Comammox and unknown ammonia oxidizers contribute to nitrite accumulation in an integrated A-B stage process that incorporates side-stream EBPR (S2EBPR).

A novel integrated pilot-scale A-stage high rate activated sludge, B-stage short-cut biological nitrogen removal and side-stream enhanced biological phosphorus removal (A/B-shortcut N-S2EBPR) process for treating municipal wastewater was demonstrated with the aim to achieve simultaneous and carbon- and energy-efficient N and P removal. In this studied period, an average of 7.62 ± 2.17 mg-N/L nitrite accumulation was achieved through atypical partial nitrification without canonical known NOB out-selection. Network analysis confirms the central hub of microbial community as Nitrospira, which was one to two orders of magnitude higher than canonical aerobic oxidizing bacteria (AOB) in a B-stage nitrification tank. The contribution of comammox Nitrospira as AOB was evidenced by the increased amoB/nxr ratio and higher ammonia oxidation activity. Furthermore, oligotyping analysis of Nitrospira revealed two dominant sub-clusters (microdiveristy) within the Nitrospira. The relative abundance of oligotype II, which is phylogenetically close to Nitrospira_midas_s_31566, exhibited a positive correlation with nitrite accumulation in the same operational period, suggesting its role as comammox Nitrospira. Additionally, the phylogenetic investigation suggested that heterotrophic organisms from the family Comamonadacea and the order Rhodocyclaceae embedding ammonia monooxygenase and hydroxylamine oxidase may function as heterotrophic nitrifiers. This is the first study that elucidated the impact of integrating the S2EBPR on nitrifying populations with implications on short-cut N removal. The unique conditions in the side-stream reactor, such as low ORP, favorable VFA concentrations and composition, seemed to exert different selective forces on nitrifying populations from those in conventional biological nutrient removal processes. The results provide new insights for integrating EBPR with short-cut N removal process for mainstream wastewater treatment.

Side-Stream Enhanced Biological Phosphorus Removal (S2EBPR) enables effective phosphorus removal in a pilot-scale A-B stage shortcut nitrogen removal system for mainstream municipal wastewater treatment.

While the adsorption/bio-oxidation (A/B) process has been widely studied for carbon capture and shortcut nitrogen (N) removal, its integration with enhanced biological phosphorus (P) removal (EBPR) has been considered challenging and thus unexplored. Here, full-scale pilot testing with an integrated system combining A-stage high-rate activated sludge with B-stage partial (de)nitrification/anammox and side-stream EBPR (HRAS-P(D)N/A-S2EBPR) was conducted treating real municipal wastewater. The results demonstrated that, despite the relatively low influent carbon load, the B-stage P(D)N-S2EBPR system could achieve effective P removal performance, with the carbon supplement and redirection of the A-stage sludge fermentate to the S2EBPR. The novel process configuration design enabled a system shift in carbon flux and distribution for efficient EBPR, and provided unique selective factors for ecological niche partitioning among different key functionally relevant microorganisms including polyphosphate accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs). The combined nitrite from B-stage to S2EBPR and aerobic-anoxic conditions in our HRAS-P(D)N/A-S2EBPR system promoted DPAOs for simultaneous internal carbon-driven denitrification via nitrite and P removal. 16S rRNA gene-based oligotyping analysis revealed high phylogenetic microdiversity within the Accumulibacter population and discovered coexistence of certain oligotypes of Accumulibacter and Competibacter correlated with efficient P removal. Single-cell Raman micro-spectroscopy-based phenotypic profiling showed high phenotypic microdiversity in the active PAO community and the involvement of unidentified PAOs and internal carbon-accumulating organisms that potentially played an important role in system performance. This is the first pilot study to demonstrate that the P(D)N-S2EBPR system could achieve shortcut N removal and influent carbon-independent EBPR simultaneously, and the results provided insights into the effects of incorporating S2EBPR into A/B process on metabolic activities, microbial ecology, and resulted system performance.

Impact of instrumentation reliability on mainstream suspended partial denitrification-anammox (PdNA).

This study successfully revealed the importance of probe reliability and sensitivity with ion sensitive electrode (ISE) probes on achieving high partial denitrification (PdN) efficiency; and decreasing carbon overdosing events that cause the decline of microbial populations and performance of PdNA. In a mainstream integrated hybrid granule-floc system, an average PdN efficiency of 76% was achieved with acetate as the carbon source. Thauera was identified as the dominant PdN species; its presence in the system was analogous to instrumentation reliability and PdN selection and was not a consequence of bioaugmentation. Up to 27-121 mg total inorganic nitrogen/L/d, an equivalent of 18-48% of the overall total inorganic nitrogen removed, was achieved through the PdNA pathway. Candidatus Brocadia was the main anoxic ammonium oxidizing bacteria species that was seeded from sidestream and enriched and retained in the mainstream system with observed growth rates of 0.04-0.13 day-1 . Moreover, there was no direct negative impact of methanol's use for post-polishing on anoxic ammonium oxidizing bacteria activity and growth. PRACTITIONER POINTS: Stress testing with ISE sensors revealed the importance of probe reliability and sensitivity on PdN selection and PdNA performance. Up to 121 mg TIN/L/d was achieved via PdNA in a mainstream suspended hybrid granule-floc partial denitrification-anammox (PdNA) system. Candidatus Brocadia was the dominant AnAOB species with observed growth rates of 0.04-0.13 day-1. There was no direct negative impact of methanol's use for post-polishing on AnAOB activity and growth.

Introducing bioflocculation boundaries in process control to enhance effluent quality of high-rate contact-stabilization systems.

High-rate activated sludge (HRAS) systems suffer from high variability of effluent quality, clarifier performance, and carbon capture. This study proposed a novel control approach using bioflocculation boundaries for wasting control strategy to enhance effluent quality and stability while still meeting carbon capture goals. The bioflocculation boundaries were developed based on the oxygen uptake rate (OUR) ratio between contactor and stabilizer (feast/famine) in a high-rate contact stabilization (CS) system and this OUR ratio was used to manipulate the wasting setpoint. Increased oxidation of carbon or decreased wasting was applied when OUR ratio was <0.52 or >0.95 to overcome bioflocculation limitation and maintain effluent quality. When no bioflocculation limitations (OUR ratio within 0.52-0.95) were detected, carbon capture was maximized. The proposed control concept was shown for a fully automated OUR-based control system as well as for a simplified version based on direct waste flow control. For both cases, significant improvements in effluent suspended solids level and stability (<50-mg TSS/L), solids capture over the clarifier (>90%), and COD capture (median of 32%) were achieved. This study shows how one can overcome the process instability of current HRAS systems and provide a path to achieve more reliable outcomes. PRACTITIONER POINTS: Online bioflocculation boundaries (upper and lower limit) were defined by the OUR ratio between contactor and stabilizer (feast/famine). To maintain effluent quality, carbon oxidation was minimized when bioflocculation was not limited (0.52-0.95 OUR ratio) and increased otherwise. A fully automated control concept was piloted, also a more simplified semiautomated option was proposed. Wasting control strategies with bioflocculation boundaries improved effluent quality while meeting carbon capture goals. Bioflocculation boundaries are easily applied to current wasting control schemes applied to HRAS systems (i.e., MLSS, SRT, and OUR controls).

Mainstream partial denitrification-anammox in sand and expanded clay deep-bed polishing filters under practical loading rates and backwashing conditions.

This study focused on evaluating the feasibility of expanded clay and sand as media types for mainstream partial denitrification-anammox (PdNA) in deep-bed single-media polishing filters under nitrogen and solids loading rates as well as backwash conditions similar to conventional denitrification filters. The surface roughness and iron content of the expanded clay were hypothesized to allow for enhanced anammox retention, nitrogen removal rates, and runtimes. However, under the tested loading rates and backwash conditions, no clear benefit of expanded clay was observed compared with conventional sand. This study showed the feasibility of PdNA in filters with both sand and expanded clay with PdN efficiencies of 76% and 77%, PdNA rates of 840 and 843 g N/m3 /d and TIN removal rates of 960 and 964 g N/m3 /d, respectively. Glycerol demands were 1.5-1.6 g COD added per g TIN removed , thus indicating potential carbon savings up to 75% compared with conventional denitrification. Overall, this study showed for the first time PdNA filters performing at nitrogen removal rates double that of previous PdNA studies under realistic conditions while providing insights into the media choice and backwashing conditions. Future research on expanded clay backwash conditions is needed to provide its full potential in PdNA filters. PRACTITIONER POINTS: Hydraulic and TSS loading rates similar to conventional denitrification can be applied in PdNA filters. Conventional sand can be used when retrofitting conventional denitrification filters into PdNA filters. Carbon savings up to 75% can be achieved with glycerol when retrofitting conventional filters into PdNA filters.

Media selection for anammox-based polishing filters: Balancing anammox enrichment and retention with filtration function.

Retrofitting conventional denitrification filters into partial denitrification-anammox (PdNA)- or anammox (AnAOB)-based filters will reduce the needs for external carbon addition. The success of AnAOB-based filters depends on anammox growth and retention within such filters. Studies have overlooked the importance of media selection and its impact on AnAOB capacity, head loss progression dynamics, and shear conditions applied onto the AnAOB biofilm. The objective of this study was to evaluate viable media types (10 types) that can enhance AnAOB rates for efficient nitrogen removal in filters. Given the higher backwash requirement and lower AnAOB capacity of the conventionally used sand, expanded clay (3-5 mm) was recommended for AnAOB-based filters in this study. Owing to its surface characteristics, expanded clay had higher AnAOB activity (304- vs. 104-g NH4 + -N/m2 /day) and higher AnAOB retention (43% more) than sand. Increasing the iron content of expanded clay to 37% resulted in an increase in zeta potential, which led to 56% more anammox capacity compared to expanded clay with 7% iron content. This work provides insight into the importance of media types in the growth and retention of AnAOB in filters, and this knowledge could be used as basis in the development of PdNA filters. PRACTITIONER POINTS: Expanded clay showed the lowest head loss buildup and most likely will result in longer runtime for full-scale PdNA applications The highest AnAOB rates were achieved in expanded clay types and sand compared with smaller media typically used in biofiltration Expanded clay resulted in better AnAOB retention under shear, whereas sand could not withstand shear and required more frequent backwashing Expanded clay iron coating enhanced AnAOB enrichment and retention, most likely due to increased surface roughness and/or positive charge.

Partial denitrification-anammox (PdNA) application in mainstream IFAS configuration using raw fermentate as carbon source.

This research examined the feasibility of raw fermentate for mainstream partial denitrification-anammox (PdNA) in a pre-anoxic integrated fixed-film activated sludge (IFAS) process. Fermentate quality sampled from a full-scale facility was highly dynamic, with 360-940 mg VFA-COD/L and VFA/soluble COD ratios ranging from 24% to 48%. This study showed that PdNA selection could be achieved even when using low quality fermentate. Nitrate residual was identified as the main factor driving the PdN efficiency, while management of nitrate conversion rates was required to maximize overall PdNA rates. AnAOB limitation was never observed in the IFAS system. Overall, this study showed PdN efficiencies up to 38% and PdNA rates up to 1.2 ± 0.7 g TIN/m2 /d with further potential for improvements. As a result of both PdNA and full denitrification, this concept showed the potential to save 48-89% methanol and decrease the carbon footprint of water resource recovery facilities (WRRF) by 9-15%. PRACTITIONER POINTS: Application of PdNA with variable quality fermentate is feasible when the nitrate residual concentration is increased to enhance PdN selection. To maximize nitrogen removed through PdNA, nitrate conversion rates need enhancement through optimization of upstream aeration and PdN control setpoints. The IFAS PdNA process was never anammox limited; success depended on the degree of PdN achieved to make nitrite available. Application of PdNA with fermentate can yield 48-89% savings in methanol or other carbon compared with conventional nitrification and denitrification. Integrating PdNA upstream from polishing aeration and anoxic zones guarantees that stringent limits can be met (<5 mg N/L).

Primary sludge fermentate as carbon source for mainstream partial denitrification-anammox (PdNA).

Primary sludge fermentate, a concentrated hydrolyzed wastewater carbon, was evaluated for use as an alternative carbon source for mainstream partial denitrification-anammox (PdNA) in a suspended growth activated sludge process in terms of partial denitrification (PdN) efficiency, PdNA nitrogen removal contributions, and final effluent quality. Fermenter operation at a 2-day sludge retention time (SRT) resulted in the maximum achievable yield of 0.14 ± 0.05 g sCOD/g VSS without release of excessive ammonia and phosphorus to the system. Based on the results of batch experiments, fermentate addition led to PdN efficiency of 93 ± 14%, which was similar to acetate at a nitrate residual of 2-3 mg N/L. In the pilot-scale mainstream deammonification reactor, PdN efficiency using fermentate was 49 ± 24%, which was lower than acetate (66 ± 24% during acetate period I and 70 ± 21% during acetate period II), most probably due to lower nitrate and ammonium kinetics in the PdN zone. Methanol cost-saving potential for the application of PdNA as the main short-cut nitrogen pathway was estimated to be 30% to 55% depending on the PdN efficiency achieved. PRACTITIONER POINTS: Primary sludge fermentate was evaluated as an alternative carbon source for mainstream partial denitrification-anammox (PdNA). Fermenter operated at a 1 to 2 day SRT resulted in the maximum achievable yield without the release of excessive ammonia and phosphorus to the system. Although 93% partial denitrification efficiency was achieved with fermentate in batch experiments, around 49% PdN efficiency was achieved in pilot studies. Application of PdNA with fermentate can result in significant methanol cost savings.

Nitrate residual as a key parameter to efficiently control partial denitrification coupling with anammox.

Despite the increased research efforts, full-scale implementation of shortcut nitrogen removal strategies has been challenged by the lack of consistent nitrite-oxidizing bacteria out-selection. This paper proposes an alternative path using partial denitrification (PdN) selection coupled with anaerobic ammonium-oxidizing bacteria (AnAOB). A nitrate residual concentration (>2 mg N/L) was identified as the crucial factor for metabolic PdN selection using acetate as a carbon source, unlike the COD/N ratio which was often suggested. Therefore, a novel and simple acetate dosing control strategy based on maintaining a nitrate concentration was tested in the absence and presence of AnAOB, achieving PdN efficiencies above 80%. The metabolic-based PdN selection allowed for flexibility to move between PdN and full denitrification when required to meet effluent nitrate levels. Due to the independence of this strategy on species selection and management of nitrite competition, this novel approach will guarantee nitrite availability for AnAOB under mainstream conditions unlike shortcut nitrogen removal approaches based on NOB out-selection. Overall, a COD addition of only 2.2 g COD/g TIN removed was needed for the PdN-AnAOB concept showing its potential for significant savings in external carbon source needs to meet low TIN effluent concentrations making this concept a competitive alternative. PRACTITIONER POINTS: Nitrate residual is the key control parameter for partial denitrification selection. Metabolic selection allowed for flexibility of moving from partial to full denitrification. 2.2 g COD/g TIN removed was needed for partial denitrification-anammox process.

'Hot topic' - combined energy and process modeling in thermal hydrolysis systems.

The thermal hydrolysis process (THP) is applied to enhance biogas production in anaerobic digestion (AD), reduce viscosity for improved mixing and dewatering and to reduce and sterilize cake solids. Large heat demands for steam production rely on dynamic effects like sludge throughput, gas availability and THP process parameters. Here, we propose a combined energy and process model suitable to describe the dynamic behaviour of THP in a full-plant context. The process model addresses interactions of THP with operational conditions covered by the AD model obeying mass continuity. Energy conservation is considered in balancing and converting various energy species dominated by thermal heat and calorific energy. The combined energy and process model was then applied on the THP at Blue Plains advanced WWTP (DC Water) to analyse the process and assess potential energy optimizations. It was found that dynamic effects like mismatched steam production and consumption, temporary gas shortages and underloaded units are responsible for energy inefficiencies with losses in electricity-production up to 29%.

Impact of carbon source and COD/N on the concurrent operation of partial denitrification and anammox.

In this study, concurrent operation of anammox and partial denitrification within a nonacclimated mixed culture system was proposed. The impact of carbon sources (acetate, glycerol, methanol, and ethanol) and COD/NO3- -N ratio on partial denitrification selection under both short- and long-term operations was investigated. Results from short-term testing showed that all carbon sources supported partial denitrification. However, acetate and glycerol were preferred due to their display of efficient partial denitrification selection, which may be related to their different electron transport pathways in comparison with methanol. Long-term operation confirmed results of batch tests by showing the contribution of partial denitrification to nitrate removal above 90% after acclimation in both acetate and glycerol reactors. In contrast, methanol showed challenges of maintaining efficient partial denitrification. COD/NO3- -N ratio mainly controlled the rate of nitrate reduction and not directly partial denitrification selection; thus, it should be used to balance between denitrification rate and anammox rate. PRACTITIONER POINTS: The authors aimed to investigate the impact of carbon sources and COD/NO3-N ratio on partial denitrification selection. All the carbon sources supported partial denitrification as long as the nitrite sink was available. 90% partial denitrification could be achieved with both acetate and glycerol in long-term operations. COD/NO3-N ratio did not directly control partial denitrification but can be used to balance between denitrification rate and anammox rate.

Supernatant organics from anaerobic digestion after thermal hydrolysis cause direct and/or diffusional activity loss for nitritation and anammox.

Treatment of sewage sludge with a thermal hydrolysis process (THP) followed by anaerobic digestion (AD) enables to boost biogas production and minimize residual sludge volumes. However, the reject water can cause inhibition to aerobic and anoxic ammonium-oxidizing bacteria (AerAOB & AnAOB), the two key microbial groups involved in the deammonification process. Firstly, a detailed investigation elucidated the impact of different organic fractions present in THP-AD return liquor on AerAOB and AnAOB activity. For AnAOB, soluble compounds linked to THP conditions and AD performance caused the main inhibition. Direct inhibition by dissolved organics was also observed for AerAOB, but could be overcome by treating the filtrate with extended aerobic or anaerobic incubation or with activated carbon. AerAOB additionally suffered from particulate and colloidal organics limiting the diffusion of substrates. This was resolved by improving the dewatering process through an optimized flocculant polymer dose and/or addition of coagulant polymer to better capture the large colloidal fraction, especially in case of unstable AD performance. Secondly, a new inhibition model for AerAOB included diffusion-limiting compounds based on the porter-equation, and achieved the best fit with the experimental data, highlighting that AerAOB were highly sensitive to large colloids. Overall, this paper for the first time provides separate identification of organic fractions within THP-AD filtrate causing differential types of inhibition. Moreover, it highlights the combined effect of the performance of THP, AD and dewatering on the downstream autotrophic nitrogen removal kinetics.

A greenhouse gas source of surprising significance: anthropogenic CO2 emissions from use of methanol in sewage treatment.

The impact of methanol (CH3OH) as a source of anthropogenic carbon dioxide (CO2) in denitrification at wastewater treatment plants (WWTPs) has never been quantified. CH3OH is the most commonly purchased carbon source for sewage denitrification. Until recently, greenhouse gas (GHG) reporting protocols consistently ignored the liberation of anthropogenic CO2 attributable to CH3OH. This oversight can likely be attributed to a simplifying notion that CO2 produced through activated-sludge-process respiration is biogenic because most raw-sewage carbon is un-sequestered prior to entering a WWTP. Instead, a biogenic categorization cannot apply to fossil-fuel-derived carbon sources like CH3OH. This paper provides a summary of how CH3OH use at DC Water's Blue Plains Advanced Wastewater Treatment Plant (AWTP; Washington, DC, USA) amounts to 60 to 85% of the AWTP's Scope-1 emissions. The United States Environmental Protection Agency and Water Environment Federation databases suggest that CH3OH CO2 likely represents one quarter of all Scope-1 GHG emissions attributable to sewage treatment in the USA. Finally, many alternatives to CH3OH use exist and are discussed.

Method to identify potential phosphorus rate-limiting conditions in post-denitrification biofilm reactors within systems designed for simultaneous low-level effluent nitrogen and phosphorus concentrations.

Water-quality standards requiring simultaneous low level effluent N and P concentrations are increasingly common in Europe and the United States of America. Moving bed biofilm reactors (MBBRs) and biologically active filters (BAFs) have been used as post-denitrification biofilm reactors in processes designed and operated for this purpose (Boltz et al., 2010a). There is a paucity of information describing systematic design and operational protocols that will minimize the potential for phosphorus rate-limited conditions as well as a lack of information describing the interaction between these post-denitrification biofilm reactors and unit processes that substantially alter phosphorus speciation (e.g., chemically enhanced clarification). In this paper, a simple mathematical model for estimating the threshold below which P becomes rate-limiting, and the model is presented and evaluated by comparing its predictions with operational data from post-denitrification MBBRs and BAFs. Ortho-phosphorus (PO(4)-P), which is the dissolved reactive component of total phosphorus, was a primary indicator of P rate-limiting conditions in the evaluated post-denitrification biofilm reactors. The threshold below which PO(4)-P becomes the rate-limiting substrate is defined: S(PO4-P):S(NOx-N) = 0.0086 g P/g N and S(PO4-P):S(M) = 0.0013 g P/g COD. Additional analyses indicate J(NOx-N)(avg) =0.48 g/m2/d when S(PO4-P):S(NOx-N) > 0.0086, and J(NOx-N)(avg) = 0.06 g/m2/d when S(PO4-P):S(NOx-N) < 0.0086. Effluent nitrate-nitrogen plus nitrite-nitrogen concentration (S(NOx-N)) from the evaluated post-denitrification biofilm reactors began to rapidly increase when S(PO4-P):S(NOx-N) was 0.01, approximately (consistent with the rate-limitation threshold of S(PO4-P):S(NOx-N) < 0.0086 predicted by the mathematical model described in this paper). Depending on the processes used at a given WWTP, optimizing chemically enhanced clarification to increase the amount of PO(4)-P that remains in the clarifiers effluent stream, dosing phosphoric acid in the MBBR or BAF influent stream, and/or optimizing secondary process EBPR may overcome phosphorus rate-limitations in the biofilm-based post-denitrification process.