Temporal assessment of SARS-CoV-2 detection in wastewater and its epidemiological implications in COVID-19 case dynamics.

This research evaluated the relationship between daily new Coronavirus Disease 2019 (COVID-19) cases and Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) concentrations in wastewater, followed by effects of differential SARS-CoV-2 shedding loads across various COVID-19 outbreaks. Linear regression analyses were utilized to examine the lead time of the SARS-CoV-2 signal in wastewater relative to new COVID-19 clinical cases. During the Delta wave, no lead time was evident, highlighting limited predictive capability of wastewater monitoring during this phase. However, significant lead times were observed during the Omicron wave, potentially attributed to testing capacity overload and subsequent case reporting delays or changes in shedding patterns. During the Post-Omicron wave (Febuary 23 to May 19, 2022), no lead time was discernible, whereas following the lifting of the COVID-19 state of emergency (May 30, 2022 to May 30, 2023), the correlation coefficient increased and demonstrated the potential of wastewater surveillance as an early warning system. Subsequently, we explored the virus shedding in wastewater through feces, operationalized as the ratio of SARS-CoV-2 concentrations to daily new COVID-19 cases. This ratio varied significantly across the Delta, Omicron, other variants and post-state-emergency phases, with the Kruskal-Wallis H test confirming a significant difference in medians across these stages (P < 0.0001). Despite its promise, wastewater surveillance of COVID-19 disease prevalence presents several challenges, including virus shedding variability, data interpretation complexity, the impact of environmental factors on viral degradation, and the lack of standardized testing procedures. Overall, our findings offer insights into the correlation between COVID-19 cases and wastewater viral concentrations, potential variation in SARS-CoV-2 shedding in wastewater across different pandemic phases, and underscore the promise and limitations of wastewater surveillance as an early warning system for disease prevalence trends.

A mass balance approach for quantifying the role of natural decay and fate mechanisms on SARS-CoV-2 genetic marker removal during water reclamation.

The recent SARS-CoV-2 outbreak yielded substantial data regarding virus fate and prevalence at water reclamation facilities (WRFs), identifying influential factors as natural decay, adsorption, light, pH, salinity, and antagonistic microorganisms. However, no studies have quantified the impact of these factors in full scale WRFs. Utilizing a mass balance approach, we assessed the impact of natural decay and other fate mechanisms on genetic marker removal during water reclamation, through the use of sludge and wastewater genetic marker loading estimates. Results indicated negligible removal of genetic markers during P/PT (primary effluent (PE) p value: 0.267; preliminary and primary treatment (P/PT) accumulation p value: 0.904; and thickened primary sludge (TPS) p value: 0.076) indicating no contribution of natural decay and other fate mechanisms toward removal in P/PT. Comparably, adsorption and decomposition was found to be the dominant pathway for genetic marker removal (thickened waste activated sludge (TWAS) log loading 9.75 log10 GC/day); however, no estimation of log genetic marker accumulation could be carried out due to high detections in TWAS. PRACTITIONER POINTS: The mass balance approach suggested that the contribution of natural decay and other fate mechanisms to virus removal during wastewater treatment are negligible compared with adsorption and decomposition in P/PT (p value: 0.904). During (P/PT), a higher viral load remained in the (PE) (14.16 log10 GC/day) compared with TPS (13.83 log10 GC/day); however, no statistical difference was observed (p value: 0.280) indicting that adsorption/decomposition most probably did not occur. In secondary treatment (ST), viral genetic markers in TWAS were consistently detected (13.41 log10 GC/day) compared with secondary effluent (SE), indicating that longer HRT and the potential presence of extracellular polymeric substance-containing enriched biomass enabled adsorption/decomposition. Estimations of total solids and volatile solids for TPS and TWAS indicated that adsorption affinity was different between solids sampling locations (p value: <0.0001).

Per- and poly-fluoroalkyl substances removal in multi-barrier advanced water purification system for indirect potable reuse.

The study evaluated the removal efficacy of per- and poly-fluoroalkyl substances (PFAS) across various advanced water treatment (AWT) processes in a field-scale AWT train using secondary effluent samples from a full-scale water reclamation facility (WRF). Samples collected from April to October 2020 revealed PFCAs as the dominant PFAS compounds in the WRF secondary effluent, with PFPeA having the highest average concentration and PFSAs in notably lower amounts. Temporal fluctuations in total PFAS concentrations peaked in September 2020, which may reflect the seasonality in PFAS discharges related to applications like AFFFs and pesticides. In assessing AWT processes, coagulation-flocculation-clarification-filtration system showed no notable PFAS reduction, while ozonation resulted in elevated PFBS and PFBA concentrations. Biological activated carbon (BAC) filtration effectively removed long-chain PFAS like PFOS and PFHxS but saw increased concentrations of short-chain PFAS post-treatment. Granular activated carbon (GAC) filtration was the most effective treatment, reducing all PFSAs below the detection limits and significantly decreasing most PFCAs, though short-chain PFCAs persisted. UV treatment did not remove short-chain PFCAs such as PFBA, PFPeA, and PFHxA. The findings highlight the efficacy of AWT processes like GAC in PFAS reduction for potable reuse, but also underscore the challenge presented by short-chain PFAS, emphasizing the need for tailored treatment strategies. PRACTITIONER POINTS: Secondary effluents showed higher concentrations of PFCAs compared to PFSAs. Advanced water treatment effectively removes long-chain PFAS but not short-chain. Ozonation may contribute to formation of short-chain PFAS. BAC is less effective on short-chain PFAS, requiring further GAC treatment.

Contaminants of emerging concern reduction and microbial community characterization across a three-barrier advanced water treatment system.

This research investigated the removal of contaminants of emerging concern (CECs) and characterized the microbial community across an advanced water treatment (AWT) train consisting of Coagulation/Flocculation/Clarification/Granular Media Filtration (CFCGMF), Ozone-Biological Activated Carbon Filtration (O3/BAC), Granular Activated Carbon filtration, Ultraviolet Disinfection, and Cartridge Filtration (GAC/UV/CF). The AWT train successfully met the goals of CECs and bulk organics removal. The microbial community at each treatment step of the AWT train was characterized using 16S rRNA sequencing on the Illumina MiSeq platform generated from DNA extracted from liquid and solid (treatment media) samples taken along the treatment train. Differences in the microbial community structure were observed. The dominant operational taxonomic units (OTU) decreased along the treatment train, but the treatment steps did impact the microbial community composition downstream of each unit process. These results provide insights into microbial ecology in advanced water treatment systems, which are influenced and shaped by each treatment step, the microbial community interactions, and their potential metabolic contribution to CECs degradation.

Understanding the influence of dissolved organic nitrogen characteristics on enhanced coagulation performance for water reuse.

With the recent focus on using advanced water treatment processes for water reuse, interest is growing for utilizing enhanced coagulation to remove dissolved chemical species. Up to 85% of the nitrogen in wastewater effluent is made up of dissolved organic nitrogen (DON), but there is a knowledge gap regarding its removal during coagulation, which can be influenced by DON characteristics. To address this issue, tertiary-treated wastewater samples were analyzed before and after coagulation with polyaluminum chloride and ferric chloride. Samples were size-fractionated into four molecular weight fractions (0.45 µm, 0.1 µm, 10 kDa, and 3 kDa) using vacuum filtration and ultrafiltration. Each fraction was further evaluated by coagulating it separately to assess DON removal during enhanced coagulation. The size fractionated samples were also separated into hydrophilic and hydrophobic fractions using C18 solid phase extraction disks. Fluorescence excitation-emission matrices were used to investigate the characteristics of dissolved organic matter contributing to DON during the coagulation process. The results showed that DON compounds of size <3 kDa constituted a majority of the total DON. Coagulation removed more than 80% DON from size fractions 0.45 µm-0.1 µm and 0.1 µm-10 kDa, but less than 20% was removed from 10 kDa to 3 kDa and <3 kDa fractions. Coagulation on pre-filtered samples removed 19% and 25% of the <3 kDa DON fraction using polyaluminum chloride and ferric chloride, respectively. In all molecular weight fractions, hydrophilic DON compounds were found to be dominant (>90%), and enhanced coagulation was not effective in removing hydrophilic DON compounds. LMW fractions respond poorly to enhanced coagulation due to their hydrophilic nature. Enhanced coagulation effectively removes humic acid-like substances, but poorly removes proteinaceous compounds such as tyrosine and tryptophan. This study's findings provide insights into DON behavior during coagulation and factors affecting its removal, potentially improving wastewater treatment strategies.

Pilot-scale demonstration of dissolved organic nitrogen removal from an advanced water reclamation facility using enhanced coagulation.

Most wastewater treatment facilities that satisfy stricter discharge restrictions for nutrients, remove dissolved inorganic nitrogen (DIN) species efficiently, leaving dissolved organic nitrogen (DON) to be present at a higher proportion (up to 85 %) of total nitrogen (TN) in the effluent. Discharged DON promotes algae growth in receiving water bodies and is a growing concern in effluent potable reuse applications considering its potential to form hazardous nitrogenous disinfection byproducts (N-DBPs). Enhanced coagulation is an established process in the advanced water treatment train for most potable reuse applications. However, so far, no information has been collected at the pilot scale to address DON removal efficiency and process implications by enhanced coagulation under real conditions. This study performed a comprehensive evaluation of DON removal from the effluent of the Truckee Meadows Water Reclamation Facility (TMWRF) by enhanced coagulation over the course of 11 months at the pilot scale. Three different coagulants (aluminum sulfate (alum), poly­aluminum chloride (PACl), ferric chloride (FC)) and a cationic polymer coagulant aid (Clarifloc) were used. Optimum doses for each coagulant and polymer and ideal pH were determined by jar tests and applied at the pilot. Alum (24 mg/L) resulted in highly variable DON removal (6 % - 40 %, 21 % on average), which was enhanced by the addition of polymer, leading to 32 % DON removal on average. PACl (40 mg/L) and FC (100 mg/L) resulted in more consistent DON removal (on average 45 % and 57 %, respectively); however, polymer addition exerted minimal enhancement for these coagulants. Overall, enhanced coagulation effectively reduced DON in the tertiary effluent at the pilot scale. The treatment showed auxiliary benefits, including dissolved organic carbon (DOC) and orthophosphate removal.

Innovative Technological Approach for the Cyclic Nutrients Adsorption by Post-Digestion Sewage Sludge-Based Ash Co-Formed with Some Nanostructural Additives under a Circular Economy Framework.

This paper presents a new, innovative technological approach, in line with Circular Economy principles, to the effective management of sludge generated during municipal wastewater treatment processes and subsequently used for biogas production. This approach allows for optimal, functional, and controlled cascade-type biotechnological thermal conversion of carbon compounds present in sewage sludge, later in solid digestate residues (after biogas production), and finally in the ash structure (after incineration, purposefully dosed nanostructural additives make the production of a useful solid product possible, especially for cyclic adsorption and slow release of nutrients (N, P, K) in the soil). The idea is generally targeted at achieving an innovative conversion cycle under a Circular Economy framework. In particular, it is based on an energy carrier (methane biogas) and direct energy production. The functionalized combustion by-products can be advantageous in agriculture. The use of ashes with nanostructural additives (halloysite, kaolinite) from combustion of sewage sludge after the anaerobic fermentation as an adsorbent of selected nutrients important in agriculture (Na+, K+, NO3-, SO42-, PO43-, Cl-) was verified at laboratory scale. The tests were carried out both for pure ash and for the ash derived from combustion with the purposeful addition of kaolinite or halloysite. The equilibrium conditions for nitrate, potassium, sodium, phosphate(V), sulphate(VI), and chloride ions from aqueous solutions with the use of the three adsorbent structures were determined. The obtained innovative results were interpreted theoretically with adsorption isotherm models (Langmuir, Freundlich, Temkin, Jovanovic). The most spectacular and clearly favorable results related to the influence of nanostructural additives in the process of sludge combustion, and formation of sorption surfaces under high temperature conditions were identified in the case of sorption-based separation of phosphate(V) ions (an increase from 1.13% to 61.24% with the addition of kaolinite, and even up to 76.19% with addition of halloysite).

Microbial community characterization in advanced water reclamation for potable reuse.

This study investigated the microbial community structure and composition across two treatment steps used in advanced water reclamation for potable reuse applications, namely Coagulation/Flocculation/Clarification/Granular Media Filtration (CFCGMF) and Ozone-Biological Activated Carbon filtration (O3/BAC). The study examined the richness, variations, and similarities of the microorganisms involved at each treatment step to better understand the role of ecology and the dynamics on unit process performance and the microbial community developed within it. The bacterial microbiomes at each treatment step were independently characterized using 16S metagenomic sequencing. Combining both treatment steps, a total of 3801 species were detected. From the total species detected, 38% and 98% were identified at CFCGMF and O3/BAC, respectively. The most abundant phyla were Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes in both treatment steps. The identified species were classified based on their preferences to free-living style (59%) vs attached-living style (22%) showing a relatively low richness in the BAC media, but higher diversities. At the taxonomic class level, Betaproteobacteria was the predominant in both system processes. Additionally, a list of eight genera were identified as potential bacterial pathogens present in both process effluents. They are Aeromonas, Clostridium, Enterobacter, Escherichia, Flavobacterium, Legionella, Mycobacterium, and Pseudomonas. CFCGMF effluent yielded less pathogenic bacteria than both the ozone and BAC filter effluent from the O3/BAC process unit; their relative abundance accounted for about 2% and 8% for CFCGMF and O3/BAC, respectively. Detailed studies to characterize the microbial communities are crucial in interpreting the mechanisms and synergies between processes performance and microorganisms by identifying the needs and best practices to ensure public health protection. Key points • Microbial communities of two treatment processes are characterized using 16S rRNA sequencing. • Organisms that can tolerate ozone and form biofilms define microbial community in subsequent biofilters. • In relatively low abundances, potential pathogenic bacteria are detected in the treated water.

Longitudinal monitoring of SARS-CoV-2 in wastewater using viral genetic markers and the estimation of unconfirmed COVID-19 cases.

In this study, wastewater-based surveillance was carried out to establish the correlation between SARS-CoV-2 viral RNA concentrations in wastewater and the incidence of corona virus disease 2019 (COVID-19) from clinical testing. The influent wastewater of three major water reclamation facilities (WRFs) in Northern Nevada, serving a population of 390,750, was monitored for SARS-CoV-2 viral RNA gene markers, N1 and N2, from June 2020 through September 2021. A total of 614 samples were collected and analyzed. The SARS-CoV-2 concentrations in wastewater were observed to peak twice during the study period. A moderate correlation trend between coronavirus disease 2019 (COVID-19) incidence data from clinical testing and SARS-CoV-2 viral RNA concentrations in wastewater was observed (Spearman r = 0.533). This correlation improved when using weekly average SARS-CoV-2 marker concentrations of wastewater and clinical case data (Spearman r = 0.790), presumably by mitigating the inherent variability of the environmental dataset and the effects of clinical testing artifacts (e.g., reporting lags). The research also demonstrated the value of wastewater-based surveillance as an early warning signal for early detection of trends in COVID-19 incidence. This was accomplished by identifying that the reported clinical cases had a stronger correlation to SARS-CoV-2 wastewater monitoring data when they were estimated to lag 7-days behind the wastewater data. The results aided local decision makers in developing strategies to manage COVID-19 in the region and provide a framework for how wastewater-based surveillance can be applied across localities to enhance the public health monitoring of the ongoing pandemic.

Investigation of direct waste-activated sludge dewatering benefits and costs in a water resource recovery facility.

Dewatering of anaerobic digested (AD) sludge containing waste-activated sludge (WAS) from enhanced biological phosphorus removal (EBPR) poses numerous challenges including poor dewaterability, struvite scale formation, and recycling of high N and P levels in the sludge liquor to the treatment process. A full-scale water reclamation facility was investigated to mitigate these problems by experimenting with direct dewatering of EBPR WAS, bypassing the AD step. The investigations experimented with various blends of AD primary sludge with undigested thickened WAS to achieve dewatering performance improvements and overall operational cost savings. Direct thickened WAS dewatering has had many positive impacts including enhanced sludge cake solids concentration, reduced chemical use for facility operations, reduced struvite scaling, reduced biogas conditioning media servicing, eliminated need for centrate treatment, recovered capacity of existing unit operations including anaerobic digesters, and eliminated several proposed capital improvement projects that were previously deemed necessary. Although bypassing of WAS to AD reduced total biogas production, the specific gas yield increased to meet all of the facility's biogas demands and minimized excess gas flaring. The overall biosolids production mass increased causing increased transportation costs for disposal and caused notable odors, both of which are being currently investigated. PRACTITIONER POINTS: Direct WAS dewatering bypassing anaerobic digestion yields operational and process benefits in an EBPR water resource recovery facility Dewatered cake solids were increased compared with combined primary and WAS anaerobic digestion and dewatering Nutrient loads in sludge processing returns streams and operational costs are reduced by direct WAS dewatering.

The Discovery of Vitreoscilla Hemoglobin and Early Studies on Its Biochemical Functions, the Control of Its Expression, and Its Use in Practical Applications.

In 1986, the surprising identification of a hemoglobin (VHb) in the bacterium Vitreoscilla greatly extended the range of taxa in which this oxygen binding protein functions. Elucidation of many of its biochemical properties and relation to overall cell physiology, as well as the sequence of the gene encoding it and aspects of control of its expression were determined in the following years. In addition, during the early years following its discovery, strategies were developed to use its expression in heterologous microbial hosts to enhance processes of practical usefulness. The VHb discovery also served as the foundation for what has become the fascinatingly rich field of bacterial hemoglobins. VHb's position as the first known bacterial hemoglobin and its extensive use in biotechnological applications, which continue today, make a review of the early studies of its properties and uses an appropriate and interesting topic thirty-five years after its discovery.

Microbial community in biofilters for water reuse applications: A critical review.

The combination of ozonation (O3) and biofiltration processes has become practical and desirable in advanced water reclamation for water reuse applications. However, the role of microbial community and its characteristics (source, abundance, composition, viability, structure) on treatment performance has not received the same attention in water reclamation biofilters as in other applications, such as in drinking water biofilters. Microbial community characterization of biofilters used in water reuse applications will add evidence to better understand the potential microorganisms, consequent risks, and mechanisms that will populate drinking water sources and ultimately influence public health and the environment. This critical review provides insights into O3-biofiltration as a treatment barrier with a focus on development, structure, and composition of the microbial community characteristics involved in the process. The effect of microorganism seeding by the influent before and after the biofilter and ozone oxidation effects are explored to capture the microbial ecology interactions and environmental factors affecting the media ecosystem. The findings of reviewed studies concurred in identifying Proteobacteria as the most dominant phylum. However, Proteobacteria and other phyla relative abundance differ substantially depending upon environmental factors (e.g., pH, temperature, nutrients availability, among others) gradients. In general, we found significant gaps to relate and explain the biodegradation performance and metabolic processes within the biofilter, and hence deserve future attention. We highlighted and identified key challenges and future research ideas to assure O3-biofiltration reliability as a promising barrier in advanced water treatment applications.

Critical review of effluent dissolved organic nitrogen removal by soil/aquifer-based treatment systems.

Soil/aquifer-based treatment systems improve wastewater effluent quality by removing trace contaminants in the soil and/or aquifer during groundwater recharge. This paper critically reviews these systems with a focus on removing nitrogen, particularly low levels of dissolved organic nitrogen (DON) present in the wastewater effluent. DON in wastewater effluent is a concern because of its contribution to nitrogen concentration in surface or groundwater and its role as a precursor of nitrogenous disinfection by-products, which are harmful to human health. Biodegradation and sorption are the main DON removal mechanisms in the subsurface environment where most of the removal happens in the vadose zone. Different factors such as temperature, redox conditions, retention time, indigenous microbial community, and soil type affect DON removal in soil/aquifer-based treatment systems. Lack of sufficient current knowledge underlines the need for designing lab/field scale systems for further determination of the relative contribution of biodegradation and sorption, optimal hydraulic loading rate, and the relationship between DON characteristics such as functional groups and physiochemical processes and its removal. Future research needs for DON removal in soil/aquifer-based treatment systems are identified.

Fate and reduction of bromate formed in advanced water treatment ozonation systems: A critical review.

Disinfection in water treatment and reclamation systems eliminates the potential health risks associated with waterborne pathogens, however it may produce disinfection by-products (DBPs) harmful to human health. Potentially carcinogenic bromate is a DBP formed during the ozonation of bromide-containing waters. To mitigate the problem of bromate formation, different physical/chemical or biological reduction methods of bromate have been investigated. Until now, adsorption-based physical method has proven to be more effective than chemical methods in potable water treatment. Though several studies on biological reduction methods have been carried out in a variety of bioreactor systems, such as in biologically active carbon filters and denitrifying bioreactors, the microbiological mechanisms or biochemical pathways of bromate minimization have not been clearly determined to date. Genetic analysis could provide a broader picture of microorganisms involved in bromate reduction which might show cometabolic or respiratory pathways, and affirm the synergy functions between different contributing groups. The hypothesis established from the diffusion coefficients of different electron donor and acceptors, illustrates that some microorganisms preferring bromate over oxygen contain specific enzymes which lower the activation energy required for bromate reduction. In addition, considering microbial bromate reduction as an effective treatment strategy; field scale investigations are required to observe quantitative correlations of various influencing parameters such as pH, ozone dose, additives or constituents such as ammonia, hydrogen peroxide, and/or chloramine, dissolved organic carbon levels, dissolved oxygen gradient within biofilm, and empty bed contact time on bromate removal or reduction.

Full-scale N removal from centrate using a sidestream process with a mainstream carbon source.

An innovative approach to treat centrate for rapid nitrogen load discharge reduction was investigated and applied at the Truckee Meadows Water Reclamation Facility (TMWRF) in Reno, NV. This process allowed TMWRF to circumvent an anticipated exceedance of the individual waste load allocation in 2018. Existing infrastructure and equipment were re-purposed in a full-scale suspended growth biological centrate treatment system, attaining simultaneous nitrification, and denitrification with no additional capital investment. Functioning within a few days of start-up, the average ammonia reduction was 81% (1,106 kg/day) and the average total nitrogen reduction in the sidestream was 53% (757 kg/day) using primary effluent as carbon source. Alkalinity and carbon limitations were both anticipated and observed; however, adaptive operations allowed for balancing of nitrification and denitrification processes, providing pH stability and success in meeting treatment goals. Immediately after the sidestream treatment system was placed into service, nitrogen in the mainstream facility was measured at concentrations significantly lower than typical and was sustained at historically low concentrations throughout the operation. This translated into a significant methanol cost savings of $1,500 per day (USD). The system has become a critical supplemental treatment process during upcoming rehabilitation projects to address aging infrastructure of existing nitrogen treatment facilities. PRACTITIONER POINTS: Full scale demonstration of sidestream N removal using a hybrid process. Integration of the sidestream N process to reduce N effluent load without alkalinity or supplementary carbon augmentation. Operational solution to reduce operating costs without new infrastructure.

Use of biochar to produce reclaimed water for irrigation use.

Emerging contaminants, especially, pharmaceutical and personal care products (PPCPs) are not removed well during conventional wastewater treatment and hence pose water quality risk to the environment and potentially to public health. Long-term use of reclaimed wastewater for irrigation can lead to accumulation of trace contaminants in the soil, ground water and their subsequent uptake by plants and potentially can enter human food chain. This paper uses biochar as an adsorbent to remove emerging contaminants from treated wastewater by performing fixed bed experiments. Ten emerging contaminants namely, carbamazepine (CBZ), caffeine, diethyltoluamide (DEET), diphenhydramine (DPH), meprobamate (MPB), primidone (PMD), sulfamethoxazole (SMX), fluoxetine (FXT), perfluorooctanoic acid (PFOA) and trimethoprim (TMP) were monitored during lab scale experiments. Results from the continuous flow runs showed that the breakthrough curve for compounds caffeine, CBZ, DEET and PFOA follow second order Thomas model with adsorption capacities of 396 µg g-1, 392 µg g-1, 1160 µg g-1 and 32 µg g-1 biochar, respectively. Whereas compounds such as DPH, TMP and FXT were completely removed throughout the column runs by biochar. Results for rest of the compounds were interfered by leaching of these compounds from biochar. It was observed that commercially available GAC performed much better than biochar for all the compounds considered. Even at 1% of obtained capacity, biochar amendment to soils where reclaimed water is used for irrigation can reduce the uptake of these compounds by plants.

Phosphorus Recovery by Methods Beyond Struvite Precipitation.

Unintentional phosphate precipitation at water resource recovery facilities (WRRFs) causes operation and maintenance challenges. With global phosphorus (P) scarcity looming and receiving water eutrophication caused by excess discharges of P, intentional P recovery at WRRFs has been gaining traction. To date, struvite recovery as slow release fertilizer has been the focus of P recovery. However, struvite recovery is not always the most cost-effective approach, especially when phosphate fertilizers cost considerably less than the cost to recover struvite as fertilizer. The aim of this state-of-an-art review paper is to discuss P recovery as calcium phosphate, which could be a fertilizer feedstock and incurs less chemical costs to produce. Calcium phosphate also offers broader applications for other industrial uses beyond fertilizers since the composition is close to mined phosphate rock. A strategic approach for a regional reclaimed phosphate reserve is proposed to secure the most economical future supply of P.

Biomass density-function relationships in suspended growth biological processes - A critical review.

Good settling performance in suspended growth biomass systems, for example in activated sludge (AS) process, leads to efficient wastewater and sludge treatment. Factors that cause the differences in settleablility of AS include the morphology of bacteria, microbial community structure, and the density of bacteria and flocs. Density of AS at three levels, namely, cell, floc, and process, have been discussed here to explain the variations in AS settleability. Dense materials, inside or outside the cell, significantly increase density of AS bacteria or flocs. Functional bacteria, defined as those performing N and P removal and recovery such as phosphate accumulating organisms, nitrifiers, and anammox contain cellular inclusions that increase their density, and consequently a dense and well-settling biomass results at the process level in those systems. A density based selector of AS can be used to enrich functional bacteria in the process through the wasting and sludge age control operations in AS process. This paper critically reviews the latest literature to elucidate mechanisms of density enhancement from cell to process level, and identifies needs/strategies to improve the AS process through a biomass density selector.

Engineering of Nitrosomonas europaea to express Vitreoscilla hemoglobin enhances oxygen uptake and conversion of ammonia to nitrite.

Nitrosomonas europaea was transformed with a recombinant plasmid bearing the gene (vgb) encoding the hemoglobin (VHb) from the bacterium Vitreoscilla under control of the N. europaea amoC P1 promoter. Vgb was maintained stably and appeared to be expressed in the transformants at VHb levels of about 0.75 nmol/g wet weight. Expression of VHb in the N. europaea transformants was correlated with an approximately 2 fold increase in oxygen uptake rate by whole cells at oxygen concentrations in the range of 75-100% saturation, but no change in oxygen uptake rate at oxygen concentrations below 25% saturation. VHb expression was also correlated with an increase of as much as about 30% in conversion of ammonia to nitrite by growing cells. The results suggest that engineering of key aerobic wastewater bacteria to express bacterial hemoglobins may be a useful strategy to produce species with enhanced respiratory abilities.

Directed evolution to produce sludge communities with improved oxygen uptake abilities.

Two activated sludge cultures, seeded with activated sludge from the same source, were cultivated for 370 days in synthetic wastewater. Both cultures were transferred weekly to fresh medium; one culture was operated at high dissolved oxygen (DO) (near saturation) and the other at low DO (0.25 mg O2/L). There were significant changes in the abundances of bacterial species and phyla present in each culture throughout the 370-day operational period. In the low DO culture, over time, there was a continuously increasing proportion of cells of species known to encode truncated hemoglobins (Hbs). These are the types of Hbs which may enhance delivery of oxygen to the respiratory chain, to enhance ATP production, especially under low aeration conditions. The levels of heme b, the heme found in Vitreoscilla hemoglobin, increased in parallel to the increase in Hb-encoding species, to much higher levels in the low DO culture than in the high DO culture. Specific oxygen uptake rates increased by 3 % for the high DO culture near the end of the 370-day period, while those for the low DO culture increased steadily to a level 28 % higher than that of the starting culture. Thus, imposition of low DO conditions may, due to selection for Hb-expressing species, be useful in developing bacterial communities with enhanced ability to function efficiently in aerobic wastewater treatment, especially under low aeration conditions.