Optimizing nitrogen removal in PD/A reactors: Effects of influent composition and temperature on system stability and microbial dynamics.

This study investigates the performance and microbial community dynamics in two partial denitrification/anammox (PD/A) reactors with different influent wastewater compositions (differ in the presence/absence of NO2-) subjected to a controlled temperature gradient reduction from mesophilic (30 °C) to room temperature (20.92 °C) over 76 days. Two lab-scale PD/A reactors (R1 and R2), both operated with a total inorganic nitrogen (TIN) concentrations of 70 mg N/L. R1 maintained a NH4+/NO2-/NO3- ratio of 3:3:1 and a COD/NO3- ratio of 2.0, while R2 had an NH4+/NO3- ratio of 3:4, and COD/NO3- ratios of 2.0 and 2.5. Our findings reveal distinct responses to the temperature transitions: the optimization of the NH4+/NO2-/NO3- ratio at 3:3:1 facilitated more stable nitrogen removal as temperatures decreased. This stability can be attributed to the enhanced synchronization between anammox bacteria and denitrifiers, promoting a balanced bioconversion process that is less susceptible to temperature-induced disruptions. Notably, the specific anammox activity (SAA) in both reactors declined linearly with the decrease in temperature, but the relative abundance of anammox bacteria (Ca. Brocadia) in R1 increased from 2.1 % to 9.7 %. Furthermore, the percentage of anammox-related key genes was higher in R1 than in R2, suggesting a microbial mechanism underlying the stable performance of R1. These results underscore the significant impact of influent nitrogen composition on PD/A performance amid temperature gradients and highlight the critical role of optimizing influent ratios for maintaining efficient nitrogen removal. This study offers valuable insights into enhancing the stability of PD/A systems under varying thermal conditions.

Starvation resilience in anammox-based bioreactors: A stable nitrogen removal route on partial denitrification/anammox (PD/A).

This study investigates the performance, resilience and microbial community dynamics of two anaerobic processes, i.e. pure anammox (R1) and partial denitrification/anammox (PD/A) (R2), following a 30-day starvation period. The tolerance to starvation was assessed by comparing nitrogen removal efficiency and microbial activity across both reactors. Results show that the PD/A process recovery to pre-starvation performance levels within just one day, as compared to the pure anammox process. Notably, although the activity of anammox bacteria decreased in both processes during starvation, the decay rate in R1 was 69.59 % higher than in R2, potentially explaining the quicker recovery of R2. Furthermore, enhanced secretion of extracellular polymeric substance (EPS) during starvation served as a protective mechanism. The potential functions and genes in microorganisms, as well as the pathway of nitrogen cycling, were demonstrated through analyses using the KEGG database. This research reveals essential mechanistic insights and strategic guidance for the effective implementation of anammox-based biological nitrogen removal processes.

Influence of residual anaerobic granular sludge (AnGS) from anaerobically digested molasses wastewater in aerobic granular sludge reactor.

This study investigated the impact of residual anaerobic granular sludge (AnGS) from anaerobic digesters treating molasses wastewater on ammonium reduction in a downstream aerobic granular sludge (AGS) reactor. Two conditions were tested: raw (high AnGS concentration) and settled (low AnGS concentration) anaerobically digested molasses wastewaters were fed into the AGS reactor. With the introduction of raw wastewater, enhanced nitrite accumulation at 30 % and improved total inorganic nitrogen (TIN) removal at 11 % were observed compared to 1 % nitrite accumulation and 8 % TIN removal with the introduction of settled wastewater. However, AnGS adversely affected other aspects of reactor performance, increasing effluent solid content and decreasing soluble chemical oxygen demand removal efficiency from 20 % in the low AnGS condition to 11 % in the high AnGS condition. Despite the observed retention of AnGS in the reactor, no significant bioaugmentation effects on the microbial community of the AGS were observed. Aerobic granular sludge was consistently observed in both conditions. The study suggests that AnGS may act as a nucleus for granule formation, helping to maintain granule stability in a disturbed environment. This study offers a systematic understanding of the impact of AnGS on subsequent nitrogen removal process using AGS, aiding in the decision making in the treatment of high solid anaerobic digestate.

Mechanisms of anammox granular sludge reactor effluent as biostimulant: Shaping microenvironment for anammox metabolism.

Effluent from anammox granular sludge (AnGS) bioreactor contains microbes and microbial products. This study explored mechanisms of utilizing AnGS-effluent as biostimulant for anammox process enhancement. Compared with no AnGS-effluent supplemented control reactor, 5.0 and 1.3 times higher ammonium nitrogen and total inorganic nitrogen removal rates, respectively were obtained with continuous AnGS-effluent supplementation after 98 days' operation. Anammox bacteria from Candidatus Brocadia accounted for 0.1 % (DNA level) and 1.3 %-1.5 % (RNA level) in control reactor, and 2.9 % (DNA level) and 54.5 %-55.4 % (RNA level) in the AnGS-effluent-fed reactor. Influent microbial immigration evaluation showed that bacterial immigration via AnGS-effluent supplementation was not the main contributor to active anammox community development. Amino acids biosynthesis, B-vitamins and coenzymes metabolism related pathways were facilitated by AnGS-effluent supplementation. AnGS-effluent supplementation aided anammox metabolic activity by shaping microenvironment and microbial interactions. This study provides insights into enhancing anammox bacterial metabolism with AnGS-effluent microbial products as biostimulant.

Treatment of high ammonia anaerobically digested molasses wastewater using aerobic granular sludge reactor.

This study addressed the treatment of high ammonia, low biodegradable chemical oxygen demand (bCOD) anaerobically digested molasses wastewater, utilizing an aerobic granular sludge (AGS) reactor. The AGS achieved 99 % ammonia removal regardless of the bCOD supplementation. By adding low ammonia (<60 mg/L), high bCOD raw molasses wastewater (before anaerobic digestion) as a carbon source, enhanced nitrogen removal, increasing from 10 % to 97 %, and improved sludge settleability via bio-induced calcite precipitation were observed. Functional genes prediction suggested two potential denitrification pathways, including heterotrophic denitrification by Paracoccus and Thauera, and autotrophic denitrification, specifically sulfide-oxidizing autotrophic denitrification by Thiobacillus. An increase in the relative abundance of microorganisms involved in heterotrophic denitrification was observed with the addition of high bCOD raw molasses wastewater. Consequently, incorporating raw molasses wastewater into the AGS presents a sustainable approach to achieve mixotrophic denitrification, maintain stable granular sludge and ensure stable treatment performance when treating anaerobically digested molasses wastewater.

Efficient nitrogen removal from ammonia rich wastewater using aerobic granular sludge (AGS) reactor: Selection and enrichment of effective microbial community.

Anaerobically digested sludge supernatant, characterized by its high ammonia and low biodegradable chemical oxygen demand (COD) content, has raised concerns when returned to mainstream treatment lines due to potential impacts on effluent quality. Addressing this, an aerobic granular sludge (AGS) reactor adopted nitritation/denitritation with external COD addition was utilized and achieved a considerable nitrogen treatment capacity of 4.2 kg N/m3/d, reaching over 90% removal efficiencies for both ammonia and total inorganic nitrogen. This study applied progressively increased nitrogen loading to select for a microbial community that exhibited high nitrogen oxidation and reduction rates, demonstrating peak rates of 0.5 g N/g VSS/d and 3 g N/g VSS/d, respectively. The enrichment of highly efficient microbial community was achieved along with the increased biomass density peaked at 17 g/L MLVSS, with the system retaining small-sized granular sludge at 0.5 mm. The primary ammonia oxidizing bacteria was Nitrosomonas, while Thauera was the dominated denitrifiers. Quantitative polymerase chain reaction analyses reinforced the enhanced nitrogen removal capacity based on the progressively increased abundance of nitrogen cycling functional genes. The high nitrogen treatment capacity, synergistic attributes of high specific microbial activities and the substantial biomass retention, suggest the AGS's efficacy and capacity in ammonia rich wastewater treatment.

Enhancing anammox process with granular activated carbon: A study on Microbial Extracellular Secretions (MESs).

Granular activated carbon (GAC), a porous carbon-based material, provides increased attachment space for functional microorganisms and enhances nitrogen removal by facilitating extracellular electron transfer in the anammox process. This study investigates the effects of GAC on the biosynthesis of microbial extracellular secretions (MESs) and explores the roles of these secretions in anammox activities. Four lab-scale reactors were operated: two downstream UASB reactors (D1 and D2) receiving effluents from the upstream UASB reactors (U1: no-GAC, U2: yes-GAC). Our results indicate that MESs were enhanced with the addition of GAC. The effluent from U2 exhibited a 59.62 % higher amino acid content than that from U1. These secretions contributed to an increase in the nitrogen loading rate (NLR) in the downstream reactors. Specifically, NLR in D1 increased from 130.5 to 142.7 g N/m3/day, and in D2, it escalated from 137.5 to 202.8 g N/m3/day, likely through acting as cross-feeding substrates or vital nutrients. D2 also showed increased anammox bacterial activity, enriched Ca. Brocadia population and hao gene abundance. Furthermore, this study revealed that D2 sludge has significantly higher extracellular polymeric substances (EPS) (48.71 mg/g VSS) and a larger average granule size (1.201 ± 0.119 mm) compared to D1 sludge. Overall, GAC-stimulated MESs may have contributed to the enhanced performance of the anammox process.

Deciphering the role of granular activated carbon (GAC) in anammox: Effects on microbial succession and communication.

Anaerobic ammonium oxidation (anammox) offered an energy-efficient option for nitrogen removal from wastewater. Granular activated carbon (GAC) addition has been reported that improved biomass immobilization, but the role of GAC in anammox reactors has not been sufficiently revealed. In this study, it was observed that GAC addition in an upflow anaerobic sludge blanket (UASB) reactor led to the significantly shortened anammox enrichment time (shortened by 45 days) than the reactor without GAC addition. The nitrogen removal rate was 0.83 kg N/m3/day versus 0.76 kg N/m3/day in GAC and non-GAC reactors, respectively after 255 days' operation. Acyl-homoserine lactone (AHL) quorum sensing signal molecule C8-HSL had comparable concentrations in both anammox reactors, whereas the signal molecule C12-HSL was more pervasive in the reactor containing GAC than the reactor without GAC. Microbial analysis revealed distinct anammox development in both reactors, with Candidatus Brocadia predominant in the reactor that did not contain GAC, and Candidatus Kuenenia predominant in the reactor that contained GAC. Denitrification bacteria likely supported anammox metabolism in both reactors. The analyses of microbial functions suggested that AHL-dependent quorum sensing was enhanced with the addition of GAC, and that GAC possibly augmented the extracellular electron transfer (EET)-dependent anammox reaction.

[Long-term Performance and Bacterial Community Composition Analysis of AGS-SBR Treating the Low COD/N Sewage at Low DO Concentration Condition].

This study utilized the sequencing batch activated sludge reactor (SBR) inoculated aerobic granular sludge (AGS) to treat the low COD/N ratio (<4.0) domestic wastewater under low DO (0.5-1.0 mg·L-1) concentration condition. Long-term performance of simultaneous nitrogen and phosphorus removal and bacterial community composition of AGS-SBR were studied. The results showed that the AGS-SBR system had good and stable decontamination abilities in its 180-day operation. The average removal rates of COD, NH4+-N, TN and TP were 87.17%, 95.21%, 77.05%, and 91.11%, respectively. At the same time, the AGS showed good settling performance, and always kept its integrated and compact structure. No obvious granular sludge disintegration phenomenon occurred in 180 days. Meanwhile, by using Illumina 16S rRNA gene MiSeq sequencing, we investigated the bacterial abundance in AGS-SBR reactor. Proteobacteria, Firmicutes, Chlorobi, Chloroflex, and Bacteroidetes were the dominant microbial communities in the simultaneous nitrogen and phosphorus removal reactor. Denitratisoma, Planctomycetaceae, Thauera, Comamonas, Nitrosomonas and Nitrospira were suggested to be the primary organisms responsible for the nitrogen removal. Clostridium and Anaerolinea were the main bacterial communities of phosphorus removal.