Search

Microalgae have the potential to exceed current nutrient recovery limits from wastewater, enabling water resource recovery facilities (WRRFs) to achieve increasingly stringent effluent permits. The use of photobioreactors (PBRs) and the separation of hydraulic retention and solids residence time (HRT/SRT) further enables increased biomass in a reduced physical footprint while allowing operational parameters (e.g., SRT) to select for desired functional communities. However, as algal technology transitions to full-scale, there is a need to understand the effect of operational and environmental parameters on complex microbial dynamics among mixotrophic microalgae, bacterial groups, and pests (i.e., grazers and pathogens) and to implement robust process controls for stable long-term performance. Here, we examine a full-scale, intensive WRRF utilizing mixed microalgae for tertiary treatment in the US (EcoRecover, Clearas Water Recovery Inc.) during a nine-month monitoring campaign. We investigated the temporal variations in microbial community structure (18S and 16S rRNA genes), which revealed that stable system performance of the EcoRecover system was marked by a low-diversity microalgal community (D INVSIMPSON = 2.01) dominated by Scenedesmus sp. (MRA = 55 %-80 %) that achieved strict nutrient removal (effluent TP < 0.04 mg·L-1) and steady biomass concentration (TSS monthly avg. = 400-700 mg·L -1 ). Operational variables including pH, alkalinity, and influent ammonium (NH 4 + ), correlated positively (p < 0.05, method = Spearman) with algal community during stable performance. Further, the use of these parameters as operational controls along with N/P loading and SRT allowed for system recovery following upset events. Importantly, the presence or absence of bacterial nitrification did not directly impact algal system performance and overall nutrient recovery, but partial nitrification (potentially resulting from NO 2 - accumulation) inhibited algal growth and should be considered during long-term operation. The microalgal communities were also adversely affected by zooplankton grazers (ciliates, rotifers) and fungal parasites (Aphelidium), particularly during periods of upset when algal cultures were experiencing culture turnover or stress conditions (e.g., nitrogen limitation, elevated temperature). Overall, the active management of system operation in order to maintain healthy algal cultures and high biomass productivity can result in significant periods (>4 months) of stable system performance that achieve robust nutrient recovery, even in winter months in northern latitudes (WI, USA)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Mixed community microalgal wastewater treatment technologies have the potential to advance the limits of technology for biological nutrient recovery while producing a renewable carbon feedstock, but a deeper understanding of their performance is required for system optimization and control. In this study, we characterized the performance of a 568 m 3 ·day -1 Clearas EcoRecover system for tertiary phosphorus removal (and recovery as biomass) at an operating water resource recovery facility (WRRF). The process consists of a (dark) mix tank, photobioreactors (PBRs), and a membrane tank with ultrafiltration membranes for the separation of hydraulic and solids residence times. Through continuous online monitoring, long-term on-site monitoring, and on-site batch experiments, we demonstrate (i) the importance of carbohydrate storage in PBRs to support phosphorus uptake under dark conditions in the mix tank and (ii) the potential for polyphosphate accumulation in the mixed algal communities. Over a 3-month winter period with limited outside influences (e.g., no major upstream process changes), the effluent total phosphorus (TP) concentration was 0.03 ± 0.03 mg-P·L -1 (0.01 ± 0.02 mg-P·L -1 orthophosphate). Core microbial community taxa included Chlorella spp. , Scenedesmus spp. , and Monoraphidium spp. , and key indicators of stable performance included near-neutral pH, sufficient alkalinity, and a diel rhythm in dissolved oxygen.

Graphene and zero-valent-iron based nanohybrid (rGO-nZVI NH) with oxidant H 2 O 2 can remove perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) through adsorption-degradation in a controlled aquatic environment. In this study, we evaluated how and to what extent different environmental and operational parameters, such as initial PFAS concentration, H 2 O 2 dose, pH, ionic strength, and natural organic matter (NOM), influenced the removal of PFOS and PFOA by rGO-nZVI. With the increase in initial PFAS concentration (from 0.4 to 50 ppm), pH (3 to 9), ionic strength (0 to 100 mM), and NOM (0 to 10 ppm), PFOS removal reduced by 20%, 30%, 2%, and 6%, respectively, while PFOA removal reduced by 54%, 76%, 11%, and 33% respectively. In contrast, PFOS and PFOA removal increased by 10% and 41%, respectively, with the increase in H 2 O 2 (from 0 to 1 mM). Overall, the effect of changes in environmental and operational parameters was more pronounced for PFOA than PFOS. Mechanistically, •OH radical generation and availability showed a profound effect on PFOA removal. Also, the electrostatic interaction between rGO-nZVI NH and deprotonated PFAS compounds was another key factor for removal. Most importantly, our study confirms that rGO-nZVI in the presence of H 2 O 2 can degrade both PFOS and PFOA to some extent by identifying the important by-products such as acetate, formate, and fluoride., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Per and polyfluoroalkyl substances (PFAS) present a unique challenge to remediation techniques because their strong carbon-fluorine bonds make them difficult to degrade. This review explores the use of in silico enzymatic design as a potential PFAS degradation technique. The scope of the enzymes included is based on currently known PFAS degradation techniques, including chemical redox systems that have been studied for perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) defluorination, such as those that incorporate hydrated electrons, sulfate, peroxide, and metal catalysts. Bioremediation techniques are also discussed, namely the laccase and horseradish peroxidase systems. The redox potential of known reactants and enzymatic radicals/metal-complexes are then considered and compared to potential enzymes for degrading PFAS. The molecular structure and reaction cycle of prospective enzymes are explored. Current knowledge and techniques of enzyme design, particularly radical-generating enzymes, and application are also discussed. Finally, potential routes for bioengineering enzymes to enable or enhance PFAS remediation are considered as well as the future outlook for computational exploration of enzymatic in situ bioremediation routes for these highly persistent and globally distributed contaminants.

The sequencing of human virus genomes from wastewater samples is an efficient method for tracking viral transmission and evolution at the community level. However, this requires the recovery of viral nucleic acids of high quality. We developed a reusable tangential-flow filtration system to concentrate and purify viruses from wastewater for genome sequencing. A pilot study was conducted with 94 wastewater samples from four local sewersheds, from which viral nucleic acids were extracted, and the whole genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was sequenced using the ARTIC V4.0 primers. Our method yielded a high probability (0.9) of recovering complete or near-complete SARS-CoV-2 genomes (>90% coverage at 10× depth) from wastewater when the COVID-19 incidence rate exceeded 33 cases per 100 000 people. The relative abundances of sequenced SARS-CoV-2 variants followed the trends observed from patient-derived samples. We also identified SARS-CoV-2 lineages in wastewater that were underrepresented or not present in the clinical whole-genome sequencing data. The developed tangential-flow filtration system can be easily adopted for the sequencing of other viruses in wastewater, particularly those at low concentrations., Competing Interests: The authors declare no competing financial interest.

The use of wastewater-grown microalgae has the potential to reduce the cost of algae-derived biofuels while simultaneously advancing nutrient recovery at water resource recovery facilities (WRRFs). However, a significant barrier has been the low yield and high protein content of phototrophic biomass. Here, we examine the use of solids residence time (SRT) as a selective pressure in driving biochemical composition, yield, biofuel production, and WRRF nutrient management cost. We cultivated mixed phototrophic communities in controlled, laboratory-scale photobioreactors on the local WRRF secondary effluent to link SRT with biochemical composition and techno-economic analysis to yield insights into biomass composition and downstream processing effects on minimum fuel selling price. SRT significantly impacted biochemical composition, with total and dynamic carbohydrates the highest at low SRT (total carbohydrates being 0.60 and 0.32 mg-carbohydrate·mg-protein -1 at SRT 5 and 15 days, respectively). However, there were distinct differences between extant, steady-state performance and intrinsic potential, and longer SRT communities were able to accumulate significant fractions (51% on an ash-free dry weight basis, AFDW %) of carbohydrate reserves under nutrient starvation. Overall, hydrothermal liquefaction (HTL) was found to be more suitable than lipid extraction for hydrotreating (LEH) and combined algal processing (CAP) for conversion of biomass to fuels, but LEH and CAP became more competitive when intrinsic carbon storage potential was realized. The results suggest that the use of algae for nutrient recovery could reduce the nutrient management cost at WRRFs through revenue from algal biofuels, with HTL resulting in a net revenue.

Suspended growth, mixed community phototrophic wastewater treatment systems (including high-rate algal ponds and photobioreactors) have the potential to achieve biological nitrogen and phosphorus recovery with effluent nutrient concentrations below the current limit-of-technology. In order to achieve reliable and predictive performance, it is necessary to establish a thorough understanding of how design and operational decisions influence the complex community structure governing nutrient recovery in these systems. Solids residence time (SRT), a critical operational parameter governing growth rate, was leveraged as a selective pressure to shape microbial community structure in laboratory-scale photobioreactors fed secondary effluent from a local wastewater treatment plant. In order to decouple the effects of SRT and hydraulic retention time (HRT), nutrient loading was fixed across all experimental conditions and the effect of changing SRT on microbial community structure, diversity, and stability, as well as its impact on nutrient recovery, was characterized. Reactors were operated at distinct SRTs (5, 10, and 15 days) with diurnal lighting over long-term operation (>6 SRTs), and in-depth examination of the eukaryotic and bacterial community structure was performed using amplicon-based sequencing of the 18S and 16S rRNA genes, respectively. In order to better represent the microalgal community structure, this study leveraged improved 18S rRNA gene primers that have been shown to provide a more accurate representation of the wastewater process-relevant algal community members. Long-term operation resulted in distinct eukaryotic communities across SRTs, independent of the relative abundance of Operational Taxonomic Units (OTUs) in the inoculum. The longest SRT (15 days, SRT 15) resulted in a more stable algal community along with stable bacterial nitrification, while the shortest SRT (5 days, SRT 5) resulted in a less stable, more dynamic community. Although SRT was not strongly associated with overall bacterial diversity, the eukaryotic community of SRT 15 was significantly less diverse and less even than SRT 5, with a few dominant OTUs making up a majority of the eukaryotic community structure in the former. Overall, although longer SRTs promote stable bacterial nitrification, short SRTs promote higher eukaryotic diversity, increased functional stability, and better total N removal via biomass assimilation. These results indicate that SRT may be a key factor in not only controlling microalgal community membership, but community diversity and functional stability as well. Ultimately, the efficacy and reliability of NH 4 + removal may be in tension with TN removal in mixed phototrophic systems given that lower SRTs may achieve better total N removal (via biomass assimilation) through increased eukaryotic diversity, biomass productivity, and functional stability., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The capacity of microalgae to advance the limit of technology of nutrient recovery and accumulate storage carbon make them promising candidates for wastewater treatment. However, the extent to which these capabilities are influenced by microbial community composition remains poorly understood. To address this knowledge gap, 3 mixed phototrophic communities sourced from distinct latitudes within the continental United States (28° N, Tampa, FL; 36° N, Durham, NC; and 40° N, Urbana, IL) were operated in sequencing batch reactors (8 day solids residence time, SRT) subjected to identical diel light cycles with media addition at the start of the nighttime period. Despite persistent differences in community structure as determined via 18S rRNA (V4 and V8-V9 hypervariable regions) and 16S rRNA (V1-V3) gene amplicon sequencing, reactors achieved similar and stable nutrient recovery after 2 months (8 SRTs) of operation. Intrinsic carbohydrate and lipid storage capacity and maximum specific carbon storage rates differed significantly across communities despite consistent levels of observed carbon storage across reactors. This work supports the assertion that distinct algal communities cultivated under a common selective environment can achieve consistent performance while maintaining independent community structures and intrinsic carbon storage capabilities, providing further motivation for the development of engineered phototrophic processes for wastewater management.

Microalgal treatment systems could advance nutrient recovery from wastewater by achieving effluent nitrogen (N) and phosphorus (P) levels below the current limit of technology, but their successful implementation requires an understanding of how design decisions influence nutrient uptake over daily (i.e., diel) cycles. This work demonstrates the ability to influence microalgal N:P recovery ratio via solids residence time (SRT) while maintaining complete nutrient removal across day/night cycles through carbon storage and mobilization. Experiments were conducted with two microalgal species, Scenedesmus obliquus and Chlamydomonas reinhardtii, in photobioreactors (PBRs) operated as cyclostats (chemostats subjected to simulated natural light cycles) with retention times of 6-22 days (S. obliquus) and 7-13 days (C. reinhardtii). Nutrient loading and all other factors were fixed across all experiments. Elevated SRTs (>8 days) achieved limiting nutrient concentrations (either N or P) below the detection limit throughout the diel cycle. N:P mass ratio in algal biomass was linearly correlated with SRT, varying from 9.9:1 to 5.0:1 (S. obliquus) and 4.7:1 to 4.3:1 (C. reinhardtii). Carbohydrate content of biomass increased in high irradiance and decreased in low irradiance and darkness across all experiments, whereas lipid dynamics were minimal over 24-h cycles. Across all nutrient-limited cultures, specific (i.e., protein-normalized) dynamic carbohydrate generally decreased with increasing SRT. Nighttime consumption of stored carbohydrate fueled uptake of nutrients, enabling complete nutrient limitation throughout the night. Dynamic carbohydrate consumption for nutrient assimilation was consistent with dark protein synthesis but less than that of heterotrophic growth, underscoring the need for algal process models to decouple growth from nutrient uptake in periods of low/no light. The ability to tailor microalgal N:P uptake ratio and target an optimal energy storage metabolism with traditional engineering process controls (such as SRT) may enable advanced nutrient recovery facilities to target continuous and reliable dual limitation of nitrogen and phosphorus., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Unlabelled: The use of high-throughput sequencing technologies with the 16S rRNA gene for characterization of bacterial and archaeal communities has become routine. However, the adoption of sequencing methods for eukaryotes has been slow, despite their significance to natural and engineered systems. There are large variations among the target genes used for amplicon sequencing, and for the 18S rRNA gene, there is no consensus on which hypervariable region provides the most suitable representation of diversity. Additionally, it is unclear how much PCR/sequencing bias affects the depiction of community structure using current primers. The present study amplified the V4 and V8-V9 regions from seven microalgal mock communities as well as eukaryotic communities from freshwater, coastal, and wastewater samples to examine the effect of PCR/sequencing bias on community structure and membership. We found that degeneracies on the 3' end of the current V4-specific primers impact read length and mean relative abundance. Furthermore, the PCR/sequencing error is markedly higher for GC-rich members than for communities with balanced GC content. Importantly, the V4 region failed to reliably capture 2 of the 12 mock community members, and the V8-V9 hypervariable region more accurately represents mean relative abundance and alpha and beta diversity. Overall, the V4 and V8-V9 regions show similar community representations over freshwater, coastal, and wastewater environments, but specific samples show markedly different communities. These results indicate that multiple primer sets may be advantageous for gaining a more complete understanding of community structure and highlight the importance of including mock communities composed of species of interest., Importance: The quantification of error associated with community representation by amplicon sequencing is a critical challenge that is often ignored. When target genes are amplified using currently available primers, differential amplification efficiencies result in inaccurate estimates of community structure. The extent to which amplification bias affects community representation and the accuracy with which different gene targets represent community structure are not known. As a result, there is no consensus on which region provides the most suitable representation of diversity for eukaryotes. This study determined the accuracy with which commonly used 18S rRNA gene primer sets represent community structure and identified particular biases related to PCR amplification and Illumina MiSeq sequencing in order to more accurately study eukaryotic microbial communities., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

This study evaluated the role of physical and biological filter characteristics on the reduction of MS2 bacteriophage in biosand filters (BSFs). Three full-scale concrete Version 10 BSFs, each with a 55 cm sand media depth and a 12 L charge volume, reached 4 log10 reduction of MS2 within 43 days of operation. A consistently high reduction of MS2 between 4 log10 and 7 log10 was demonstrated for up to 294 days. Further examining one of the filters revealed that an average of 2.8 log10 reduction of MS2 was achieved within the first 5 cm of the filter, and cumulative virus reduction reached an average of 5.6 log10 after 240 days. Core sand samples from this filter were taken for protein, carbohydrate, and genomic extraction. Higher reduction of MS2 in the top 5 cm of the sand media (0.56 log10 reduction per cm vs 0.06 log10 reduction per cm for the rest of the filter depth) coincided with greater diversity of microbial communities and increased concentrations of carbohydrates. In the upper layers, "Candidatus Nitrosopumilus maritimus" and "Ca. Nitrospira defluvii" were found as dominant populations, while significant amounts of Thiobacillus-related OTUs were detected in the lower layers. Proteolytic bacterial populations such as the classes Sphingobacteria and Clostridia were observed over the entire filter depth. Thus, this study provides the first insight into microbial community structures that may play a role in MS2 reduction in BSF ecosystems. Overall, besides media ripening and physical reduction mechanisms such as filter depth and long residence time (45 min vs 24 ± 8.5 h), the establishment of chemolithotrophs and proteolytic bacteria could greatly enhance the reduction of MS2.

The negative energy balance of wastewater treatment could be reversed if anaerobic technologies were implemented for organic carbon oxidation and phototrophic technologies were utilized for nutrient recovery. To characterize the potential for energy positive wastewater treatment by anaerobic and phototrophic biotechnologies we performed a comprehensive literature review and analysis, focusing on energy production (as kJ per capita per day and as kJ m(-3) of wastewater treated), energy consumption, and treatment efficacy. Anaerobic technologies included in this review were the anaerobic baffled reactor (ABR), anaerobic membrane bioreactor (AnMBR), anaerobic fluidized bed reactor (AFB), upflow anaerobic sludge blanket (UASB), anaerobic sequencing batch reactor (ASBR), microbial electrolysis cell (MEC), and microbial fuel cell (MFC). Phototrophic technologies included were the high rate algal pond (HRAP), photobioreactor (PBR), stirred tank reactor, waste stabilization pond (WSP), and algal turf scrubber (ATS). Average energy recovery efficiencies for anaerobic technologies ranged from 1.6% (MFC) to 47.5% (ABR). When including typical percent chemical oxygen demand (COD) removals by each technology, this range would equate to roughly 40-1200 kJ per capita per day or 110-3300 kJ m(-3) of treated wastewater. The average bioenergy feedstock production by phototrophic technologies ranged from 1200-4700 kJ per capita per day or 3400-13 000 kJ m(-3) (exceeding anaerobic technologies and, at times, the energetic content of the influent organic carbon), with usable energy production dependent upon downstream conversion to fuels. Energy consumption analysis showed that energy positive anaerobic wastewater treatment by emerging technologies would require significant reductions of parasitic losses from mechanical mixing and gas sparging. Technology targets and critical barriers for energy-producing technologies are identified, and the role of integrated anaerobic and phototrophic bioprocesses in energy positive wastewater management is discussed.

Morphological alterations of the cerebral vasculature as related to the permeability of plasma proteins and angiographic contrast media following unilateral cerebral angiography were studied. Both Evans blue albumin and horseradish peroxidase were employed as protein tracers for light and electron microscopy investigation respectively. Grey matter regions of the cerebral cortex, cerebellum corpus striatum, hippocampus and midbrain showed the most extensive and consistent leakage of these protein tracers. The most extensive penetration of EBA was noted at 1 hr following cerebral angiography as compared to the 5 or 30 min sample times. Permeability changes were noted in small venules and arterioles as well as capillaries. The extent of permeability, however, was appreciably greater in the capillaires as evidences by rapid extravasation of HRP into the surrounding neuropil extracellular spaces. The glial basement membrane surrounding the perivascular spaces of small venules and arterioles precluded rapid penetration of HRP into the neuropil interstitium. Opening of the tight junctions between the endothelial cells was primarily responsible for the extravasation of HRP in all vessel types. Furthermore, it is out opinion that the hyperosmolar nature of the contrast medium is responsible for opening of these tight junctions.

The object of this study was to determine the type of cerebral vessel affected by injection of radiopaque contrast agents used in cerebral angiography. Seventeen rabbits were prepared surgically for a left intracarotid injection of methylglucamine iothalamate (Conray 60) or methylglucamine diatrizoate (Reno-M-60). Extravasations of the tracers, Evans blue and horseradish peroxidase, occurred in the left half of the brain and occasionally in the right half. Within those areas of blood-brain barrier breakdown, the frequency of leakage was 60% for arterioles, 25% for venules, and 12% for capillaries. The leakage appeared to be primarily intercellular, rather than intracellular. This study provides evidence that greater blood-brain barrier alterations occur in arterioles and venules than in capillaries following cerebral angiography.