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Interactions among biomacromolecules, predominantly noncovalent, underpin biological processes. However, recent advancements in biospecific chemistry have enabled the creation of specific covalent bonds between biomolecules, both in vitro and in vivo. This Review traces the evolution of biospecific chemistry in proteins, emphasizing the role of genetically encoded latent bioreactive amino acids. These amino acids react selectively with adjacent natural groups through proximity-enabled bioreactivity, enabling targeted covalent linkages. We explore various latent bioreactive amino acids designed to target different protein residues, ribonucleic acids, and carbohydrates. We then discuss how these novel covalent linkages can drive challenging protein properties and capture transient protein-protein and protein-RNA interactions in vivo. Additionally, we examine the application of covalent peptides as potential therapeutic agents and site-specific conjugates for native antibodies, highlighting their capacity to form stable linkages with target molecules. A significant focus is placed on proximity-enabled reactive therapeutics (PERx), a pioneering technology in covalent protein therapeutics. We detail its wide-ranging applications in immunotherapy, viral neutralization, and targeted radionuclide therapy. Finally, we present a perspective on the existing challenges within biospecific chemistry and discuss the potential avenues for future exploration and advancement in this rapidly evolving field.

Purpose: 3D cell culture and hypoxia have been demonstrated to increase the therapeutic effects of mesenchymal stem/stromal cells (MSCs)-derived extracellular vesicles (EVs). In this study, a process for the production of MSC-EVs in a novel 3D bioreactor system under normoxic and hypoxic conditions was established and the resulting EVs were characterized. Methods: Human adipose-derived MSCs were seeded and cultured on a 3D membrane in the VITVO® bioreactor system for 7 days. Afterwards, MSC-EVs were isolated and characterized via fluorescence nanoparticle tracking analysis, flow cytometry with staining against annexin V (Anx5) as a marker for EVs exposing phosphatidylserine, as well as CD73 and CD90 as MSC surface markers. Results: Cultivation of MSC in the VITVO® bioreactor system demonstrated a higher concentration of MSC-EVs from the 3D bioreactor (9.1 × 109 ± 1.5 × 109 and 9.7 × 109 ± 3.1 × 109 particles/mL) compared to static 2D culture (4.2 × 109 ± 7.5 × 108 and 3.9 × 109 ± 3.0 × 108 particles/mL) under normoxic and hypoxic conditions, respectively. Also, the particle-to-protein ratio as a measure for the purity of EVs increased from 3.3 × 107 ± 1.1 × 107 particles/µg protein in 2D to 1.6 × 108 ± 8.3 × 106 particles/µg protein in 3D. Total MSC-EVs as well as CD73−CD90+ MSC-EVs were elevated in 2D normoxic conditions. The EV concentration and size did not differ significantly between normoxic and hypoxic conditions. Conclusion: The production of MSC-EVs in a 3D bioreactor system under hypoxic conditions resulted in increased EV concentration and purity. This system could be especially useful in screening culture conditions for the production of 3D-derived MSC-EVs.

Microalgae are vital microorganisms due to their role in oxygen production, marine ecosystems, and as a source of various high-value products such as vitamins, biofuel, livestock feed, cosmetics, and more. Within some applications, challenges arise in large-scale microalgae production, characterized by high costs and growth rates that fall short of meeting commercial production demands. Therefore, there is still a need for further research to be conducted on microalgae strains and cultivation systems to improve costs and production rates. The focus of this study was to research a novel, patent-pending 360 Air-Loop Series Bioreactor (360-ASBR) and compare it to a Bubble Column Bioreactor (BCBR) that serves as the control. The specific objectives were to (1) determine and compare the hydrodynamic variables of the 360-ASBR and the BCBR including mixing time, residence time, and oxygen liquid mass transfer coefficient (KLa) and (2) determine the effect of the amount and placement of spargers in the BCBR. The results of this study showed that: (1) the mixing time of 198 seconds for the high flow rate and 247 seconds for the low flow rate for the ASBR significantly exceeded that of 25.1 seconds and 38.7 seconds respectively for the BCBR (2) the average residence time of 829 seconds was not significantly less than 1048 seconds for the BCBR at the high flow rate whereas the average residence time of 698 seconds for the ASBR was significantly less than 1053 seconds for the BCBR at the low flow rate, (3) the KLa of 0.012 s-1 for the ASBR was not significantly higher than 0.011 s-1 for the BCBR at the high flow rate whereas 0.009 s-1 for the ASBR significantly exceeded the KLa of 0.002 s-1 for the BCBR at the low flow rate, and (4) the mixing times of 25.1, 25.0, and 29.5 s for each sparger arrangement at the high flow rate and 38.7, 29.8, and 39.2 s for each sparger arrangement at the low flow rate did not result in significant differences. The principal conclusions drawn fro

Lower back pain continues to be a global epidemic, limiting quality of life and ability to work, due in large part to symptomatic disc degeneration. Development of more effective and less invasive biological strategies are needed to treat disc degeneration. In vitro models such as macro- or micro-bioreactors or mechanically active organ-chips hold great promise in reducing the need for animal studies that may have limited clinical translatability, due to harsher and more complex mechanical loading environments in human discs than in most animal models. This review highlights the complex loading conditions of the disc in situ, evaluates state-of-the-art designs for applying such complex loads across multiple length scales, from macro-bioreactors that load whole discs to organ-chips that aim to replicate cellular or engineered tissue loading. Emphasis was placed on the rapidly evolving more customizable organ-chips, given their greater potential for studying the progression and treatment of symptomatic disc degeneration. Lastly, this review identifies new trends and challenges for using organ-chips to assess therapeutic strategies.

One of the major challenges mankind faces nowadays is combating climate change. A substantial fraction of greenhouse gases are released by industrial processes, as steelmaking, (oil)refinery and waste processing. Emissions from these processes can partly be prevented with a recently developed technology called gas fermentation. Within this process, synthesis gas – amixture containing CO, CO2 and H2 – is converted into ethanol and acetic acid by bacteria such as Clostridium autoethanogenum. These products could be used in a wide range of applications, like fuels, plastics and cosmetics. Whilst gas fermentation is already applied at commercial-scale, challenges in scale-up persists due to complex multi-scale interactions among the bioreactor, gas bubbles, and bacteria. The poor solubility of CO and H2 alongside gas bubble coalescence, leads to low gas-to-liquid mass transfer rates (typically denoted via kLa). Slow mixing in industrial bioreactors (500m3), and high gas conversion rates, result in large spatial variations in dissolved gas concentrations. Bacteria experiencing concentration fluctuations have often been related to decreased process performance..., BT/Bioprocess Engineering

A major source of CO2 emissions is the flaring of steel mill gas. This work demonstrated the enrichment of carboxydotrophic bacteria for converting steel mill gas into volatile fatty acids and H2, via gas fermentation. Several combinations of pure and mixed anaerobic cultures were used as inoculum in 0.5-L reactors, operated at 30 and 60 °C. The process was then scaled up in a 4-L membrane bioreactor, operated for 20 days, at 48 °C. The results showed that the enriched microbiomes can oxidize CO completely to produce H2/H+ which is subsequently used to fix the CO2. At 30 °C, a mixture of acetate, isobutyrate and propionate was obtained while H2 and acetate were the main products at 60 °C. The highest CO conversion and H2 production rate observed in the membrane bioreactor were 29 and 28 mL/LR/h, respectively. The taxonomic diversity of the bacterial community increased and the dominant species was Pseudomonas.

Volatile fatty acids (VFAs) appear to be an economical carbon feedstock for the cost-effective production of polyhydroxyalkanoates (PHAs). The use of VFAs, however, could impose a drawback of substrate inhibition at high concentrations, resulting in low microbial PHA productivity in batch cultivations. In this regard, retaining high cell density using immersed membrane bioreactor (iMBR) in a (semi-) continuous process could enhance production yields. In this study, an iMBR with a flat-sheet membrane was applied for semi-continuous cultivation and recovery of Cupriavidus necator in a bench-scale bioreactor using VFAs as the sole carbon source. The cultivation was prolonged up to 128 h under an interval feed of 5 g/L VFAs at a dilution rate of 0.15 (d−1), yielding a maximum biomass and PHA production of 6.6 and 2.8 g/L, respectively. Potato liquor and apple pomace-based VFAs with a total concentration of 8.8 g/L were also successfully used in the iMBR, rendering the highest PHA content of 1.3 g/L after 128 h of cultivation. The PHAs obtained from both synthetic and real VFA effluents were affirmed to be poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with a crystallinity degree of 23.8 and 9.6%, respectively. The application of iMBR could open an opportunity for semi-continuous production of PHA, increasing the feasibility of upscaling PHA production using waste-based VFAs.

The definitive treatment for end-stage renal disease is kidney transplantation, which remains limited by organ availability and post-transplant complications. Alternatively, an implantable bioartificial kidney could address both problems while enhancing the quality and length of patient life. An implantable bioartificial kidney requires a bioreactor containing renal cells to replicate key native cell functions, such as water and solute reabsorption, and metabolic and endocrinologic functions. Here, we report a proof-of-concept implantable bioreactor containing silicon nanopore membranes to offer a level of immunoprotection to human renal epithelial cells. After implantation into pigs without systemic anticoagulation or immunosuppression therapy for 7 days, we show that cells maintain >90% viability and functionality, with normal or elevated transporter gene expression and vitamin D activation. Despite implantation into a xenograft model, we find that cells exhibit minimal damage, and recipient cytokine levels are not suggestive of hyperacute rejection. These initial data confirm the potential feasibility of an implantable bioreactor for renal cell therapy utilizing silicon nanopore membranes.

This study evaluated a tertiary wastewater treatment technology using cyanobacteria to recover value-added phycobiliproteins. The presence of contaminants of emerging concern (CECs) in wastewater, cyanobacterial biomass and pigments recovered were also analyzed. For this, a wastewater-borne cyanobacterium (Synechocystis sp. R2020) was used to treat secondary effluent from a municipal wastewater treatment plant, with and without nutrients supplementation. Then, the stability of phycobiliprotein production was assessed by operating the photobioreactor in semicontinuous mode. Results showed similar biomass productivity with and without nutrients supplementation (153.5 and 146.7 mg L- 1 d- 1 , respectively). Upon semi-continuous operation, the phycobiliprotein content was stable and reached up to 74.7 mg gDW- 1 . The phycocyanin purity ratio ranged from 0.5 to 0.8, corresponding to food grade (>0.7). Out of 22 CECs detected in secondary effluent, only 3 were present in the phycobiliprotein extracts. In order to identify applications, prospective research should focus on CECs removal during pigment purification., This research was funded by the R + D + I projects AL4BIO (RTI2018-099495-B-C21) and Cyan2Bio (PID2021-126564OB-C32), funded by MCIN/AEI/ 10.13039/501100011033 and “ERDF A way of making Europe’’. Marta Bellver acknowledges her grant PRE2019-091552 funded by MCIN/AEI/ 10.13039/501100011033 and by “ESF Investing in your future”. Rubén Díez-Montero acknowledges his grant IJC2019-042069-I funded by MCIN/AEI/ 10.13039/501100011033. Mònica Escolà is grateful to the Beatriu de Pinós 2018 grant-programme (MSCA grant agreement number 801370)., Peer Reviewed, Objectius de Desenvolupament Sostenible::6 - Aigua Neta i Sanejament::6.3 - Per a 2030, millorar la qualitat de l’aigua mitjançant la reducció de la contaminació, l’eliminació dels abocaments i la reducció al mínim de la descàrrega de materials i productes químics perillosos, la reducció a la meitat del percentatge d’aigües residuals sense tractar, i un augment substancial a escala mundial del reciclat i de la reutilització en condicions de seguretat, Objectius de Desenvolupament Sostenible::6 - Aigua Neta i Sanejament, Objectius de Desenvolupament Sostenible::12 - Producció i Consum Responsables, Objectius de Desenvolupament Sostenible::12 - Producció i Consum Responsables::12.2 - Per a 2030, assolir la gestió sostenible i l’ús eficient dels recursos naturals, Postprint (published version)

A major source of CO2 emissions is the flaring of steel mill gas. This work demonstrated the enrichment of carboxydotrophic bacteria for converting steel mill gas into volatile fatty acids and H2, via gas fermentation. Several combinations of pure and mixed anaerobic cultures were used as inoculum in 0.5-L reactors, operated at 30 and 60 °C. The process was then scaled up in a 4-L membrane bioreactor, operated for 20 days, at 48 °C. The results showed that the enriched microbiomes can oxidize CO completely to produce H2/H+ which is subsequently used to fix the CO2. At 30 °C, a mixture of acetate, isobutyrate and propionate was obtained while H2 and acetate were the main products at 60 °C. The highest CO conversion and H2 production rate observed in the membrane bioreactor were 29 and 28 mL/LR/h, respectively. The taxonomic diversity of the bacterial community increased and the dominant species was Pseudomonas.

Volatile fatty acids (VFAs) appear to be an economical carbon feedstock for the cost-effective production of polyhydroxyalkanoates (PHAs). The use of VFAs, however, could impose a drawback of substrate inhibition at high concentrations, resulting in low microbial PHA productivity in batch cultivations. In this regard, retaining high cell density using immersed membrane bioreactor (iMBR) in a (semi-) continuous process could enhance production yields. In this study, an iMBR with a flat-sheet membrane was applied for semi-continuous cultivation and recovery of Cupriavidus necator in a bench-scale bioreactor using VFAs as the sole carbon source. The cultivation was prolonged up to 128 h under an interval feed of 5 g/L VFAs at a dilution rate of 0.15 (d−1), yielding a maximum biomass and PHA production of 6.6 and 2.8 g/L, respectively. Potato liquor and apple pomace-based VFAs with a total concentration of 8.8 g/L were also successfully used in the iMBR, rendering the highest PHA content of 1.3 g/L after 128 h of cultivation. The PHAs obtained from both synthetic and real VFA effluents were affirmed to be poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with a crystallinity degree of 23.8 and 9.6%, respectively. The application of iMBR could open an opportunity for semi-continuous production of PHA, increasing the feasibility of upscaling PHA production using waste-based VFAs.

Computational models are increasingly used to investigate and predict the complex dynamics of biological and biochemical systems. Nevertheless, governing equations of a biochemical system may not be (fully) known, which would necessitate learning the system dynamics directly from, often limited and noisy, observed data. On the other hand, when expensive models are available, systematic and efficient quantification of the effects of model uncertainties on quantities of interest can be an arduous task. This paper leverages the notion of flow-map (de)compositions to present a framework that can address both of these challenges via learning data-driven models useful for capturing the dynamical behavior of biochemical systems. Data-driven flow-map models seek to directly learn the integration operators of the governing differential equations in a black-box manner, irrespective of structure of the underlying equations. As such, they can serve as a flexible approach for deriving fast-to-evaluate surrogates for expensive computational models of system dynamics, or, alternatively, for reconstructing the long-term system dynamics via experimental observations. We present a data-efficient approach to data-driven flow-map modeling based on polynomial chaos Kriging. The approach is demonstrated for discovery of the dynamics of various benchmark systems and a coculture bioreactor subject to external forcing, as well as for uncertainty quantification of a microbial electrosynthesis reactor. Such data-driven models and analyses of dynamical systems can be paramount in the design and optimization of bioprocesses and integrated biomanufacturing systems.

This research investigates a highly efficient compact tubular ceramic-supported carbon-based membrane reactor integrated with anaerobic biodegradation to decolorize the azo dyes. Two carbon-based membranes, produced using Matrimid 5218 polyimide and graphene oxide solutions, are evaluated for the comparative color removal of three structurally different azo dyes, Acid Orange 7 (AO7), Reactive Black 5 (RB5), and Direct Blue 71 (DB71). Based on FESEM microscopic images, the average pore size of the tubular ceramic-supported carbonized membrane (TCSCM) was approximately 25 nm, while for the tubular ceramic-supported graphene oxide membrane (TCSGOM), it was 12 nm. Additionally, TCSCM had a thinner layer at only 1.10 µm, while TCSGOM was slightly thicker at 2.11 µm. These features influenced the permeate flux of the membrane, in which the TCSGOM exhibited lower permeate flux (18.2 L·m-2·h-1) than the TCSCM (45.6 L·m-2·h-1). However, the anaerobic decolorization results indicated that the TCSGOM bioreactor (B-TCSGOM) was more efficient and effective at removing color from all dye solutions than the TCSCM bioreactor (B-TCSCM) over a wide range of feed concentrations. In both reactors, the highest decolorization was achieved at low feed concentration (50 mg·L-1), and removal was 94 % for AO7, 90 % for RB5, and 88 % for DB71 in B-TCSGOM, whereas 88 %, 85 %, and 69 %, respectively, in B-TCSCM. These suggest that the robust conductive nanoporous surface of B-TCSGOM makes it more effective at removing different azo dye solutions from wastewater., This project has been supported by the European Union’s Horizon 2020 Research and Innovation Programme under the Marie Skłodowska-Curie grant agreement No. 713679 and by the Universitat Rovira i Virgili (URV), contract 2017MFP-COFUND-18. This article was possible thanks to the grants RTI2018-096467-B-I00 and PID2021-126895OB-I00 funded by MCIN/AEI/ 10.13039/501100011033 and “ERDF A way of making Europe”. The authors research group is recognized by the Comissionat per a Universitats i Recerca, DIUE de la Generalitat de Catalunya (2017 SGR 396 and 2021 SGR 00200), and supported by the Universitat Rovira i Virgili (2021PFR-URV-87)., Peer Reviewed, Postprint (published version)

Extracellular vesicles (EVs) are implicated as promising therapeutics and drug delivery vehicles in various diseases. However, successful clinical translation will depend on the development of scalable biomanufacturing approaches, especially due to the documented low levels of intrinsic EV-associated cargo that may necessitate repeated doses to achieve clinical benefit in certain applications. Thus, here the effects of a 3D-printed scaffold-perfusion bioreactor system are assessed on the production and bioactivity of EVs secreted from bone marrow-derived mesenchymal stem cells (MSCs), a cell type widely implicated in generating EVs with therapeutic potential. The results indicate that perfusion bioreactor culture induces an ≈40-80-fold increase (depending on measurement method) in MSC EV production compared to conventional cell culture. Additionally, MSC EVs generated using the perfusion bioreactor system significantly improve wound healing in a diabetic mouse model, with increased CD31+ staining in wound bed tissue compared to animals treated with flask cell culture-generated MSC EVs. Overall, this study establishes a promising solution to a major EV translational bottleneck, with the capacity for tunability for specific applications and general improvement alongside advancements in 3D-printing technologies.

My master's thesis focuses on the effect of adding pure glycerol as a co-substrate during the anaerobic digestion of sewage sludge. This study is divided into two interconnected parts. The first part concerns the molecular and bioinformatic characterization of the communities in the bioreactors. Microbial populations involved in anaerobic digestion for biogas production were investigated through genomic analyses in three different phases: parallel operation, serial operation of reactor pairs, and during the addition of glycerol as a co-substrate in serial mode. The bioinformatic analysis revealed species composition and variations in alpha and beta diversity among the different phases, with glycerol addition reversing the trend. Co-digestion promoted the development of specific populations contributing to substrate degradation. Metagenomic analysis identified numerous unclassified genomic species at lower taxonomic levels, opening up new research perspectives. The second part of the study concerns the physicochemical characterization of the system with the aim of maximizing biomethane production from sewage sludge by utilizing the priming effect of 1% (v/v) pure glycerol as a co-substrate in a serial system. Serial operation and glycerol addition increased biogas production without overloading the reactors. Further analysis is needed to confirm the specific biomethane production effects. Overall, this approach holds promise for enhancing biomethane production from sewage sludge. Overall, this study suggests that the combination of two-stage digestion in serial bioreactors and glycerol co-digestion is an effective approach for increasing biomethane production from sewage sludge without overloading the system., Masterarbeit Universität Innsbruck 2023

Eiwitten maken van ammonium uit afvalwater door bacteriën in een bioreactor te voeden met waterstof. Dat is het idee achter het Power-to-Protein project. Waterinstituut KWR onderzoekt hoe het reactorontwerp verbeterd kan worden met behulp van simulaties. Berekend wordt hoe de stofoverdracht van het slecht oplosbare waterstof samen met zuurstof en koolzuurgas in een bellenkolomreactor geoptimaliseerd kan worden. Het Power-to-Protein project onderzoekt de productie van een waardevolle grondstof, eiwit, uit een restproduct, ammonium, dat bijvoorbeeld vrijkomt bij de vergisting van aëroob slib uit de rioolwaterzuivering. Daarvoor wordt energie, in de vorm van waterstof, toegevoerd aan de ammoniumrijke afvalwaterstroom. Waterstof-oxiderende bacteriën kunnen in een bioreactor het ammonium omzetten in eiwit voor menselijke of dierlijke consumptie. Bij deze biosynthese zijn ook zuurstof en koolzuurgas nodig. Als de waterstof op ‘groene’ wijze is opgewekt met wind- of zonne-energie, is sprake van een duurzaam en circulair proces. Interview met B.A. Wols, KWR.

The future of large-scale kelp aquaculture is standing at a crossroad, with the diverging paths being characterized by two fundamentally different cultivation methods that differ on how well gametophyte reproduction can be controlled. The cultivation method that does not directly control gametophyte reproduction is more widely utilized at the moment, but interest in better controlling gametophyte reproduction is growing steadily. Here, we validate a bioreactor system that overcomes a number of implementation challenges for this controlled reproductive method, expanding the possibility of clonal gametophyte cultivation outside of expensive laboratory settings. The main goals of this system include (i) the maintenance of clean gametophyte clonal cultures in non-sterile environments over prolonged periods of time, (ii) the production of large numbers of juvenile sporophytes, and (iii) effective transportation of gametophytes and sporophytes. The “SeaCoRe system” consists out of three parts that correspond to these three challenges: (1) clone-reactors, (2) a clone-inducer, and (3) a transporter. The validation of the system showed that delayed Saccharina latissima and Alaria esculenta gametophytes can grow reliably for 75 days in the clone-reactors. Initial gametophyte densities of 0.4 mg DW and 0.6 mg DW gametophtyes mL−1 were optimal for S. latissima and A. esculenta, resulting in reproductive successes of 604 and 422 sporophytes mL−1, respectively. Lastly, gametophyte transport was simulated, with high reproductive success still achieved within 19 days in ~ 20 °C environments. The SeaCoRe system helps unlock the full potential of large-scale kelp cultivation using multiannual delayed clonal.

Producción Científica, The influence of suspended solids and pH in anaerobically digested piggery wastewater on membrane-based NH3 extraction was evaluated in batch tests. The increase in pH in the anaerobic broth from 8 to 9 resulted in an increase in NH3 removal efficiencies from 15.8 % ± 0.1 % to 20.9 % ± 0.4 % regardless of the suspended solids. The influence of membrane based NH3 extraction on piggery wastewater treatment was also assessed in a CSTR interconnected with PTFE membrane modules. The decrease in TKN concentrations mediated by membrane operation induced an increase in CH4 yield from 380.4 ± 84.9 up to 566.1 ± 7.8 NmLCH4 g VS fed−1. Likewise, COD and VS removal efficiencies significantly increased from 33.0 % ± 2.0 % and 25.7 % ± 2.3 % up to 61.8 % ± 1.3 % and 37.9 % ± 1.8 %, respectively. Interestingly, the decrease in NH3 concentration entailed a complete assimilation of VFA., Junta de Castilla y Leon - Fondo Europeo de Desarrollo Regional (grants VA088G19, CL-EI-2021-07, UIC 315 and UIC 082), Ministerio de Ciencia, Innovación y Universidades (grant PID2019-109403RB-C21/AEI/10.13039/501100011033)

Producción Científica, The application of biogas as a low-priced substrate for the production of ectoines constitutes an opportunity to decrease their production costs and to enhance the viability of anaerobic digestion. The influence of operational conditions on CH4-biogas biodegradation and on ectoines production yields was assessed in continuous pilot bubble column bioreactors. The rise in biomass concentration from 1 to 3 g L-1 resulted in a decrease in the specific ectoine content from 42±8 to 30±4 mgectoine gVSS-1. The concentration of Cu2+ and Mg2+ did not impact process performance, while the use of ammonium as N source resulted in low CH4 biodegradation and ectoine yields (13±7 mgectoine gVSS-1). The increase in CH4 content from 4.5 to 9 %v·v-1 enhanced CH4 removal efficiency. Process operation at NaCl concentrations of 3 %w·w-1 instead of 6 %w·w-1 decreased the ectoine yield to 17 mgectoine gVSS-1. Finally, Methylomicrobium buryatense was identified as the dominant species., Junta de Castilla y León - Fondo Europeo de Desarrollo Regional (projects CLU-2017-09 and UIC-315)

Producción Científica, The application of biogas as a low-priced substrate for the production of ectoines constitutes an opportunity to decrease their production costs and to enhance the viability of anaerobic digestion. The influence of operational conditions on CH4-biogas biodegradation and on ectoines production yields was assessed in continuous pilot bubble column bioreactors. The rise in biomass concentration from 1 to 3 g L-1 resulted in a decrease in the specific ectoine content from 42±8 to 30±4 mgectoine gVSS-1. The concentration of Cu2+ and Mg2+ did not impact process performance, while the use of ammonium as N source resulted in low CH4 biodegradation and ectoine yields (13±7 mgectoine gVSS-1). The increase in CH4 content from 4.5 to 9 %v·v-1 enhanced CH4 removal efficiency. Process operation at NaCl concentrations of 3 %w·w-1 instead of 6 %w·w-1 decreased the ectoine yield to 17 mgectoine gVSS-1. Finally, Methylomicrobium buryatense was identified as the dominant species., Junta de Castilla y León - Fondo Europeo de Desarrollo Regional (projects CLU-2017-09 and UIC-315)

Engineered tissues provide an alternative to graft material, circumventing the use of donor tissue such as autografts or allografts and non-physiological synthetic implants. However, their lack of vasculature limits the growth of volumetric tissue more than several millimeters thick which limits their success post-implantation. Perfused bioreactors enhance nutrient mass transport inside lab-grown tissue but remain poorly customizable to support the culture of personalized implants. Here, a multiscale framework of computational fluid dynamics (CFD), additive manufacturing, and a perfusion bioreactor system are presented to engineer personalized volumetric tissue in the laboratory. First, microscale 3D printed scaffold pore geometries are designed and 3D printed to characterize media perfusion through CFD and experimental fluid testing rigs. Then, perfusion bioreactors are custom-designed to combine 3D printed scaffolds with flow-focusing inserts in patient-specific shapes as simulated using macroscale CFD. Finally, these computationally optimized bioreactor-scaffold assemblies are additively manufactured and cultured with pre-osteoblast cells for 7, 20, and 24 days to achieve tissue growth in the shape of human calcaneus bones of 13 mL volume and 1 cm thickness. This framework enables an intelligent model-based design of 3D printed scaffolds and perfusion bioreactors which enhances nutrient transport for long-term volumetric tissue growth in personalized implant shapes. The novel methods described here are readily applicable for use with different cell types, biomaterials, and scaffold microstructures to research therapeutic solutions for a wide range of tissues.

Tissue biomanufacturing aims to produce lab-grown stem cell grafts and biomimetic drug testing platforms but remains limited in its ability to recapitulate native tissue mechanics. The emerging field of soft robotics aims to emulate dynamic physiological locomotion, representing an ideal approach to recapitulate physiologically complex mechanical stimuli and enhance patient-specific tissue maturation. The kneecap's femoropopliteal artery (FPA) represents a highly flexible tissue across multiple axes during blood flow, walking, standing, and crouching positions, and these complex biomechanics are implicated in the FPA's frequent presentation of peripheral artery disease. We developed a soft pneumatically actuated (SPA) cell culture platform to investigate how patient-specific FPA mechanics affect lab-grown arterial tissues. Silicone hyperelastomers were screened for flexibility and biocompatibility, then additively manufactured into SPAs using a simulation-based design workflow to mimic normal and diseased FPA extensions in radial, angular, and longitudinal dimensions. SPA culture platforms were seeded with mesenchymal stem cells, connected to a pneumatic controller, and provided with 24 h multi-axial exercise schedules to demonstrate the effect of dynamic conditioning on cell alignment, collagen production, and muscle differentiation without additional growth factors. Soft robotic bioreactors are promising platforms for recapitulating patient-, disease-, and lifestyle-specific mechanobiology for understanding disease, treatment simulations, and lab-grown tissue grafts.

Industrial cell culture processes are inherently expensive, time-consuming, and variable. These limitations have become a critical bottleneck for the industrial translation of human cell and tissue biomanufacturing, as few human cell culture products deliver sufficient benefit, value, and consistency to offset their high manufacturing costs and produce useful clinical or biomedical solutions. Recent advances in biomedical image analysis and computational modelling can enhance the design and operation of high-efficiency tissue biomanufacturing platforms, as well as the high-content characterisation and monitoring of culture performance, to enable bioprocess control, optimisation, and automation. These computational technologies aim to maximize culture outcomes while minimizing variability and process development expense. In this review, we outline current resources and approaches which harness biomedical imaging and image-based computational models to design and operate efficient and robust human tissue biomanufacturing platforms.

Producción Científica, The influence of suspended solids and pH in anaerobically digested piggery wastewater on membrane-based NH3 extraction was evaluated in batch tests. The increase in pH in the anaerobic broth from 8 to 9 resulted in an increase in NH3 removal efficiencies from 15.8 % ± 0.1 % to 20.9 % ± 0.4 % regardless of the suspended solids. The influence of membrane based NH3 extraction on piggery wastewater treatment was also assessed in a CSTR interconnected with PTFE membrane modules. The decrease in TKN concentrations mediated by membrane operation induced an increase in CH4 yield from 380.4 ± 84.9 up to 566.1 ± 7.8 NmLCH4 g VS fed−1. Likewise, COD and VS removal efficiencies significantly increased from 33.0 % ± 2.0 % and 25.7 % ± 2.3 % up to 61.8 % ± 1.3 % and 37.9 % ± 1.8 %, respectively. Interestingly, the decrease in NH3 concentration entailed a complete assimilation of VFA., Junta de Castilla y Leon - Fondo Europeo de Desarrollo Regional (grants VA088G19, CL-EI-2021-07, UIC 315 and UIC 082), Ministerio de Ciencia, Innovación y Universidades (grant PID2019-109403RB-C21/AEI/10.13039/501100011033)

In recent years, the mathematical and computational sciences have developed novel methodologies and insights that can aid in designing advanced bioreactors, microfluidic setups or organ-on-chip devices, in optimizing culture conditions, or predicting long-term behavior of engineered tissues in vivo. In this review, we introduce the concept of computational models and how they can be integrated in an interdisciplinary workflow for Tissue Engineering and Regenerative Medicine (TERM). We specifically aim this review of general concepts and examples at experimental scientists with little or no computational modeling experience. We also describe the contribution of computational models in understanding TERM processes and in advancing the TERM field by providing novel insights. Although in recent years the use of mathematical and computational sciences has increased in the Tissue Engineering and Regenerative Medicine (TERM) field, we believe that a further integration of experimental and computational approaches has a huge potential for advancing the field due to the ability of models to explain and predict experimental results and efficiently optimize TERM product and process designs. By providing an overview of existing computational models, how they have contributed to the field, as well as a future perspective, this review represents an important step to help realize TERM's ultimate goal: a cure instead of care.

Microbial community structure is influenced by the environment and in turn exerts control on many environmental parameters. We applied this concept in a bioreactor study to test whether microbial community structure contains information sufficient to predict the concentration of H2S as the product of sulfate reduction. Microbial sulfate reduction is a major source of H2S in many industrial and environmental systems and is often influenced by the existing physicochemical conditions. Production of H2S in industrial systems leads to occupational hazards and adversely affects the quality of products. A long-term (148 days) experiment was conducted in upflow bioreactors to mimic sulfidogenesis, followed by inhibition with nitrate salts and a resumption of H2S generation when inhibition was released. We determined microbial community structure in 731 samples across 20 bioreactors using 16S rRNA gene sequencing and applied a random forest algorithm to successfully predict different phases of sulfidogenesis and mitigation (accuracy = 93.17%) and sessile and effluent microbial communities (accuracy = 100%). Similarly derived regression models that also included cell abundances were able to predict H2S concentration with remarkably high fidelity (R2 > 0.82). Metabolic profiles based on microbial community structure were also found to be reliable predictors for H2S concentration (R2 = 0.78). These results suggest that microbial community structure contains information sufficient to predict sulfidogenesis in a closed system, with anticipated applications to microbially driven processes in open environments. IMPORTANCE Microbial communities control many biogeochemical processes. Many of these processes are impractical or expensive to measure directly. Because the taxonomic structure of the microbial community is indicative of its function, it encodes information that can be used to predict biogeochemistry. Here, we demonstrate how a machine learning technique can be used to p

Producción Científica, The influence of suspended solids and pH in anaerobically digested piggery wastewater on membrane-based NH3 extraction was evaluated in batch tests. The increase in pH in the anaerobic broth from 8 to 9 resulted in an increase in NH3 removal efficiencies from 15.8 % ± 0.1 % to 20.9 % ± 0.4 % regardless of the suspended solids. The influence of membrane based NH3 extraction on piggery wastewater treatment was also assessed in a CSTR interconnected with PTFE membrane modules. The decrease in TKN concentrations mediated by membrane operation induced an increase in CH4 yield from 380.4 ± 84.9 up to 566.1 ± 7.8 NmLCH4 g VS fed−1. Likewise, COD and VS removal efficiencies significantly increased from 33.0 % ± 2.0 % and 25.7 % ± 2.3 % up to 61.8 % ± 1.3 % and 37.9 % ± 1.8 %, respectively. Interestingly, the decrease in NH3 concentration entailed a complete assimilation of VFA., Junta de Castilla y Leon - Fondo Europeo de Desarrollo Regional (grants VA088G19, CL-EI-2021-07, UIC 315 and UIC 082), Ministerio de Ciencia, Innovación y Universidades (grant PID2019-109403RB-C21/AEI/10.13039/501100011033)

Producción Científica, The application of biogas as a low-priced substrate for the production of ectoines constitutes an opportunity to decrease their production costs and to enhance the viability of anaerobic digestion. The influence of operational conditions on CH4-biogas biodegradation and on ectoines production yields was assessed in continuous pilot bubble column bioreactors. The rise in biomass concentration from 1 to 3 g L-1 resulted in a decrease in the specific ectoine content from 42±8 to 30±4 mgectoine gVSS-1. The concentration of Cu2+ and Mg2+ did not impact process performance, while the use of ammonium as N source resulted in low CH4 biodegradation and ectoine yields (13±7 mgectoine gVSS-1). The increase in CH4 content from 4.5 to 9 %v·v-1 enhanced CH4 removal efficiency. Process operation at NaCl concentrations of 3 %w·w-1 instead of 6 %w·w-1 decreased the ectoine yield to 17 mgectoine gVSS-1. Finally, Methylomicrobium buryatense was identified as the dominant species., Junta de Castilla y León - Fondo Europeo de Desarrollo Regional (projects CLU-2017-09 and UIC-315)

In this review, we focus on the holistic continuous enzymatic production and put special emphasis on process intensification by up- and downstream processing in continuous flow biocatalysis. After a brief introduction, we provide an overview of current examples of enzyme immobilization as an upstream process for flow biocatalysis. Thereafter, we provide an overview of unit operations as downstream processing strategies, namely continuous (i) liquid–liquid extraction, (ii) adsorptive downstream processing, and (iii) crystallization and precipitation. Eventually, we present our perspectives on future trends in this research field.

Monoterpene indole alkaloids (MIAs) are a diverse family of complex plant secondary metabolites with many medicinal properties, including the essential anti-cancer therapeutics vinblastine and vincristine1. As MIAs are difficult to chemically synthesize, the world's supply chain for vinblastine relies on low-yielding extraction and purification of the precursors vindoline and catharanthine from the plant Catharanthus roseus, which is then followed by simple in vitro chemical coupling and reduction to form vinblastine at an industrial scale2,3. Here, we demonstrate the de novo microbial biosynthesis of vindoline and catharanthine using a highly engineered yeast, and in vitro chemical coupling to vinblastine. The study showcases a very long biosynthetic pathway refactored into a microbial cell factory, including 30 enzymatic steps beyond the yeast native metabolites geranyl pyrophosphate and tryptophan to catharanthine and vindoline. In total, 56 genetic edits were performed, including expression of 34 heterologous genes from plants, as well as deletions, knock-downs and overexpression of ten yeast genes to improve precursor supplies towards de novo production of catharanthine and vindoline, from which semisynthesis to vinblastine occurs. As the vinblastine pathway is one of the longest MIA biosynthetic pathways, this study positions yeast as a scalable platform to produce more than 3,000 natural MIAs and a virtually infinite number of new-to-nature analogues.

Environmental concerns are becoming increasingly relevant during a period of hemorrhaging ecosystem goods and services. Restoring these would result in positive outcomes for public health and economic benefit. This thesis seeks to address two environmental concerns: (1) accurate soil mapping and (2) treatment of nitrogen to affect water quality change.The current method of soil mapping, SSURGO (USDA‐NRCS Soil survey), is often erroneous and misleading. Two studies in this dissertation are conducted to evaluate the potential that different resolution digital elevation models (DEMs) have to distribute soil characteristics successfully. These studies are conducted in southwest Virginia and western Vermont. The aforementioned studies evaluated 36 and 59 soil samples, respectively. Spatial characteristics, including slope, catchment area, and topographic wetness, are derived from several DEMs. In chapter 2, these characteristics are spatially compared, and we found that small resolution rasters result in narrow flow paths relative to coarser rasters. In chapter 3, we isolate the analysis to focus on resolution size, instead of a mix of both resolution size and generation method. This is done by recursively coarsening small rasters, deriving spatial attributes from said rasters and evaluating their potential to fit the soil characteristics of interest. Here we found that slopes generated from resolutions smaller than 11m were poor predictors of soil characteristics. Both chapters are finished by proposing and evaluating a soil map. Proposed regressions beat SSURGO in all investigated properties. Furthermore, proposed maps consistently beat out uninformed smallest resolution derived maps.Chesapeake bay water quality managers are struggling to achieve targets for nitrogen loading. This is in part due to the widespread presence of legacy nitrogen. Legacy nitrogen is an emerging issue, and springs exporting high levels of nitrogen are not uncommon in northern Virginia. This t

Producción Científica, The influence of suspended solids and pH in anaerobically digested piggery wastewater on membrane-based NH3 extraction was evaluated in batch tests. The increase in pH in the anaerobic broth from 8 to 9 resulted in an increase in NH3 removal efficiencies from 15.8 % ± 0.1 % to 20.9 % ± 0.4 % regardless of the suspended solids. The influence of membrane based NH3 extraction on piggery wastewater treatment was also assessed in a CSTR interconnected with PTFE membrane modules. The decrease in TKN concentrations mediated by membrane operation induced an increase in CH4 yield from 380.4 ± 84.9 up to 566.1 ± 7.8 NmLCH4 g VS fed−1. Likewise, COD and VS removal efficiencies significantly increased from 33.0 % ± 2.0 % and 25.7 % ± 2.3 % up to 61.8 % ± 1.3 % and 37.9 % ± 1.8 %, respectively. Interestingly, the decrease in NH3 concentration entailed a complete assimilation of VFA., Junta de Castilla y Leon - Fondo Europeo de Desarrollo Regional (grants VA088G19, CL-EI-2021-07, UIC 315 and UIC 082), Ministerio de Ciencia, Innovación y Universidades (grant PID2019-109403RB-C21/AEI/10.13039/501100011033)

Producción Científica, The application of biogas as a low-priced substrate for the production of ectoines constitutes an opportunity to decrease their production costs and to enhance the viability of anaerobic digestion. The influence of operational conditions on CH4-biogas biodegradation and on ectoines production yields was assessed in continuous pilot bubble column bioreactors. The rise in biomass concentration from 1 to 3 g L-1 resulted in a decrease in the specific ectoine content from 42±8 to 30±4 mgectoine gVSS-1. The concentration of Cu2+ and Mg2+ did not impact process performance, while the use of ammonium as N source resulted in low CH4 biodegradation and ectoine yields (13±7 mgectoine gVSS-1). The increase in CH4 content from 4.5 to 9 %v·v-1 enhanced CH4 removal efficiency. Process operation at NaCl concentrations of 3 %w·w-1 instead of 6 %w·w-1 decreased the ectoine yield to 17 mgectoine gVSS-1. Finally, Methylomicrobium buryatense was identified as the dominant species., Junta de Castilla y León - Fondo Europeo de Desarrollo Regional (projects CLU-2017-09 and UIC-315)

Cultured meat has potential to diversify methods for protein production, but innovations in production efficiency will be required to make cultured meat a feasible protein alternative. Microcarriers provide a strategy to culture sufficient volumes of adherent cells in a bioreactor that are required for meat products. However, cell culture on inedible microcarriers involves extra downstream processing to dissociate cells prior to consumption. Here, we present edible microcarriers that can support the expansion and differentiation of myogenic cells in a single bioreactor system. To fabricate edible microcarriers with a scalable process, we used water-in-oil emulsions as templates for gelatin microparticles. We also developed a novel embossing technique to imprint edible microcarriers with grooved topology in order to test if microcarriers with striated surface texture can promote myoblast proliferation and differentiation in suspension culture. In this proof-of-concept demonstration, we showed that edible microcarriers with both smooth and grooved surface topologies supported the proliferation and differentiation of mouse myogenic C2C12 cells in a suspension culture. The grooved edible microcarriers showed a modest increase in the proliferation and alignment of myogenic cells compared to cells cultured on smooth, spherical microcarriers. During the expansion phase, we also observed the formation of cell-microcarrier aggregates or microtissues for cells cultured on both smooth and grooved microcarriers. Myogenic microtissues cultured with smooth and grooved microcarriers showed similar characteristics in terms of myotube length, myotube volume fraction, and expression of myogenic markers. To establish feasibility of edible microcarriers for cultured meat, we showed that edible microcarriers supported the production of myogenic microtissue from C2C12 or bovine satellite muscle cells, which we harvested by centrifugation into a cookable meat patty that maintained it

In this work, the early-stage response of six lab-scale biogas bioreactors fed with different amounts of a sulfate-rich organic agro-industrial effluent was investigated. Biogas characterization, gas chromatography selective for sulfur compounds and high-throughput sequencing of 16S rRNA gene were performed. Hydrogen sulfide (H2S) yield went from transient to steady state in ~ 2 weeks for all the studied conditions. In addition, volatile sulfur compounds (VSCs), like methanethiol (MeSH) and dimethyl sulfide (DMS), were generated at high sulfate loads. Changes were evidenced in the microbial community structures, with a higher abundance of genes involved in the dissimilatory sulfate-reduction pathway in high loaded sulfate bioreactors, as determined by PICRUSt analysis. Principal component analysis (PCA) and correlation analyses evidenced strong relationships between H2S, VSCs and the microbial community. Sulfate-reducing bacteria (SRB) like Desulfocarbo, Desulfocella and Desulfobacteraceae might be possibly linked with methylation processes of H2S, Peer Reviewed, Postprint (author's final draft)

The future of large-scale kelp aquaculture is standing at a crossroad, with the diverging paths being characterized by two fundamentally different cultivation methods that differ on how well gametophyte reproduction can be controlled. The cultivation method that does not directly control gametophyte reproduction is more widely utilized at the moment, but interest in better controlling gametophyte reproduction is growing steadily. Here, we validate a bioreactor system that overcomes a number of implementation challenges for this controlled reproductive method, expanding the possibility of clonal gametophyte cultivation outside of expensive laboratory settings. The main goals of this system include (i) the maintenance of clean gametophyte clonal cultures in non-sterile environments over prolonged periods of time, (ii) the production of large numbers of juvenile sporophytes, and (iii) effective transportation of gametophytes and sporophytes. The “SeaCoRe system” consists out of three parts that correspond to these three challenges: (1) clone-reactors, (2) a clone-inducer, and (3) a transporter. The validation of the system showed that delayed Saccharina latissima and Alaria esculenta gametophytes can grow reliably for 75 days in the clone-reactors. Initial gametophyte densities of 0.4 mg DW and 0.6 mg DW gametophtyes mL−1 were optimal for S. latissima and A. esculenta, resulting in reproductive successes of 604 and 422 sporophytes mL−1, respectively. Lastly, gametophyte transport was simulated, with high reproductive success still achieved within 19 days in ~ 20 °C environments. The SeaCoRe system helps unlock the full potential of large-scale kelp cultivation using multiannual delayed clonal.

[Abstract] Current breeding programs aim to increase the number of ink-tolerant chestnut trees using vegetative propagation of selected genotypes. However, the commercial vegetative propagation of chestnut species is still a bottleneck for the forest industry, mainly due to problems in the rooting and acclimation of propagules. This study aimed to explore the potential benefits of decreasing sucrose supplementation during chestnut micropropagation. Explants were cultured with high light intensity and CO2-enriched air in temporary or continuous immersion bioreactors and with different sucrose supplementation to evaluate the impact of these treatments on growth, rooting and physiological status (monosaccharide content, soluble phenolics and antioxidant activity). The proliferation and rooting performance of shoots cultured by continuous immersion decreased sharply with sucrose concentrations lower than 1%, whereas shoots cultured by temporary immersion grew and rooted successfully with 0.5% sucrose. These results suggest this system is appropriate to culture chestnut with low sucrose concentration and to explore photoautotrophic propagation of this species.

Ieder huis een microbiële moestuin? De dag beginnen met een verse algenshake, afgetapt uit de bioreactor op de vensterbank? Daarmee is het mogelijk het equivalent van een krop sla per dag te produceren, met minder energie dan nodig is voor een krop uit de tuinbouwkas. Zo ver is het nog niet, maar bezoekers van de Floriade Expo kunnen er alvast aan wennen: algen worden in de toekomst steeds belangrijker. We gaan ze overal tegenkomen: in ons voedsel, veevoer, onze cosmetica en biobrandstoffen.

The Anaerobic Fluidized Membrane Bioreactor (AnFMBR) is a membrane–based hybrid technology that can overcome the limitations of conventional anaerobic sewage treatment. Although previous studies have demonstrated excellent performance in the removal of conventional organic pollutants, further research into the removal paths of emerging contaminants (ECs) under various operating conditions is required for proper design and development of the AnFMBR technology. Regarding this, the fate of four ECs in a lab-scale AnFMBR filled with thermoplastic gel for biofilm growth was investigated under various Hydraulic Retention Time (HRT) conditions. When the HRT was 13 h, diclofenac and 17ß-estradiol were efficiently removed at 93% and 72% respectively. Even after an HRT reduction to 6.5 h, the system was still able to maintain high ECs removals (74% for diclofenac and 69% for 17ß-estradiol). However, irrespective of HRT operational condition, smaller removals of 17a-ethinylestradiol (37–52%) were observed, while only marginal removals of amoxicillin were achieved (5–29%). Biotransformation was attributed as the main route for ECs removal. The results obtained in this study indicate that the membrane-based hybrid AnFMBR can be used to treat the target ECs without influence on anaerobic process. The technology had better removal efficiency for diclofenac and 17ß-estradiol. However, the AnFMBR system exhibits high variability in EC removal and low capacity for amoxicillin removal, implying that a combination of other processes is still required to properly avoid the release of these contaminants into the environment., The authors wish to thank the São Paulo Research Foundation (FAPESP) (Grant number: 2016/23684–0) and the National Council for Scientific and Technological Development (CNPq) (Grant number: 456619/2014–3). The authors are grateful to the Laboratory of Urban Wastewater Treatment and Water Reuse (LabTAUS) and the Multiuser Experimental Central of UFABC (CEM-UFABC) for the experimental support., Peer Reviewed, Postprint (author's final draft)

In recent years, the mathematical and computational sciences have developed novel methodologies and insights that can aid in designing advanced bioreactors, microfluidic setups or organ-on-chip devices, in optimizing culture conditions, or predicting long-term behavior of engineered tissues in vivo. In this review, we introduce the concept of computational models and how they can be integrated in an interdisciplinary workflow for Tissue Engineering and Regenerative Medicine (TERM). We specifically aim this review of general concepts and examples at experimental scientists with little or no computational modeling experience. We also describe the contribution of computational models in understanding TERM processes and in advancing the TERM field by providing novel insights. Although in recent years the use of mathematical and computational sciences has increased in the Tissue Engineering and Regenerative Medicine (TERM) field, we believe that a further integration of experimental and computational approaches has a huge potential for advancing the field due to the ability of models to explain and predict experimental results and efficiently optimize TERM product and process designs. By providing an overview of existing computational models, how they have contributed to the field, as well as a future perspective, this review represents an important step to help realize TERM's ultimate goal: a cure instead of care.

Current breeding programs aim to increase the number of ink-tolerant chestnut trees using vegetative propagation of selected genotypes. However, the commercial vegetative propagation of chestnut species is still a bottleneck for the forest industry, mainly due to problems in the rooting and acclimation of propagules. This study aimed to explore the potential benefits of decreasing sucrose supplementation during chestnut micropropagation. Explants were cultured with high light intensity and CO-enriched air in temporary or continuous immersion bioreactors and with different sucrose supplementation to evaluate the impact of these treatments on growth, rooting and physiological status (monosaccharide content, soluble phenolics and antioxidant activity). The proliferation and rooting performance of shoots cultured by continuous immersion decreased sharply with sucrose concentrations lower than 1%, whereas shoots cultured by temporary immersion grew and rooted successfully with 0.5% sucrose. These results suggest this system is appropriate to culture chestnut with low sucrose concentration and to explore photoautotrophic propagation of this species.

Indoor air treatment in different reactor configurations, Indoor air treatment in different reactor configurations

Carbon-based membranes integrated with anaerobic biodegradation are presented as a unique wastewater treatment approach to deal with dye effluents. This study explores the scope of ceramic-supported carbon membrane bioreactors (B-CSCM) and ceramic-supported graphene oxide membrane bioreactors (B-CSGOM) to decolorize azo dye mixtures (ADM) and other dyes. The mixture was prepared using an equimolar composition of monoazo Acid Orange 7, diazo Reactive Black 5, and triazo Direct Blue 71 dye aqueous solution. Afterwards, as in the ADM experiment, both compact units were investigated for their ability in the biodecolorization of Methylene Blue (MB) and Rhodamine B (RhB) dye solutions, which do not belong to the azo family. The obtained outcomes revealed that the conductive surface of the graphene oxide (GO) membrane resulted in a more efficient and higher color removal of all dye solutions than B-CSCM under a wide feed concentration and permeate flux ranges. The maximum color removal at low feed concentration (50 mg·L-1) and permeate flux (0.05 L·m-2·h-1) was 96% for ADM, 98% for MB and 94% for RhB, whereas it was 89%, 94% and 66%, respectively, for B-CSCM. This suggests that the robust, cost-effective, efficient nanostructures of B-CSGOM can successfully remove diverse azo dye solutions from wastewater better than the B-CSCM does., This project has been supported by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 713679 and by the Universitat Rovira i Virgili (URV), contract 2017MFP-COFUND-18. This article was possible thanks to the grant RTI2018-096467-B-I00 funded by MCIN/AEI/10.13039/501100011033 and “ERDF A way of making Europe”. The authors research group is recognized by the Comissionat per a Universitats i Recerca, DIUE de la Generalitat de Catalunya (2017 SGR 396), and supported by the Universitat Rovira i Virgili (2021PFR-URV-87)., Peer Reviewed, Postprint (published version)

Producción Científica, Biogas contain minor concentration of volatile methyl siloxanes (VMS), responsible for severe damages in turbines or internal combustion engines. Sustainable biological processes for VMS abatement are limited by the low aqueous solubility of VMS. In order this limitation, the siloxanes (D4, D5, L2 and L3) removal performance of a two-phase partitioning biotrickling filter (TP-BTF) was study in terms of the empty bed residence time (EBRT) and the fraction of the organic phase (silicone oil). A decrease in the total VMS removal from 76 to 49% was observed when the EBRT was reduced from 60 to 15 min. The highest removals were achieved for D4 (53–84%) and D5 (69–87%), compared to the lower values recorded for L2 (19–45%) and L3 (31–81%). The increase in the share of silicone oil in the recycling mineral medium from 5 to 45% resulted in an improvement of the total VMS abatement from 35 to 52%. This enhancement was observed for L3 (21–50%), D4 (26–64%) and D5 (58–78%), whereas L2 removals remained < 25%. A highly specialized bacterial community dominated by the genus KCM-B-112 was retrieved at the end of the experiment., Bio Based Industries Joint Undertaking (Horizon 2020 grant 745785), Junta de Castilla y León (grants CLU 2017-09 and UIC 071), European Commission-H2020-MSCA–IF–2019 (project 897284)

[ES] Los mini-biorreactores son una herramienta de gran interés en los laboratorios de biología sintética. Estos mini-biorreactores se encargan de monitorizar y controlar las condiciones de los cultivos de bacterias para los experimentos (entrada y salida de medio, agitacíon del cultivo, aireación, etc.). En este proyecto, el objetivo principal consiste en la implementación de un bucle de control de temperatura mediante un sensor de infrarrojos. Se diseñarán e implementarán leyes de control novedosas basadas en condicionamiento de referencia para hacer frente a la fuerte diferencia entre la dinámica de calentamiento y la de enfriamiento. Objetivos adicionales serán: - Implementar derivadores software para estimar la derivada de la temperatura en el biorreactor. - Implementar un simulador embebido del biorreactor. - Utilización de Github para el control de versiones del código desarrollado. - Calibración del sistema: bomba peristáltica, sensor de densidad óptica, sensor de fluorescencia y del sensor de temperatura basado en infrarrojos. El sensor de densidad óptica y el de fluorescencia ya están implementados, asíi como su software asociado. - Pruebas experimentales para validar el control de temperatura, [EN] Mini-bioreactors are a tool of great interest in synthetic biology laboratories. These mini-bioreactors are in charge of monitoring and controlling the conditions of the bacteria cultures for the experiments (inlet and outlet of medium, shaking the culture, aeration, etc.). In this project, the main objective is to implement a temperature control loop using an infrared sensor. Novel control laws based on reference conditioning will be designed and implemented to cope with the strong difference between heating and cooling dynamics. As secondary objectives, the calibration of the sensor, experimental tests to validate the temperature control will be carried out and, finally, work will be carried out towards the start-up of the mini-bioreactor. Additional objectives will be: - Implement software derivatives to estimate the derivative of the temperature in the bioreactor. - Implement an embedded simulator of the bioreactor. - Use of Github for version control of the developed code. - Calibration of the system: peristaltic pump, optical density sensor, fluorescence sensor and infrared-based temperature sensor. The optical density sensor and the fluorescence sensor are already implemented, as well as their associated software. - Experimental tests to validate the temperature control

[ES] Con la ayuda de la biotecnología, es posible producir metabolitos de interés (por ejemplo, aminoácidos, biocombustibles, fármacos) en células cultivadas en biorreactores. Este proyecto busca diseñar e implementar un sistema de placas de milifluídica para automatizar las tareas de alimentación de medio fresco, control de volumen y muestreo para un sistema de biorreactores de laboratorio de pequeña escala (10 ml a 100 ml) para su uso en aplicaciones de biología sintética. Tras revisar el estado del arte, se diseñan e implementan prototipos de validación para probar el proceso de fabricación y las funciones básicas de las placas de milifluídica. El proceso de fabricación consiste en la preparación del material con un adhesivo, el procesamiento mediante una cortadora láser y el montaje de las placas. Tras la calibración de la bomba peristáltica, se realizan pruebas experimentales de los prototipos de validación para validar el sistema y mejorar el diseño y el proceso de fabricación. Posteriormente, se diseña y fabrica un prototipo para controlar cuatro biorreactores en paralelo. Se desarrolla un simulador para comparar diferentes estrategias y parámetros de control para los experimentos que utilizan el sistema milifluídico. El simulador permite elegir cualquier número de biorreactores a controlar, lo que lo hace aplicable a una amplia gama de configuraciones experimentales. Finalmente, se discuten los aspectos de escalabilidad del sistema., [EN] With the help of biotechnology, it is possible to produce metabolites of interest (e.g. amino acids, biofuel, drugs) in cells that are cultured in bioreactors. This project seeks to design and implement a millifluidic board system to automate the tasks of fresh medium feeding, volume control and sampling for a small scale (10 ml to 100 ml) laboratory bioreactor system for use in synthetic biology applications. After reviewing the state of the art, validation prototypes are designed and implemented to test the manufacturing process and the basic functions of the millifluidic boards. The manufacturing process consists of preparation of the material with adhesive, laser processing and the assembly. After calibration of the peristaltic pump, experimental tests of the validation prototypes are carried out to validate the system and to improve the design and manufacturing process. Subsequently, a prototype for controlling four bioreactors in parallel is designed and manufactured. A simulator is developed to compare different control strategies and parameters for experiments which use the millifluidic system. The simulator allows choosing any number of bioreactors to control thus making it applicable to a wide range of experimental setups. Finally, the scalability aspects of the system are discussed.

[ES] Con la ayuda de la biotecnología, es posible producir metabolitos de interés (por ejemplo, aminoácidos, biocombustibles, fármacos) en células cultivadas en biorreactores. Este proyecto busca diseñar e implementar un sistema de placas de milifluídica para automatizar las tareas de alimentación de medio fresco, control de volumen y muestreo para un sistema de biorreactores de laboratorio de pequeña escala (10 ml a 100 ml) para su uso en aplicaciones de biología sintética. Tras revisar el estado del arte, se diseñan e implementan prototipos de validación para probar el proceso de fabricación y las funciones básicas de las placas de milifluídica. El proceso de fabricación consiste en la preparación del material con un adhesivo, el procesamiento mediante una cortadora láser y el montaje de las placas. Tras la calibración de la bomba peristáltica, se realizan pruebas experimentales de los prototipos de validación para validar el sistema y mejorar el diseño y el proceso de fabricación. Posteriormente, se diseña y fabrica un prototipo para controlar cuatro biorreactores en paralelo. Se desarrolla un simulador para comparar diferentes estrategias y parámetros de control para los experimentos que utilizan el sistema milifluídico. El simulador permite elegir cualquier número de biorreactores a controlar, lo que lo hace aplicable a una amplia gama de configuraciones experimentales. Finalmente, se discuten los aspectos de escalabilidad del sistema., [EN] With the help of biotechnology, it is possible to produce metabolites of interest (e.g. amino acids, biofuel, drugs) in cells that are cultured in bioreactors. This project seeks to design and implement a millifluidic board system to automate the tasks of fresh medium feeding, volume control and sampling for a small scale (10 ml to 100 ml) laboratory bioreactor system for use in synthetic biology applications. After reviewing the state of the art, validation prototypes are designed and implemented to test the manufacturing process and the basic functions of the millifluidic boards. The manufacturing process consists of preparation of the material with adhesive, laser processing and the assembly. After calibration of the peristaltic pump, experimental tests of the validation prototypes are carried out to validate the system and to improve the design and manufacturing process. Subsequently, a prototype for controlling four bioreactors in parallel is designed and manufactured. A simulator is developed to compare different control strategies and parameters for experiments which use the millifluidic system. The simulator allows choosing any number of bioreactors to control thus making it applicable to a wide range of experimental setups. Finally, the scalability aspects of the system are discussed.

Biofilms play an important role in degradation, transformation and assimilation of anthropogenic pollutants in aquatic ecosystems. In this study, we assembled a tubular bioreactor containing a biofilm substrate and aeration device, which was introduced into mesocosms to explore the effects of bioreactor on physicochemical and microbial characteristics of a hypereutrophic urban river. The biofilm bioreactor greatly improved water quality, especially by decreasing dissolved inorganic nitrogen (DIN) concentrations, suggesting that biofilms were the major sites of nitrification and denitrification with an oxygen concentration gradient. The biofilm bioreactor increased the abundance of planktonic bacteria, whereas diversity of the planktonic microbial community decreased. Sequencing revealed that Proteobacteria, Bacteroidetes, Planctomycetes, and Actinobacteria were the four predominant phyla in the planktonic microbial community, and the presence of the biofilm bioreactor increased the relative abundance of Proteobacteria. Variations in microbial communities were most strongly affected by the presence of the biofilm bioreactor, as indicated by principal component analysis (PCA) and redundancy analysis (RDA). This study provides valuable insights into changes in ecological characteristics associated with self-purification processes in hypereutrophic urban rivers, and may be of important for the application of biofilm bioreactor in natural urban river.

Microorganisms play vital roles in modulating organic matter decomposition and nutrient cycling in soil ecosystems. The enzyme latch paradigm posits microbial degradation of polyphenols is hindered in anoxic peat leading to polyphenol accumulation, and consequently diminished microbial activity. This model assumes that polyphenols are microbially unavailable under anoxia, a supposition that has not been thoroughly investigated in any soil type. Here, we use anoxic soil reactors amended with and without a chemically defined polyphenol to test this hypothesis, employing metabolomics and genome-resolved metaproteomics to interrogate soil microbial polyphenol metabolism. Challenging the idea that polyphenols are not bioavailable under anoxia, we provide metabolite evidence that polyphenols are depolymerized, resulting in monomer accumulation, followed by the generation of small phenolic degradation products. Further, we show that soil microbiome function is maintained, and possibly enhanced, with polyphenol addition. In summary, this study provides chemical and enzymatic evidence that some soil microbiota can degrade polyphenols under anoxia and subvert the assumed polyphenol lock on soil microbial metabolism.

Retinal degeneration is a leading cause of vision impairment and blindness worldwide and medical care for advanced disease does not exist. Stem cell-derived retinal organoids (RtOgs) became an emerging tool for tissue replacement therapy. However, existing RtOg production methods are highly heterogeneous. Controlled and predictable methodology and tools are needed to standardize RtOg production and maintenance. In this study, we designed a shear stress-free micro-millifluidic bioreactor for nearly labor-free retinal organoid maintenance. We used a stereolithography (SLA) 3D printer to fabricate a mold from which Polydimethylsiloxane (PDMS) was cast. We optimized the chip design using in silico simulations and in vitro evaluation to optimize mass transfer efficiency and concentration uniformity in each culture chamber. We successfully cultured RtOgs at three different differentiation stages (day 41, 88, and 128) on an optimized bioreactor chip for more than 1 month. We used different quantitative and qualitative techniques to fully characterize the RtOgs produced by static dish culture and bioreactor culture methods. By analyzing the results from phase contrast microscopy, single-cell RNA sequencing (scRNA seq), quantitative polymerase chain reaction (qPCR), immunohistology, and electron microscopy, we found that bioreactor-cultured RtOgs developed cell types and morphology comparable to static cultured ones and exhibited similar retinal genes expression levels. We also evaluated the metabolic activity of RtOgs in both groups using fluorescence lifetime imaging (FLIM), and found that the outer surface region of bioreactor cultured RtOgs had a comparable free/bound NADH ratio and overall lower long lifetime species (LLS) ratio than static cultured RtOgs during imaging. To summarize, we validated an automated micro-millifluidic device with significantly reduced shear stress to produce RtOgs of comparable quality to those maintained in conventional static culture.