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.

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 were 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. © 2018 Water Environment Federation • 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.

Primary sludge fermentate, a concentrated hydrolyzed wastewater carbon, was evaluated for use as an alternative carbon source for mainstream partial denitrificationanammox (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. © 2020 Water Environment Federation • 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.

Background Wastewater treatment plants are an essential part of maintaining the health and safety of the general public. However, they are also an anthropogenic source of antibiotic resistance genes. In this study, we characterized the resistome, the distribution of classes 1–3 integron-integrase genes (intI1, intI2, and intI3) as mobile genetic element biomarkers, and the bacterial and phage community compositions in the North End Sewage Treatment Plant in Winnipeg, Manitoba. Samples were collected from raw sewage, returned activated sludge, final effluent, and dewatered sludge. A total of 28 bacterial and viral metagenomes were sequenced over two seasons, fall and winter. Integron-integrase genes, the 16S rRNA gene, and the coliform beta-glucuronidase gene were also quantified during this time period. Results Bacterial classes observed above 1% relative abundance in all treatments were Actinobacteria (39.24% ± 0.25%), Beta-proteobacteria (23.99% ± 0.16%), Gamma-proteobacteria (11.06% ± 0.09%), and Alpha-proteobacteria (9.18 ± 0.04%). Families within the Caudovirales order: Siphoviridae (48.69% ± 0.10%), Podoviridae (23.99% ± 0.07%), and Myoviridae (19.94% ± 0.09%) were the dominant phage observed throughout the NESTP. The most abundant bacterial genera (in terms of average percent relative abundance) in influent, returned activated sludge, final effluent, and sludge, respectively, includes Mycobacterium (37.4%, 18.3%, 46.1%, and 7.7%), Acidovorax (8.9%, 10.8%, 5.4%, and 1.3%), and Polaromonas (2.5%, 3.3%, 1.4%, and 0.4%). The most abundant class of antibiotic resistance in bacterial samples was tetracycline resistance (17.86% ± 0.03%) followed by peptide antibiotics (14.24% ± 0.03%), and macrolides (10.63% ± 0.02%). Similarly, the phage samples contained a higher prevalence of macrolide (30.12% ± 0.30%), peptide antibiotic (10.78% ± 0.13%), and tetracycline (8.69% ± 0.11%) resistance. In addition, intI1 was the most abundant integron-integrase gene throughout treatment (1.14 × 104 gene copies/mL) followed by intI3 (4.97 × 103 gene copies/mL) while intI2 abundance remained low (6.4 × 101 gene copies/mL). Conclusions Wastewater treatment successfully reduced the abundance of bacteria, DNA phage and antibiotic resistance genes although many antibiotic resistance genes remained in effluent and biosolids. The presence of integron-integrase genes throughout treatment and in effluent suggests that antibiotic resistance genes could be actively disseminating resistance between both environmental and pathogenic bacteria.

We investigated the potential use and quantification of human enteric viruses in municipal wastewater samples of Winnipeg (Manitoba, Canada) as alternative indicators of contamination and evaluated the processing stages of the wastewater treatment plant. During the fall 2019 and winter 2020 seasons, samples of raw sewage, activated sludge, effluents, and biosolids (sludge cake) were collected from the North End Sewage Treatment Plant (NESTP), which is the largest wastewater treatment plant in the City of Winnipeg. DNA (Adenovirus and crAssphage) and RNA enteric viruses (Pepper mild mottle virus, Norovirus genogroups GI and GII, Rotavirus Astrovirus, and Sapovirus) as well as the uidA gene found in Escherichia coli were targeted in the samples collected from the NESTP. Total nucleic acids from each wastewater treatment sample were extracted using a commercial spin-column kit. Enteric viruses were quantified in the extracted samples via quantitative PCR using TaqMan assays. Overall, the average gene copies assessed in the raw sewage were not significantly different (p-values ranged between 0.1023 and 0.9921) than the average gene copies assessed in the effluents for DNA and RNA viruses and uidA in terms of both volume and biomass. A significant reduction (p-value ≤ 0.0438) of Adenovirus and Noroviruses genogroups GI and GII was observed in activated sludge samples compared with those for raw sewage per volume. Higher GCNs of enteric viruses were observed in dewatered sludge samples compared to liquid samples in terms of volume (g of sample) and biomass (ng of nucleic acids). Enteric viruses found in gene copy numbers were at least one order of magnitude higher than the E. coli marker uidA, indicating that enteric viruses may survive the wastewater treatment process and viral-like particles are being released into the aquatic environment. Viruses such as Noroviruses genogroups GI and GII, and Rotavirus were detected during colder months. Our results suggest that Adenovirus, crAssphage, and Pepper mild mottle virus can be used confidently as complementary viral indicators of human fecal pollution.

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