Biofilm Surface Area Research Articles

Kinetics of ferrihydrite reduction in a biofilm system

Microorganisms can exist in both suspended and attached forms in natural and engineered systems. While extensive research has been performed to investigate the bioreduction kinetics of iron oxides by suspended microorganisms, the kinetics of iron oxide reduction by biofilms has not been well studied. This study investigated the rates of ferrihydrite bioreduction by biofilms of Shewanella oneidensis MR-1 with variable concentration ratios of electron donor to acceptor, and different biofilm thickness. Results indicated that the bioreduction rate of ferrihydrite by biofilms increased with increasing biofilm surface area and was faster than that by suspended microorganisms for the same biomass amount, apparently as a result of the biofilm surface area facilitating the direct contact between ferrihydrite and microorganisms for electron transfer. Increasing biofilm thickness decreased bioreduction rate for the same total biomass as a result of the decrease of the ratio of the biofilm surface area to total biomass. The bioreduction rate increased with increasing electron donor and ferrihydrite inputs, consistent with the saturation type of kinetic models. The different rates of ferrihydrite reduction in the biofilm system resulted in different rates of Fe(II) production as a function of electron donor, acceptor, and biofilm surface area, which in turn affected the type and concentrations of the secondary iron minerals, such as lepidocrocite and goethite. A kinetic model was developed that integrates biofilm surface area, and electron donor and acceptor concentrations that well described the measured rates of ferrihydrite bioreduction. The results emphasized the importance of the biofilm surface area in understanding and modeling iron redox biogeochemistry and iron-related biogeochemical processes.

Evaluating the biocompatibility of ceramic materials for constructing artificial reefs

IntroductionCoastal ecosystems, including reefs, are becoming increasingly threatened as anthropogenic development continues to encroach on intertidal habitats with little initiative to establish ecologically considerate infrastructure. Submerging human-made, shelter-providing structures known as artificial reefs (AR) can contribute to the preservation of these ecosystems. ARs are historically used for promoting the abundance and biodiversity of marine species for aquaculture, conservation, and ecotourism; and are typically made of concretes or metal structures. An AR’s success correlates to its ability to establish a surface layer of microorganisms, such as microalgae and bacteria, known as a biofilm. The productivity of the biofilm can be influenced by material surface properties. It is hypothesized that material pH and porosity affect the rate of biofilm formation.MethodsHere - a range of concrete mixtures were cast and submerged in circulating seawater and mass per surface area of biofilm accumulation was measured to evaluate this theory. These mixtures included standard Portland Cement (PC), PC with admixtures of diatomaceous earth (PDC) and limestone (PLC), fine-aggregate high-performance concrete (DUC), and terra cotta (TER). ARs were manufactured as 38mm tall cylinders, 76mm in diameter, and submerged in circulating seawater to evaluate mass per surface area of biofilm accumulation.ResultsOur results indicate that biofilm formation is directly affected by surface porosity and less-so by pH, as determined by measuring material properties after submersion. We found that the PDC samples were most successful in forming a biofilm despite being more fragile than other concrete samples.DiscussionThis preliminary study provides insight into how different material properties influence the accumulation of biofilm as a starting point for designing ARs. Future work will investigate the long-term performance of such samples in relevant conditions.

Biofilm Formation and Genetic Diversity of Microbial Communities in Anaerobic Batch Reactor with Polylactide (PLA) Addition.

In this paper, an anaerobic digestion (AD) study was conducted on confectionery waste with granular polylactide (PLA) as a cell carrier. Digested sewage sludge (SS) served as the inoculum and buffering agent of systems. This article shows the results of the analyses of the key experimental properties of PLA, i.e., morphological characteristics of the microstructure, chemical composition and thermal stability of the biopolymer. The evaluation of quantitative and qualitative changes in the genetic diversity of bacterial communities, performed using the state-of-the-art next generation sequencing (NGS) technique, revealed that the material significantly enhanced bacterial proliferation; however, it does not change microbiome biodiversity, as also confirmed via statistical analysis. More intense microbial proliferation (compared to the control sample, without PLA and not digested, CW-control, CW-confectionery waste) may be indicative of the dual role of the biopolymer-support and medium. Actinobacteria (34.87%) were the most abundant cluster in the CW-control, while the most dominant cluster in digested samples was firmicutes: in the sample without the addition of the carrier (CW-dig.) it was 68.27%, and in the sample with the addition of the carrier (CW + PLA) it was only 26.45%, comparable to the control sample (CW-control)-19.45%. Interestingly, the number of proteobacteria decreased in the CW-dig. sample (17.47%), but increased in the CW + PLA sample (39.82%) compared to the CW-control sample (32.70%). The analysis of biofilm formation dynamics using the BioFlux microfluidic system shows a significantly faster growth of the biofilm surface area for the CW + PLA sample. This information was complemented by observations of the morphological characteristics of the microorganisms using fluorescence microscopy. The images of the CW + PLA sample showed carrier sections covered with microbial consortia.

A promising innovative technique for mycotoxin detoxification from beverages using biofilms of lactic acid bacteria

This study assessed the removal of aflatoxin M1 (AFM1) and ochratoxin A (OTA) from artificially contaminated whole UHT milk and red grape juice, respectively, using biofilms from Lactobacillus rhamnosus GG. Using ELISA, the level of AFM1 and OTA removal from beverages was determined depending on various factors. Biofilms of various ages demonstrated varying degrees of AFM1 removal capacity from phosphate-buffered saline (PBS). Different levels of AFM1 contaminated whole UHT milk (0.1, 0.2, and 1 μg/L) and OTA contaminated red grape juice (2 and 4 μg/L) were tested in the detoxification process. The binding ability of mycotoxins was improved by increasing the biofilm surface area up to 70 cm2. L. rhamnosus GG biofilm was effective in removing mycotoxins within a short contact time ranging from 1 to 10 min. The proportion of bound AFM1 and OTA by L. rhamnosus GG biofilm was 64.6 and 98.3% respectively. A new machine has been proposed and used as a trial for detoxication purposes which would be a promising application in liquid food industries.

Biofilm-responsive encapsulated-phage coating for autonomous biofouling mitigation in water storage systems

Biofilms in water storage systems may harbor pathogens that threaten public health. Chemical disinfectants are marginally effective in eradicating biofilms due to limited penetration, and often generate harmful disinfection byproducts. To enhance biofouling mitigation in household water storage tanks, we encapsulated bacteriophages (phages) in chitosan crosslinked with tri-polyphosphate and 3-glycidoxypropyltrimethoxysilane. Phages served as self-propagating green biocides that exclusively infect bacteria. This pH-responsive encapsulation (244 ± 11 nm) enabled autonomous release of phages in response to acidic pH associated with biofilms (corroborated by confocal microscopy with pH-indicator dye SNARF-4F), but otherwise remained stable in pH-neutral tap water for one month. Encapsulated phages instantly bind to plasma-treated plastic and fiberglass surfaces, providing a facile coating method that protects surfaces highly vulnerable to biofouling. Biofilm formation assays were conducted in tap water amended with 200 mg/L glucose to accelerate growth and attachment of Pseudomonas aeruginosa, an opportunistic pathogen commonly associated with biofilms in drinking water distribution and storage systems. Biofilms formation on plastic surfaces coated with encapsulated phages decreased to only 6.7 ± 0.2% (on a biomass basis) relative to the uncoated controls. Likewise, biofilm surface area coverage (4.8 ± 0.2 log CFU/mm2) and live/dead fluorescence ratio (1.80) were also lower than the controls (6.6 ± 0.2 log CFU/mm2 and live/dead ratio of 11.05). Overall, this study offers proof-of-concept of a chemical-free, easily implementable approach to control problematic biofilm-dwelling bacteria and highlights benefits of this bottom-up biofouling control approach that obviates the challenge of poor biofilm penetration by biocides.

Unsweetened and sucrose-sweetened black and green tea modifies the architecture of in vitro oral biofilms

ObjectiveIt is unclear whether tea infusions with or without sucrose supplementation alter oral biofilm development, so we evaluated the effect of unsweetened and sucrose-sweetened black and green tea infusions on in vitro saliva-derived biofilms. DesignBiofilms were developed from human saliva for 20 h in cell-free 25% human saliva within static glass-bottom microplates. During biofilm development, biofilms were treated with either (i) unsweetened black tea, (ii) unsweetened green tea, (iii) 10% sucrose-sweetened black tea, (iv) 10% sucrose-sweetened green tea (v) deionized water (negative control), or (vi) 10% sucrose (positive control). Biofilms were incubated at 37 °C in 5% CO2. After 20 h of development, biofilms were imaged using a CLSM, and biofilm architecture and viability were evaluated. ResultsAll the tea infusions reduced biofilm biomass and altered some other biofilm architectural outcomes (e.g., biofilm surface area) compared to the control groups. Statistically significant differences in biofilm biomass, number of objects, surface area, and convex-hull porosity were observed between biofilms treated with green and black tea. The addition of sugar to tea did not significantly modify the ability of tea to alter biofilm architecture. Only the treatment of biofilms with unsweetened black tea significantly reduced bacterial viability. ConclusionsWhile both teas reduced biofilm biomass and altered biofilm architecture, black tea had an enhanced effect that may relate to this tea's observed antimicrobial activity. The addition of sucrose to tea infusions did not appear to reduce the impact of either tea in modifying oral biofilm architecture

Ammonia Removal in Free-Surface Constructed Wetlands Employing Synthetic Floating Islands

Free water surface constructed wetlands (FSCWs) can be used to complement conventional waste water treatment but removal efficiencies are often limited by a high ratio of water volume to biofilm surface area (i.e. high water depth). Floating treatment wetlands (FTWs) consist of floating matrices which can enhance the surface area available for the development of fixed microbial biofilms and provide a platform for plant growth (which can remove pollutants by uptake). In this study the potential of FTWs for ammoniacal nitrogen (AN) removal was evaluated using experimental mesocosms operated under steady-state flow conditions with ten different treatments (two water depths, two levels of FTW mat coverage, two different plant densities and a control, all replicated three times). A simple model was constructed as a framework for understanding N dynamics in each treatment. The model was calibrated using data obtained from one treatment and validated independently for the other treatments. Specifically, we hypothesized that the nitrification and volatilization rate constants are inversely proportional to water depth and proportional to mat surface area. This allowed the relative magnitude of different removal mechanisms to be estimated. The model was able to predict steady-state concentrations of AN and total oxidized nitrogen (TON) across the different treatments well (values for correlation in the regression between measured and predicted steady-state concentrations and RMSE were 0.88 and 0.40 mg N L-1 for AN, and 0.63 and 1.75 mg N L-1 for TON). The results confirm that nitrification is the principal AN removal process, with maximum removal occurring in shallow systems with high matrix cover (i.e. a high ratio of biofilm surface area to water volume). Plant uptake was a relatively minor loss process compared to nitrification. Integrated experimental and model-based approach was found to be a useful tool to improve mechanistic understanding AN dynamics in FSCWs and system performance.

Design of 3D microbial anodes for microbial electrolysis cells (MEC) fuelled by domestic wastewater. Part I: Multiphysics modelling

The performance of a microbial electrolysis cell (MEC) supplied with domestic wastewater (dWW) is essentially limited by the kinetics of the anodic bioelectrochemical reactions and the low ionic conductivity of the electrolyte. A strategy to boost-up the anodic bioelectrochemical kinetics is to use three-dimensional (3D) microbial anodes that offer a high total anodic surface area and volume density of electroactive biofilm. In this work, a 3D multiphysics model was designed to simulate the current generation and resulting hydrogen production in double and triple-compartment MECs fed continuously with dWW. Simulations indicated that optimised 3D microbial anode geometries could simultaneously increase current and chemical oxygen demand (COD) removal by 86% compared to a 2D planar graphite electrode. At a constant CEM voltage, the current produced increased with the thickness of the 3D microbial anode up to a limiting thickness of 20 mm. Beyond this value, the current was stagnant due to the predominant ohmic drop. Current generation and COD removal could be further increased by designing 3D anode geometrical arrangements that force the dWWs to flow through the porosity of the 3D microbial anode. A gain of 20% was calculated by substituting a monolithic 3D graphite anode with a 3D anode of the same thickness (20 mm) but constructed of plates stacked on top of each other and spaced 2.5 mm apart. Finally, hydrogen production performance was additionally optimised by a further + 20% by switching from a two-compartment MEC design (anode-cathode) to a three-compartment MEC design (cathode-anode-cathode).

Model-based analysis of sulfur-based denitrification in a moving bed biofilm reactor

ABSTRACT In this study, a biofilm model was developed for sulfur-based denitrification in a moving bed biofilm reactor (MBBR), including mass transport as well as the conversion kinetics of sulfur-oxidizing bacteria (SOB). The experimental reactor simulated received a synthetic wastewater containing nitrate, sulfide and thiosulfate. The substrate affinity of SOB for intermediary elemental sulfur (S0) was found the most sensitive parameter. After estimating this single parameter, the model could adequately describe the steady state performance of the experimental MBBR. The experimental and simulated mass balances indicated that a fraction of influent sulfur accumulated into intermediate S0. Furthermore, the simulations showed that SOB were active over the entire thickness of a 200 µm biofilm. The simulation results allowed to quantify the extent of diffusion and substrate limitation. Scenario analyses indicated that the specific nitrogen loading rate could be increased from 0.05 to 0.20 kg N.kg−1 VSS.day−1 (corresponding to 0.22–0.86 kg N.m−2.day−1 expressed per biofilm surface area) while maintaining nitrogen removal efficiencies above 70%. An increasing specific nitrogen loading rate in this range resulted in an almost linearly increasing specific nitrogen removal rate, independent from whether it was realized through a decreasing HRT, carrier filling ratio or biofilm thickness.

THE EVOLVING WRINKLE PATTERN OF THE BACILLUS SUBTILIS BIOFILM PROVIDING MORE LIVING SPACE FOR CELLS

The biofilm wrinkle evolution is the growth mechanism by which bacteria regulate their physiological state in response to the environmental change. We use the parameter of surface complexity to describe different wrinkle patterns. The surface complexity is defined that the biofilm surface area contact with the air is divided by the projected area of the biofilm. We find that the biofilm surface complexity variation is positively proportional to the number of spores. Although each wrinkle pattern has various wrinkle thickness and width, surface complexities of some patterns are almost same, which guarantees cells have enough living space. Through the observation of the growth of the damaged biofilm, we further find that the biofilm expansion along the circumferential direction is faster than that along radial direction, which means that the internal stress along the circumferential direction contributes the wrinkle formation. Our work provides a new perspective to study biofilm morphologies, and relates the morphology evolution with phenotypes in the Bacillus subtilis biofilm.

Performance analysis and optimization of ammonium removal in a new biological folded non-aerated filter reactor

A new type of biological folded non-aerated filter (BFNAF) was found to be superior and feasible for the treatment of NH4+-N wastewater. It was constructed with the folded structure suitable for the nylon biomass carrier. The advantages of the BFNAF included low energy consumption, long reaction path, large biofilm surface area and non-clogging compared to the traditional biological aerated filter. In this study, the effects of hydraulic retention time (HRT), and the influent NH4+-N concentration on the performance of BFNAF were investigated and optimized by the response surface methodology. Under the optimal operating condition (HRT, 10 h; NH4+-N concentration, 52 mg/L), the removal efficiency and removal rate were 94.62 ± 0.63% and 0.106 kg-NH4+ m−3 day−1, respectively. The results showed that the BFNAF reactor could remove NH4+-N from wastewater and realized the nitrification process effectively under natural ventilation conditions.

Marine Bacteria, A Source for Alginolytic Enzyme to Disrupt Pseudomonas aeruginosa Biofilms.

Pseudomonas aeruginosa biofilms are typically associated with the chronic lung infection of cystic fibrosis (CF) patients and represent a major challenge for treatment. This opportunistic bacterial pathogen secretes alginate, a polysaccharide that is one of the main components of its biofilm. Targeting this major biofilm component has emerged as a tempting therapeutic strategy for tackling biofilm-associated bacterial infections. The enormous potential in genetic diversity of the marine microbial community make it a valuable resource for mining activities responsible for a broad range of metabolic processes, including the alginolytic activity responsible for degrading alginate. A collection of 36 bacterial isolates were purified from marine water based on their alginolytic activity. These isolates were identified based on their 16S rRNA gene sequences. Pseudoalteromonas sp. 1400 showed the highest alginolytic activity and was further confirmed to produce the enzyme alginate lyase. The purified alginate lyase (AlyP1400) produced by Pseudoalteromonas sp. 1400 showed a band of 23 KDa on a protein electrophoresis gel and exhibited a bifunctional lyase activity for both poly-mannuronic acid and poly-glucuronic acid degradation. A tryptic digestion of this gel band analyzed by liquid chromatography-tandem mass spectrometry confirmed high similarity to the alginate lyases in polysaccharide lyase family 18. The purified alginate lyase showed a maximum relative activity at 30 °C at a slightly acidic condition. It decreased the sodium alginate viscosity by over 90% and reduced the P. aeruginosa (strain PA14) biofilms by 69% after 24 h of incubation. The combined activity of AlyP1400 with carbenicillin or ciprofloxacin reduced the P. aeruginosa biofilm thickness, biovolume and surface area in a flow cell system. The present data revealed that AlyP1400 combined with conventional antibiotics helped to disrupt the biofilms produced by P. aeruginosa and can be used as a promising combinational therapeutic strategy.

Zirconium Nitride Coating Reduced Staphylococcus epidermidis Biofilm Formation on Orthopaedic Implant Surfaces: An In Vitro Study.

One of the most commonly identified pathogens responsible for orthopaedic implant infection is Staphylococcus epidermidis, which can form biofilms on surfaces. Currently, orthopaedic implants made of various surface materials are available, each with features influencing osseointegration, biocompatibility, and adherence of bacteria to the surface, which is the first step in biofilm formation. The aim of this experimental study was to investigate the effect of a high tribologic-resistant 2.5-µm zirconium nitride top coat on an antiallergic multilayer ceramic-covered cobalt-chromium-molybdenum surface on the formation of S. epidermidis biofilm compared with other commonly used smooth and rough orthopaedic implant surface materials. (1) When evaluating the surfaces of a cobalt-chromium-molybdenum (CoCrMo) alloy with a zirconium (Zr) nitride coating, a CoCrMo alloy without a coating, titanium alloy, a titanium alloy with a corundum-blasted rough surface, and stainless steel with a corundum-blasted rough surface, does a Zr coating reduce the number of colony-forming units of S. epidermidis in an in vitro setting? (2) Is there quantitatively less biofilm surface area on Zr-coated surfaces than on the other surfaces tested in this in vitro model? To determine bacterial adhesion, five different experimental implant surface discs were incubated separately with one of 31 different S. epidermidis strains each and subsequently sonicated. Twenty test strains were obtained from orthopaedic patients undergoing emergency hip prosthesis surgeries or revision of implant infection and 10 further strains were obtained from the skin of healthy individuals. Additionally, one reference strain, S. epidermidis DSM 3269, was tested. After serial dilutions, the number of bacteria was counted and expressed as colony-forming units (CFUs)/mL. For biofilm detection, discs were stained with 0.1% Safranin-O for 15 minutes, photographed, and analyzed with computer imaging software. The lowest bacterial count was found in the CoCrMo + Zr surface disc (6.6 x 10 CFU/mL ± 4.6 x 10 SD) followed by the CoCrMo surface (1.1 x 10 CFU/mL ± 1.9 x 10 SD), the titanium surface (1.36 x 10 CFU/mL ± 1.8 x 10 SD), the rough stainless steel surface (2.65 x 10 CFU/mL ± 3.8 x 10 SD), and the rough titanium surface (2.1 x 10 CFU/mL ± 3.0 x 10 SD). The mean CFU count was lower for CoCrMo + Zr discs compared with the rough stainless steel surface (mean difference: 2.0 x 10, p = 0.021), the rough titanium alloy surface (mean difference: 1.4 x 10, p = 0.002), and the smooth titanium surface (mean difference: 7.0 x 10, p = 0.016). The results of biofilm formation quantification show that the mean covered area of the surface of the CoCrMo + Zr discs was 19% (± 16 SD), which was lower than CoCrMo surfaces (35% ± 23 SD), titanium alloy surface (46% ± 20 SD), rough titanium alloy surface (66% ± 23 SD), and rough stainless steel surface (58% ± 18 SD). These results demonstrate that a multilayer, ceramic-covered, CoCrMo surface with a 2.5-µm zirconium nitride top coat showed less S. epidermidis biofilm formation compared with other surface materials used for orthopaedic implants. CoCrMo with a 2.5-µm zirconium nitride top coat seems to be a promising surface modification technology able to reduce bacterial attachment on the surface of an implant and, hence, may further prevent implant infection with S. epidermidis biofilm formation.

A marine biofilm flow cell for in situ screening marine fouling control coatings using optical coherence tomography

A novel fouling marine flow cell was designed and fitted with a clear plastic lid to allow real-time imaging of biofilms using optical coherence tomography (OCT). Marine biofilms were grown under controlled shear flow on coupons coated with 6 different biocidal antifouling coatings (SPC1, SPC2, SPC3, SPC4, CDP1 and CDP2, AkzoNobel) and one inert coating which contained no biocidal actives (NB-D) for 8 weeks. One set of coupons coated with NB was statically immersed in sea water during the same time period (NB-S). Biofilm removal was assessed by increasing the flow velocity while OCT simultaneously measured the biofilm cross-sectional area (CSA). The highest initial removal rates were observed for NB-S, NB-D, and SPC2. Percent biofilm cross-sectional area reduction (%CSAred) was higher on SPCs (>60%) compared to CDPs (<50%). SPCs had the highest percent reduction in biofilm surface area coverage (%SACred >60%) compared to the CDPs (<20%). The marine biofilm flow cell combined with OCT can be used to screen for coating-specific differences in biofilm growth and removal in real time rather than traditional before and after surface area coverage measurements. Future testing will focus on how the biofilm-coatings interactions interact with biofilm mechanical and structural properties to produce drag.

The role of interactions of effective biofilm surface area and mass transfer in nitrogen removal efficiency of an integrated fixed-film activated sludge system

A reaction-diffusion biofilm model was implemented to simulate the nitrification/denitrification performance of a lab-scale integrated fixed-film activated sludge (IFAS) reactor. The model was capable of representing the system performance, i.e. changes in organic load and decrease in sludge age. Furthermore, nitrification batch tests with sludge and carrier material could also be simulated successfully with the model. Model simulation revealed that the diffusive fluxes into biofilm depended strongly on substrate loading as well as sludge age. The microbial composition in the biofilm matrix was mainly influenced by the diffusive flux of chemical oxygen demand (COD) into biofilm. When COD removal started to switch to biofilm, heterotrophic bacteria quickly replaced the previously dominating autotrophic bacteria. Running a set of simulations with a range of effective biofilm surface area and different mass transfer coefficients revealed the strong influence of these two parameters on the IFAS performance. The analysis showed that both parameters were dominating factors for ammonium removal. The optimum mass transfer coefficient was in the range of 3–4 m d−1 and the effective biofilm surface was around 63–88% of the theoretical carrier surface.

Middle ear irrigation using a hydrodebrider decreases biofilm surface area in an animal model of otitis media.

ObjectiveTo compare the safety and efficacy of manual and powered irrigation of the middle ear using saline or 1% baby shampoo to treat biofilm‐forming bacterial middle ear infections.BackgroundBiofilms play a major role in recalcitrant otitis media and are challenging to treat. Many therapeutic strategies have been attempted and the role of topical therapies is still being investigated. Topical irrigation using saline or 1% baby shampoo and the use of a hydrodebrider have been investigated in biofilms involved in chronic rhinosinusitis and their role within the middle ear is yet to be determined.MethodsTwenty‐two adult chinchillas underwent bilateral trans‐bullar inoculation of non‐typable biofilm forming Haemophilus influenza followed by unilateral middle ear irrigation 5 days later using saline administered via a powered hydrodebrider or manual irrigation of saline or 1% baby shampoo. Contralateral inoculated ears served as control and were not irrigated. Two days following irrigation, the bullae were harvested and processed for scanning electron microscopy to assess biofilm surface area. Auditory brainstem responses were performed before bacterial inoculation and prior to euthanasia.ResultsManual and powered irrigation were effective in reducing the surface area of biofilm when compared to the control group. The hydrodebrider demonstrated to be more effective at eradicating biofilm than manual irrigation, especially in areas of difficult access, such as the ventral portion of the chinchillas' bullae. There was no difference in manual irrigation of saline when compared to 1% baby shampoo. Irrigations either manually or using the hydrodebrider did not affect hearing, the vestibular system or facial function.ConclusionMiddle ear biofilms can be treated safely and effectively with rinses using either normal saline or 1% baby shampoo administered manually or with a powered hydrodebrider.Level of EvidenceNA.

Structure and cell viability analysis of Actinobacillus succinogenes biofilms as biocatalysts for succinic acid production

Biofilms of Actinobacillus succinogenes have demonstrated exceptional capabilities as biocatalysts for high productivity, titre, and yield production of succinic acid. This work gauges the effectiveness of these biofilms through analysis of the structure and cell viability of biofilms by using confocal scanning laser microscopy, scanning electron microscopy and image analysis software. The structure of the mature six-day-old biofilms of A. succinogenes was characterized by amorphous cell microcolonies of variable area and shape, with a mean thickness of 92±35μm and low surface area to volume ratios of 0.25±0.07μm2μm−3. Biofilm surface area coverage was low near the base (33±13%) but increased towards the outer layer. Despite the presence of water channels at the deeper portions of the biofilm, a greater portion of inactive cells was closest to the attachment surface and 65±2% of the biofilm consisted of dead cells. In this way, a cell viability gradient existed where the outer layer exhibits a greater fraction of active cells compared to the base layer. Biofilm cells underwent a phenotypic change, expressing a filamentous cell morphology different from rod-like morphology of suspended cells. The filamentous morphology permits extensive cell entanglements within a microcolony which may add to the intactness of microcolonies.

Influence of Different Sewer Biofilms on Transformation Rates of Drugs.

To estimate drug consumption more reliably, wastewater-based epidemiology would benefit from a better understanding of drug residue stability during in-sewer transport. We conducted batch experiments with real, fresh wastewater and sewer biofilms. Experimental conditions mimic small to medium-sized gravity sewers with a relevant ratio of biofilm surface area to wastewater volume (33 m2 m-3). The influences of biological, chemical, and physical processes on the transformation of 30 illicit drug and pharmaceutical residues were quantified. Rates varied among locations and over time. Three substances were not stable-that is, >20% transformation, mainly due to biological processes-at least for one type of tested biofilm for a residence time ≤2 h: amphetamine, 6-acetylcodeine, and 6-monoacetylmorphine. Cocaine, ecgonine methyl ester, norcocaine, cocaethylene, and mephedrone were mainly transformed by chemical hydrolysis and, hence, also unstable in sewers. In contrast, ketamine, norketamine, O-desmethyltramadol, diclofenac, carbamazepine, and methoxetamine were not substantially affected by in-sewer processes under all tested conditions and residence times up to 12 h. Our transformation rates include careful quantification of uncertainty and can be used to identify situations in which specific compounds are not stable. This will improve accuracy and uncertainty estimates of drug consumption when applied to the back-calculation.

Submerged bed versus unsaturated flow reactor: A pressurized hydrogenotrophic denitrification reactor as a case study

The paper compares the main features of a submerged bed reactor (SuBR) with bubbling and recirculation of gas to those of an unsaturated flow reactor (uSFR) with liquid recirculation. A novel pressurized closed-headspace hydrogenotrophic denitrification system characterized by safe and economic utilization of H2 gas was used for the comparison.Under similar conditions, denitrification rates were lower in the SuBR as a result of a lower effective biofilm surface area and overall gas-liquid mass transfer coefficient kLa. Similar values of effluent DOC were achieved for both reactors, although effluent suspended solids concentration of the SuBR were substantially higher. On the other hand, the required cleaning frequency in the SuBR was 2.5 times lower. Moreover, the SuBR is expected to reduce the recirculation energy consumption by 0.35 kWh/m3 treated.

CHARACTERISATION OF SULPHATE-REDUCING BACTERIAL BIOFILMS FROM CRUDE OIL AND PRODUCED WATER AT LAMINAR FLOW

Characterisation of sulfate-reducing bacterial biofilms from crude oil and producedwater at laminar flow was carried out with the aid of a flow-cell set-up, using In situimage microscopy. Biofilms of sulfate-reducing bacteria (SRB) indigenous to crudeoil (sample A) and produced water (sample B), were grown in square section glasscapillary flow cells at 55 ?C. Image analysis revealed predominantly clusters of cellsmade up of both coccoid and rod-shaped bacteria, ranging in length from 1-3 ?m,and some short chain and single cells with a variety of cell sizes demonstrating thevaried number of consortia in the mixed biofilm. Biofilm surface area cover showedincrease in surface area covered with time. Statistical significance of the means ofDifferential Interference Contrast (DIC) microscopy images of biofilm from bothsamples was determined and a p-value of less than 0.05 was considered significant.Results from statistical analysis showed that there was significant differencebetween biofilms of both samples, while there was no significant difference betweenmeans within each sample. A significant difference in biofilm cover and possiblegene expression between both samples at 55 ?C during laminar flow was thereforeimplied