipment is critical for performance and reliability. While conventional biocides are effective in biofouling control, they are potentially hazardous to the environment and in some cases corrosive to materials. Enzymatic approaches have been shown to be effective and can overcome the disadvantages of traditional biocides, however they are typically uneconomic for routine biofouling control. The aim of this study was to design a robust and reusable enzyme-functionalized nano-bead system having biofilm dispersion properties. This work describes the biochemical covalent functionalization of silica-based nanobeads (hereafter referred to as Si-NanoB) with Proteinase K (PK). Results showed that PK-functionalized Si-NanoB are effective in dispersing both protein-based model biofilms and structurally altering Pseudomonas fluorescens biofilms, with significant decreases in surface coverage and thickness of 30.1% and 38.85%, respectively, while increasing surface roughness by 19 % following 24 h treatments on bacterial biofilms. This study shows that enzyme-functionalized nanobeads may potentially be an environmentally friendly and cost effective alternative to pure enzyme and chemical treatments.
330Scopus© Citations 15 - PublicationDisinfection of a polyamide nanofiltration membrane using ethanol(Elsevier, 2013-12-15)
; ; ; ; ; ; It is imperative that nanofiltration membranes are disinfected before they are used for laboratory-scale bacterial adhesion or biofouling experiments, yet currently no suitable disinfection protocol exists. This study aimed to determine if an ethanol treatment at a minimum inhibitory concentration (MIC) could be used to effectively disinfect nanofiltration membranes without altering membrane properties which could affect research. Two strains of bacteria, Pseudomonas fluorescens and Staphylococcus sp., were exposed to a range of ethanol concentrations to determine the MIC required for a 4log10 reduction in bacteria. In parallel, ethanol's effects on the filtration, surface and mechanical properties of a Dow Filmtec NF90 membrane were analysed. A 1.5 hour treatment with 40% ethanol was shown to effectively disinfect the membrane without significantly affecting any of the membranes properties tested. This treatment protocol can now be safely used to disinfect the studied membrane prior to bacterial adhesion or biofouling experiments. This study also acts as a guideline for researchers using other membranes to determine a suitable disinfection protocol for their needs.820Scopus© Citations 27