Development Of Biofilm Communities Research Articles

Bacterial communities of planktonic bacteria and mature biofilm in service lines and premise plumbing of a Megacity: Composition, Diversity, and influencing factors

Although simulated studies have provided valuable knowledge regarding the communities of planktonic bacteria and biofilms, the lack of systematic field studies have hampered the understanding of microbiology in real-world service lines and premise plumbing. In this study, the bacterial communities of water and biofilm were explored, with a special focus on the lifetime development of biofilm communities and their key influencing factors. The 16S rRNA gene sequencing results showed that both the planktonic bacteria and biofilm were dominated by Proteobacteria. Among the 15,084 observed amplicon sequence variants (ASVs), the 33 core ASVs covered 72.8 %, while the 12 shared core ASVs accounted for 62.2 % of the total sequences. Remarkably, it was found that the species richness and diversity of biofilm communities correlated with pipe age. The relative abundance of ASV2 (f_Sphingomonadaceae) was lower for pipe ages 40–50 years (7.9 %) than for pipe ages 10–20 years (59.3 %), while the relative abundance of ASV10 (f_Hyphomonadaceae) was higher for pipe ages 40–50 years (19.5 %) than its presence at pipe ages 20–30 years (1.9 %). The community of the premise plumbing biofilm had significantly higher species richness and diversity than that of the service line, while the steel-plastics composite pipe interior lined with polyethylene (S-PE) harbored significantly more diverse biofilm than the galvanized steel pipes (S-Zn). Interestingly, S-PE was enriched with ASV27 (g_Mycobacterium), while S-Zn pipes were enriched with ASV13 (g_Pseudomonas). Moreover, the network analysis showed that five rare ASVs, not core ASVs, were keystone members in biofilm communities, indicating the importance of rare members in the function and stability of biofilm communities. This manuscript provides novel insights into real-world service lines and premise plumbing microbiology, regarding lifetime dynamics (pipe age 10–50 years), and the influences of pipe types (premise plumbing vs. service line) and pipe materials (S-Zn vs. S-PE).

Convergence of biofilm successional trajectories initiated during contrasting seasons.

Biofilm communities play a major role in explaining the temporal variation of biogeochemical conditions in freshwater ecosystems, and yet we know little about how these complex microbial communities change over time (aka succession), and from different initial conditions, in comparison to other stream communities. This has resulted in limited knowledge on how biofilm community structure and microbial colonization vary over relevant time scales to become mature biofilms capable of significant alteration of the freshwater environment in which they live. Here, we monitored successional trajectories of biofilm communities from summer and winter in a headwater stream and evaluated their structural state over time by DNA high-throughput sequencing. Significant differences in biofilm composition were observed when microbial colonization started in the summer vs. winter seasons, with higher percentage of algae (Bacillariophyta) and Bacteroidetes in winter-initiated samples but higher abundance of Proteobacteria (e.g., Rhizobiales, Rhodobacterales, Sphingomonadales, and Burkholderiales), Actinobacteria, and Chloroflexi in summer-initiated samples. Interestingly, results showed that despite seasonal effects on early biofilm succession, biofilm community structures converged after 70 days, suggesting the existence of a stable, mature community in the stream that is independent of the environmental conditions during biofilm colonization. Overall, our results show that algae are important in the early development of biofilm communities during winter, while heterotrophic bacteria play a more critical role during summer colonization and development of biofilms.

Polymicrobial Aggregates in Human Saliva Build the Oral Biofilm.

ABSTRACTBiofilm community development has been established as a sequential process starting from the attachment of single cells on a surface. However, microorganisms are often found as aggregates in the environment and in biological fluids. Here, we conduct a comprehensive analysis of the native structure and composition of aggregated microbial assemblages in human saliva and investigate their spatiotemporal attachment and biofilm community development. Using multiscale imaging, cell sorting, and computational approaches combined with sequencing analysis, a diverse mixture of aggregates varying in size, structure, and microbial composition, including bacteria associated with host epithelial cells, can be found in saliva in addition to a few single-cell forms. Phylogenetic analysis reveals a mixture of complex consortia of aerobes and anaerobes in which bacteria traditionally considered early and late colonizers are found mixed together. When individually tracked during colonization and biofilm initiation, aggregates rapidly proliferate and expand tridimensionally, modulating population growth, spatial organization, and community scaffolding. In contrast, most single cells remain static or are incorporated by actively growing aggregates. These results suggest an alternative biofilm development process whereby aggregates containing different species or associated with human cells collectively adhere to the surface as “growth nuclei” to build the biofilm and shape polymicrobial communities at various spatial and taxonomic scales.

Effects of Methyl, Ester, and Amine Surface Groups on Microbial Activity and Communities in Nitrifying Biofilms.

The objective of this study was to determine how different attachment surface chemistries affected the initial and long-term performance and microbial populations of nitrifying biofilms under well-controlled hydrodynamic mixing conditions. While much previous research has focused on the effects of surface properties such as hydrophobicity on bacterial attachment in pure cultures, this study evaluated the effects of specific functional groups on mixed culture composition and functional behavior. Three surfaces with varying hydrophobicity and charge were evaluated for biofilm community development and performance: unmodified poly(dimethylsiloxane) (PDMS), which included terminal methyl groups and was relatively hydrophobic (P-Methyl), PDMS silanized with ester groups (P-Ester), which was uncharged and relatively hydrophilic, and PDMS modified with amine groups (P-Amine), which possessed a positive charge and was the most hydrophilic. The surface chemistries of the three attachment surfaces were characterized by contact angle goniometry, Fourier transform infrared (FTIR) spectroscopy, and X-ray photoelectron spectroscopy (XPS). These surfaces were inoculated with dilute activated sludge, and biofilms were grown in rotating annular bioreactors for 80 days, with experimental triplicates. Nitrification rates increased most rapidly in P-Amine biofilm reactors, and their biofilm communities contained significantly more Nitrosomonas (p < 0.05) than those on the other surfaces in early growth stages (days 40-50). From days 50-60, the P-Amine surface biofilm had significantly higher nitrate production rates than the P-Methyl and P-Ester biofilms. The biofilms grown on the P-Amine and P-Methyl surfaces were significantly (p < 0.05) more diverse than the P-Ester biofilms, containing higher relative abundances of the order Rhizobiales, including a significantly higher abundance of the nitrifying genus Nitrobacter (p < 0.05), which coincided with higher rates of nitrate generation. Conversely, biofilms grown on the uncharged hydrophilic P-Ester surface were consistently less productive and had lower diversity than biofilms on the other surfaces. These results indicate that surface chemistry may be a useful design parameter to improve the performance of nitrifying biofilm systems for wastewater treatment and that surface chemistry affects mixed biofilm community composition.

Bedform characteristics and biofilm community development interact to modify hyporheic exchange

The physical and biological attributes of riverine ecosystems interact in a complex manner which can affect the hydrodynamic behaviour of the system. This can alter the mixing characteristics of a river at the sediment-water interface. Research on hyporheic exchange has increased in recent years driven by a greater appreciation for the importance of this dynamic ecotone in connecting and regulating river systems. An understanding of process-based interactions driving hyporheic exchange is still limited, specifically the feedbacks between the physical and biological controlling factors. The interplay between bed morphology and sediment size on biofilm community development and the impact on hyporheic exchange mechanisms, was experimentally considered. Purpose built recirculating flume systems were constructed and three profiles of bedform investigated: i) flat, ii) undulating λ = 1 m, ii) undulating λ = 0.2 m, across two different sized sediments (0.5 mm and 5 mm). The influence of biofilm growth and bedform interaction on hyporheic exchange was explored, over time, using discrete repeat injections of fluorescent dye into the flumes. Hyporheic exchange rates were greatest in systems with larger sediment sizes (5 mm) and with more bedforms (undulating λ = 0.2). Sediment size was a dominant control in governing biofilm growth and hyporheic exchange in systems with limited bedform. In systems where bedform was prevalent, sediment size and biofilm appeared to no longer be a control on exchange due to the physical influence of advective pumping. Here, exchange rates within these environments were more consistent overtime, despite greater microbial growth. As such, bedform has the potential to overcome the rate limiting effects of biotic factors on hyporheic exchange and sediment size on microbial penetration. This has implications for pollutant and nutrient penetration; bedforms increase hydrological connectivity, generating the opportunity to support microbial communities at depth and as such, improve the self-purification ability of river systems.

Seasonal diversity of the bacterial communities associated with petroglyphs sites from the Negev Desert, Israel

Rock art sites in the form of petroglyphs are found in the Negev Desert in southern Israel. These petroglyph sites include thousands of images depicting the life and culture of people in the region over the years. As they are part of the desert environment, these petroglyphs are exposed to natural and anthropogenic weathering processes. In natural rock sites, it is known that climatic conditions influence weathering and biofilm community development (Friedmann in Orig Life 10:223–235, 1980; Li et al. in PloS one 11:(9),163–287, 2016) Therefore, in order to develop tools for preservation and remediation, it is imperative to ascertain possible seasonal diversity of surface-inhabiting bacterial communities. To do so, we undertook to study and characterize the microbial communities inhabiting the rock sites during winter and summer. Next-generation sequencing (NGS) technologies using 16S rRNA gene were implemented on rock-surface slab samples to identify the microbial communities. 16S rRNA gene sequencing data revealed the presence of five main bacterial phyla, among them Actinobacteria, Cyanobacteria, Proteobacteria, Bacteroidetes, and Chloroflexi across the analyzed sites. The predominant phylum was Cyanobacteria (10–60% relative abundance), followed by Actinobacteria (20–60% relative abundance), Proteobacteria (5–35%), Bacteroidetes (0.02–35%), and Chloroflexi (1–12%). There was no significant difference in the bacterial diversity between samples from winter and summer. Five main phyla were observed in samples from the two seasons, winter (December) 2017 and summer (July) 2018. This study provides an important first step in understanding the possible seasonal dynamic of bacterial communities associated with the petroglyphs.

High-Speed "4D" Computational Microscopy of Bacterial Surface Motility.

Bacteria exhibit surface motility modes that play pivotal roles in early-stage biofilm community development, such as type IV pili-driven "twitching" motility and flagellum-driven "spinning" and "swarming" motility. Appendage-driven motility is controlled by molecular motors, and analysis of surface motility behavior is complicated by its inherently 3D nature, the speed of which is too fast for confocal microscopy to capture. Here, we combine electromagnetic field computation and statistical image analysis to generate 3D movies close to a surface at 5 ms time resolution using conventional inverted microscopes. We treat each bacterial cell as a spherocylindrical lens and use finite element modeling to solve Maxwell's equations and compute the diffracted light intensities associated with different angular orientations of the bacterium relative to the surface. By performing cross-correlation calculations between measured 2D microscopy images and a library of computed light intensities, we demonstrate that near-surface 3D movies of Pseudomonas aeruginosa translational and rotational motion are possible at high temporal resolution. Comparison between computational reconstructions and detailed hydrodynamic calculations reveals that P. aeruginosa act like low Reynolds number spinning tops with unstable orbits, driven by a flagellum motor with a torque output of ∼2 pN μm. Interestingly, our analysis reveals that P. aeruginosa can undergo complex flagellum-driven dynamical behavior, including precession, nutation, and an unexpected taxonomy of surface motility mechanisms, including upright-spinning bacteria that diffuse laterally across the surface, and horizontal bacteria that follow helicoidal trajectories and exhibit superdiffusive movements parallel to the surface.

Genetic Characterization of Periphyton Communities Associated with Selenium Bioconcentration and Trophic Transfer in a Simple Food Chain.

A major source of uncertainty in predicting selenium (Se) distribution in aquatic food webs lies in the enrichment factor (EF), the ratio of Se bioconcentration in primary producers and microorganisms relative to the concentration of Se in the surrounding water. It has been well demonstrated that EFs can vary dramatically among individual algal taxa, but data are lacking regarding the influence of periphyton community composition on EFs for a given geochemical form of Se. Therefore, the goals of this study were first to assess whether different periphyton communities could be established in aquaria with the same starting inoculum using different light and nutrient regimes, and second, to determine if the periphyton assemblage composition influences the uptake of waterborne Se (as selenite) and subsequent Se transfer to a model macroinvertebrate primary consumer. Periphyton biofilms were grown in aquaria containing filtered pond water (from Saskatoon, SK) spiked with approximately 20 μg Se/L (mean measured concentration 21.0 ± 1.2 μg Se/L), added as selenite. Five different light and nutrient regimes were applied to the aquaria (three replicates per treatment) to influence biofilm community development. After 6 weeks of biofilm maturation, 40 to 80 immature cultured snails (Stagnicola elodes) were added to each aquarium. The bacterial and algal members of the periphyton community were characterized by targeted metagenomic analyses before and after addition of snails to ensure the snails themselves did not significantly alter the biofilm community. Samples were collected for Se analysis of water, periphyton, and whole-body snail. The nutrient and light treatments resulted in substantially different compositions of the periphytic biofilms, with each being relatively consistent across replicates and throughout the study. Although the aqueous concentration of dissolved Se administered to treatments was constant, uptake by the different periphytic biofilms differed significantly. Both the low-light (61.8 ± 12.1 μg Se/g d.w.) and high-light (30.5 ± 4.7 μg Se/g d.w.) biofilms, which were found to have high proportions of cyanobacteria, contained statistically higher concentrations of Se relative to the other treatments. Furthermore, the concentration of Se in bulk periphyton was predictive of Se bioaccumulation in grazing snails but as an inverse relationship, opposite to expectations. The trophic transfer factor was inversely correlated with periphyton enrichment factor (r = -0.841). A number of different bacterial and algal taxa were correlated (either positively or negatively) with Se accumulation in periphyton biofilm and snails. Recent advancements in genetic methods make it possible to conduct detailed characterization of periphyton assemblages and begin to understand the influence that periphyton composition has on Se biodynamics in aquatic systems.

Hydraulic and biotic impacts on neutralisation of high-pH waters

The management of alkaline (pH11–12.5) leachate is an important issue associated with the conditioning, afteruse or disposal of steel slags. Passive in-gassing of atmospheric CO2 is a low cost option for reducing Ca(OH)2 alkalinity, as Ca(OH)2 is neutralised by carbonic acid to produce CaCO3. The relative effectiveness of such treatment can be affected by both the system geometry (i.e. stepped cascades versus settlement ponds) and biological colonization. Sterilized mesocosm experiments run over periods of 20days showed that, due to more water mixing and enhanced CO2 dissolution at the weirs, the cascade systems (pH11.2→9.6) are more effective than settlement ponds (pH11.2→11.0) for lowering leachate alkalinity in all the tested conditions. The presence of an active microbial biofilm resulted in significantly more pH reduction in ponds (pH11.2→9.5), but had a small impact on the cascade systems (pH11.2→9.4). The pH variation in biofilm colonized systems shows a diurnal cycle of 1 to 1.5pH units due to CO2 uptake and release associated with respiration and photosynthesis. The results demonstrate that, where gradient permits, aeration via stepped cascades are the best option for neutralisation of steel slag leachates, and where feasible, the development of biofilm communities can also help reduce alkalinity.

Identification and characterization of a novel Fusobacterium nucleatum adhesin involved in physical interaction and biofilm formation with Streptococcus gordonii.

To successfully colonize the oral cavity, bacteria must directly or indirectly adhere to available oral surfaces. Fusobacterium nucleatum plays an important role in oral biofilm community development due to its broad adherence abilities, serving as a bridge between members of the oral biofilm that cannot directly bind to each other. In our efforts to characterize the molecular mechanisms utilized by F. nucleatum to physically bind to key members of the oral community, we investigated the involvement of F. nucleatum outer membrane proteins in its ability to bind to the pioneer biofilm colonizer, Streptococcus gordonii. Here, we present evidence that in addition to the previously characterized fusobacterial adhesin RadD, the interaction between F. nucleatum ATCC 23726 and S. gordonii V288 involves a second outer membrane protein, which we named coaggregation mediating protein A (CmpA). We also characterized the role of CmpA in dual‐species biofilm formation with S. gordonii V288, evaluated growth‐phase‐dependent as well as biofilm expression profiles of radD and cmpA, and confirmed an important role for CmpA, especially under biofilm growth conditions. Our findings underscore the complex set of specific interactions involved in physical binding and thus community integration of interacting bacterial species. This complex set of interactions could have critical implications for the formation and maturation of the oral biofilms in vivo, and could provide clues to the mechanism behind the distribution of organisms inside the human oral cavity.

Role of Candida albicans secreted aspartyl protease Sap9 in interkingdom biofilm formation.

The fungus Candida albicans colonizes oral cavity surfaces and is carried by up to 60% of human populations. Biofilm development by C.albicans may be modulated by oral streptococci, such as Streptococcus gordonii, S. oralis or S. mutans, so as to augment pathogenicity. In this study we sought to determine if the cell wall-associated secreted aspartyl proteinase Sap9 was necessary for hyphal adhesin functions associated with biofilm community development. A sap9Δ mutant of C.albicans SC5314 formed biofilms that were flatter, and contained fewer blastospores and more hyphal filaments than the parent strain. This phenotypic difference was accentuated under flow (shear) conditions and in the presence of S. gordonii. Dual-species biofilms of C.albicans sap9Δ with S. oralis, S. sanguinis, S. parasanguinis, S. mutans and Enterococcus faecalis all contained more matted hyphae and more bacteria bound to substratum compared to C.albicans wild type. sap9Δ mutant hyphae showed significantly increased cell surface hydrophobicity, ∼25% increased levels of binding C.albicans cell wall protein Als3, and reduced interaction with Eap1, implicating Sap9 in fungal cell-cell recognition. These observations suggest that Sap9 is associated with protein-receptor interactions between fungal cells, and with interkingdom communication in the formation of polymicrobial biofilm communities.

Pseudomonas aeruginosa PAO1 exopolysaccharides are important for mixed species biofilm community development and stress tolerance.

Pseudomonas aeruginosa PAO1 produces three polysaccharides, alginate, Psl, and Pel that play distinct roles in attachment and biofilm formation for monospecies biofilms. Considerably less is known about their role in the development of mixed species biofilm communities. This study has investigated the roles of alginate, Psl, and Pel during biofilm formation of P. aeruginosa in a defined and experimentally informative mixed species biofilm community, consisting of P. aeruginosa, Pseudomonas protegens, and Klebsiella pneumoniae. Loss of the Psl polysaccharide had the biggest impact on the integration of P. aeruginosa in the mixed species biofilms, where the percent composition of the psl mutant was significantly lower (0.06%) than its wild-type (WT) parent (2.44%). In contrast, loss of the Pel polysaccharide had no impact on mixed species biofilm development. Loss of alginate or its overproduction resulted in P. aeruginosa representing 8.4 and 18.11%, respectively, of the mixed species biofilm. Dual species biofilms of P. aeruginosa and K. pneumoniae were not affected by loss of alginate, Pel, or Psl, while the mucoid P. aeruginosa strain achieved a greater biomass than its parent strain. When P. aeruginosa was grown with P. protegens, loss of the Pel or alginate polysaccharides resulted in biofilms that were not significantly different from biofilms formed by the WT PAO1. In contrast, overproduction of alginate resulted in biofilms that were comprised of 35–40% of P. aeruginosa, which was significantly higher than the WT (5–20%). Loss of the Psl polysaccharide significantly reduced the percentage composition of P. aeruginosa in dual species biofilms with P. protegens (<1%). Loss of the Psl polysaccharide significantly disrupted the communal stress resistance of the three species biofilms. Thus, the polysaccharide composition of an individual species significantly impacts mixed species biofilm development and the emergent properties of such communities.

Rhodobacteraceaeare the key members of the microbial community of the initial biofilm formed in Eastern Mediterranean coastal seawater

The formation of biofilms and biofouling is a common feature in aquatic environments. The aim of this study was to identify the primary colonizers of biofilm formed in Eastern Mediterranean Coastal water at different seasons and follow early dynamics of biofilm community development. Pre-treated coastal seawater and biofilm samples were collected from six different sampling events of 2 weeks' duration each during 1 year. The microbial community composition and specific abundance were estimated by 16S rRNA gene clone libraries and fluorescence in situ hybridization-confocal laser scanning microscopy (FISH-CLSM), respectively. The biofilm formed over the course of the year was fairly consistent in terms of community composition and overall abundance with the exception of spring season. Alphaproteobacteria (30-70% of total bacteria), in particular Rhodobacteraceae, were the dominant bacteria in the biofilm, regardless of season, followed by Bacteroidetes (5-35%) and Gammaproteobacteria (6-35%). There was a decrease in relative abundance of Alphaproteobacteria and an increase in the abundance of Bacteroidetes between the initial and 2-week-old biofilm. This observation may aid man-made facilities that have to deal with biofilm formation and help the development of appropriate strategies to control those biofilms.

Implications of colonizing biofilms and microclimate on west stucco masks at North Acropolis, Tikal, Guatemala

Introduction: The Mayan archaeological sites belong to the World cultural heritage. The porous nature of limestone and stucco (calcareous coating) along with the high humidity and temperature typical of Southern Mexico and Central America, make these monuments prone to stone biodeterioration. The Mayan masks and stelae of the North Acropolis Complex (Tikal, Guatemala) are one of the most emblematic and valuable items at this site. As a common practice to keep these items from weathering, archaeologists and restorers build palm roofs over them. A field survey undertaken in august 2006 has shown that the West Mask (WM) semi-protected under a roof was heavily colonized by biofilms and display decayed feature. The East Mask (EM) located in a vault and kept from the exterior environment appears dry and sound. The apparent correlation of biofilm coverage and substratum deterioration led us to hypothesize that biofilm coating the WM had deteriogenic activity. Results: The purpose of this study was to characterize the biofilms colonizing WM, determine efflorescence chemistry and monitor microclimate under the palm roof in order to shed light on the ongoing deterioration phenomena. Contrasting microclimates were observed associated with WM and EM, with varying levels of relative humidity, sunlight and temperature being linked to the former. These conditions allowed the development of biofilm communities dominated by cyanobacteria only on the WM. Some of the detected taxa are recognized endolithic organisms (i.e. Gloecapsa). Fourier-transformed infrared spectroscopy showed that polysaccharides dominated the chemistry of biofilms. X-ray diffraction analyses (XRD) confirmed the presence of halite, gypsum and weddellite in efflorescences associated with the WM. Our results suggest that the joint impact of a varying microclimatic regime and the presence of biofilms promoted the deterioration. Conclusions: The use of palm roofs as protective practice in the Mayan area should be reassessed. Although they prevent deterioration stucco masks and stelae from direct exposure to the environment, they also induce a microclimate suitable for phototrophic biofilms capable of deteriorating directly and indirectly stucco materials. Management of microclimate and biofilm control should be further investigated as additional means to conserve these valuable items.

The relationship between epilithic biofilm stability and its associated meiofauna under two patterns of flood disturbance

Habitat stability is an important driver of ecological community composition and development. River epilithic biofilms are particularly unstable habitats for the establishment of benthic communities because they are regularly disturbed by floods. Our aim was to determine the influence of habitat instability on meiobenthic organisms. We hypothesized that hydrologic variables are the most important predictors of meiofauna distribution. We monitored epilithic communities (meiofauna and microalgae) with a high sampling frequency during 2 sampling periods with contrasting hydrodynamic patterns in a temperate river (the Garonne, France). Nematodes and rotifers dominated meiofaunal assemblages. The critical flow velocity threshold for their maintenance in the biofilm was ,30 cm/s, a result suggesting that meiofauna can resist higher flow velocity within the biofilm than within sediments. Nematode distribution was primarily influenced by the duration of undisturbed periods, whereas rotifer distribution was also correlated with the thickness of the biofilm. During the periods after floods, rotifers were faster colonizers than nematodes. Collectively, our results show that flow regime was an essential driver for biofilm community development.

Streptococcus gordonii Modulates Candida albicans Biofilm Formation through Intergeneric Communication

The fungus Candida albicans colonizes human oral cavity surfaces in conjunction with a complex microflora. C. albicans SC5314 formed biofilms on saliva-coated surfaces that in early stages of development consisted of approximately 30% hyphal forms. In mixed biofilms with the oral bacterium Streptococcus gordonii DL1, hyphal development by C. albicans was enhanced so that biofilms consisted of approximately 60% hyphal forms. Cell-cell contact between S. gordonii and C. albicans involved Streptococcus cell wall-anchored proteins SspA and SspB (antigen I/II family polypeptides). Repression of C. albicans hyphal filament and biofilm production by the quorum-sensing molecule farnesol was relieved by S. gordonii. The ability of a luxS mutant of S. gordonii deficient in production of autoinducer 2 to induce C. albicans hyphal formation was reduced, and this mutant suppressed farnesol inhibition of hyphal formation less effectively. Coincubation of the two microbial species led to activation of C. albicans mitogen-activated protein kinase Cek1p, inhibition of Mkc1p activation by H(2)O(2), and enhanced activation of Hog1p by farnesol, which were direct effects of streptococci on morphogenetic signaling. These results suggest that interactions between C. albicans and S. gordonii involve physical (adherence) and chemical (diffusible) signals that influence the development of biofilm communities. Thus, bacteria may play a significant role in modulating Candida carriage and infection processes in the oral cavity.

Modeling Growth and Quorum Sensing in Biofilms Grown in Microfluidic Chambers

Biofilms are highly organized structures coordinately formed by multiple species of bacteria. Quorum sensing (QS) is one cell-cell communication mechanism that is used by bacteria during biofilm formation. Biofilm formation is widely acknowledged to occur through a sequence of spatially and temporally regulated colonization events. While several mathematical models exist for describing biofilm development, these have been developed for open systems and are not applicable to closed systems where biofilm development and hydrodynamics are interlinked. Here, we report the development of a mathematical model describing QS and biofilm formation in a closed system such as a microfluidic channel. The model takes into account the effect of the external environment viz the mass and momentum transport in the microfluidic channel on QS and biofilm development. Model predictions of biofilm thickness were verified experimentally by developing Pseudomonas aeruginosa PA14 biofilms in microfluidic chambers and reflect the interplay between the dynamics of biofilm community development, mass transport, and hydrodynamics. Our QS model is expected to guide the design of experiments in closed systems to address spatio-temporal aspects of QS in biofilm development and can lead to novel approaches for controlling biofilm formation through disruption of QS spatio-temporal dynamics.

Manganese removal from mine waters – investigating the occurrence and importance of manganese carbonates

Manganese is a common contaminant of mine water and other waste waters. Due to its high solubility over a wide pH range, it is notoriously difficult to remove from contaminated waters. Previous systems that effectively remove Mn from mine waters have involved oxidising the soluble Mn(II) species at an elevated pH using substrates such as limestone and dolomites. However it is currently unclear what effect the substrate type has upon abiotic Mn removal compared to biotic removal by in situ micro-organisms (biofilms). In order to investigate the relationship between substrate type, Mn precipitation and the biofilm community, net-alkaline Mn-contaminated mine water was treated in reactors containing one of the pure materials: dolomite, limestone, magnesite and quartzite. Mine water chemistry and Mn removal rates were monitored over a 3-month period in continuous-flow reactors. For all substrates except quartzite, Mn was removed from the mine water during this period, and Mn minerals precipitated in all cases. In addition, the plastic from which the reactor was made played a role in Mn removal. Manganese oxyhydroxides were formed in all the reactors; however, Mn carbonates (specifically kutnahorite) were only identified in the reactors containing quartzite and on the reactor plastic. Magnesium-rich calcites were identified in the dolomite and magnesite reactors, suggesting that the Mg from the substrate minerals may have inhibited Mn carbonate formation. Biofilm community development and composition on all the substrates was also monitored over the 3-month period using denaturing gradient gel electrophoresis (DGGE). The DGGE profiles in all reactors showed no change with time and no difference between substrate types, suggesting that any microbiological effects are independent of mineral substrate. The identification of Mn carbonates in these systems has important implications for the design of Mn treatment systems in that the provision of a carbonate-rich substrate may not be necessary for successful Mn precipitation.

Biofilms as a Mode of Existence of Bacteria in External Environment and Host Body: The Phenomenon, Genetic Control, and Regulation Systems of Development

Studies of the last decade have shown that most bacteria exist in natural ecosystems as specifically organized, attached to substrates biofilms rather than as freely floating plankton cells. The formation of these biofilms is a complex and highly regulated process. The development of biofilm communities is a primary strategy of bacterial survival not only in the external environment but also in the bodies of infected macroorganisms. In these organisms, bacteria are joined by complicated cell-cell associations, which makes them functionally similar to multicellular organisms. In the present review, we consider the structural organization of biofilms, factors affecting initiation of the biofilm formation, differential expression of bacterial genes at various stages of the biofilm development and their regulation. The significance of studies in this field for medicine, in particular, for prevention and protection against pathogenic bacteria, is discussed.