A novel microbial community restructuring strategy for enhanced hydrogen production using multiple pretreatments and CSTR operation.

To achieve rapid enrichment of the targeted hydrogen-producing bacterial population and reconstruction of the microbial community in the biological hydrogen-producing reactor, the activated sludge underwent multiple pretreatments using micro-aeration, alkaline treatment, and heat treatment. The activated sludge obtained from the multiple pretreatments was inoculated into the continuous stirred tank reactor (CSTR) for continuous operations. The community structure alteration and hydrogen-producing capability of the activated sludge were analyzed throughout the operation of the reactor. We found that the primary phyla in the activated sludge population shifted to Proteobacteria, Firmicutes, and Bacteroidetes, which collectively accounted for 96.69% after undergoing several pretreatments. This suggests that the multiple pretreatments facilitated in achieving the selective enrichment of the fermentation hydrogen-producing microorganisms in the activated sludge. The CSTR start-up and continuous operation of the biological hydrogen production reactor resulted in the reactor entering a highly efficient hydrogen production stage at influent COD concentrations of 4000 mg/L and 5000 mg/L, with the highest hydrogen production rate reaching 8.19 L/d and 9.33 L/d, respectively. The main genus present during the efficient hydrogen production stage in the reactor was Ethanoligenens, accounting for up to 33% of the total population. Ethanoligenens exhibited autoaggregation capabilities and a superior capacity for hydrogen production, leading to its prevalence in the reactor and contribution to efficient hydrogen production. During high-efficiency hydrogen production, flora associated with hydrogen production exhibited up to 46.95% total relative abundance. In addition, redundancy analysis (RDA) indicated that effluent pH and COD influenced the distribution of the primary hydrogen-producing bacteria, including Ethanoligenens, Raoultella, and Pectinatus, as well as other low abundant hydrogen-producing bacteria in the activated sludge. The data indicates that the multiple pretreatments and reactor's operation has successfully enriched the hydrogen-producing genera and changed the community structure of microbial hydrogen production.

Use of waste marble powder for the synthesis of novel calcium-rich biochar: Characterization and application for phosphorus recovery in continuous stirring tank reactors.

This study investigates-for the first time-the synthesis of a novel Ca-rich biochar (N-Ca-B) and its potential use for phosphorus (P) recovery from both synthetic solutions (SS) and treated urban wastewater (TUW) in a continuous stirring tank reactor (CSTR) mode. The novel biochar was synthesized by pyrolysis at 900 °C of a mixture composed of three different materials: animal biomass (poultry manure; PM), lignocellulosic waste (date palm fronds; DPFs), and abundant mineral waste (waste marble powder; WMP). Characterization of N-Ca-B showed that it has good textural properties: well-developed porosity, and high specific surface area. Furthermore, high calcium hydroxide (Ca(OH)2) and calcium oxides (CaO) nanoparticle loads were observed on the biochar surface. The dynamic CSTR assays indicated that the P recovery efficiency mainly depended on the biochar mass, P influent concentration, and, especially, the Ca content of the feeding solution. Owing to its richness in Ca cations, TUW exhibited the highest adsorbed P amount (109.2 mg g-1), i.e., about 14% larger than the SS. P recovery occurs through precipitation as hydroxyapatite, surface complexation, and electrostatic interactions with positively charged biochar particles. In real-world scenarios, CSTR systems can be applied as a tertiary treatment step in existing wastewater treatment plants (WWTPs). Decanted P-loaded biochar can be used in agriculture as a slow-release fertilizer instead of commercial products.

Phenanthrene removal from a spent sediment washing solution in a continuous-flow stirred-tank reactor.

The issue of polycyclic aromatic hydrocarbons (PAHs) is widespread in marine sediments involving ecological systems and human health. Sediment washing (SW) has proven to be the most effective remediation approach for sediments polluted by PAHs, such as phenanthrene (PHE). However, SW still raises waste handling concerns due to a considerable amount of effluents generated downstream. In this context, the biological treatment of a PHE- and ethanol-containing spent SW solution can represent a highly efficient and environmentally-friendly strategy, but its knowledge is still scarce in scientific literature and no studies have so far been conducted in continuous mode. Therefore, a synthetic PHE-polluted SW solution was biologically treated in a 1 L aerated continuous-flow stirred-tank reactor for 129 days by evaluating the effect of different pH values, aeration flowrates and hydraulic retention times as operating parameters over five successive phases. A PHE removal efficiency of up to 75-94% was achieved by an acclimated PHE-degrading consortium mainly composed of Proteobacteria, Bacteroidota and Firmicutes phyla through biodegradation following the adsorption mechanism. PHE biodegradation, mainly occurring via the benzoate route due to the presence of PAH-related-degrading functional genes and a phthalate accumulation up to 46 mg/L, was also accompanied by a reduction of dissolved organic carbon and ammonia nitrogen above 99% in the treated SW solution.

Simple Rate Expression for Catalyzed Ammonia Decomposition for Fuel Cells.

This paper examines NH3 decomposition rates based on a literature-proven six-step elementary catalytic (Ni-BaZrO3) mechanism valid for 1 × 105 Pa pressure in a 650-950 K range. The rates are generated using a hypothetical continuous stirred tank catalytic reactor model running the literature mechanism. Excellent correlations are then obtained by fitting these rates to a simple overall kinetic expression based on an assumed slow step, with the remaining steps in fast pseudo-equilibria. The robust overall simple rate expression is then successfully demonstrated in various packed bed reactor applications. This expression facilitates engineering calculations without the need for a complex, detailed mechanism solver package. The methodology used in this work is independent of the choice of catalyst. It relies on the availability of a previously published and validated elementary reaction mechanism.

Analytical solution for a hybrid Logistic-Monod cell growth model in batch and continuous stirred tank reactor culture.

Monod and Logistic growth models have been widely used as basic equations to describe cell growth in bioprocess engineering. In the case of the Monod equation, the specific growth rate is governed by a limiting nutrient, with the mathematical form similar to the Michaelis-Menten equation. In the case of the Logistic equation, the specific growth rate is determined by the carrying capacity of the system, which could be growth-inhibiting factors (i.e., toxic chemical accumulation) other than the nutrient level. Both equations have been found valuable to guide us build unstructured kinetic models to analyze the fermentation process and understand cell physiology. In this work, we present a hybrid Logistic-Monod growth model, which accounts for multiple growth-dependent factors including both the limiting nutrient and the carrying capacity of the system. Coupled with substrate consumption and yield coefficient, we present the analytical solutions for this hybrid Logistic-Monod model in both batch and continuous stirred tank reactor (CSTR) culture. Under high biomass yield (Yx/s ) conditions, the analytical solution for this hybrid model is approaching to the Logistic equation; under low biomass yield condition, the analytical solution for this hybrid model converges to the Monod equation. This hybrid Logistic-Monod equation represents the cell growth transition from substrate-limiting condition to growth-inhibiting condition, which could be adopted to accurately describe the multi-phases of cell growth and may facilitate kinetic model construction, bioprocess optimization, and scale-up in industrial biotechnology.

Novel, linked bioreactor system for continuous production of biologics.

A novel, alternative intensified cell culture process comprised of a linked bioreactor system is presented. An N-1 perfusion bioreactor maintained cells in a highly proliferative state and provided a continuous inoculum source to a second bioreactor operating as a continuous-flow stirred-tank reactor (CSTR). An initial study evaluated multiple system steady-states by varying N-1 steady-state viable cell densities, N-1 to CSTR working volume ratios, and CSTR dilution rates. After identifying near optimum system steady-state parameters yielding a relatively high volumetric productivity while efficiently consuming media, a subsequent lab-scale experiment demonstrated the startup and long-term operation of the envisioned manufacturing process for 83 days. Additionally, to compensate for the cell-specific productivity loss over time due to cell line instability, the N-1 culture was also replaced with younger generation cells, without disturbing the steady-state of the system. Using the model cell line, the system demonstrated a two-fold volumetric productivity increase over the commercial-ready, optimized fed-batch process.

Metagenomic analysis and characterization of acidogenic microbiome and effect of pH on organic acid production.

Organic acid production including lactate and acetate is an economically attractive technology that has gained momentum worldwide over the past years. These series of action need to be performed by an esoteric and complex microbial community, in which different members have distinct roles in the establishment of a collective organization. In this study, we analyzed the bioma from bioreactors with various pH conditions of 4.0, 5.0 and 6.0 (R1, R2 and R3), respectively, involved in acidogenic digestion for stable production of various organic acids by means of high-throughput Illumina sequencing, disclosing thousands of genes and extracting more than 53 microbial genomes. At pH 5.0, the hydrolysis reaction was enhanced and thus the lactic acid fermentation was stably improved to 45.96 mm/L and acetic acid to 73.77 mm/L. R2 was found with the most suitable pH condition for stable organic acids production as Lactobacilli and Bifidobacteria were the major members. Both the members have the key roles in heterofermentation and produce higher transcripts of key encoding enzymes involved in the dominant heterofermentation pathways.

Modeling and simulation of enzymatic gluconic acid production using immobilized enzyme and CSTR-PFTR circulation reaction system.

OBJECTIVES: Production of gluconic acid by using immobilized enzyme and continuous stirred tank reactor-plug flow tubular reactor (CSTR-PFTR) circulation reaction system. RESULTS: A production system is constructed for gluconic acid production, which consists of a continuous stirred tank reactor (CSTR) for pH control and liquid storage and a plug flow tubular reactor (PFTR) filled with immobilized glucose oxidase (GOD) for gluconic acid production. Mathematical model is developed for this production system and simulation is made for the enzymatic reaction process. The pH inhibition effect on GOD is modeled by using a bell-type curve. CONCLUSIONS: Gluconic acid can be efficiently produced by using the reaction system and the mathematical model developed for this system can simulate and predict the process well.

A general and atom-efficient continuous-flow approach to prepare amines, amides and imines via reactive N-chloramines.

Chloramines are an important class of reagents, providing a convenient source of chlorine or electrophilic nitrogen. However, the instability of these compounds is a problem which makes their isolation and handling difficult. To overcome these hazards, a continuous-flow approach is reported which generates and immediately reacts N-chloramines directly, avoiding purification and isolation steps. 2-Chloramines were produced from the reaction of styrenes with N-alkyl-N-sulfonyl-N-chloramines, whilst N-alkyl or N,N'-dialkyl-N-chloramines reacted with anisaldehyde in the presence of t-BuO2H oxidant to afford amides. Primary and secondary imines were produced under continuous conditions from the reaction of N-chloramines with base, with one example subsequently reduced under asymmetric conditions to produce a chiral amine in 94% ee.

Adaptation of continuous biogas reactors operating under wet fermentation conditions to dry conditions with corn stover as substrate.

Corn stover (CS) is the agricultural by-product of maize cultivation. Due to its high abundance and high energy content it is a promising substrate for the bioenergy sector. However, it is currently neglected in industrial scale biogas plants, because of its slow decomposition and hydrophobic character. To assess the maximum biomethane potential of CS, long-term batch fermentations were carried out with various substrate concentrations and particle sizes for 72 days. In separate experiments we adapted the biogas producing microbial community in wet fermentation arrangement first to the lignocellulosic substrate, in Continuous Stirred Tank Reactor (CSTR), then subsequently, by continuously elevating the feed-in concentration, to dry conditions in solid state fermenters (SS-AD). In the batch tests, the <10 mm fraction of the grinded and sieved CS was amenable for biogasification, but it required 10% more time to produce 90% of the total biomethane yield than the <2 mm sized fraction, although in the total yields there was no significant difference between the two size ranges. We also observed that increasing amount of substrate added to the fermentation lowered the specific methane yield. In the CSTR experiment, the daily substrate loading was gradually increased from 1 to 2 gvs/L/day until the system produced signs of overloading. Then the biomass was transferred to SS-AD reactors and the adaptation process was studied. Although the specific methane yields were lower in the SS-AD arrangement (177 mL CH4/gvs in CSTR vs. 105 mL in SS-AD), the benefits of process operational parameters, i.e. lower energy consumption, smaller reactor volume, digestate amount generated and simpler configuration, may compensate the somewhat lower yield.

Oxidative degradation of tetramethylammonium hydroxide (TMAH) by UV/persulfate and associated acute toxicity assessment.

Tetramethylammonium hydroxide (TMAH) is widely used in high-tech industries as a developing agent. Ultraviolet (UV) light-activated persulfate (PS, S2O82-) can be used to generate strongly oxidative sulfate radicals, and it also exhibits the potential to treat TMAH-containing wastewater. This study initially investigated the effect of S2O82- concentration and UV strength on the UV/S2O82- process for the degradation of TMAH in a batch reactor. The results suggested that 15 watts (W) of UV-activated S2O82- at concentrations of 10 or 50 mM resulted in pseudo-first-order TMAH degradation rate constants of 3.1-4.2 × 10-2 min-1, which was adopted for determining the hydraulic retention time (HRT) in a continuous stirred tank reactor (CSTR). The operating conditions (15 W UV/10 mM S2O82-) with a HRT of 129 min resulted in stable residual concentrations of S2O82- and TMAH at approximately 2.6 mM and 20 mg L-1 in effluent, respectively. Several TMAH degradation intermediates including trimethylamine, dimethylamine, and methylamine were also detected. The effluent was adjusted to a neutral pH and evaluated for its biological acute toxicity using Cyprinus carpio as a bioassay organism. The "bio-acute toxicity unit" (TUa) was determined to be 1.41, which indicated that the effluent was acceptable for being discharged into an aquatic ecosystem.

Metabolomic analysis of human fecal microbiota: a comparison of feces-derived communities and defined mixed communities.

The extensive impact of the human gut microbiota on its human host calls for a need to understand the types of communication that occur among the bacteria and their host. A metabolomics approach can provide a snapshot of the microbe-microbe interactions occurring as well as variations in the microbes from different hosts. In this study, metabolite profiles from an anaerobic continuous stirred-tank reactors (CSTR) system supporting the growth of several consortia of bacteria representative of the human gut were established and compared. Cell-free supernatant samples were analyzed by 1D (1)H nuclear magnetic resonance (NMR) spectroscopy, producing spectra representative of the metabolic activity of a particular community at a given time. Using targeted profiling, specific metabolites were identified and quantified on the basis of NMR analyses. Metabolite profiles discriminated each bacterial community examined, demonstrating that there are significant differences in the microbiota metabolome between each cultured community. We also found unique compounds that were identifying features of individual bacterial consortia. These findings are important because they demonstrate that metabolite profiles of gut microbial ecosystems can be constructed by targeted profiling of NMR spectra. Moreover, examination of these profiles sheds light on the type of microbes present in the gut and their metabolic interactions.

Modeling the competition between PHA-producing and non-PHA-producing bacteria in feast-famine SBR and staged CSTR systems.

Although the enrichment of specialized microbial cultures for the production of polyhydroxyalkanoates (PHA) is generally performed in sequencing batch reactors (SBRs), the required feast-famine conditions can also be established using two or more continuous stirred-tank reactors (CSTRs) in series with partial biomass recirculation. The use of CSTRs offers several advantages, but will result in distributed residence times and a less strict separation between feast and famine conditions. The aim of this study was to investigate the impact of the reactor configuration, and various process and biomass-specific parameters, on the enrichment of PHA-producing bacteria. A set of mathematical models was developed to predict the growth of Plasticicumulans acidivorans-as a model PHA producer-in competition with a non-storing heterotroph. A macroscopic model considering lumped biomass and an agent-based model considering individual cells were created to study the effect of residence time distribution and the resulting distributed bacterial states. The simulations showed that in the 2-stage CSTR system the selective pressure for PHA-producing bacteria is significantly lower than in the SBR, and strongly affected by the chosen feast-famine ratio. This is the result of substrate competition based on both the maximum specific substrate uptake rate and substrate affinity. Although the macroscopic model overestimates the selective pressure in the 2-stage CSTR system, it provides a quick and fairly good impression of the reactor performance and the impact of process and biomass-specific parameters.

Biohydrogen and Bioethanol Production from Biodiesel-Based Glycerol by Enterobacter aerogenes in a Continuous Stir Tank Reactor.

Crude glycerol from the biodiesel manufacturing process is being produced in increasing quantities due to the expanding number of biodiesel plants. It has been previously shown that, in batch mode, semi-anaerobic fermentation of crude glycerol by Enterobacter aerogenes can produce biohydrogen and bioethanol simultaneously. The present study demonstrated the possible scaling-up of this process from small batches performed in small bottles to a 3.6-L continuous stir tank reactor (CSTR). Fresh feed rate, liquid recycling, pH, mixing speed, glycerol concentration, and waste recycling were optimized for biohydrogen and bioethanol production. Results confirmed that E. aerogenes uses small amounts of oxygen under semi-anaerobic conditions for growth before using oxygen from decomposable salts, mainly NH4NO3, under anaerobic condition to produce hydrogen and ethanol. The optimal conditions were determined to be 500 rpm, pH 6.4, 18.5 g/L crude glycerol (15 g/L glycerol) and 33% liquid recycling for a fresh feed rate of 0.44 mL/min. Using these optimized conditions, the process ran at a lower media cost than previous studies, was stable after 7 days without further inoculation and resulted in yields of 0.86 mol H2/mol glycerol and 0.75 mol ethanol/mole glycerol.

Continuous formation of N-chloro-N,N-dialkylamine solutions in well-mixed meso-scale flow reactors.

The continuous flow synthesis of a range of organic solutions of N,N-dialkyl-N-chloramines is described using either a bespoke meso-scale tubular reactor with static mixers or a continuous stirred tank reactor. Both reactors promote the efficient mixing of a biphasic solution of N,N-dialkylamine in organic solvent, and aqueous sodium hypochlorite to achieve near quantitative conversions, in 72-100% in situ yields, and useful productivities of around 0.05 mol/h with residence times from 3 to 20 minutes. Initial calorimetric studies have been carried out to inform on reaction exotherms, rates and safe operation. Amines which partition mainly in the organic phase require longer reaction times, provided by the CSTR, to compensate for low mass transfer rates in the biphasic system. The green metrics of the reaction have been assessed and compared to existing procedures and have shown the continuous process is improved over previous procedures. The organic solutions of N,N-dialkyl-N-chloramines produced continuously will enable their use in tandem flow reactions with a range of nucleophilic substrates.

Influence of organic loading rate on methane production in a CSTR from physicochemical sludge generated in a poultry slaughterhouse.

The influence of the increase of the organic loading rate (OLR) on methane production in a continuous stirred-tank reactor (CSTR) from physicochemical sludge generated in a poultry slaughterhouse was evaluated. Total solid (TS) to obtain OLR of 1, 5, 10 and 15 g VS L(-1) day(-1), with hydraulic retention times of 29, 6, 6 and 4, respectively, were conditioned. The results showed a decrease in pH levels and an increase in the theoretical volatile fatty acids (VFA). While the yield of methane production decreased from 0.48 to 0.10 LCH4/g VSremoved, respectively, the OLR-10 managed on average 38% removal of volatile solids (VS) and a yield biogas production of 0.81 Lbiogas g(-1) VSremoved and 1.35 L day(-1). This suggests that the OLR increases in an anaerobic system from physicochemical sludge only inhibits the methanogenic metabolism, because there is still substrate consumption and biogas production.

Analysis of ammonia-nitrogen removal kinetics by stage in pilot scale vertical flow wetlands treating landfill leachate in series.

This study explored the effect stage number and plant type have on ammonia-nitrogen (NH3-N) removal kinetics in a two-stage pilot-scale vertical flow constructed wetland (VFCW) system treating landfill leachate. Half of the VFCW columns were planted with Typha latifolia and the other half Scirpus californicus, and half of the columns were loaded with municipal solid waste landfill leachate (diluted to 1 part leachate to 2 parts total) with the effluent from these columns was collected in two separate barrels. The remaining columns were loaded with the effluent collected from the first columns, creating a two-stage VFCW system with four unique pairs to be tested. The leachate used here experienced no prior pre-treatment, and average influent concentrations of NH3-N for the first- and second-stage VFCWs were 508 and 321 mg L-1, respectively- much higher than many other VFCW treatment systems. Some reduction in chemical oxygen demand was observed, as well as generation of nitrate and nitrite, evidence of nitrification. No apparent correlation between aboveground biomass and removal of NH3-N was observed. Overall removal efficiency of NH3-N through two stages of VFCWs was 53.7% for columns planted with T. latifolia and 58.3% for those planted with S. Californicus. Average NH3-N removal efficiencies for the first stage VFCWs were 32.7% and 34.3%, while those in the second stage were 31.3% and 36.5%; no significant difference was observed between the first and second stage, suggesting that stage number does not have a significant effect on the removal efficiency of NH3-N in the primary treatment of landfill leachate via VFCWs. However, average mass removal rates of NH3-N in the first stage were 166 and 175 mg L-1 d-1; the second stage was significantly lower at 99.4 and 112 mg L-1 d-1, indicating that the first stage removed more pollutants overall.

Continuous H-B-E fermentation by Clostridium carboxidivorans: CO vs syngas.

Leveraging renewable carbon-based resources for energy and chemical production is a promising approach to decrease reliance on fossil fuels. This entails a thermo/biotechnological procedure wherein bacteria, notably Clostridia, ferment syngas, converting CO or CO2 + H2 into Hexanol, Butanol and Ethanol (H-B-E fermentation). This work reports of Clostridium carboxidivorans performance in a stirred tank reactor continuously operated with respect to the gas and the cell/liquid phases. The primary objective was to assess acid and solvent production at pH 5.6 by feeding pure CO or synthetic syngas under gas flow differential conditions. Fermentation tests were conducted at four different dilution rates (DL) of the fresh medium in the range 0.034-0.25 h-1. The fermentation pathways of C. carboxidivorans were found to be nearly identical for both CO and syngas, with consistent growth and metabolite production at pH 5.6 within a range of dilution rates. Wash-out conditions were observed at a DL of 0.25 h-1 regardless of the carbon source. Ethanol was the predominant solvent produced, but a shift towards butanol production was observed with CO as the substrate and towards hexanol production with synthetic syngas. In particular, the maximum cell concentration (0.5 gDM/L) was obtained with pure CO at DL 0.05 h-1; the highest solvent productivity (60 mg/L*h of total solvent) was obtained at DL 0.17 h-1 by using synthetic syngas as C-source. The findings highlight the importance of substrate composition and operating conditions in syngas fermentation processes. These insights contribute to the optimization of syngas fermentation processes for biofuel and chemical production.

Silver Recovery from Crystalline Silicon Photovoltaic Solar Cells Using Continuous Stirred-Tank Reactors.

Silver can be recycled from the end-of-life crystalline silicon photovoltaic (PV), yet the recycling and its technology scale-up are still at an early stage especially in continuously operations e.g., continoursely stirred tank reactors (CSTR). Here, the silver recovery from the solar cells is technically understood and optimized in the CSTR system from the point of view of silver recovery efficiency, through integrating experimental and numerical investigations. Specifically, based on the experiments, a kinetics model is developed and scanning electron microscopy surface morphology is characterized; and a computational fluid dynamics-discrete element method (CFD-DEM) particle-scale model is integrated with the kinetics model and validated against the fluid-flow pattern and silver leaching performance results from lab measurements. The validated CFD-DEM model is then applied to understand the particle-scale behavior of silver leaching in the CSTR system in terms of hydrodynamics and AgNO3 distribution under different impeller speeds. The simulation results show that the silver leaching performance is improved in an improved CSTR design with a lower impeller position and doubled impeller layers. This work reveals the effectiveness and underlying hydrodynamics of silver leaching in CSTR systems and lays a foundation for improving silver recovery in PV recycling.

Rapid start-up and operational characteristics of partial denitrification coupled with anammox driven by innovative strategies.

The Partial Denitrification-Anammox (PD/A) process established a low-consumption, efficient and sustainable pathway for complete nitrogen removal, which is of great interest to the industry. Rapid initiation and stable operation of the PD/A systems were the main issues limiting its engineering application in wastewater nitrogen removal. A PD/A system was initiated in a continuous stirred-tank reactors (CSTRs) in the presence of low concentration of organic matter, and the effects of organic matter types and COD/NO3--N ratios on the performance of the PD/A system, and microbial community characteristics were explored. The results showed that low concentrations of organic matter could promote the rapid initiation of the Anammox process and then the strategy of gradually replacing NO2--N with NO3--N could successfully initiate the PD/A system at 70 days. The type of organic matter had a significant effect on the initiation of the Anammox and the establishment of the PD/A system. Compared to glucose, sodium acetate was more favorable for rapid start-up and the synergy among microorganisms, and organic matter was lower, with an optimal COD/NO3--N ratio of 3.0. Microorganisms differed in their sensitivity to environmental factors. The relative abundance of Planctomycetota and Proteobacteria in R2 was 51 %, with the presence of three typical anammox bacteria, Candidatus_Brocadia, Candidatus_Kuenenia, and Candidatus_Jettenia in the system. This study provides a new strategy for the rapid initiation and stable operation of the PD/A process.