Ammonia-oxidizing bacteria and archaea exhibit differential nitrogen source preferences.

Ammonia-oxidizing microorganisms (AOM) contribute to one of the largest nitrogen fluxes in the global nitrogen budget. Four distinct lineages of AOM: ammonia-oxidizing archaea (AOA), beta- and gamma-proteobacterial ammonia-oxidizing bacteria (ß-AOB and γ-AOB) and complete ammonia oxidizers (comammox), are thought to compete for ammonia as their primary nitrogen substrate. In addition, many AOM species can utilize urea as an alternative energy and nitrogen source through hydrolysis to ammonia. How the coordination of ammonia and urea metabolism in AOM influences their ecology remains poorly understood. Here we use stable isotope tracing, kinetics and transcriptomics experiments to show that representatives of the AOM lineages employ distinct regulatory strategies for ammonia or urea utilization, thereby minimizing direct substrate competition. The tested AOA and comammox species preferentially used ammonia over urea, while ß-AOB favoured urea utilization, repressed ammonia transport in the presence of urea and showed higher affinity for urea than for ammonia. Characterized γ-AOB co-utilized both substrates. These results reveal contrasting niche adaptation and coexistence patterns among the major AOM lineages.

Partitioning of nutrient removal contribution between granules and flocs in a hybrid granular activated sludge system.

Sludge granulation in continuous-flow systems is an emerging technology to intensify existing activated sludge infrastructure for nutrient removal. In these systems, the nutrient removal contributions and partitioning of microbial functions between granules and flocs can offer insights into process implementations. To this end, a reactor system that simulates the continuous-flow environment using an equal amount of initial granule and floc biomass was investigated. The two operational strategies for maintaining granule growth in the continuous-flow system were (a) the higher solids retention time (SRT) for the granules versus flocs, as well as (b) selective feeding of carbon to the granules. The SRT of the large granule fractions (>425 µm, LG) and floc/small granule fractions (<425 µm, FSG) were controlled at 20 and 2.7-6.0 days, respectively. Long term operation of the hybrid granule/floc system achieved high PO43- and NH4+ removal efficiencies. Higher polyphosphate-accumulating organisms (PAO) activity was observed in the FSG than LG, while ammonia-oxidizing bacteria (AOB) activities were similar in the two biomass fractions. Nitrite shunt was observed in the FSG, possibly due to out-competition by the high NOB activity in LG. More importantly, washing out the FSG caused a reduction in LG's AOB and PAO activity, indicating a possible dependency of LG on FSG for maintaining its nutrient removal capacity. Our findings highlighted the partitioning and potential competition/cooperation of key microbial functional groups between LG and FSG, facilitating nutrient removal in a hybrid granular activated sludge system, as well as implications for practical application of the treatment platform.

Application of aerobic kenaf granules for biological nutrient removal in a full-scale continuous flow activated sludge system.

Aerobic granular sludge (AGS) is a biofilm technology that offers more treatment capacity in comparison to activated sludge. The integration of AGS into existing continuous-flow activated sludge systems is of great interest as process intensification can be achieved without the use of plastic-based biofilm carriers. Such integration should allow good separation of granules/flocs and ideally with minor retrofitting, making it an ongoing challenge. This study utilized an all-organic media carrier made of porous kenaf plant stalks with high surface areas to facilitate biofilm attachment and granule development. A 5-stage Bardenpho plant was upgraded with the addition of kenaf media and a rotary drum screen to retain the larger particles from the secondary clarifier underflow whereas flocs were selectively wasted. Startup took 5 months with a sludge volume index (SVI) reduction from >200 to 50 mL g-1. Most of the kenaf granules fell in the size range of 600-1400 µm and had a clear biofilm layer. The wet biomass density, SVI30, and SVI30/SVI5 of the kenaf granules were 1035 g L-1, 30.6 mL g-1, and 1.0, respectively, which met the standards of aerobic granules. Improved stability of biological phosphorus removal performance enabled a 25% reduction in sodium aluminate usage. Microbial activities of kenaf granules were compared with aerobic granules, showing comparable N and P removal rates and presence of ammonium-oxidizing bacteria and polyphosphate-accumulating organisms in the outer 50-60 µm layer of the granule. This work is the first viable example for integrating fully organic biofilm particles in existing continuous-flow systems.

Aerobic granular sludge: Impact of size distribution on nitrification capacity.

The relationship between ammonia oxidation rate, nitrifiers population, and modelled aerobic zone volume in different granule sizes was investigated using aerobic granular sludge from a pilot-scale reactor. The pilot was fed with centrate and secondary effluent amended with acetate as the main carbon source. The maximum specific ammonia oxidation rates and community composition of different aerobic granular sludge size fractions were evaluated by batch tests, quantitative PCR, and genomic analysis. Small (331µm) granules had a 4.72 ± 0.09 times higher maximum specific ammonia oxidizing rate per 1 gVSS, and a 4.05 ± 0.17 times higher specific amoA gene copy number than large (2225µm) granules per 1 gram of wet biomass. However, when related to surface area, small granules had 1.43 ± 0.01 times lower maximum specific ammonia oxidation rate and a 1.66 ± 0.04 times lower specific amoA gene copy number per unit surface than large granules. Experimental results aligned with modeling results in which smaller granules had a higher specific aerobic zone volume to biomass and lower specific aerobic zone volume to surface area. Aerobic granular sludge reactors having the same average diameter of granules may have very different proportions of granule size fractions and hence possess different nitrification rates. Therefore, instead of the commonly reported average granule diameter, a new method was proposed to determine the aerobic volume density per sample, which correlated well with the nitrification rate. This work provides a roadmap to control nitrification capacity by two methods: (a) crushing larger granules into smaller fractions, or (b) increasing the mixed liquor suspended solid concentration to increase the total aerobic zone volume of the system.

Flocs in disguise? High granule abundance found in continuous-flow activated sludge treatment plants.

To date, high performance of full-scale aerobic granular sludge (AGS) technology has been demonstrated on a global scale. Its further integration with existing continuous flow activated sludge (CFAS) treatment plants is the next logical step. All granular sludge reactors operated in sequencing batch reactors (SBR) mode with anaerobic feeding conditions select for growth of phosphorus and glycogen accumulating organisms (PAO and GAO, respectively), which are known to enhance sludge settling characteristics. Therefore, we hypothesized that AGS are commonly present at full-scale CFAS processes with enhanced biological phosphorus removal (EBPR) and low sludge volume index (SVI). This hypothesis was confirmed at 13 EBPR plants, where granules were found present (at plants where SVI was lower than 100 ml/g) with a strong correlation between high granule abundance and low SVI. A wide range of granule abundance was found among the plants, ranging from 0.5% to as high as 80%. Evaluations of the EBPR plant process configurations showed that high granule abundances may be related to selector design features such as high anaerobic food to mass (F/M) ratios, unmixed in-line fermentation, and high influent soluble COD fraction. Granules were also observed at a non-EBPR plant with an aerobic selector receiving high F/M feeds. Quantitative PCR and 16S rRNA gene sequencing analyses revealed higher relative gene abundance of Accumulibacter PAO and Competibacter GAO in the granules over flocs, as well as a correlation between granule abundance and some possible EPS producers such as Flavobacterium and Competibacter. Our results indicated that process configurations that select for slow-growing or EPS-producing heterotrophs play an important role for granule formation in full-scale CFAS systems as previously shown in SBR configurations.

Recovery of phosphorus and nitrogen from human urine by struvite precipitation, air stripping and acid scrubbing: A pilot study.

Sustainable and closed-loop nutrient cycling require the recovery of valuable resources from wastewater. Resource recovery from diluted wastewater streams is limited by diluted concentrations and unfavorable reaction kinetics. In comparison, source separated urine allows resource recovery from a highly concentrated nutrient stream, resulting in a more sustainable and efficient recovery practice. Different nutrient recovery methods from urine have been studied in lab-scale, but pilot or full-scale process evaluations remain sparse. In this study, recovery of struvite and ammonium sulfate from urine of pregnant women was demonstrated at a pilot-scale treatment facility by means of precipitation and air stripping/acid scrubbing. The system achieved 94% struvite precipitation efficiency but merely 55% of the crystals were removed and recovered. The low phosphorus recovery was due to the washout of small crystals that escaped the sieve and settling tank, hence requiring an improved method for crystals capture. The removal and recovery efficiencies for nitrogen were 93% and 85%, respectively. Composition analysis of the produced fertilizers indicated that struvite was the dominated precipitate and quality of the ammonium sulfate met European standards. Carbamazepine and diclofenac were added in the urine to measure the fate of pharmaceuticals in the treatment system. Very little of the spiked pharmaceuticals (<0.01%) accumulated in the produced struvite and ammonium sulfate. The overall energy demand of the pilot system was 1066 MJ per m3 urine processed or 198 MJ per kg N removed. Energy efficiency was not optimized and can be improved in many ways.