Mesenchymal stem and stromal cells (MSCs) hold potential to treat a broad range of clinical indications, but clinical translation has been limited to date due in part to challenges with batch-to-batch reproducibility of potential critical quality attributes (pCQAs) that can predict potency/efficacy. Here, we designed and implemented a microcarrier-microbioreactor approach to cell therapy manufacturing, specific to anchorage-dependent cells such as MSCs. We sought to assess whether increased control of the biochemical and biophysical environment had the potential to create product with consistent presentation and elevated expression of pCQAs relative to established manufacturing approaches in tissue culture polystyrene (TCPS) flasks. First, we evaluated total cell yield harvested from dissolvable, gelatin microcarriers within a microbioreactor cassette (Mobius Breez) or a flask control with matched initial cell seeding density and culture duration. Next, we identified 24 genes implicated in a therapeutic role for a specific motivating indication, acute respiratory distress syndrome (ARDS); expression of these genes served as our pCQAs for initial in vitro evaluation of product potency. We evaluated mRNA expression for three distinct donors to assess inter-donor repeatability, as well as for one donor in three distinct batches to assess within-donor, inter-batch variability. Finally, we assessed gene expression at the protein level for a subset of the panel to confirm successful translation. Our results indicated that MSCs expanded with this microcarrier-microbioreactor approach exhibited reasonable donor-to-donor repeatability and reliable batch-to-batch reproducibility of pCQAs. Interestingly, the baseline conditions of this microcarrier-microbioreactor approach also significantly improved expression of several key pCQAs at the gene and protein expression levels and reduced total media consumption relative to TCPS culture. This proof-of-concept study illustrates key benefits of this approach to therapeutic cell process development for MSCs and other anchorage-dependent cells that are candidates for cell therapies.

Over the recent years, ever-increasing population growth and higher wastewater production has been a challenge for decentralized wastewater treatment plants (WWTPs). In addition, sludge treatment due to high cost for equipment and place make authorities to find a sustainable approach in both of economical and technical perspectives. One of the proposed solutions is transferring the sludge produced from decentralized WWTP to centralized WWTP. However, the appropriate proportional ratio of raw sludge to raw sewage is a challenge, otherwise, it make anaerobic conditions and sewage rotting along the sewer network based on permissible limit of dihydrogen sulfide (H2S) gas (5 ppm). In the present study, seven reactors with different ratios of sludge to raw sewage (0, 15, 20, 25, 50, 75, 100) were used to stimulate the feasibility of transferring Shahrake Gharb WWTP sludge along the wastewater transfer pipe to the centralized sewage treatment south Tehran WWTP plant in Tehran, Iran. The septic situation and H2S emission of different reactors within 7 h (Time to reach the compound in the south treatment plant) was analyzed by gas meter. The results indicated that the optimum ratio of sludge to raw sewage was 15% without H2S production during 7 h. In addition, due to the high volume of sludge produced by the Shahrake Gharb WWTP, the optimal ratio of lime to total solids (TS) in sludge (gr/gr) (0.6) increased the sludge loading rate from 15 to 30% without any H2S emission during the stimulation study period. Therefore, the lime stabilization and transfer of sludge from a decentralized WWTP to a centralized WWTP is a feasible way to manage the sludge and enhance the treatment capacity in local WWTP.

Biological approaches and coagulation are frequently used to reduce the chemical oxygen demand (COD) for treatment of ceramic effluent water. The technology known as the moving bed biofilm reactor (MBBR) can accomplish this goal. Further, the process of emulsification-aided innovative MBBR using biosurfactants can be proposed for ceramic effluent treatment. In a step-by-step upgrading scheme, biosurfactants and a consortia of halophilic and halotolerant microbial culture was utilized for the treatment of the effluent water. Over the course of 21 days, a progressive decrease in COD of up to 95.79% was achieved. Over the next 48 h period, the biochemical oxygen demand (BOD) was reduced by 98.3%, while total suspended solids (TSS) decreased by 79.41%. With the use of this innovative MBBR technology, biofilm formation accelerated, lowering the COD, BOD, and TSS levels. This allows treated water to be used for further research on recycling it back into the ceramics sector and repurposing it for agricultural purposes. PRACTITIONER POINTS: Implementation of modified MBBR technology for the treatment of effluent water. Biosurfactants could reduce in the organic and inorganic loads. Increase in MLSS values with COD removal observed. The plant operations without the use of chemical coagulants was effective with biosurfactants. Biofilm formation on carriers was scraped and the presence of surfactin and rhamnolipid was confirmed.

Digital transformation for the water sector has gained momentum in recent years, and many water resource recovery facilities modelers have already started transitioning from developing traditional models to digital twin (DT) applications. DTs simulate the operation of treatment plants in near real time and provide a powerful tool to the operators and process engineers for real-time scenario analysis and calamity mitigation, online process optimization, predictive maintenance, model-based control, and so forth. So far, only a few mature examples of full-scale DT implementations can be found in the literature, which only address some of the key requirements of a DT. This paper presents the development of a full-scale operational DT for the Eindhoven water resource recovery facility in The Netherlands, which includes a fully automated data-pipeline combined with a detailed mechanistic full-plant process model and a user interface co-created with the plant\'s operators. The automated data preprocessing pipeline provides continuous access to validated data, an influent generator provides dynamic predictions of influent composition data and allows forecasting 48 h into the future, and an advanced compartmental model of the aeration and anoxic bioreactors ensures high predictive power. The DT runs near real-time simulations every 2 h. Visualization and interaction with the DT is facilitated by the cloud-based TwinPlant technology, which was developed in close interaction with the plant\'s operators. A set of predefined handles are made available, allowing users to simulate hypothetical scenarios such as process and equipment failures and changes in controller settings. The combination of the advanced data pipeline and process model development used in the Eindhoven DT and the active involvement of the operators/process engineers/managers in the development process makes the twin a valuable asset for decision making with long-term reliability. PRACTITIONER POINTS: A full-scale digital twin (DT) has been developed for the Eindhoven WRRF. The Eindhoven DT includes an automated continuous data preprocessing and reconciliation pipeline. A full-plant mechanistic compartmental process model of the plant has been developed based on hydrodynamic studies. The interactive user interface of the Eindhoven DT allows operators to perform what-if scenarios on various operational settings and process inputs. Plant operators were actively involved in the DT development process to make a reliable and relevant tool with the expected added value.

Although anaerobic digestion is the mainstream technology for treating food waste (FW), the high pollutant concentration in the resultant food waste anaerobic digestate (FWAD) often poses challenges for the subsequent biochemical treatment such as activated sludge process. In this study, taking a typical FW treatment plant as an example, we analyzed the reasons behind the difficulties in treating FWAD and tested a novel process called as bio-conditioning dewatering followed by activated sludge process (BDAS) to purify FWAD. Results showed that high concentrations of suspended solids (SS) (16439 ± 475 mg/L), chemical oxygen demand (COD) (24642 ± 1301 mg/L), and ammonium nitrogen (NH4+-N) (2641 ± 52 mg/L) were main factors affecting the purification efficiency of FWAD by the conventional activated sludge process. By implementing bio-conditioning dewatering for solid-liquid separation, near 100% of SS and total phosphorus (TP), 90% of COD, 38% of total nitrogen (TN), and 37% of NH4+-N in the digestate could be effectively removed or recovered, consequently generating the transparent filtrate with relatively low pollution load and dry sludge cake (<60% of moisture content). Furthermore, after ammonia stripping and biochemical treatment, the effluent met the relevant discharge standards regulated by China, with the concentrations of COD, TN, NH4+-N, and TP ranging from 151 to 405, 10-56, 0.9-31, and 0.4-0.8 mg/L, respectively. This proposed BDAS approach exhibited stable performance and low operating costs, offering a promising solution to purify FWAD in practical engineering and simultaneously realize resource recovery.

A continuous stirred tank bioreactor (CSTB) with cell recycling combined with ceramic membrane technology and inoculated with Rhodococcus opacus PD630 was employed to treat petroleum refinery wastewater for simultaneous chemical oxygen demand (COD) removal and lipid production from the retentate obtained during wastewater treatment. In the present study, the COD removal efficiency (CODRE) (%) and lipid concentration (g/L) were predicted using two artificial intelligence models, i.e., an artificial neural network (ANN) and a neuro-fuzzy neural network (NF-NN) with a network topology of 6-25-2 being the best for NF-NN. The results revealed the superiority of NF-NN over ANN in terms of determination coefficient (R2), root mean square error (RMSE), and mean absolute percentage error (MAPE). Three learning algorithms were tested with NF-NN; among them, the Bayesian regularization backpropagation (BR-BP) outperformed others. The sensitivity analysis revealed that, if solid retention time and biomass concentrations were maintained between 35 and 75 h and 3.0 g/L and 3.5 g/L, respectively, high CODRE (93%) and lipid concentration (2.8 g/L) could be obtained consistently.

Biosolid management is becoming one of the most crucial issues for wastewater treatment plant (WWTP) operators. The application of the Oxic Settling Anaerobic (OSA) process allows the minimisation of excess sludge production. This study compares conventional activated sludge (CAS) and OSA layouts in a full-scale WWTP (namely, Corleone - Italy). Extensive monitoring campaigns were conducted to assess treatment performances regarding carbon and nutrient removal, greenhouse gas (GHG) emissions, excess sludge production, and biomass activity (by means of respirometric analysis). Results showed that the effluent quality consistently met the Italian discharge limits. However, with the implementation of the OSA process, there was a decrease in ammonium removal efficiency, which could be attributed to reduced nitrifier activity related to reduced biomass production and extended anaerobic conditions affecting the nitrification process. On the other hand, the OSA configuration significantly increased phosphorus removal, indicating a high phosphorus content in the resulting waste sludge. A worsening of the sludge settling properties was observed with the OSA configuration likely due to decreased EPS concentrations. The sludge production in the OSA configuration decreased by 17.3 % compared to CAS. Nitrous-oxide measurements did not show a variation between CAS and OSA configurations, confirming that the OSA process can be a suitable solution for reducing WWTP\'s carbon footprint.

In the current transition to intensified upstream processing, the risks of adopting traditional single-use systems for high-titer, long-duration perfusion cultures, have thus far not been considered. This case study uses the Failure Modes and Effects Analysis (FMEA) method to evaluate the risks associated with implementing upstream single-use technology. The simulated model process was used to compare the risk level of single-use technology for a traditional fed-batch cell culture with that for perfusion culture, under the same annual protein production conditions. To provide a reasonable source of potential risk for FMEA, all single-use upstream operations for both fed-batch and perfusion processes were investigated using an analytical method developed to quantify the impact of process parameters and operating conditions on single-use system specifications and to ensure objectivity. Many of the risks and their levels, were similar in long-duration perfusion cultures and fed-batch cultures. However, differences were observed for high-risk components such as daily sampling and installation. The result of this analysis indicates that the reasons for risk are different for fed-batch cultures and perfusion cultures such as larger bioreactors in fed-batch and longer runs in perfusion, respectively. This risk assessment method could identify additional control measures and be part of a holistic contamination control strategy and help visualize their effectiveness.

In recent years, Moving Bed Biofilm Reactors (MBBRs) have been preferred to conventional processes with suspended biomass. The main reason for this preference is that it can achieve better removal efficiencies than conventional systems with smaller footprints. However, unlocking the full potential of MBBRs in large-scale WWTPs remains challenging in real life. In this study, the performance of three different treatment technologies, Extended Aeration Activated Sludge (EAAS), Hybrid Fixed Bed Biofilm Reactor (HFBBR), and Hybrid Moving Bed Biofilm Reactor (HMBBR), was investigated over a year in a WWTP located in El-Gouna, Egypt. The COD removal efficiencies of the three systems were comparable, with the EAAS achieving 93.5%, HFBBR 94%, and HMBRR 95%. Nevertheless, the NH4 removal efficiency of the EAAS was slightly lower (97.5%) than that of the HFBBR and the HMBBR, that achieved a removal efficiency of 98%. BioWin Software was able to mimic the real case of the WWTP of El-Gouna and critically defined all plant limitations and operational data. Different simulations were modeled to test the hydraulic and organic loading capacities of the three systems under different scenarios and operating conditions. The HMBBR system failed to withstand the increase in organic load because of the biomass sloughing effect and subsequently high TSS loads in the settlers. Biomass sloughing overloaded the settlers and lead to biomass loss in the effluent. As the settleability of the HMBBR sludge was significantly lower than for the HFBBR the TSS loss in the effluent happened that much earlier that the moving carrier application had an adverse effect contradicting with the primary purpose of adding media carriers. Model simulations and data analysis findings were used to recommend the most suitable configuration for upgrading an existing system using the attached growth technique with all kinetic parameters and operational conditions. The recommended configuration focuses mainly on the separation of plastic media in a compartment with a very low hydraulic retention time to absorb the incoming shock load.

The study estimated the risk due to Cryptosporidium, Giardia, and Ascaris, associated with non-potable water reuse in the city of Jaipur, India. The study first determined the exposure dose of Cryptosporidium, Giardia, and Ascaris based on various wastewater treatment technologies for various scenarios of reuse for six wastewater treatment plants (WWTPs) in the city. The exposure scenarios considered were (1) garden irrigation; (2) working and lounging in the garden; and (3) consumption of crops irrigated with recycled water. The estimated annual risk of infection varied between 8.57 × 10-7 and 1.0 for protozoa and helminths, respectively. The order of treatment processes, in decreasing order of annual risk of infection, was found to be: moving-bed bioreactor (MBBR) technology > activated sludge process (ASP) technology > sequencing batch reactor (SBR) technology. The estimated annual risk was found to be in this order: Ascaris > Giardia > Cryptosporidium. The study also estimated the maximum allowable concentration (Cmax) of pathogen in the effluent for a benchmark value of annual infection of risk equal to 1:10,000, the acceptable level of risk used for drinking water. The estimated Cmax values were found to be 6.54 × 10-5, 1.37 × 10-5, and 2.89 × 10-6 (oo) cysts/mL for Cryptosporidium, Giardia, and Ascaris, respectively.