Exploring the inhibition boundaries of mixed cultures of purple phototrophic bacteria for wastewater treatment in anaerobic conditions.

The development of novel wastewater platforms should include the analysis of the most critical functional factors including the effects of toxic or inhibitory substances. Due to the novelty of purple phototrophic bacteria (PPB)-based wastewater treatment systems, this analysis has not been done yet in mixed cultures. In this work, various relevant chemical compounds, including aromatic (phenol, 2,4,6-trichlorophenol or 246TCP, 4-nitrophenol or 4CP, sulfathiazole) and aliphatic organics (methanol, trichlorethylene or TCE, oleic acid, ethanol, propionic acid), inorganic salts (ammonium, ClO3-, Na+), and metals (Fe3+, Fe2+, Cu2+, Zn2+, Ni2+, Al3+), as well as pH, are analyzed for their effect on mixed PPB cultures in anaerobic photoheterotrophic conditions using acetate as the model organic substrate. The most toxic substances detected were 246TCP, 4NP, Cu2+, Fe2+ and Ni2+, (Ki for activity: 23 ± 2, 97 ± 12, 3.1 ± 0.4, 13 ± 3, 13 ± 1 mg/L, and Ki (or toxicity threshold) for growth: 17 ± 2, (119), 3.5 ± 0.4, (4.8), (22.9) mg/L, respectively). Some substances inhibited the activity more than the growth (sulfathiazole, Ni2+ and Fe3+), or the growth more than the activity (TCE, 4NP and Fe2+). In addition, some organic substrates, such as phenol, ethanol and propionate, specifically inhibited the acetate uptake, being noncompetitive in the case of phenol and ethanol, and most likely competitive in the case of propionate. These findings are relevant for the wastewater treatment and resource recovery applications of the PPB technology, as well as for the upgrading of current models (Photo-Anaerobic Model). In addition, the data will open possibilities to promote the production of specific compounds (as PHA or single-cell proteins) by selectively inhibiting some parts of the PPB metabolism.

Assessing the effects of intra-granule precipitation in a full-scale industrial anaerobic digester.

In this paper, a multi-scale model is used to assess the multiple mineral precipitation potential in a full-scale anaerobic granular sludge system. Reactor behaviour is analysed under different operational conditions (addition/no addition of reject water from dewatering of lime-stabilized biomass) and periods of time (short/long term). Model predictions suggest that a higher contribution of reject water promotes the risk of intra-granule CaCO3 formation as a result of the increased quantity of calcium arriving with that stream combined with strong pH gradients within the biofilm. The distribution of these precipitates depends on: (i) reactor height; and (ii) granule size. The study also exposes the potential undesirable effects of the long-term addition of reject water (a decrease in energy recovery of 20% over a 100-day period), caused by loss in biomass activity (due to microbial displacement), and the reduced buffer capacity. This demonstrates how both short-term and long-term operational conditions may affect the formation of precipitates within anaerobic granules, and how it may influence methane production and consequently energy recovery.

Metabolic modelling of mixed culture anaerobic microbial processes.

Mixed culture anaerobic processes are important to environmental systems, including the global carbon cycle, and industrial and environmental biotechnology. Mixed culture metabolic modelling (MM) is an essential tool to analyse these systems. MM predicts microbial function based on knowledge or assumption of cellular metabolism. It may be developed based on observations at the process level - biochemical process modelling (BPM) or fundamental knowledge of the cell being modelled - cellular level modelling (CLM). There is a substantial gap between these two fields, with BPM not considering genetic constraints, particularly where this may be important to interspecies interactions (e.g. amino acid transfer), and CLM commonly not considering mass transfer principles, such as advection/diffusion/migration. No unified approach is useful for all applications, but there is an increasing need to consider genetic information and constraints in developing BPM, and translate BPM principles (including mass-transfer and inorganic chemistry) for application to CLM.

Evaluation of anaerobic digestion post-treatment options using an integrated model-based approach.

The objective of this paper is to present the main results of an engineering-research project dealing with model-based evaluation of waste streams treatment from a biotech company. This has been extensively done in domestic treatment systems, but is equally important, and with different challenges in industrial wastewater treatment. A new set of biological (activated sludge, anaerobic digestion), physicochemical (aqueous phase, precipitation, mass transfer) process models and model interfaces are required to describe removal of organics in an upflow anaerobic sludge blanket (UASB) reactor plus either traditional nitrification/denitrification (A1) or partial nitritation (PN)/anammox (ANX) (A2) processes. Model-based analysis shows that option A1 requires a decrease in digestion energy recovery (Erecovery) in order to have enough organic substrate for subsequent post NO3 reduction treatment (95 kWh.kg N-1). In contrast, A2 in an aerobic granular sludge reactor allows for higher UASB conversion since N removal is carried out autotrophically. The study also reveals that the addition of an aerated pre-treatment unit prior to the PN/ANX (A2) reactor promotes COD and H2S oxidation, CO2 and CH4 stripping, a pH increase (up to 8.5) and a reduction of the risk of intra-granular precipitation as well as sulfide inhibition. Simulations indicate clear differences regarding the microbial distribution/abundance within the biofilm in A2 when comparing the two operational modes. Final results show the effects of different loading and operational conditions; dissolved oxygen (DO), Total Suspended Solids (TSSop), energy recovery (Erecovery); on the overall process performance; N removal, aeration energy (Eaeration), net energy production (Erecovery); using response surfaces, highlighting the need of integrated approaches to avoid sub-optimal outcomes. The study shows the benefits of virtual plant simulation and demonstrates the potential of model-based evaluation when process engineers in industry have to decide between competing options.

Plant-wide model-based analysis of iron dosage strategies for chemical phosphorus removal in wastewater treatment systems.

Stringent phosphorus discharge standards (i.e. 0.15-0.3 g P.m-3) in the Baltic area will compel wastewater treatment practice to augment enhanced biological phosphorus removal (EBPR) with chemical precipitation using metal salts. This study examines control of iron chemical dosing for phosphorus removal under dynamic loading conditions to optimize operational aspects of a membrane biological reactor (MBR) pilot plant. An upgraded version of the Benchmark Simulation Model No. 2 (BSM2) with an improved physico-chemical framework (PCF) is used to develop a plant-wide model for the pilot plant. The PCF consists of an equilibrium approach describing ion speciation and pairing, kinetic minerals precipitation (such as hydrous ferric oxides (HFO) and FePO4) as well as adsorption and co-precipitation. Model performance is assessed against data sets from the pilot plant, evaluating the capability to describe water and sludge lines across the treatment process under steady-state operation. Simulated phosphorus differed as little as 5-10% (relative) from measured phosphorus, indicating that the model was representative of reality. The study also shows that environmental factors such as pH, as well operating conditions such as Fe/P molar ratios (1, 1.5 and 2), influence the concentration of dissolved phosphate in the effluent. The time constant of simultaneous precipitation in the calibrated model, due to a step change decrease/increase in FeSO4 dosage, was found to be roughly 5 days, indicating a slow dynamic response due to a multi-step process involving dissolution, oxidation, precipitation, aging, adsorption and co-precipitation. The persistence effect of accumulated iron-precipitates (HFO particulates) in the activated sludge seemed important for phosphorus removal, and therefore solids retention time plays a crucial role according to the model. The aerobic tank was deemed to be the most suitable dosing location for FeSO4 addition, due to high dissolved oxygen levels and good mixing conditions. Finally, dynamic model-based analyses show the benefits of using automatic control when dosing chemicals.

Humic acid inhibition of hydrolysis and methanogenesis with different anaerobic inocula.

There is increasing evidence that humic acid (HA) is hampering the performance of anaerobic digesters treating animal manures and thermally-hydrolysed waste activated sludge. In the present study, HA inhibition and inhibition resilience was examined for hydrolysis (carbohydrate and protein) and acetotrophic methanogenesis with four distinct full-scale anaerobic inocula. The aim was to further understand HA inhibition and to explore potential relationships between microbial factors and inhibition resilience. For two of the four tested inocula, cellulose degradation showed a start-up delay that lengthened as HA concentration increased from 0 to 2 g L-1. This inhibition was reversible because, after the initial delay, subsequent hydrolysis rates and methane yields were not significantly influenced by HA concentration. Cellulose hydrolysis results at HA concentrations below 2 g L-1 support a threshold inhibition mechanism, i.e. HA complexes with hydrolytic enzymes preventing them from binding with cellulose, but once all the HA had been complexed, enzymes subsequently released are free to bind with cellulose. Inocula with higher cellulose hydrolytic activity were less affected by HA inhibition, suggesting a potential link between HA inhibition resilience and microbial activity. However, above 5 gHA L-1, cellulose hydrolysis rates decreased with increasing HA concentration; indicating that the mechanisms of inhibition may change depending on some threshold HA concentration. Protein hydrolysis and acetotrophic methanogenesis were less susceptible to HA inhibition than cellulose hydrolysis, since signs of inhibition were only observed above 5 gHA L-1. Acetotrophic methanogenesis was partially inhibited at 10 gHA L-1 and completely inhibited at 20 gHA L-1. These results further support that HA inhibition is selective towards particular enzymes.

Sorbents can tailor nitrogen release from organic wastes to match the uptake capacity of crops.

Delivering nutrients from mineral or organic fertilizers out of synchrony with crop uptake causes inefficiencies and pollution. We explore methodologies for evaluating sorbents as additives to organic agricultural wastes to retain nitrogen in an exchangeable form and deliver at rates that approximate the uptake capacity of roots. Focussing on ammonium (NH4+) as the main inorganic nitrogen form in the studied wastes (sugarcane mill mud, poultry litter), we tested geo-sorbents and biochar for their ability to retain NH4+. Sorption capacity was ranked palagonite < bentonite, biochar, vermiculite < chabazite, clinoptilolite (5.7 to 24.3 mg NH4+ g-1 sorbent). Sorbent-waste formulations were analysed for sorption capacity, leaching and fluxes of NH4+. Ammonium-sorption capacity broadly translated to sorbent-waste formulations with clinoptilolite conferring the strongest NH4+ attenuation (80%), and palagonite the lowest (7%). A 1:1 ratio of sorbent:waste achieved stronger sorption than a 0.5:1 ratio, and similar sorption as a 1:1.5 ratio. In line with these results, clinoptilolite-amended wastes had the lowest in situ NH4+ fluxes, which exceeded the NH4+ uptake capacity (Imax) of sugarcane and sorghum roots 9 to 84-fold, respectively. Less efficient sorbent-waste formulations and un-amended wastes exceeded Imax of crop roots up to 274-fold. Roots preferentially colonized stronger sorbent-waste formulations and avoided weaker ones, suggesting that lower NH4+ fluxes generate a more favourable growth environment. This study contributes methodologies to identify suitable sorbents to formulate organic wastes as next-generation fertilizers with view of a crop's nutrient physiology. Efficient re-purposing of wastes can improve nutrient use efficiency in agriculture and support the circular nutrient economy.

Characterising and modelling free ammonia and ammonium inhibition in anaerobic systems.

Inhibition by ammoniacal nitrogen, consisting of free ammonia (NH3) and ammonium ion (NH4+), has been widely investigated for anaerobic digestion. However, despite the large amount of research on the subject, ammoniacal nitrogen inhibition still threatens many anaerobic digesters. This paper presents (i) a method to reliably characterise ammoniacal nitrogen inhibition and (ii) a robust inhibition modelling approach. Results showed that NH3 and NH4+ inhibition need to be jointly determined, which can only be done by performing inhibition tests at various total ammoniacal nitrogen (TAN) concentrations and pH values. These test conditions were reliably achieved using the salts NH4HCO3 and NH4Cl without pH adjustment, rather than by using NH4Cl with pH adjustment. The use of only salts showed a lower pH change during the inhibition test (∼1.5 days), thereby decreasing the uncertainty in TAN speciation and strengthening the test and model outputs. A threshold inhibition function satisfactorily described (R2 > 0.99) the joint inhibition of NH3 and NH4+ on three distinct inocula, and provided a better description of the inhibition testing results than a non-competitive inhibition function (R2 ∼0.70). The key advantage of the proposed threshold inhibition function is its capacity to identify the inhibition lower limit (concentration where inhibition starts; KImin) and upper limit (concentration where inhibition is complete; KImax). The threshold inhibition function also identifies the 50% inhibition concentration (KI50) at the midpoint of KImin and KImax. Finally, experimental and model results show that at pH 7.3-7.7 and TAN concentrations above 2000 mgN·L-1, both NH3 and NH4+ contribute significantly to overall inhibition.

Relationship between microbial community, operational factors and ammonia inhibition resilience in anaerobic digesters at low and moderate ammonia background concentrations.

The relationship between anaerobic digestion operational conditions and (i) microbial community, (ii) acetoclastic methanogenic activity and (iii) free ammonia (NH3) inhibition resilience was investigated. Thirteen inocula were obtained from full and pilot scale digesters fed with different substrates, digester configurations, operating temperatures and NH3 concentrations (0.1-241 mgN·L-1). Substrate type and temperature were the primary factors influencing microbial community composition. Methanogenic activity ranged from 0.04 to 0.14 gCOD-CH4·g-1VS·day-1, and was significantly correlated with archaeal relative abundance and archaeal community PC2. The variability of NH3 resilience among inocula was moderate, with inhibition threshold values (KI50) ranging between 32 and 175 mgNH3-N·L-1. No microbial or operational factors correlated with NH3 resilience. However, the slopes of inhibition threshold curves were influenced by some environmental factors, namely substrate type, digester temperature and NH3 concentration. Overall, these results indicate that low and moderate background NH3 concentrations is not a key determinant of microbial community nor NH3 resilience.

Modelling an industrial anaerobic granular reactor using a multi-scale approach.

The objective of this paper is to show the results of an industrial project dealing with modelling of anaerobic digesters. A multi-scale mathematical approach is developed to describe reactor hydrodynamics, granule growth/distribution and microbial competition/inhibition for substrate/space within the biofilm. The main biochemical and physico-chemical processes in the model are based on the Anaerobic Digestion Model No 1 (ADM1) extended with the fate of phosphorus (P), sulfur (S) and ethanol (Et-OH). Wastewater dynamic conditions are reproduced and data frequency increased using the Benchmark Simulation Model No 2 (BSM2) influent generator. All models are tested using two plant data sets corresponding to different operational periods (#D1, #D2). Simulation results reveal that the proposed approach can satisfactorily describe the transformation of organics, nutrients and minerals, the production of methane, carbon dioxide and sulfide and the potential formation of precipitates within the bulk (average deviation between computer simulations and measurements for both #D1, #D2 is around 10%). Model predictions suggest a stratified structure within the granule which is the result of: 1) applied loading rates, 2) mass transfer limitations and 3) specific (bacterial) affinity for substrate. Hence, inerts (XI) and methanogens (Xac) are situated in the inner zone, and this fraction lowers as the radius increases favouring the presence of acidogens (Xsu,Xaa, Xfa) and acetogens (Xc4,Xpro). Additional simulations show the effects on the overall process performance when operational (pH) and loading (S:COD) conditions are modified. Lastly, the effect of intra-granular precipitation on the overall organic/inorganic distribution is assessed at: 1) different times; and, 2) reactor heights. Finally, the possibilities and opportunities offered by the proposed approach for conducting engineering optimization projects are discussed.

A mechanistic model for anaerobic phototrophs in domestic wastewater applications: Photo-anaerobic model (PAnM).

Purple phototrophic bacteria (PPB) have been recently proposed as a key potential mechanism for accumulative biotechnologies for wastewater treatment with total nutrient recovery, low greenhouse gas emissions, and a neutral to positive energy balance. Purple phototrophic bacteria have a complex metabolism which can be regulated for process control and optimization. Since microbial processes governing PPB metabolism differ from traditional processes used for wastewater treatment (e.g., aerobic and anaerobic functional groups in ASM and ADM1), a model basis has to be developed to be used as a framework for further detailed modelling under specific situations. This work presents a mixed population phototrophic model for domestic wastewater treatment in anaerobic conditions. The model includes photoheterotrophy, which is divided into acetate consumption and other organics consumption, chemoheterotrophy (including simplified fermentation and anaerobic oxidation) and photoautotrophy (using hydrogen as an electron donor), as microbial processes, as well as hydrolysis and biomass decay as biochemical processes, and is single-biomass based. The main processes have been evaluated through targeted batch experiments, and the key kinetic and stoichiometric parameters have been determined. The process was assessed by analyzing a continuous reactor simulation scenario within a long-term wastewater treatment system in a photo-anaerobic membrane bioreactor.

Indigenous microbial capability in solid manure residues to start-up solid-phase anaerobic digesters.

Batch solid-phase anaerobic digestion is a technology for sustainable on-farm treatment of solid residues, but is an emerging technology that is yet to be optimised with respect to start-up and inoculation. In the present study, spent bedding from two piggeries (site A and B) were batch digested at total solids (TS) concentration of 5, 10 and 20% at mesophilic (37°C) and thermophilic (55°C) temperatures, without adding an external inoculum. The results showed that the indigenous microbial community present in spent bedding was able to recover the full methane potential of the bedding (140±5 and 227±6L CH4 kgVSfed-1 for site A and B, respectively), but longer treatment times were required than for digestion with an added external inoculum. Nonetheless, at high solid loadings (i.e. TS level>10%), the digestion performance was affected by chemical inhibition due to ammonia and/or humic acid. Thermophilic temperatures did not influence digestion performance but did increase start-up failure risk. Further, inoculation of residues from the batch digestion to subsequent batch enhanced start-up and achieved full methane potential recovery of the bedding. Inoculation with liquid residue (leachate) was preferred over a solid residue, to preserve treatment capacity for fresh substrate. Overall, the study highlighted that indigenous microbial community in the solid manure residue was capable of recovering full methane potential and that solid-phase digestion was ultimately limited by chemical inhibition rather than lack of suitable microbial community.

Plant-wide modelling of phosphorus transformations in wastewater treatment systems: Impacts of control and operational strategies.

The objective of this paper is to report the effects that control/operational strategies may have on plant-wide phosphorus (P) transformations in wastewater treatment plants (WWTP). The development of a new set of biological (activated sludge, anaerobic digestion), physico-chemical (aqueous phase, precipitation, mass transfer) process models and model interfaces (between water and sludge line) were required to describe the required tri-phasic (gas, liquid, solid) compound transformations and the close interlinks between the P and the sulfur (S) and iron (Fe) cycles. A modified version of the Benchmark Simulation Model No. 2 (BSM2) (open loop) is used as test platform upon which three different operational alternatives (A1, A2, A3) are evaluated. Rigorous sensor and actuator models are also included in order to reproduce realistic control actions. Model-based analysis shows that the combination of an ammonium ( [Formula: see text] ) and total suspended solids (XTSS) control strategy (A1) better adapts the system to influent dynamics, improves phosphate [Formula: see text] accumulation by phosphorus accumulating organisms (XPAO) (41%), increases nitrification/denitrification efficiency (18%) and reduces aeration energy (Eaeration) (21%). The addition of iron ( [Formula: see text] ) for chemical P removal (A2) promotes the formation of ferric oxides (XHFO-H, XHFO-L), phosphate adsorption (XHFO-H,P, XHFO-L,P), co-precipitation (XHFO-H,P,old, XHFO-L,P,old) and consequently reduces the P levels in the effluent (from 2.8 to 0.9 g P.m-3). This also has an impact on the sludge line, with hydrogen sulfide production ( [Formula: see text] ) reduced (36%) due to iron sulfide (XFeS) precipitation. As a consequence, there is also a slightly higher energy production (Eproduction) from biogas. Lastly, the inclusion of a stripping and crystallization unit (A3) for P recovery reduces the quantity of P in the anaerobic digester supernatant returning to the water line and allows potential struvite ( [Formula: see text] ) recovery ranging from 69 to 227 kg.day-1 depending on: (1) airflow (Qstripping); and, (2) magnesium ( [Formula: see text] ) addition. All the proposed alternatives are evaluated from an environmental and economical point of view using appropriate performance indices. Finally, some deficiencies and opportunities of the proposed approach when performing (plant-wide) wastewater treatment modelling/engineering projects are discussed.

CFD for wastewater treatment: an overview.

Computational fluid dynamics (CFD) is a rapidly emerging field in wastewater treatment (WWT), with application to almost all unit processes. This paper provides an overview of CFD applied to a wide range of unit processes in water and WWT from hydraulic elements like flow splitting to physical, chemical and biological processes like suspended growth nutrient removal and anaerobic digestion. The paper's focus is on articulating the state of practice and research and development needs. The level of CFD's capability varies between different process units, with a high frequency of application in the areas of final sedimentation, activated sludge basin modelling and disinfection, and greater needs in primary sedimentation and anaerobic digestion. While approaches are comprehensive, generally capable of incorporating non-Newtonian fluids, multiphase systems and biokinetics, they are not broad, and further work should be done to address the diversity of process designs. Many units have not been addressed to date. Further needs are identified throughout, but common requirements include improved particle aggregation and breakup (flocculation), and improved coupling of biology and hydraulics.

Pilot-scale testing of a leachbed for anaerobic digestion of livestock residues on-farm.

A leachbed is a relatively simple anaerobic digester suitable for high-solids residues and on-farm applications. However, performance characteristics and optimal configuration of leachbeds are not well-understood. In this study, two 200 L pilot-scale leachbeds fed with spent straw bedding from pigs/swine (methane potential, B0 = 195-218 L CH4 kg(-1) VS fed) were used to assess the effects of leachate recirculation mode (trickling vs. flood-and-drain) on the digestion performance. Results showed comparable substrate solubilisation extents (30-45% of total chemical oxygen demand fed) and methane conversion (50% of the B0) for the trickling and flood-and-drain modes, indicating that digestion performance was insensitive to the mode of leachate flow. However, the flood-and-drain leachbed mobilised more particulates into the leachate than the trickling leachbed, an undesirable outcome, because these particulates were mostly non-biodegradable. Inoculation with solid residues from a previous leachbed (inoculum-to-substrate ratio of 0.22 on a VS basis) hastened the leachbed start-up, but methane recovery remained at 50% of the B0 regardless of the leachate recirculation mode. Post-digestion testing indicated that the leachbeds may have been limited by microbial activity/inhibition. The high residual methane potential of leachate from the trickling (residual Bo = 732 ± 7 L CH4 kg(-1) VS fed) and flood-and-drain leachbeds (582 ± 8 L CH4 kg(-1) VS fed) indicated an opportunity for further processing of leachate via a separate methanogenic step. Overall, a trickling leachbed appeared to be more favourable than the flood-and-drain leachbed for treating spent bedding at farm-scale due to easier operation.

Recovery of energy and nutrient resources from cattle paunch waste using temperature phased anaerobic digestion.

Cattle paunch is comprised of partially digested cattle feed, containing mainly grass and grain and is a major waste produced at cattle slaughterhouses contributing 20-30% of organic matter and 40-50% of P waste produced on-site. In this work, Temperature Phased Anaerobic Digestion (TPAD) and struvite crystallization processes were developed at pilot-scale to recover methane energy and nutrients from paunch solid waste. The TPAD plant achieved a maximum sustainable organic loading rate of 1-1.5kgCODm(-3)day(-1) using a feed solids concentration of approximately 3%; this loading rate was limited by plant engineering and not the biology of the process. Organic solids destruction (60%) and methane production (230LCH4kg(-1) VSfed) achieved in the plant were similar to levels predicted from laboratory biochemical methane potential (BMP) testing. Model based analysis identified no significant difference in batch laboratory parameters vs pilot-scale continuous parameters, and no change in speed or extent of degradation. However the TPAD process did result in a degree of process intensification with a high level of solids destruction at an average treatment time of 21days. Results from the pilot plant show that an integrated process enabled resource recovery at 7.8GJ/dry tonne paunch, 1.8kgP/dry tonne paunch and 1.0kgN/dry tonne paunch.

Development and validation of a rapid test for anaerobic inhibition and toxicity.

Despite the importance of quantifying inhibitory capacity of compounds in anaerobic digestion, there is currently no well-defined method to assess it. Experimental methods in literature are frequently time-consuming and resource intensive. As a result, detailed inhibition testing rarely forms part of anaerobic digestion studies, despite the importance and utility of this information. This study develops and validates a simple and rapid inhibition test protocol, based on relative inhibition of acetoclastic methanogens. The inhibition potential of a compound is determined from the reduction in specific methanogenic activity as inhibitor concentration is increased. The method was successfully performed on two inoculums from different source environments and with both biostatic and biocidal inhibitors. Optimisation work indicated that: (i) sodium acetate is a preferred carbon source compared to acetic acid; (ii) an inoculum to acetate ratio of 5 g VS g(-1) acetate is preferred, and (iii) that the inoculum concentration should be normalised to 10 g L(-1) VS to reduce mass transfer problems and promote consistency. A key advantage over existing methods is that the sampling strategy has been optimised to three events over 1.5 days while effectively controlling the relative analytical error.

Mechanical and cell-to-cell adhesive properties of aggregated Methanosarcina.

The mechanical and adhesive properties as well as the turgor pressure of microbes play an important role in cell growth and aggregation. By applying AFM together with finite element modelling, one can determine the cell wall structural homogeneity, mechanical and cell-to-cell adhesive properties for aggregated Methanosarcina barkeri cells. This also allows a novel approach to determine in-aggregate turgor pressure determination. Analyzing the AFM force-indentation response of the aggregates under loads less than 10 nN, our study reveals structural inhomogeneity of the polymeric part of the cell wall material and suggests that the cell wall consists of two layers of methanochondroitin (external: with a thickness of 3 ± 1 nm and internal: with a thickness of 169 ± 30 nm). On average, the hyperelastic finite element model showed that the internal layer is more rigid (µ = 14 ± 4 MPa) than the external layer (µ = 2.8 ± 0.9 MPa). To determine the turgor pressure and adhesiveness of the cells, a specific mode of indentation (under a load of 45 nN), aimed towards the centre of the individual aggregate, was performed. By modelling the AFM induced decohesion of the aggregate, the turgor pressure and the cell-to-cell adhesive interface properties could be determined. On average, the turgor pressure is estimated to be 59 ± 22 kPa, the interface strength is 78 ± 12 kPa and the polymer network extensibility is 2.8 ± 0.9 nm. We predict that internal cell wall comprised highly compressed methanochondroitin chains and we are able to identify a conceptual model for stress dependent inner cell wall growth.

Platforms for energy and nutrient recovery from domestic wastewater: A review.

Alternative domestic wastewater treatment processes that recover energy and nutrients while achieving acceptable nutrient limits (<5mgNL(-1)) are a key challenge. Major drivers are value and availability of phosphorous, nitrogen, and potassium, and increasing energy costs. The two major platforms that can achieve this are (a) low energy mainline (LEM), with low strength anaerobic treatment, followed by mainline anaerobic nitrogen removal and chemical or adsorptive phosphorous removal and (b) partition-release-recover (PRR), in which carbon and nutrients are partitioned to solids through either heterotrophic or phototrophic microbes, followed by anaerobic digestion of these solids and recovery from the digestate. This paper reviews practical application of these processes, with a focus on energy costs. Compared to conventional processes which require 0.5kWhkL(-1) electricity (500mgCODL(-1) influent concentration), PRR requires only 0.05kWhkL(-1) electricity. LEM offers the possibility to recover 0.1kWhkL(-1) as electricity with net energy generation above 400mgCODL(-1)influent, while PRR becomes energy generating at >650mgCODL(-1). PRR offers the possibility for recovery of nitrogen and other nutrients (including potassium) through assimilative recovery. However, the energetic overhead of this is substantial, requiring 5kWhkgN(-1) as electricity, which compares to ammonia fixation costs. The lower energy costs, and near to market status of LEM treatment make it likely as a recovery platform in the shorter term, while ability to recover other elements such as nitrogen and potassium, as well as enhance favourability on concentrated wastewaters may enhance the desirability of partitioning in the longer term.

Anaerobic codigestion of sewage sludge and glycerol, focusing on process kinetics, microbial dynamics and sludge dewaterability.

Anaerobic codigestion (AcoD) is a proven option to significantly boost biogas production while utilizing existing digesters and infrastructure. The aim of the present research was to conduct an exhaustive study regarding anaerobic codigestion of mixed sewage sludge and crude glycerol considering impacts on organic load, hydraulic load, process performance and microbial community. The methane potential of crude glycerol varied from 370 mL CH4·g(-1) VS to 483 mL CH4·g(-1) VS for different samples tested. The half maximal inhibitory concentration of crude glycerol was 1.01 g VS L(-1), and the primary mechanism of inhibition was through overload from rapid fermentation rather than the presence of toxic compounds in the crude glycerol. In continuous operation over 200 days, feeding glycerol at up to 2% v/v, increased organic load by up to 70% and resulted in a 50% increase in methane production. Glycerol dosing resulted in no change in apparent dewaterability, with both codigestion and control reactors returning values of 22%-24%. Members of the phylum Thermotogae emerged as a niche population during AcoD of sewage sludge and glycerol; however there was no gross change in microbial community structure and only minimal changes in diversity. AcoD did not result in synergisms between sewage sludge and crude glycerol. Actually, at dose rate up to 2% v/v glycerol dosing is still an effective strategy to increase the organic loading rate of continuous anaerobic digesters with minimal impact of the hydraulic retention time. Nonetheless, the dose rate must be managed to: (i) prevent process inhibition and (ii) ensure sufficient degradation time to produce a stable biosolids product.