Microalgae to biodiesel: A novel green conversion method for high-quality lipids recovery and in-situ transesterification to fatty acid methyl esters.

Greenhouse gases (GHGs) emissions due to increasing energy demand have raised the need to identify effective solutions to produce clean and renewable energy. Biotechnologies are an effective platform to attain green transition objectives, especially when synergically integrated to promote health and environmental protection. In this context, microalgae-based biotechnologies are considered among the most effective tools for treating gaseous effluents and simultaneously capturing carbon sources for further biomass valorisation. The production of biodiesel is regarded as a promising avenue for harnessing value from residual algal biomass. Nonetheless, the existing techniques for extracting lipids still face certain limitations, primarily centred around the cost-effectiveness of the process.This study is dedicated to developing and optimising an innovative and cost-efficient technique for extracting lipids from algal biomass produced during gaseous emissions treatment based on algal-bacterial biotechnology. This integrated treatment technology combines a bio-scrubber for degrading gaseous contaminants and a photobioreactor for capturing the produced CO2 within valuable algal biomass. The cultivated biomass is then processed with the process newly designed to extract lipids simultaneously transesterificated in fatty acid methyl esters (FAME) via In Situ Transesterification (IST) with a Kumagawa-type extractor. The results of this study demonstrated the potential application of the optimised method to overcome the gap to green transition. Energy production was obtained from residuals produced during the necessary treatment of gaseous emissions. Using hexane-methanol (v/v = 19:1) mixture in the presence KOH in Kumagawa extractor lipids were extracted with extraction yield higher than 12% and converted in fatty acid methyl esters. The process showed the enhanced extraction of lipids converted in bio-sourced fuels with circular economy approach, broadening the applicability of biotechnologies as sustainable tools for energy source diversification.

Instrumental Odour Monitoring System Classification Performance Optimization by Analysis of Different Pattern-Recognition and Feature Extraction Techniques.

Instrumental odour monitoring systems (IOMS) are intelligent electronic sensing tools for which the primary application is the generation of odour metrics that are indicators of odour as perceived by human observers. The quality of the odour sensor signal, the mathematical treatment of the acquired data, and the validation of the correlation of the odour metric are key topics to control in order to ensure a robust and reliable measurement. The research presents and discusses the use of different pattern recognition and feature extraction techniques in the elaboration and effectiveness of the odour classification monitoring model (OCMM). The effect of the rise, intermediate, and peak period from the original response curve, in collaboration with Linear Discriminant Analysis (LDA) and Artificial Neural Networks (ANN) as a pattern recognition algorithm, were investigated. Laboratory analyses were performed with real odour samples collected in a complex industrial plant, using an advanced smart IOMS. The results demonstrate the influence of the choice of method on the quality of the OCMM produced. The peak period in combination with the Artificial Neural Network (ANN) highlighted the best combination on the basis of high classification rates. The paper provides information to develop a solution to optimize the performance of IOMS.

The evolution of compost stability and maturity during the full-scale treatment of the organic fraction of municipal solid waste.

Composting is the method most commonly applied worldwide for the recovery of the source sorted organic waste. The process aims at stabilizing the organic matter, so as to produce a material with soil improver properties, referred to as compost. The effective recovery of the organic waste fraction via composting implies compost safe use on soil. In this view, the assessment of compost characteristics, depending on both the quality of the input material and the process operation, is fundamental. At full scale, the process monitoring usually relies on parameters enabling the indirect control of its evolution, whereas the biological stability and maturity are usually evaluated on the final product. Aim of this work was in assessing both biological stability and maturity during the composting process of the organic fraction of municipal solid waste performed at industrial scale. Representative samples were collected over time in a composting facility operating in the South of Italy and analysed by the dynamic respirometric index, the content of humic substances as well as by their phytotoxicity. Results showed the key role of stability and maturity parameters in the monitoring of composting processes. Experimental outcomes further addressed wider considerations on the operational procedures for a sustainable compost production process.

Bioleaching of metals from WEEE shredding dust.

A bioleaching process developed in two separate steps was investigated for the recovery of base metals, precious metals and rare earth elements from dusts generated by Waste Electrical and Electronic Equipment (WEEE) shredding. In the first step, base metals were almost completely leached from the dust in 8 days by Acidithiobacillus thiooxidans (DSM 9463) that lowered the pH of the leaching solution from 3.5 to 1.0. During this step, cerium, europium and neodymium were mobilized at high percentages (>99%), whereas lanthanum and yttrium reached an extraction yield of 80%. In the second step, the cyanide producing Pseudomonas putida WSC361 mobilized 48% of gold within 3 h from the A. thiooxidans leached shredding dust. This work demonstrated the potential application of biohydrometallurgy for resource recovery from WEEE shredding dust, destined to landfill disposal, and its effectiveness in the extraction of valuable substances, including elements at high supply risk as rare earths.

Control of fouling formation in membrane ultrafiltration by ultrasound irradiation.

The increasing application of membrane filtration in water and wastewater treatment necessitates techniques to improve performance, especially in fouling control. Ultrasound is one promising technology for this purpose as cavitational effects facilitate continuous cleaning of the membrane. This research studied the ultrafiltration of lake water in systems with constant permeate flux under medium frequency (45 kHz) ultrasound irradiation. Fouling was investigated by monitoring transmembrane pressure (TMP) using continuous or intermittent ultrasound irradiation and dead-end or crossflow operation. Best performance was observed with continuous ultrasound irradiation in crossflow mode. Intermittent irradiation reduced the rate of TMP build-up but nevertheless allowed irreversible fouling to develop.

Enhanced ozonation of selected pharmaceutical compounds by sonolysis.

In search of new options to achieve removal of pharmaceuticals in the environment, combined ultrasound and ozonation has become a focus of intense investigation for wastewater treatment. In this study, three pharmaceuticals were selected as model compounds for degradation experiments: diclofenac (DCF), sulfamethoxazole (SMX) and carbamazepine (CBZ). Comparison of the degradation rates for both ozonation and combined ultrasound/ozonation treatments was performed on single synthetic solutions as well as on a mixture of the selected pharmaceuticals, under different experimental conditions. For single synthetic solutions, the efficiency removal for ozonation reached 73%, 51% and 59% after 40 min for DCF, SMX and CBZ, respectively. Comparable results were obtained for pharmaceuticals in mixture. However, the combined ultrasound/ozone treatment was found to increase degradation efficiencies for both DCF and SMX single solutions up to 94% and 61%, respectively, whereas lower removal yields, up to 56%, was noted for CBZ. Likewise, when the combined treatment was applied to the mixture, relatively low removal efficiencies was found for CBZ (44%) and 90% degradation yield was achieved for DCF.

Carbon capture and utilisation (CCU) solutions: Assessing environmental, economic, and social impacts using a new integrated methodology.

Carbon Capture and Utilisation (CCU) technologies play a significant role in climate change mitigation, as these platforms aim to capture and convert CO2 that would be otherwise emitted into the atmosphere. Effective and economically sustainable technologies are crucial to support the transition to renewable and low-carbon energy sources by 2030 and beyond. Currently, studies exploring the financial viability of CCU technologies besides the joint analyses of life-cycle costs and environmental and social impacts are still limited. In this context, the study developed and validated an innovative and integrated methodology, called Life Cycle Cost and Sustainability Assessment (LCC-SA) which allows the joint assessment of (i) project life-cycle costs, (ii) socio-cultural and environmental externalities. This tool was validated with an application to an algal photobioreactors (PBRs) and allowed to assess the economic and environmental sustainability besides identifying the main critical issues to be addressed during the transition from pilot-scale plant to industrial application. The methodology's implementation estimated benefits in two main areas: (i) environmental, including CO2 removal and avoidance through biodiesel production instead of fossil-derived diesel; (ii) socio-cultural, encompassing new patents, knowledge spillovers, human capital formation, and knowledge outputs. The analysis returned as main result that the present value of the social externalities amounts to around EUR 550,000 and the present value of the costs to approximately EUR 60,000. The Economic Net Present Value (ENPV) is EUR 487,394, which shows the significance of the extra-financial effects generated by the research project. At full-scale application, environmental benefits include capturing 187 to 1867 tons of CO2 per year and avoiding 1.7 to 16.7 tons of CO2 annually through biodiesel production instead of fossil-derived diesel.

Integrating artificial intelligence modeling and membrane technologies for advanced wastewater treatment: Research progress and future perspectives.

Membrane technologies have become proficient alternatives for advanced wastewater treatment, ensuring high contaminant removal and sustainable resource recovery. Despite significant progress, ongoing research efforts aim to further optimize treatment performance. Among the challenges faced, membrane fouling persists as a relevant obstacle in membrane technologies, necessitating the development of more effective mitigation strategies. Mathematical models, widely employed for predicting treatment performance, generally exhibit low accuracy and suffer from uncertainties due to the complex and variable nature of wastewater. To overcome these limitations, numerous studies have proposed artificial intelligence (AI) modeling to accurately predict membrane technologies' performance and fouling mechanisms. This approach aims to provide advanced simulations and predictions, thereby enhancing process control, optimization, and intensification. This literature review explores recent advancements in modeling membrane-based wastewater treatment processes through AI models. The analysis highlights the enormous potential of this research field in enhancing the efficiency of membrane technologies. The role of AI modeling in defining optimal operating conditions, developing effective strategies for membrane fouling mitigation, enhancing the performance of novel membrane-based technologies, and improving membrane fabrication techniques is discussed. These enhanced process optimization and control strategies driven by AI modeling ensure improved effluent quality, optimized resource consumption, and minimized operating costs. The potential contribution of this cutting-edge approach to a paradigm shift toward sustainable wastewater treatment is examined. Finally, this review outlines future perspectives, emphasizing the research challenges that require attention to overcome the current limitations hindering the integration of AI modeling in wastewater treatment plants.

Carbon neutrality in wastewater treatment plants: An integrated biotechnological-based solution for nutrients recovery, odour abatement and CO2 conversion in alternative energy drivers.

Wastewater treatment plants play a crucial role in water security and sanitation, ensuring ecosystems balance and avoiding significant negative effects on humans and environment. However, they determine also negative pressures, including greenhouse gas and odourous emissions, which should be minimized to mitigate climate changes besides avoiding complaints. The research has been focused on the validation of an innovative integrated biological system for the sustainable treatment of complex gaseous emissions from wastewater treatment plants. The proposed system consists of a moving bed biofilm reactor coupled with an algal photobioreactor, with the dual objective of: i) reducing the inlet concentration of the odourous contaminants (in this case, hydrogen sulphide, toluene and p-xylene); ii) capturing and converting the carbon dioxide emissions produced by the degradation process into exploitable algal biomass. The first reactor promoted the degradation of chemical compounds up to 99.57% for an inlet load (IL) of 22.97 g m-3 d-1 while the second allowed the capture of the CO2 resulting from the degradation of gaseous compounds, with biofixation rate up to 81.55%. The absorbed CO2 was converted in valuable feedstocks, with a maximum algal biomass productivity in aPBR of 0.22 g L-1 d-1. Dairy wastewater has been used as alternative nutrient source for both reactors, with a view of reusing wastewater while cultivating biomass, framing the proposed technology in a context of a biorefinery within a circular economy perspective. The biomass produced in the algal photobioreactor was indeed characterized by a high lipid content, with a maximum percentage of lipids per dry weight biomass of 35%. The biomass can therefore be exploited for the production of alternative and clean energy carrier. The proposed biotechnology represents an effective tool for shifiting the conventional plants in carbon neutral platform for implementing principles of ecological transition while achieving high levels of environmental protection.

Electro living membrane bioreactor for highly efficient wastewater treatment and fouling mitigation: Influence of current density on process performances.

The next generation of the self-forming dynamic membrane, referred to in this study as the "Living Membrane (LM)", is a new patented technology based on an encapsulated biological layer that self-forms on a designed coarse-pore size support material during wastewater treatment and acts as a natural membrane filter. Integrating electrochemical processes with wastewater treatment using the LM approach has also been recently studied (the reactor is referred to as the Electro-Living Membrane Bioreactor or e-LMBR). This study investigated the effects of varying current densities, i.e., 0.3, 0.5, and 0.9 mA/cm2, on the performance of an e-LMBR. The results were also compared with those of the Living Membrane Bioreactor or LMBR (without applied current density). Higher pollutant removals were observed in the presence of the electric field. However, the effect of varying applied current densities on the COD (98-99 %), NH3-N (97-99 %), and PO43-P (100 %) removals was not statistically significant. The more prominent differences (p < 0.05) were observed in the decrease of NO3--N concentrations from mixed liquor to effluent, with increasing current density resulting in lower mean NO3--N effluent concentrations (0.3 mA/cm2: 6.13 mg/L; 0.5 mA/cm2: 4.38 mg/L; 0.9 mA/cm2: 3.70 mg/L). The reduction of NO3--N concentrations as wastewater permeated through the LM layer also confirmed its role in removing nitrogen-containing compounds. Higher current densities resulted in lower concentrations of fouling substances, particularly those of microbial extracellular polymeric substances (EPS) and transparent exopolymer particles (TEPs). The average values of the temporal variation of transmembrane pressure (d(TMP)/d(t)) in the e-LMBR were extremely low, in the range of 0.013-0.041 kPa/day, throughout the operation period. The highest (d(TMP)/d(t)) was observed for the highest current density. However, the TMP values remained below 2 kPa in all the e-LMBR runs even after the initial LM formation stage.

Micro(nano)plastics from synthetic oligomers persisting in Mediterranean seawater: Comprehensive NMR analysis, concerns and origins.

The presence in seawater of low-molecular-weight polyethylene (PE) and polydimethylsiloxane (PDMS), synthetic polymers with high chemical resistance, has been demonstrated in this study for the first time by developing a novel methodology for their recovery and quantification from surface seawater. These synthetic polymer debris (SPD) with very low molecular weights and sizes in the nano- and micro-metre range have escaped conventional analytical methods. SPD have been easily recovered from water samples (2 L) through filtration with a nitrocellulose membrane filter with a pore size of 0.45 µm. Dissolving the filter in acetone allowed the isolation of the particulates by centrifugation followed by drying. The isolated SPD were analysed by 1H nuclear magnetic resonance spectroscopy (1H NMR), identifying PE and PDMS. These polymers are thus persisting on seawater because of their low density and the ponderal concentrations were quantified in mg/m3. This method was used in an actual case study in which 120 surface seawater samples were collected during two sampling campaigns in the Mediterranean Sea (from the Gulf of Salerno to the Gulf of Policastro in South Italy). The developed analytical protocol allowed achieving unprecedented simplicity, rapidity and sensitivity. The 1H and 13C NMR structural analysis of the PE debris indicates the presence of oxidised polymer chains with very low molecular weights. Additionally, the origin of those low molecular weight polymers was investigated by analysing influents and effluents from a wastewater treatment plant (WWTP) in Salerno as a hot spot for the release of SPD: the analysis indicates the presence of low molecular weight polymers compatible with wax-PE, widely used for coating applications, food industry, cosmetics and detergents. Moreover, the origin of PDMS debris found in surface seawater can be ascribed to silicone-based antifoamers and emulsifiers.

Living membrane bioreactor for highly effective and eco-friendly treatment of textile wastewater.

The treatability of synthetic textile wastewater containing model dyes, such as reactive black and direct black dye (25.0 ± 2.6 mgdye/L), with chemical oxygen demand (COD, 1000 ± 113 mg/L), ammonia­nitrogen (NH3-N, 140 ± 97 mg/L) and sulphate ions (SO42-, 1357 ± 10.86 mg/L) was investigated in this study using an innovative living membrane bioreactor (LMBR) using an encapsulated self-forming dynamic membrane (ESFDM). The key advantage of ESFDMBR is the self-forming of the biological filtering layer protected between two meshes of inert robust and inexpensive material. A laboratory scale bioreactor (BR) equipped with a filtering unit mounting polyester meshes with a pore size of 30 µm, operated at an influent flux of 30 LMH was thus used. After the formation of the biological living membrane (LM), the treatment significantly reduced COD and DOC concentrations to the average values of 34 ± 10 mg/L and 32 ± 7 mg/L, corresponding to reduction efficiencies of 96.0 ± 1.1 % and 94 ± 1.05 %, respectively. Throughout the LMBR operation, the colours were successfully removed from synthetic textile wastewater with an overall removal efficiency of about 85.0 ± 1.8 and 86.0 ± 1.9 % for direct and reactive dyes, respectively. In addition, the proposed system was also found effective in affording removal efficiency of ammonia (NH3) of 97 ± 0.5 %. Finally, this treatment afforded circa 40.7 ± 5.8 % sulphate removal, with a final concentration value of 805 ± 78.61 mg/L. The innovative living membrane, based on an encapsulated self-forming dynamic membrane allows a prolonged containment of the membrane fouling, confirmed by investigating the concentration of membrane fouling precursors and the time-course variations of turbidity and transmembrane pressure (TMP). Those final concentrations of wastewater pollutants were found to be below the limits for admission of the effluents in public sanitation networks in Italy and Tunisia, as representative countries for the regulation in force in Europe and North Africa. In conclusion, due to the low costs of plant and maintenance, the simple applicability, the rapid online implementation, the application of LMBR results in a promising method for the treatment of textile wastewater.

Biological desulfurization of biogas: A comprehensive review on sulfide microbial metabolism and treatment biotechnologies.

Hydrogen sulphide (H2S) removal from biogas is of high relevance as it damages combustion engines used for heat and power generation and causes adverse public health and environmental effects. Biological processes have been reported as a cost-effective and promising approach to desulfurize biogas. This review presents a detailed description of the biochemical foundations of the metabolic apparatus of H2S oxidizing bacteria, namely chemolithoautotrophs and anoxygenic photoautotrophs. The review focuses on the current and future applications of biological processes for biogas desulfurization and provides insights into their mechanism and main factors influencing their performance. The advantages, drawbacks, limitations, and technical improvements of the biotechnological applications currently based on chemolithoautotrophic organisms are covered extensively. Recent advances, sustainability and economical aspects of biological biogas desulfurization are also discussed. Anoxygenic photoautotrophic-bacteria-based photobioreactors were herein identified as useful tools to improve the sustainability and safety of biological biogas desulfurization. The review addresses gaps in the existing studies concerning the selection of the most suitable desulfurization techniques, their benefits and consequences. The research is useful for all stakeholders involved in the management and optimization of biogas and its findings are directly applicable in the development of new sustainable technologies for biogas upgrading processes on waste treatment plants.

Advanced treatment of toluene emissions with a cutting-edge algal bacterial photo-bioreactor: Performance assessment in a circular economy perspective.

A novel approach for the treatment of VOCs (by using toluene used as a model compound) and the simultaneous conversion of carbon dioxide into valuable biomass has been investigated by using a combination of an activated sludge moving bed bioreactor (MBBR) and an algal photo-bioreactor (PBR). The first unit (MBBR, R1) promoted toluene removal up to 99.9 % for inlet load (IL) of 119.91 g m-3 d-1. The CO2 resulting from the degradation of toluene was then fixed in PBR (R2), with a fixation rate up to 95.8 %. The CO2 uptake was promoted by algae, with average production of algal biomass in Stage VI of 1.3 g L-1 d-1. In the contest of the circular economy, alternative sources of nutrients have been assessed, using synthetic urban wastewater (UWW) and dairy wastewater (DWW) for liquid renewal. The produced biomass with DWW showed a high lipid content, with a maximum productivity of 450.25 mg of lipids L-1 d-1. The solution proposed may be thus regarded as a sustainable and profitable strategy for VOCs treatment in a circular economy perspective.

Unraveling microbial community by next-generation sequencing in living membrane bioreactors for wastewater treatment.

This study delves into the microbial community complexity and its role in self-forming dynamic membrane (SFDM) systems, designed to remove nutrients and pollutants from wastewater, by means of the analysis of Next-Generation Sequencing (NGS) data. In these systems, microorganisms are naturally incorporated into the SFDM layer, which acts as a biological and physical filter. The microorganisms present in an innovative and highly efficient aerobic, electrochemically enhanced, encapsulated SFDM bioreactor were studied to elucidate the nature of the dominant microbial communities present in sludge and in encapsulated SFDM, patented as living membrane® (LM) of the experimental setup. The results were compared to those obtained from the microbial communities found in similar experimental reactors without an applied electric field. The data gathered from the NGS microbiome profiling showed that the microbial consortia found in the experimental systems are comprised of archaeal, bacterial, and fungal communities. However, the distribution of the microbial communities found in e-LMBR and LMBR had significant differences. The results showed that the presence of an intermittently applied electric field in e-LMBR promotes the growth of some types of microorganisms (mainly electroactive microorganisms) responsible for the highly efficient treatment of the wastewater and for the mitigation of the membrane fouling found for those bioreactors.

Smart instrumental Odour Monitoring Station for the efficient odour emission management and control in wastewater treatment plants.

Odour emission assessment in wastewater treatment plants (WWTP) is a key aspect that needs to be improved in the plant management to avoid complaints and guarantee a sustainable environment. The research presents a smart instrumental odour monitoring station (SiOMS) composed of an advanced instrumental odour monitoring system (IOMS) integrated with other measurement units, for the continuous characterization and measurement of the odour emissions, with the aim of managing the potential odour annoyance causes in real time, in order to avoid negative effects. The application and on-site validation procedure of the trained IOMS is discussed. Experimental studies have been conducted at a large-scale WWTP. Fingerprint analysis has been applied to analyze and identify the principal gaseous compounds responsible for the odour annoyance. The artificial neural network has been adopted to elaborate and dynamically update the odour monitoring classification and quantification models (OMMs) of the IOMS. The results highlight the usefulness of a real-time measurement and control system to provide continuous and different information to the plant operators, thus allowing the identification of the odour sources and the most appropriate mitigation actions to be implemented. The paper provides important information for WWTP operators, as well as for the regulating bodies, authorities, manufacturers and end-users of odour monitoring systems involved in environmental odour impact management.

Factors influencing pressure-driven membrane-assisted volatile fatty acids recovery and purification-A review.

Volatile fatty acids (VFAs) are building block chemicals that can be produced through bioconversion of organic waste streams via anaerobic digestion as intermediate products. Purified VFAs are applicable in a wide range of industrial applications such as food, textiles, cosmetics, pharmaceuticals etc. production. The present review focuses on VFAs recovery methods and technologies such as adsorption, distillation, extraction, gas stripping, esterification and membrane based techniques etc., while presenting a discussion of their pros and cons. Moreover, a great attention has been given to the recovery of VFAs through membrane filtration as a promising sustainable clarification, fractionation and concentration approach. In this regard, a thorough overview of factors affecting membrane filtration performance for VFAs recovery has been presented. Filtration techniques such as nanofiltration and reverse osmosis have shown to be capable of recovering over 90% of VFAs content from organic effluent steams, proving the direct effect of membrane materials/surface chemistry, pore size and solution pH in recovery success level. Overall, this review presents a new insight into challenges and potentials of membrane filtration for VFAs recovery based on the effects of factors such as operational parameters, membrane properties and effluent characteristics.

Double-stage membrane-assisted anaerobic digestion process intensification for production and recovery of volatile fatty acids from food waste.

The potential of organic waste streams (i.e., food waste) for the sustainable production of precursor chemicals such as volatile fatty acids (VFAs) using anaerobic digestion (AD) has received significant attention in the present days. AD-derived VFAs have great market appeal if the challenges with their recovery and purification from the complex AD effluent is overcome. In this study, a microfiltration immersed membrane bioreactor (MBR) was used for the production of VFAs from food waste and simultaneously in-situ recovery of VFAs. The MBR set-up was applied for 98 days, with a maximum yield of 0.2 gVFA/gVSadded at an organic loading rate (OLR) of 4 g VS/L/d. The recovered permeate was then subjected to further purification using a side stream ultrafiltration unit. It was found that the removal rates of total solids (TS), total suspended solids (TSS), dissolved solids (DS), volatile solids (VS) and volatile suspended solids (VSS) were above 70-80% in both membranes (10 kDa and 50 kDa), and Phosphorus (P), Total Kjeldahl Nitrogen (TKN), chemical oxygen demand (COD), and NH4+-N were also removed partially. Particularly, VFAs concentration (above 6 g/L) was higher for 10 kDa at pH 5.4 in ultrafiltered solution and permeate flux decline was higher for 10 kDa at pH 5.4. These results are also supported by the measurement of UV-Vis spectra of the solution and visual appearance, providing a promising approach towards building a VFAs-based platform.

Advances in virus detection methods for wastewater-based epidemiological applications.

Wastewater-based epidemiology (WBE) is a powerful tool that has the potential to reveal the extent of an ongoing disease outbreak or to predict an emerging one. Recent studies have shown that SARS-CoV-2 concentration in wastewater may be correlated with the number of COVID-19 cases in the corresponding population. Most of the recent studies and applications of wastewater-based surveillance of SARS-CoV-2 applied the "gold standard" real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) detection method. However, this method also has its limitations. The paper aimed to present recent improvements and applications of the PCR-based methods for SARS-CoV-2 monitoring in wastewater. Furthermore, it aimed to review alternative methods utilized and/or proposed for the detection of the virus in wastewater matrices. From the review, it was found that several studies have investigated the use of reverse-transcription digital polymerase reaction (RT-dPCR), which was generally shown to have a lower limit of detection (LOD) over the RT-qPCR. Aside from this, non-PCR-based and non-RNA based methods have also been explored for the detection of SARS-CoV-2 in wastewater, with detailed attention given to the detection of SARS-CoV-2 proteins. The potential methods for protein detection include mass spectrometry, the use of immunosensors, and nanotechnological applications. In addition, the review of recent studies also revealed two types of emerging methods related to the detection of SARS-CoV-2 in wastewater: i) capsid-integrity assays to infer about the infectivity of SARS-CoV-2 present in wastewater, and ii) alternative methods for detection of SARS-CoV-2 variants of concern (VOCs) in wastewater. The recent studies on proposed methods of SARS-CoV-2 detection in wastewater have considered improving this approach in one or more of the following aspects: rapidity, simplicity, cost, sensitivity, and specificity. However, further studies are needed in order to realize the full application of these methods for WBE in the field.

Innovative encapsulated self-forming dynamic bio-membrane bioreactor (ESFDMBR) for efficient wastewater treatment and fouling control.

The concept of a novel living encapsulated self-forming dynamic bio-membranes (ESFDM) for an innovative wastewater treatment in membrane bioreactor (MBR) is presented in the current study. The active filtering membrane is encapsulated, and thus stabilized, between two support meshes with pore in micrometer size. The combination of activated sludge, the ESFDM and the cake layer formed external to the filtering module contributed to the treatment of municipal wastewater. COD concentration reductions (average value of 95.55 ± 1.44%) by ESFDM bioreactor (ESFDMBR) were comparable to those obtained with a previously reported membrane bioreactor (MBR), where a conventional membrane was studied under the same operating conditions. The ESFDMBR, compared to the conventional MBR, obtained higher reductions of NH3-N, NO3-N and PO43-P concentrations. Increased removals of nitrogen-containing nutrients were ascribed to anoxic conditions reached in the ESFDM layer protected from the aeration by the external cake layer. Rate of increase of transmembrane pressure (TMP) per day in the ESFDMBR (0.03 kPa/day) was lower than the value obtained in the previously reported conventional MBR (8.08 kPa/day). Lower concentrations of fouling precursors in combination with the effective filtration capacity of the porous living ESFDM resulted in the reduction of the fouling rate. Analysis of microbiological community revealed that the microbial community structures in the mixed liquor and ESFDM were different. The ESFDM layer promoted growth of bacteria as indicated by the higher total cell count and higher microbial diversity compared to those observed in the mixed liquor.