Heavy Metal Removal from Aqueous Solutions Using a Customized Bipolar Membrane Electrodialysis Process.

Lack of safe water availability and access to clean water cause a higher risk of infectious diseases and other diseases as well. Heavy metals (HMs) are inorganic pollutants that cause severe threats to humans, animals, and the environment. Therefore, an effective HM removal technology is urgently needed. In the present study, a customized bipolar membrane electrodialysis process was used to remove HMs from aqueous solutions. The impacts of the feed ionic strength, applied electrical potential, and the type and concentration of HMs (Cd2+, Co2+, Cr3+, Cu2+, and Ni2+) on the process performance were investigated. The results showed that feed solution pH changes occurred in four stages: it first decreased linearly before stabilizing in the acidic pH range, followed by an increase and stabilization in the basic range of the pH scale. HM speciation in the basic pH range revealed the presence of anionic HM species. The presence of HMs on anion exchange membranes confirmed the contribution of these membranes for HM removal in the base channels of the process. While no clear trend was seen in the ionic strength solution, the maximum HM removal was observed when 1.5 g/L NaCl was used. The initial HM concentration showed a linear increase in HMs removal of up to 30 mg/L. A similar trend was seen with an increase in the applied electrical potential of up to 15 V. In general, the amount of HMs removed increased in the following order: Cd2+ ˃ Ni2+ ˃ Co2+ ˃ Cu2+ ˃ Cr3+. Under some operational conditions, however, the removed amount of Cu2+, Co2+, and Ni2+ was similar. The mass balance and SEM-EDX results revealed that the removed HMs were sorbed onto the membranes. In conclusion, this process efficiently separates HMs from aqueous solutions. It showed the features of diluate pH adjustment, reduction in the overall stack electrical resistance, and contribution of anion exchange membranes in multivalent cation removal. The mechanisms involved in HMs removal were diffusion and migration from the bulk solution, followed by their sorption on both cation and anion exchange membranes.

Enrichment of phosphate-accumulating organisms (PAOs) in a microfluidic model biofilm system by mimicking a typical aerobic granular sludge feast/famine regime.

Wastewater treatment using aerobic granular sludge has gained increasing interest due to its advantages compared to conventional activated sludge. The technology allows simultaneous removal of organic carbon, nitrogen, and phosphorus in a single reactor system and is independent of space-intensive settling tanks. However, due to the microscale, an analysis of processes and microbial population along the radius of granules is challenging. Here, we introduce a model system for aerobic granular sludge on a small scale by using a machine-assisted microfluidic cultivation platform. With an implemented logic module that controls solenoid valves, we realized alternating oxic hunger and anoxic feeding phases for the biofilms growing within. Sampling during ongoing anoxic cultivation directly from the cultivation channel was achieved with a robotic sampling device. Analysis of the biofilms was conducted using optical coherence tomography, fluorescence in situ hybridization, and amplicon sequencing. Using this setup, it was possible to significantly enrich the percentage of polyphosphate-accumulating organisms (PAO) belonging to the family Rhodocyclaceae in the community compared to the starting inoculum. With the aid of this miniature model system, it is now possible to investigate the influence of a multitude of process parameters in a highly parallel way to understand and efficiently optimize aerobic granular sludge-based wastewater treatment systems.Key points• Development of a microfluidic model to study EBPR.• Feast-famine regime enriches polyphosphate-accumulating organisms (PAOs).• Microfluidics replace sequencing batch reactors for aerobic granular sludge research.

The potential of fluorescent dyes-comparative study of Nile red and three derivatives for the detection of microplastics.

During the last years, microplastics in the environment came to the fore in environmental science research. For an appropriate risk assessment, it is essential to know the levels of microplastic contamination in the environment. In the field of microplastic detection, extensive research has been carried out in recent years. While common methods such as Raman spectroscopy and pyrolysis GC-MS are time-consuming and require trained staff and expensive equipment, there is the need for a cheap and easily applicable method. Staining microplastics with the fluorescent dye Nile red (NR) has a high potential to fulfill these criteria. In our work, we tested Nile red and newly developed derivatives, with the aim of achieving greater selectivity for plastic particles and more intense fluorescence. In addition, the influence of using different solvents and water at different pH values in the dyeing process was investigated by analyzing solid sample fluorescence spectra of dyed microplastics and natural particles. Finally, the method developed from the acquired knowledge was tested for sea salt. Graphical abstract.

Evaluation of productive biofilms for continuous lactic acid production.

In white biotechnology research, the putative superiority of productive biofilms to conventional biotransformation processes based on planktonic cultures has been increasingly discussed in recent years. In the present study, we chose lactic acid production as a model application to evaluate biofilm potential. A pure culture of Lactobacillus bacteria was grown in a tubular biofilm reactor. The biofilm system was cultivated monoseptically in a continuous mode for more than 3 weeks. The higher cell densities that could be obtained in the continuous biofilm system compared with the planktonic culture led to a significantly increased space-time yield. The productivity reached 80% of the maximum value 10 days after start-up and no subsequent decline was observed, confirming the suitability of the system for long-term fermentation. The analysis of biofilm performance revealed that productivity increases with the flow velocity. This is explained by the reduced retention time of the liquid phase in the reactor, and, thus, a minor pH drop caused by the released lactic acid. At low flow velocities, the pH drops to a value where growth and production are significantly inhibited. The biofilm was visualized by magnetic resonance imaging to analyze biofilm thickness. To deepen the understanding of the biofilm system, we used a simple model for cell growth and lactic acid production.

Biofilm systems as tools in biotechnological production.

The literature provides more and more examples of research projects that develop novel production processes based on microorganisms organized in the form of biofilms. Biofilms are aggregates of microorganisms that are attached to interfaces. These viscoelastic aggregates of cells are held together and are embedded in a matrix consisting of multiple carbohydrate polymers as well as proteins. Biofilms are characterized by a very high cell density and by a natural retentostat behavior. Both factors can contribute to high productivities and a facilitated separation of the desired end-product from the catalytic biomass. Within the biofilm matrix, stable gradients of substrates and products form, which can lead to a differentiation and adaptation of the microorganisms' physiology to the specific process conditions. Moreover, growth in a biofilm state is often accompanied by a higher resistance and resilience towards toxic or growth inhibiting substances and factors. In this short review, we summarize how biofilms can be studied and what most promising niches for their application can be. Moreover, we highlight future research directions that will accelerate the advent of productive biofilms in biology-based production processes.

Development of a production chain from vegetable biowaste to platform chemicals.

BACKGROUND: A future bioeconomy relies on the development of technologies to convert waste into valuable compounds. We present here an attempt to design a biotechnological cascade for the conversion of vegetable waste into acetoin and electrical energy. RESULTS: A vegetable waste dark fermentation effluent containing mainly acetate, butyrate and propionate was oxidized in a bioelectrochemical system. The achieved average current at a constant anode potential of 0 mV against standard hydrogen electrode was 177.5 ± 52.5 µA/cm2. During this step, acetate and butyrate were removed from the effluent while propionate was the major remaining component of the total organic carbon content comprising on average 75.6%. The key players with regard to carbon oxidation and electrode reduction were revealed using amplicon sequencing and metatranscriptomic analysis. Using nanofiltration, it was possible to concentrate the propionate in the effluent. The effluent was revealed to be a suitable medium for biotechnological production strains. As a proof of principle, the propionate in the effluent of the bioelectrochemical system was converted into the platform chemical acetoin with a carbon recovery of 86%. CONCLUSIONS: To the best of our knowledge this is the first report on a full biotechnological production chain leading from vegetable waste to the production of a single valuable platform chemical that integrates carbon elimination steps leading to the production of the valuable side product electrical energy.

Treatment of thermophilic hydrolysis reactor effluent with ceramic microfiltration membranes.

For an undisturbed operation of two-stage high-pressure fermentation up to 100 bar, a particle-free hydrolysate appears to be necessary. This is even more important if the second stage, i.e., the methane reactor, is designed as fixed bed. Here, we present the potential of microfiltration membranes as separation unit after the first stage, which is the hydrolysis. The study included the selection of membrane material, membrane performance investigations, and long-term-behavior during the filtration period. In a series of experiments, the optimum type of membrane material and the mode of operation [either crossflow (CF) or submerged (S)] were determined. Ceramic membranes proved to be the better option to treat the process stream due to their chemical and temperature resistance. The crossflow filtration achieved a sustainable flux of up to 33 L/(m2 h), while long-term experiments with the submerged membranes confirmed a critical flux of 7 L/(m2 h). Comparative analyses of hydrolysate and permeate showed that the rejected chemical oxygen demand (COD) as well as total organic carbon (TOC) fraction and thereby the loss of organic carbon in the permeate does not reduce the methane yield.

Optical coherence tomography in biofilm research: A comprehensive review.

Imaging of biofilm systems is a prerequisite for a better understanding of both structure and its function. The review aims to critically discuss the use of optical coherence tomography (OCT) for the visualization of the biofilm structure as well as its dynamic behavior. A short overview on common and well-known, established imaging techniques for biofilms such as scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), Raman microscopy (RM), and magnetic resonance imaging (MRI) paves the way to imaging biofilms at the mesoscale, which is perfectly covered by means of OCT. Principle, resolution, imaging velocity, and limitations of OCT are subsequently presented and discussed in the context of biofilm applications. Examples are provided showing the strength of this technique with respect to the visualization of the mesoscopic biofilm structure as well as the estimation of flow profiles and shear rates. Common and new structural parameters derived from OCT datasets are presented. Additionally, the review shows the importance of OCT with respect to a better description of mechanical biofilm properties. Finally, the implementation of multi-dimensional OCT datasets in biofilm modelling is shown by several examples aiming on an improved understanding of mass transfer at the bulk-biofilm interface. Biotechnol. Bioeng. 2017;114: 1386-1402. © 2017 Wiley Periodicals, Inc.

NMR investigation of water diffusion in different biofilm structures.

Mass transfer in biofilms is determined by diffusion. Different mostly invasive approaches have been used to measure diffusion coefficients in biofilms, however, data on heterogeneous biomass under realistic conditions is still missing. To non-invasively elucidate fluid-structure interactions in complex multispecies biofilms pulsed field gradient-nuclear magnetic resonance (PFG-NMR) was applied to measure the water diffusion in five different types of biomass aggregates: one type of sludge flocs, two types of biofilm, and two types of granules. Data analysis is an important issue when measuring heterogeneous systems and is shown to significantly influence the interpretation and understanding of water diffusion. With respect to numerical reproducibility and physico-chemical interpretation, different data processing methods were explored: (bi)-exponential data analysis and the Γ distribution model. Furthermore, the diffusion coefficient distribution in relation to relaxation was studied by D-T2 maps obtained by 2D inverse Laplace transform (2D ILT). The results show that the effective diffusion coefficients for all biofilm samples ranged from 0.36 to 0.96 relative to that of water. NMR diffusion was linked to biofilm structure (e.g., biomass density, organic and inorganic matter) as observed by magnetic resonance imaging and to traditional biofilm parameters: diffusion was most restricted in granules with compact structures, and fast diffusion was found in heterotrophic biofilms with fluffy structures. The effective diffusion coefficients in the biomass were found to be broadly distributed because of internal biomass heterogeneities, such as gas bubbles, precipitates, and locally changing biofilm densities. Thus, estimations based on biofilm bulk properties in multispecies systems can be overestimated and mean diffusion coefficients might not be sufficiently informative to describe mass transport in biofilms and the near bulk.

Water quality and daily temperature cycle affect biofilm formation in drip irrigation devices revealed by optical coherence tomography.

Drip irrigation is a water-saving technology. To date, little is known about how biofilm forms in drippers of irrigation systems. In this study, the internal dripper geometry was recreated in 3-D printed microfluidic devices (MFDs). To mimic the temperature conditions in (semi-) arid areas, experiments were conducted in a temperature controlled box between 20 and 50°C. MFDs were either fed with two different treated wastewater (TWW) or synthetic wastewater. Biofilm formation was monitored non-invasively and in situ by optical coherence tomography (OCT). 3-D OCT datasets reveal the major fouling position and illustrate that biofilm development was influenced by fluid dynamics. Biofilm volumetric coverage of the labyrinth up to 60% did not reduce the discharge rate, whereas a further increase to 80% reduced the discharge rate by 50%. Moreover, the biofilm formation rate was significantly inhibited in daily temperature cycle independent of the cultivation medium used.

Community structure of a sulfate-reducing consortium in lead-contaminated wastewater treatment process.

This study evaluated the capacity to remove lead by an indigenous consortium of five sulfate-reducing bacteria (SRB): Desulfobacterium autotrophicum, Desulfomicrobium salsugmis, Desulfomicrobium escambiense, Desulfovibrio vulgaris, and Desulfovibrio carbinolicus, using continuous moving bed biofilm reactor systems. Four continuous moving bed biofilm reactors (referred as R1-R4) were run in parallel for 40 days at lead loading rates of 0, 20, 30 and 40 mg l-1 day-1, respectively. The impact of lead on community structure of the SRB consortium was investigated by dsrB gene-based denaturing gradient gel electrophoresis (dsrB-based DGGE), fluorescence in situ hybridization (FISH) and chemical analysis. These results indicated that D. escambiense and D. carbinolicus were dominant in all analyzed samples and played a key role in lead removal in R2 (20 mg l-1 day-1) and R3 (30 mg l-1 day-1). However, in R4 (40 mg l-1 day-1), these two strains were barely detected by FISH and dsrB-based DGGE. As a result, SRB activity was severely affected by lead toxicity. High lead removal efficiencies of lead (99-100%) were observed in R2 and R3 throughout the operation, whereas that in R4 was significantly decreased (91%) after 40 days of operation. This data strongly implied that the investigated SRB consortium might have potential application for lead removal. Moreover, to improve the efficiency of the lead treatment process, the lead loading rates below the inhibitory level to SRB activity should be selected.

Assessing the influence of biofilm surface roughness on mass transfer by combining optical coherence tomography and two-dimensional modeling.

Imaging and modeling are two major approaches in biofilm research to understand the physical and biochemical processes involved in biofilm development. However, they are often used separately. In this study we combined these two approaches to investigate substrate mass transfer and mass flux. Cross-sectional biofilm images were acquired by means of optical coherence tomography (OCT) for biofilms grown on carriers. A 2D biofilm model was developed incorporating OCT images as well as a simplified biofilm geometry serving as structural templates. The model incorporated fluid flow, substrate transfer and biochemical conversion of substrates and simulated the hydrodynamics surrounding the biofilm structure as well as the substrate distribution. The method allowed detailed analysis of the hydrodynamics and mass transfer characteristics at the micro-scale. Biofilm activity with respect to substrate fluxes was compared among different combinations of flow, substrate availability and biomass density. The combined approach revealed that higher substrate fluxes at heterogeneous biofilm surface under two conditions: pure diffusion and when high flow velocity along the biofilms surface renders the whole liquid-biofilm interface to be highly active. In-between the two conditions the substrate fluxes across the surface of smooth biofilm geometry were higher than that of the heterogeneous biofilms.

Modelling the influence of total suspended solids on E. coli removal in river water.

Following sewer overflows, fecal indicator bacteria enter surface waters and may experience different lysis or growth processes. A 1D mathematical model was developed to predict total suspended solids (TSS) and Escherichia coli concentrations based on field measurements in a large-scale flume system simulating a combined sewer overflow. The removal mechanisms of natural inactivation, UV inactivation, and sedimentation were modelled. For the sedimentation process, one, two or three particle size classes were incorporated separately into the model. Moreover, the UV sensitivity coefficient α and natural inactivation coefficient kd were both formulated as functions of TSS concentration. It was observed that the E. coli removal was predicted more accurately by incorporating two particle size classes. However, addition of a third particle size class only improved the model slightly. When α and kd were allowed to vary with the TSS concentration, the model was able to predict E. coli fate and transport at different TSS concentrations accurately and flexibly. A sensitivity analysis revealed that the mechanisms of UV and natural inactivation were more influential at low TSS concentrations, whereas the sedimentation process became more important at elevated TSS concentrations.

Time-resolved biofilm deformation measurements using optical coherence tomography.

The interaction of shear stress with the biofilm leads to a dynamic deformation, which is related to the structural and material characteristics of biofilms. We show how optical coherence tomography can be used as an imaging technique to investigate the time-resolved deformation on the biofilm mesoscale as well as to estimate mechanical properties of the biofilm. For the first time time-resolved deformation from cross-sectional views of the inner biofilm structure could be shown. Changes in the biofilm structure and rheological properties were calculated from cross sections in real-time and time-lapsed measurements. Heterotrophic biofilms were grown in a flow cell set-up at low shear stress of τw = 0.01 Pa. By applying higher shear stress elastic and viscoelastic behavior of biofilms were quantified. Deformation led to a change in biofilm conformation and allowed to estimate rheological properties. Assuming an ideal wall shear stress calculation, the shear modulus G = 29.7 ± 1.7 Pa and the Young's modulus E = 36.0 ± 2.6 Pa were estimated.

Determining the flow regime in a biofilm carrier by means of magnetic resonance imaging.

Biofilms on cylindrical carrier material originating from a lab-scale moving bed biofilm reactor (MBBR) were investigated by means of Magnetic Resonance Imaging (MRI). The aim of this study was to determine the local flow velocities at the inner face of the biofilm carrier. To get an insight into the mass transport processes, flow velocity maps of blank and with biofilm cultivated carriers were measured. A single carrier was placed in a tube in three different orientations and exposed to flow velocities of 0.21, 0.42, and 0.64 mm/s. The interplay of the biofilm morphology and the local flow pattern was then analyzed including the effect of the orientation of the carrier in relation to the upstream flow angle. Within this study, the biofilm carrier can be understood as an interconnected system of four sections in which the incoming fluid volume will be distributed depending on the biomass occupation and morphology. In sections with high biofilm occupation, the flow resistance is increased. Depending on the orientation of the carrier in the flow field, this effect leads to flow evasion through less covered sections showing higher flow velocities and consequently the risk of biofilm detachment. However, there was no clear correlation between biofilm coverage and flow ratio.

Systematic suspect screening and identification of sulfonamide antibiotic transformation products in the aquatic environment.

The formation of environmental sulfonamide transformation products (TPs) has only been investigated for very selected compounds, and their transformation pathways are incompletely elucidated. Given their homologous series, it is likely that similar transformation reactions occur for all sulfonamides. It has recently been demonstrated for sulfamethoxazole (SMX) that its derivative TPs retain antibiotic activity. Unfortunately, TP reference standards are often not available and a number of TPs are still unknown. Therefore, in the present study, a generic scheme was developed to predict 29 potential TPs of sulfonamide antibiotics via identification of the major transformation and breakdown reactions. Mass shifts were calculated for each transformation allowing for a suspect screening using LC/QTOF. Based on the structural elucidation of the parent sulfonamide product ion spectra, five characteristic product ions could be predicted for each of 15 derivative TPs. The predictions were confirmed with reference standards of SMX TPs as well as with data of TPs of further sulfonamides reported in literature. In a final step, samples of activated sludge biotransformation experiments were screened for suspect TPs of two sulfonamides. In total, 13 ecotoxicologically relevant TPs could be tentatively identified by use of the product ion predictions and LC/QTOF.

Optimization of sulfide production by an indigenous consortium of sulfate-reducing bacteria for the treatment of lead-contaminated wastewater.

Biological treatment with sulfate-reducing bacteria (SRB) is considered to be an excellent option to remove heavy metals from wastewater. In this study, the optimization of sulfide production for an enhanced removal of lead by a consortium of SRB was carried out based on central composite design and analyzed using response surface methodology (RSM). The sulfide production process was investigated as a function of three independent variables: solution pH (6.5-8.5), lactate concentration (32-96 mM), and sulfate concentration (16-32 mM). RSM analysis showed that the optimum conditions for a high sulfide concentration (14.2 mM) occurred at a pH of 7.5 and at lactate and sulfate concentrations of 53.4 mM and 22.6 mM, respectively. The lead removal efficiency of the SRB consortium using optimum conditions was determined in four parallel anaerobic continuous moving bed biofilm reactors (V = 2 L) that were fed with synthetic wastewater containing dissolved lead at concentrations of 0, 100, 150, 200 mg L(-1) and operated with a hydraulic retention time of 5 days. 99-100 % was removed from synthetic wastewater with lead concentrations of 100 and 150 mg L(-1) during 40 days of operation. For the highest lead concentration of 200 mg L(-1), a decrease in efficiency of removal (96 %) was observed at the end of the experiment.

Influence of seasonal temperature fluctuations on two different partial nitritation-anammox reactors treating mainstream municipal wastewater.

Partial nitritation-anammox (PN-A) has gained increasing interest for municipal wastewater treatment in recent years due to its high energy-saving potential. Moving the PN-A technology from side- to mainstream exhibited a set of challenges. Conditions are quite different, with much lower ammonium concentrations and temperatures. Biomass retention becomes highly important due to the even lower growth rates. This study compared two laboratory-scale reactors, a sequencing batch reactor (SBR) and a moving bed biofilm reactor (MBBR), employing realistic seasonal temperature variations over a 1-year period. The results revealed that both systems had to face decreasing ammonium conversion rates and nitrite accumulation at temperatures lower than 12°C. The SBR did not recover from the loss in anammox activity even when the temperature increased again. The MBBR only showed a short nitrite peak and recovered its initial ammonium turnover when the temperature rose back to >15°C. The SBR had higher biomass specific rates, indicating that suspended sludge is less diffusion-limited but also more susceptible to biomass wash-out. However, the MBBR showed the more stable performance also at low temperatures and managed to recover. Ex situ batch activity tests supported reactor operation data by providing additional insight with respect to specific biomass activities.

Start-up of a full-scale deammonification SBR-treating effluent from digested sludge dewatering.

This study shows the start-up and operation of a full-scale sequencing batch reactor (SBR) with a volume of 550 m³ for deammonification of reject water from sludge dewatering over the first 650 days of operation. The SBR was operated with discontinuous aeration and achieved an optimum of around 85% of ammonium removal at a load of 0.17 kg m⁻³ d⁻¹. The application of batch tests for the activity measurement of aerobic ammonium and nitrite oxidizing bacteria and anaerobic ammonium oxidizing bacteria were proven to support the identification of setbacks in reactor operation. Furthermore, the calculation of the oxygen uptake rates from online oxygen measurements helped to explain the overall reactor performance. The aeration regime is a key parameter for stable operation of such an SBR for deammonification. At aeration/non-aeration time ranges from 6-9 min, the best results with respect to turnover rates and low nitrate production were achieved. Compared with the nitrification/denitrification SBR operated in parallel with methanol as the carbon source, a significant reduction in costs for energy and chemicals was achieved. The costs for maintenance slightly increased.

Antibacterial activity of sulfamethoxazole transformation products (TPs): general relevance for sulfonamide TPs modified at the para position.

Sulfonamide antibiotics undergo transformation in the aquatic environment through biodegradation, photolysis, or hydrolysis. In this study, the residual antibacterial activity of 11 transformation products (TPs) of sulfamethoxazole (SMX) was investigated with regard to their in vitro growth and luminescence inhibition on Vibrio fischeri (30 min and 24 h exposure). Two transformation products, 4-hydroxy-SMX and N(4)-hydroxy-acetyl-SMX, were synthesized in-house and confirmed by nuclear magnetic resonance and high-resolution mass spectrometry. Results of individual compound experiments showed that TPs modified at the para amino group still exhibit clear antibacterial effects, whereas TPs resulting from breakdown of the SMX structure lost this mechanism of action. 4-NO2- and 4-OH-SMX were found to inhibit growth to a clearly greater extent than the parent compound, SMX. In contrast, the N(4)-acetyl- and N(4)-hydroxy-acetyl-derivatives retain less than 10 and 5% of the effect of SMX on growth and luminescence inhibition, respectively. The effect of a mixture of para-modified TPs was observed to be additive. Considering the homologous series of sulfa drugs widely prescribed and their common mechanism of action, the potential environmental impact must consider the total amount of sulfonamide antibiotics and their derivative TPs, which might end up in a water body. Extrapolating the results obtained here for the para TPs of SMX to other sulfa drugs and determining the persistence and occurrence of these compounds in the aquatic environment is required for improved risk assessment.