Expression of the nirS, hzsA, and hdh genes and antibiotic resistance genes in response to recovery of anammox process inhibited by oxytetracycline.

The inhibitory effects of oxytetracycline (OTC) on the anaerobic ammonium oxidation (anammox) performance were relieved by employing bio-augmentation (BA) tactics. However, the recovery mechanism was vague. The response of specific anammox activity (SAA), heme c, functional genes, extracellular polymeric substance (EPS) and antibiotics resistance genes (ARGs) to OTC inhibition and BA aid were traced in the present study. The results indicated that response of SAA, heme c content and functional genes, such as nirS, hzsA and hdh to OTC inhibition were not synchronous. The presence of the tetC, tetG, tetX, and intI1 genes enhanced the resistance of anammox sludge to OTC, thus accelerating the performance recovery when aided by BA. A significant correlation existed between number of anammox 16S rRNA gene copies and protein level in the soluble microbial products (SMP), between tetG gene relative abundance and polysaccharose in SMP and between tetG gene relative abundance and protein in bound EPS (EPSs). In nutshell, the current findings provide the first description of a recovery mechanism regarding OTC-inhibited anammox performance aided by BA based on functional genes and highlights the contribution of ARGs and the self-resistance ability of EPS.

Susceptibility, resistance and resilience of anammox biomass to nanoscale copper stress.

The increasing use of engineered nanoparticles (NPs) poses an emerging challenge to biological wastewater treatment. The long-term impact of CuNPs on anaerobic ammonium oxidation (anammox) process was firstly investigated in this study. The nitrogen removal capacity of anammox reactor was nearly deprived within 30days under the stress of 5.0mgL-1 CuNPs and the relative abundance of anammox bacteria (Ca. Kuenenia) was decreased from 29.59% to 17.53%. Meanwhile, copper resistance genes associated with the Cus, Cop and Pco systems were enriched to eliminate excess intracellular copper. After the withdrawal of CuNPs from the influent, the nitrogen removal capacity of anammox biomass recovered completely within 70days. Overall, anammox biomass showed susceptibility, resistance and resilience to the stress of CuNPs. Therefore, the potential impacts of ENPs on anammox-based processes should be of great concern.

Successful start-up of the anammox process: Influence of the seeding strategy on performance and granule properties.

Successful start-up of the anaerobic ammonium oxidation (anammox) process in up-flow sludge blanket reactor was achieved by seeding denitrifying (R0) or mixed denitrifying-anammox granular sludge at a 3:1 volume ratio (R1). The results demonstrated that R1 was successfully started-up on day 40 and had a nitrogen removal rate (NRR) of 0.55kgNm(-3)d(-1). By contrast, it took 98days to start up R0 (0.54kgNm(-3)d(-1)). R0 and R1 achieved maximum NRRs of 5.70 and 12.02kgNm(-3)d(-1), respectively. The biogranules from R1 generally possessed greater specific anammox activity (SAA), higher extracellular polymeric substance (EPS) and heme c content, larger granules and greater settling velocity. Quantitative PCR (qPCR) and high-throughput sequencing techniques were used to monitor the evolution of the bacterial community. These results demonstrated the feasibility of seeding mixed denitrifying-anammox granular sludge to start-up an anammox reactor.

Insight into the short- and long-term effects of Cu(II) on denitrifying biogranules.

This study aimed to investigate the short- and long-term effects of Cu(2+) on the activity and performance of denitrifying bacteria. The short-term effects of various concentrations of Cu(2+) on the denitrifying bacteria were evaluated using batch assays. The specific denitrifying activity (SDA) decreased from 14.3 ± 2.2 (without Cu(2+)) to 6.1 ± 0.1 mg N h(-1)g(-1) VSS (100 mg Cu(2+)L(-1)) when Cu(2+) increased from 0 to 100 mg L(-1) with an increment of 10 mg Cu(2+)L(-1). A non-competitive inhibition model was used to calculate the 50% inhibition concentration (IC50) of Cu(2+) on denitrifying sludge (30.6 ± 2.5 mg L(-1)). Monod and Luong models were applied to investigate the influence of the initial substrate concentration, and the results suggested that the maximum substrate removal rate would be reduced with Cu(2+) supplementation. Pre-exposure to Cu(2+) could lead to an 18.2-46.2% decrease in the SDA and decreasing percentage of the SDA increased with both exposure time and concentration. In the continuous-flow test, Cu(2+) concentration varied from 1 to 75 mg L(-1); however, no clear deterioration was observed in the reactor, and the reactor was kept stable, with the total nitrogen removal efficiency and total organic carbon efficiency greater than 89.0 and 85.0%, respectively. The results demonstrated the short-term inhibition of Cu(2+) upon denitrification, and no notable adversity was observed during the continuous-flow test after long-term acclimation.

Insight into the short- and long-term effects of inorganic phosphate on anammox granule property.

The short- and long-term effects of inorganic phosphate on property of anaerobic ammonium oxidation (anammox) granule were investigated in this study. Acute exposure to high-level phosphate (⩾50 mM) induced the cytoplasm leakage. During a 195-day continuous-flow operation, the gradually increasing phosphate (up to 500 mgP L(-1)) slightly affected the specific anammox activity, hardly impacted the heme c content, remarkably decreased the extracellular polymeric substances production and significantly stimulated the dehydrogenase activity of anammox granules. Microbial community analysis showed no shift in the dominant anammox strain and higher population but lower relative abundance of anaerobic ammonium-oxidizing bacteria compared to the control granules. Interestingly, novel anammox granules with a hydroxyapatite core were cultivated, which possessed excellent settleability, huge granule diameter and superior mechanical strength. This study supported the application of granule-based anammox process as a pre-processing treatment in phosphate-containing and ammonia-rich wastewaters.

Inhibition of the partial nitritation by roxithromycin and Cu(II).

To facilitate the application of partial nitritation (PN) - anaerobic ammonium oxidation process in nitrogen removal from livestock wastewater, the inhibition of roxithromycin (ROX) and Cu(II) on the PN sludge was examined using a respirometric method. The results showed that the IC50 of ROX and Cu(II) on PN sludge were 346 and 74.3mgL(-1), respectively. The relative specific respiration rate (SRR) of ammonia-oxidizing bacteria (AOB) decreased from 87.4% to 17.7% with the ROX concentration increased from 0 to 500mgL(-1). When the concentration of Cu(II) increased from 0 to 160mgL(-1), the SRRs of AOB and nitrite-oxidizing bacteria decreased by 85.5% and 11.2%, respectively. According to the isobole plots analysis, combined suppression by ROX and Cu(II) was synergistic. Fourier transform infrared spectroscopy analyses showed that ROX exposure altered the positions of CO bonds, and the intensity of the absorption peak at 2100cm(-1) changed under Cu(II) exposure.