Pongsak (Lek) Noophan

Anaerobic ammonium oxidation by Nitrosomonas spp. and anammox bacteria in a sequencing batch reactor.

A sequencing batch reactor (SBR) was inoculated with mixed nitrifying bacteria from an anoxic tank at the conventional activated sludge wastewater treatment plant in Nongkhaem, Bangkok, Thailand. This enriched nitrifying culture was maintained under anaerobic conditions using ammonium (NH(4)(+)) as an electron donor and nitrite (NO(2)(-)) as an electron acceptor. Autotrophic ammonium oxidizing bacteria survived under these conditions. The enrichment period for anammox culture was over 100 days. Both ammonium and nitrite conversion rates were proportional to the biomass of ammonium oxidizing bacteria; rates were 0.08 g N/gV SS/d and 0.05 g N/g VSS/d for ammonium and nitrite, respectively, in a culture maintained for 3 months at 42 mg N/L ammonium. The nitrogen transformation rate at a ratio of NH(4)(+)-N to NO(2)(-)-N of 1:1.38 was faster, and effluent nitrogen levels were lower, than at ratios of 1:0.671, 1:2.18, and 1:3.05. Fluorescent in situ hybridization (FISH) was used to identify specific autotrophic ammonium oxidizing bacteria (Nitrosomonas spp., Candidatus Brocadia anammoxidans, and Candidatus Kuenenia stuttgartiensis). The ammonium oxidizing culture maintained at 42 mg N/L ammonium was enriched for Nitrosomonas spp. (30%) over Candidati B. anammoxidans and K. stuttgartiensis (2.1%) while the culture maintained at 210 mg N/L ammonium was dominated by Candidati B. anammoxidans and K. stuttgartiensis (85.6%). The specific nitrogen removal rate of anammox bacteria (0.6 g N/g anammox VSS/d) was significantly higher than that of ammonium oxidizing bacteria (0.4 g N/g Nitrosomonas VSS/d). Anammox bacteria removed up to 979 mg N/L/d of total nitrogen (ammonium:nitrite concentrations, 397:582 mg N/L). These results suggest significant promise of this approach for application to wastewater with high nitrogen but low carbon content, such as that found in Bangkok.

Effects of oxytetracycline on anammox activity

Batch experiments were conducted to investigate the effects of oxytetracycline on anaerobic ammonium oxidation (anammox) process. The short-and long-term effects on anammox activity were studied by measuring ammonium (NH4 +), nitrite (NO2 −), and nitrate (NO3 −) concentrations over time. Experiments were conducted at NH4 +and NO2 − concentrations of 60–90 mg N/L and 60–190 mg N/L, respectively (NH4 +:NO2 − ratio from 1:1–1:2.25), oxytetracycline concentrations of 10–100 mg/L, and biomass concentrations of 300–800 mg/L. In the short-term study, anammox activity was inhibited by all oxytetracycline concentrations studied. However, daily addition of oxytetracycline to a concentration of 5 ± 3.5 mg/L in the anammox sequencing batch reactor completely inhibited anammox activity in the fifth week. Fluorescent in situ hybridization was used to identify autotrophic ammonium oxidizing bacteria (Nitrosomonas spp., Nitrobacter spp., Nitrospira spp., Candidatus Brocadia anammoxidans, and Candidatus Kuenenia stuttgartiensis). The population of anammox culture was significantly decreased while Nitrosomonas spp. and Nitrospira spp. increased in the fifth week compared with the first week of experiment. A not-competitive model fit the anammox inhibition data at oxytetracycline concentrations of 0–100 mg/L quite well with Vmax of 0.0435 mg N/mg VSS-hr and Ki of 54.66 mg/L.

Autotrophic nitrogen removal in sequencing batch biofilm reactors at different oxygen supply modes

Conventional nitrification-denitrification treatment is a common way to treat nitrogen in wastewater, but this process is costly for low COD/N wastewaters due to the addition of air and external carbon-source. However, ammonia may alternatively be converted to dinitrogen gas by autotrophic bacteria utilizing aerobically autotrophically produced nitrite as an electron acceptor under anoxic conditions. Lab-scale sequencing batch biofilm reactors (SBBRs) inoculated with normal nitrifying sludge were employed to study the potential of an oxygen-limited autotrophic nitrification-denitrification process initiated with typical nitrifying sludge for treating a synthetic ammonia wastewater devoid of organic carbon in one step. The ring-laced fibrous carrier (length 0.32 m, surface area 3.4 m2/m) was fixed vertically in a 3 L reactor. Two different air supply modes were applied:continuous aeration to control dissolved oxygen at 1.5 mg/L and intermittent aeration. High nitrogen removals of more than 50% were obtained in both SBBRs. At an ammonia loading of 0.882 gm N/m2-day [hydraulic retention time (HRT) of 24 hr], the SBBR continuously aerated to 1.5 mg DO/L had slightly higher nitrogen removal (64%) than the intermittently alternated SBBR (55%). The main form of residual nitrogen in the effluent was ammonia, at concentrations of 25 mg/L and 37 mg N/L in continuous and intermittent aeration SBBRs, respectively. Ammonia was completely consumed when ammonia loading was reduced to 0.441 gm N/m2-day [HRT extended to 48 hr]. The competitive use of nitrite by aerobic nitrite oxidizing bacteria (ANOB) with anaerobic ammonia-oxidizing bacteria (anammox bacteria) during the expanded aeration period under low remaining ammonia concentration resulted in higher nitrate production and lower nitrogen loss in the continuous aeration SBBR than in the intermittent aeration SBBR. The nitrogen removal efficiencies in SBBRs with continuous and alternating aerated were 80% and 86% respectively. Specific microorganisms in the biofilm were characterized using fluorescence in situ hybridization. Aerobic ammonia-oxidizing bacteria (AAOB) occurred side by side with putative anammox bacteria (cells hybridizing with probe AMX820) throughout the biofilm, though ANOB were rarely detected.

Identification and quantification of bacteria and archaea responsible for ammonia oxidation in different activated sludge of full-scale wastewater treatment plants

In this study, the abundance and sequences of the amoA gene in ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA) were defined in three wastewater treatment plants using activated sludge with biological nitrogen removal in different countries: Thailand, United States of America (USA), and Japan. Quantitative real-time polymerase chain reaction (PCR) and PCR coupled with denaturing gradient gel electrophoresis were used to find the comparative abundance and identity of AOB and AOA. The conditions at the Phuket WWTP in Thailand promoted the dominance of AOA amoA genes over AOB amoA genes, while conditions at the WWTPs in Japan and USA promoted growth of AOB. Three parameters that may have contributed to the AOA dominance in Phuket were longer SRT, higher temperature, and higher pH. The Phuket WWTP is a unique system that can be used to better understand the conditions that promote AOA growth and dominance over AOB. In addition, analysis of operational data in conjunction with AOA and AOB community structure from the Phuket WWTP may elucidate advantages of AOA in meeting stricter treatment standards.

ANAMMOX-like performances for nitrogen removal from ammonium-sulfate-rich wastewater in an anaerobic sequencing batch reactor

ABSTRACT Ammonium removal by the ANaerobic AMonium OXidation (ANAMMOX) process was observed through the Sulfate-Reducing Ammonium Oxidation (SRAO) process. The same concentration of ammonium (100 mg N L−1) was applied to two anaerobic sequencing batch reactors (AnSBRs) that were inoculated with the same activated sludge from the Vermicelli wastewater treatment process, while nitrite was fed in ANAMMOX and sulfate in SRAO reactors. In SRAO-AnSBR, in substrates that were fed with a ratio of NH4+/SO42− at 1:0.4 ± 0.03, a hydraulic retention time (HRT) of 48 h and without sludge draining, the Ammonium Removal Rate (ARR) was 0.02 ± 0.01 kg N m−3.d−1. Adding specific ANAMMOX substrates to SRAO-AnSBR sludge in batch tests results in specific ammonium and nitrite removal rates of 0.198 and 0.139 g N g−1 VSS.d, respectively, indicating that the ANAMMOX activity contributes to the removal of ammonium in the SRAO process using the nitrite that is produced from SRAO. Nevertheless, the inability of ANAMMOX to utilize sulfate to oxidize ammonium was also investigated in batch tests by augmenting enriched ANAMMOX culture in SRAO-AnSBR sludge and without nitrite supply. The time course of sulfate in a 24-hour cycle of SRAO-AnSBR showed an increase in sulfate after 6 h. For enriched SRAO culture, the uptake molar ratio of NH4+/SO42− at 8 hours in a batch test was 1:0.82 lower than the value of 1:0.20 ± 0.09 as obtained in an SRAO-AnSBR effluent, while the stoichiometric ratio of 1:0.5 that includes the ANAMMOX reaction was in this range. After a longer operation of more than 2 years without sludge draining, the accumulation of sulfate and the reduction of ammonium removal were observed, probably due to the gradual increase in the sulfur denitrification rate and the competitive use of nitrite with ANAMMOX. The 16S rRNA gene PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and PCR cloning analyses resulted in the detection of the ANAMMOX bacterium (Candidatus Brocadia sinica JPN1) Desulfacinum subterraneum belonging to the genus Desulfacinum and bacteria that are involved in sulfur metabolism (Pseudomonas aeruginosa strain SBTPe-001 and Paracoccus denitrificans strain IAM12479) in SRAO-AnSBR.

Comparison of nitrogen removal rates and nitrous oxide production from enriched anaerobic ammonium oxidizing bacteria in suspended and attached growth reactors

Attached growth-systems for the anaerobic ammonium oxidation (anammox) process have been postulated for implementation in the field. However, information about the anammox process in attached growth-systems is limited. This study compared nitrogen removal rates and nitrous oxide (N2O) production of enriched anammox cultures in both suspended and attached growth sequencing batch reactors (SBRs). Suspended growth reactors (SBR-S) and attached growth reactors using polystyrene sponge as a medium (SBR-A) were used in these experiments. After inoculation with an enriched anammox culture, significant nitrogen removals of ammonium (NH4 +) and nitrite (NO2 −) were observed under NH4 +:NO2 − ratios ranging from 1:1 to 1:2 in both types of SBRs. The specific rates of total nitrogen removal in SBR-S and SBR-A were 0.52 mg N/mg VSS-d and 0.44 mg N/mg VSS-d, respectively, at an NH4 +:NO2 − ratio of 1:2. N2O production by the enriched anammox culture in both SBR-S and SBR-A was significantly higher at NH4 +:NO2 − ratio of 1:2 than at NH4 +:NO2 − ratios of 1:1 and 1:1.32. In addition, N2O production was higher at a pH of 6.8 than at pH 7.3, 7.8, and 8.3 in both SBR-S and SBR-A. The results of this investigation demonstrate that the anammox process may avoid N2O emission by maintaining an NH4 +:NO2 − ratio of less than 1:2 and pH higher than 6.8.

Nitrogen removal of anammox cultures under different enrichment conditions.

Anammox bacteria in sludge from an anoxic tank of a municipal wastewater treatment plant at Nongkhaem, Bangkok, Thailand were enriched in two sequencing batch reactors (SBRs; SBR-1 and SBR-2), under different conditions. SBR-1 was open to the atmosphere, while SBR-2 was closed and flushed with a mixture of 95% argon and 5% CO2 during the fill period in order to provide strict anaerobic conditions. The specific nitrogen removal rates of SBR-1 and SBR-2 were 0.43 g N/g VSS-d and 2.59 g N/g VSS-d, respectively. Denaturing gradient gel electrophoresis (DGGE) analyses showed differences in band patterns among the Nongkhaem sludge and the two enrichment cultures. Based on fluorescent in situ hybridization (FISH), the anammox bacteria in both systems were either “Candidatus Brocadia anammoxidans” or “Candidatus Kuenenia stuttgartiensis”. The results from this study demonstrate the potential of alternative anammox systems for nitrogen removal and provide information on the microbial communities of anammox cultures under different enrichment conditions.

Resuscitation of starved suspended- and attached-growth anaerobic ammonium oxidizing bacteria with and without acetate

Anammox application for nutrient removal from wastewater is increasing, though questions remain about anammox resilience to fluctuating conditions. Resuscitation of anammox suspended- and attached-growth cultures after 3 months of starvation was studied with and without acetate dosing. Without acetate, the attached-growth culture recovered more quickly than the suspended-growth culture. Suspended-growth cultures recovered more quickly (within 60 days) with weekly and daily acetate dosing than without, but anammox activity and copy numbers decreased with continued acetate addition. All attached-growth cultures recovered within 60 days, but after that activity with acetate dosing was consistently at least 20% lower than that without acetate addition. Ca. Jettenia caeni, Ca. Anammoxoglobus sp., Ca. Brocadia fulgida, Ca. Brocadia anammoxidans, Ca. Brocadia fulgida and Ca. Jettenia asiatica were identified. Acetate addition can significantly accelerate short-term resuscitation of enriched anammox suspended-growth cultures after starvation but may reduce anammox activity over the longer term in suspended- and attached-growth cultures.

Improvement of TiO2/LDPE Composite Films for Photocatalytic Oxidation of Acetone

Titanium dioxide with coupling agent (ETES) was applied as a photocatalyst for a synthesis of the TiO2/LDPE composite film. The physical properties of TiO2/LDPE composite film were analyzed by a Scanning Electron Microscope (SEM). TiO2 particles were impregnated into the polymer matrix film as a LDPE composite film. The results from the X-ray Diffraction (XRD) technique revealed that the structure of TiO2/LDPE composite film were anatase crystalline. The chemical structure of the TiO2/ LDPE composite films were analyzed by an ATR-Fourier transforms infrared (ATR-FTIR) spectrometer. Wavenumber of FTIR spectra at 719 cm1 indicated the Ti-O-Ti bond. Band gap energies of the films ranged from 3.19-3.29 eV. The photocatalytic activity of the film was tested for removal of gaseous acetone in a closed chamber. Experimental conditions were set as follows: a UV light intensity of approximately 2.7 mW.cm-2, flow rate of 2 L.min-1, and an initial acetone concentration of about 435±20 ppm. While the catalyst dosage was varied from 3% to 15% (wt. cat/wt. film).The degradation rate of acetone increased when increasing dosage of TiO2 from 3% to 10%, then decreased a little bit when increasing the dosage to 15%. The TiO2/LDPE composite film at the dosage of 10% yielded the highest removal efficiency of 75%, followed by the film at the dosage of 15%, 5%, and 3%, respectively.